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The Microbiology Manual

A member company of International Diagnostics Group plc

© 2002 IDG (UK) Limited idg, LAB M, Captivate and Harlequin are trademarks of the International Diagnostics Group plc group of companies.

CONTENTS
Contents
Introduction ISO 9000:2001 Certification Customised media service i iv v Coliforms / Enterobacteriaceae E. coli presence or absence enumeration using membranes enumeration without membranes O157:H7 Yeasts and moulds Staphylococcus aureus Pseudomonas spp. Bacillus cereus Clostridium perfringens Listeria FDA method modified USDA method Salmonella semi-solid method conventional method Campylobacter xxiii xxiv xxiv xxiv xxiv xxv xxv xxv xxv xxvi xxvi xxvi xxvi xxvii xxvii

Manufacturing Process
The process outline Quality criteria The application of growth rate analysers

pages v- viii
v vi pages vii-viii

Preparing Culture Media
Dehydrated Media storage weighing out pH of culture media glassware addition of powder to water heat dispensing prior to sterilisation Autoclaving Culture Media total heat input measurement points volume of medium heat conductance of agar load distribution / composition agitation equipment Sterility Indicators adhesive tape Browne’s tubes Spore indicators Molten Media re-melting agar pouring plates drying plates Sterile Supplements Storage of Prepared Media Troubleshooting Guide

pages ix-xiii
ix ix ix ix x x x x x x xi xi xi xi xi xi xii xii xii xii xii xii xii xii xiii xiii xiii

Culture Media Information
Format and abbreviation guide Dehydrated media guide

pages xxix – 3.6
xxix xxix

Media Range
Alphabetical Listing of Products

1.1 1.1.1 2.1 3.1 4.1 5.1 6.1 7.1 8.1
8.1 8.4 8.8 Last page

New Media Products Harlequin™ Chromogenic Media Biomolecular Products Captivate™ Selective Supplements Agars, Peptones Extracts and other Media Constituents Sterile Additives and Ready Prepared Media Indexes
By product code By product name By organism Request a copy of the manual

Quality Control of Culture Media
The ecometric technique Productivity ratio Liquid media Templates / suggested records Preservation of stock cultures Laboratory accreditation HACCP

pages xiv – xxi
xiv xv xv pages xvi-xviii xix xx xxi

Microbiology Methods
Sources of information TVC

pages xxiii-xxvii
xxiii xxiii

iii

iv .

Each component is individually tested for suitability. What do we offer? • Individually designed media. • Assurance of compliance with regulatory requirements. Each container is immediately labelled with product details. Weighings are double checked before the components are blended. safeguarding your products/process. • Commitment to on-going production. • Intellectual Property options. From time to time. Dyes.Customised Media Service The LAB M Customised Media Service Since its inception. Results are recorded and a reference sample stored. • Quality products and service. Milling. Blending A production batch is made from raw materials of specified batch number which have been pre-tested for compatibility. Comparison with previous batch and competition Quality control first checks that the batch is completely blended. engineered to meet specific customer requirements. then a series of physical and biological tests are performed to ensure the product meets the exacting standards required by our customers. This has evolved into a more proactive approach to resolving customer needs: The LAB M Customised Media Service. QUALITY CONTROL Physical. however. LAB M has been asked to ‘design’ media to meet specific customer applications. through product trials and into routine production. This is a unique service that differentiates us from our competitors and our Technical Support Group would be happy to discuss your needs on a confidential basis. • Development programmes that are both cost and time efficient. BOTTLING Into 500g sealed containers. Comparisons with previous and competitor’s batches are made. commencing with feasibility tests. v . • Total client confidentiality. code and batch number. CUSTOMERS LAB M products are dispatched all over the world to microbiologists in all types of laboratory. The components are individually milled to ensure uniform particle size. Biological parameters. • Post-development consultancy. • Customised packaging and labelling. • Close client collaboration. LAB M Culture Media: The Process Outline RAW MATERIALS Agars. The increasing importance of regulatory compliance has also been a significant factor in this process of innovation. Peptones. LAB M has been committed to customer needs by offering a wide range of quality media products for global markets. Extracts. or bulk containers at request Automated equipment delivers pre-weighed amounts into containers which are hermetically sealed. Strict batch traceability in accordance with ISO9001 ensures we can recall all products if necessary. • A process approach to development. Growth promoting components are selected with the help of an automated growth rate analyser (Malthus Instruments) PRODUCTION Weighing. Chemicals etc. Its continued programme of product innovation and development has ensured that the Company has been responsive to market trends and changes in custom and practice.

gel strength. Mg++) compatibility with other components. comparison with previous batch and competition. productivity ratio. Biological – Growth characteristics. Bile Salts – Clarity. freedom from toxicity. we can select those peptones that trigger the exponential growth of inoculated microorganisms in the shortest possible time. colour. moisture. For example. Agar – Clarity. pH. thin layer chromatography. growth promotion inhibition. We were the first company to utilise automated growth rate analysers to ensure only the most suitable ingredients available are chosen. pH. heat stability. with the use of growth rate analysis. chemical parameters. pH. properties after incorporation into culture media. compatibility with other components. setting point. Choosing peptones for enrichment media . Selection of ingredients LAB M pioneered the use of impedance technology for the selection of Culture Media ingredients. Raw materials Peptones and Extracts – Clarity. clarity. The survival time of the test organisms is also taken into consideration. All components weighed accurately and checked. redox. Compatibility with other components. this technology is also useful in minimising batch to batch variation. Components milled to uniform particle size.LAB M Culture Media – The quality criteria Production All components from specified batch numbers. Components blended for specified time. heavy metal content (particularly Ca++.peptone A is chosen because of a faster response time. multiple samples taken to ensure thorough blending. gel strength. Quality control Physical – pH. chemical reactions and colour changes. vi . haemolysis patterns. viscosity. Clarity on re-melt. melting point. growth promoting properties with Gram positive and Gram negative organisms aerobically and anaerobically. Further information on this technique is on pages 5 and 6. antibiotic antagonists. Dyes & Chemicals – pH.

meat. infusions and extracts that are used by microbiologists are manufactured from biological raw materials and these are subject to variations and inconsistency (e. and G5. Introduction One of the fundamental performance parameters of a bacteriological culture medium is its ability to promote the early and rapid growth of micro-organisms. Denton Peptones The peptones. and both performance and comparison with ‘stock’ material will be taken into account before a decision to purchase is made. and against a competitor’s product.The application of growth rate analysers in the manufacture of bacteriological culture media by W. (Figure 1) Because of this variation it is standard practice for culture media manufacturers to test samples of these raw materials from many sources in order to maintain a supply of products with suitable performance characteristics. The base lines for each channel are set on the chart recorder and the growth curves for each test are recorded. Normally pre-shipment samples will be evaluated. The main electrical parameters measured. Full quality control of the bulk material is then also carried out. The tubes are then placed in an accurately controlled water bath and connections are made between the electrodes and the analyser. are conductance and impedance. The curves produced on the equipment could be directly related to the total viable count.c still suffer from giving a point source of information in what is a dynamic analysis. Unfortunately this technique is both laborious and inaccurate. Impedance/conductance techniques In 1898. The speed of the chart and the sensitivity of the instrument can be adjusted in order to accommodate the growth characteristics of various organisms. are much more variable in performance than reagent grade chemicals. Using this technique it is possible with some organisms and substrates to obtain a result in less than half a working day. The viable count gives information on the state of the culture at a single moment in time but gives no indication of the growth rate. c) The novel spiral plater technique4. with five tubes counted from each dilution. Since this time. There have been many methods devised to assess parameters. finally crystallising in the 1970’s with papers by Ur and Brown (1975)6 and Cady (1975)7 describing the use of continuous impedance monitoring as a tool with wide potential in microbiology. variation can be from -70 to +260 per cent! Other variations on the Lister technique for assessing substrate performance are: a) Agar dilution techniques. This has enabled closer control of the performance characteristics of the individual peptones and culture media formulations available from the company. eg the Malthus system9. The introduction of electrical methods into the field of culture media performance analysis has provided microbiologists with the means to accurately record development of a culture over a large section of the growth curve. Figure 1 vii . Techniques a . The nephelometric/photometric techniques are relatively insensitive and give little information about the log phase. R. unless serial viable counts are performed. Multi-channel growth rate analysers are commercially available. and they cannot be used with materials which produce turbidity. Halverston and Ziegler in 19331 showed the technique to be subject to very large experimental errors. There are advantages in measuring either of these parameters.g. Method of use The eight channel machine is used to compare the performance of two substrates inoculated with three to four organisms or when testing finished product to compare the new batch of product against a stock batch. that products described as ‘meat peptone’. for example. LAB M introduced an eight channel Malthus Growth Rate Analyser into its raw material selection and quality control procedures in 1982. d) Photometric and nephelometric techniques. both alone and made up into typical formulations. These are manufactured by acid or enzyme digestion processes which are cumbersome and difficult to control. workers at the Torry Research Station in Aberdeen produced equipment capable of accurately plotting the growth curve of bacteria in various fluid culture media using impedance techniques8. plants). Using this method he was able to plot the classic growth rate curve of a bacterial culture.A. It is not surprising. b) Surface inoculation techniques such as the drop method of Miles and Misra2 and the ecometric technique of Mossel et al3. several workers have made use of conductance and impedance measurements in the analysis of bacterial growth. For example. Methods based on the classical method of Lister are used for estimating growth rate. milk. therefore. Stewart showed that bacterial growth in fluid could be detected by changes in the electrical properties of that fluid. The organisms are inoculated into 2ml or 10ml volumes of the sterile substrate in tubes which contain the electrodes. and these are outlined in the excellent book by Eden. In 1977. most of which rely on the viable count. Hyde and K.

a new aspect of culture media quality control is opened up.. 26.S... D. (1933a) J. 732 3. Miles. N. however. Hobbs. J. (1975) (edited by C. (1977) J. Bact. J.. First published in International Labmate 1987 Vol.. Malthus Instruments Ltd.12 Issue 2 pp 36-37 Figure 3 viii . hyg..M. sterility test media. Cady. Maximum peak of the curve .This is related to the maximum ‘number’ of bacteria achieved in the culture. New York 6. 244-252 5. D.A. M. Ziegler. M. Gilchrist. I.. John Wiley and Sons.. (1980) J. Eden. Verdouden M. C. D. b) For culture media that demand standardisation of performance from batch to batch. Impedance Microbiology (1984) Research Studies Press. J.T.O. 7. c) The maximum peak is of interest when choosing a raw material for production of large numbers of bacteria such as for vaccines. J.A..73-99. G. 3 9. (1938) J. P.F. Halvorson.The log phase gradient is a measure of the rate of metabolism in the multiplying bacteria. Koopmans. Detection time or take off – When the curve shifts from the baseline lag phase into the log phase. Wash Lane.. Appl. 439-454 4. 38.Illeni. S.. Delany. Camb. 559 2.g. New York). In culture media manufacture the Growth Rate Analyser provides information that makes a significant contribution to improving both the performance and standardisation of products manufactured from variable raw materials. G.. Bury. Heden and T... Microbiol. F. Lancs. (1933b) I.G. blood culture media. (Figure 3) References 1. (1975) New Approaches to the Identification of Microorganisms (edited by C.. and needs to be tested for. Bact. Firstenberg. Ur. New York 8. (1973) Appl.G. Mossel.Growth curve information There are several parameters which can be assessed from the growth curve produced by the Malthus system... for example: a) The peptone with the shortest lag time can be chosen for those media that are required to give rapid growth from a small inoculum. 331. 49. H.R. this information can be used as a measure of the number of organisms present in a sample with the aid of suitable calibration graphs (Figure 2). Heden and T.A. Appl. Donelly. R. Topley House. Peeler. The information can be used in a number of ways.E.. 101. Rapid metabolism causing changes in electrical properties may be detected before cell division begins. Bid. Illeni) P. 43. can performance of the culture media on similar equipment in customers’ laboratories be assured. e. Eelderink. These include: Lag time – The time that elapses before a change in electrical properties can be detected. Figure 2 Conclusion Electrical methods of monitoring bacterial growth have been known since the latter part of the 19th century. New York.61-71 John Wiley and Sons. and fermentation media. M. these are the electrical properties of the product. it is possible to choose peptones with a specified lag time growth rate. Log phase . only in the last few years with the development of micro-processors and sophisticated electronics are these methods beginning to reach their full potential in Growth Rate Analysers. A. Eden. 25. As equipment based on impedance methods became established in the microbiological quality control procedures of the food industry. A. Only by the use of impedance/conductance measuring equipment in quality control. Biol. Gibson. John Wiley and Sons.B. Brown. P.. 25. Misra. Hendriks. Van Rossem.

A 1. gel filled unbreakable. The components of some formulations can be irritant so the wearing of a face mask at this stage is advisable.e. dry conditions Loose cap. and sterilisation is carried out precisely to manufacturer’s instructions. stored in light on bench Loose cap. Use pH 4.0. if possible. but some problems have been found by users. Similarly a 4. of media e. Time allowance must be made for various volume sizes of media to ensure that overheating leading to acid formation and caramelisation does not occur. It is advisable to use only fresh purified water with a conductivity of less than 10 microsiemens. Dehydrated media should not be used if it shows any sign of moisture gain i. D. Stored water tends to become acidic because it absorbs atmospheric CO2. Storage in a refrigerator is generally not recommended as there is the risk of condensation on the container when it is brought out of the refrigerator. Electrodes Dehydrated Culture Media Storage Dehydrated media stored unopened under optimal conditions have a shelf life of 3-5 years but once the container is opened the contents should be used within six months.5 units. Further details are given under Autoclaving Culture Media. This assumes 1 litre volume produced strictly according to media manufacturer’s instructions. Do not tip the media out of the container as this will cause excess dust which may be irritant and will certainly need cleaning up. A. 38 No. sugar fermentation. The following section outlines the correct procedures which will ensure high quality reconstituted products. even temperature away from any sources of moisture such as washing up areas or laboratory autoclaves and away from strong light. Temperature compensator Weighing Out Using a top-pan balance with an accuracy of ±0. macrolides). pH culture media Acceptable pH variations ± 0.0 and pH 7. Tech. 7. Media difficult to Ringers. Wash in deionised water.2 is usual. Measure conductivity < 10 micro Siemens. pH 10. Adjustment of pH Should not be necessary if all systems correct.4 moisture gain % Short-term storage pH 4. To check pH add 0.4% gain in moisture will result in a 78% reduction in isolation rate. pH calibrating or checking attachment. dark.g. Mixed volume loads should be avoided. ix . This demonstrates the importance of ensuring the container lid is tightly closed and the pot stored in cool.). Calomel. Am. The lid on the container should be replaced quickly after media has been taken out and closed tightly. Tap water is not recommended because of the potential presence of heavy metal ions which can cause inhibition and precipitation problems. but not predictable. Deionised or double-distilled water is only weakly dissociated. Water Purification by distillation or deionisation is advisable.3ml saturated KCl to 100ml water.0 buffer of electrode Electrode faults Indicated by slow or erratic readings and inability to obtain two points on the meter without adjustment. antibiotics less active (fusidin. dry. dark conditions.0. become lumpy or discoloured. (1972).0 buffer for 2 hours.5% pepsin in N HCl for 1-2 hours. balance. tetracycline. stored in light in autoclave room 0 1. volume etc. combination electrodes shielded with detachable shield for cleaning.Preparing Culture Media Quality Assured Before each batch of LAB M Culture media is passed for sale it undergoes a rigorous quality control procedure to ensure it gives maximum recovery and reproducibility. L. They DO NOT compensate for changes in ionisation of solution at high temperatures. cephalosporins. (Available from BDH. Cleaning of electrode Daily checks Testing meter Water Culture media Acidity: Bile salts precipitated. G. Optimum 20 . soak in pH 7.1 gram the powder should be spooned onto a weighing boat or clean beaker. pH of culture media Meter Sensitive to one decimal place. Storage vessels for purified water must also be monitored for microbial colonisation. TABLE 1 – Deterioration of SS Agar stored in various conditions for 6 months.25˚C. Alkalinity: Potentiates aminoglycocides. A review of some common sources of error in the Preparation of Agar Media. It is important that the equipment is properly maintained. Maximal Recovery Diluent pH because of low ionic strength Effect of autoclaving or irradiation Effect of volume of media on pH Autoclaving or irradiation will lower the pH about 0. and Fay. the output of ion resins need to be electronically monitored and microbial colonisation of the resin and tubing must be avoided. Storage conditions Unopened bottle stored in cool. In-house quality control by the user will help determine the condition of product in opened containers. These adjust for changes within the electrode only. The best conditions for storing dehydrated media are in a cool. sugar fermentation. or according to media temperature to manufacturer’s instructions. Flat electrodes may be used. antibiotics less by pH active (aminoglycocides. Reconstitution of media in the user’s laboratory must be done with care to ensure the same high standards of performance. The effect of the moisture gain on the performance of the agar can be quite dramatic. 0.1% gain in moisture on storage will lead to a 53% reduction in the numbers of Salmonella isolated. Med. water quality.2 . and suggests simple quality control techniques that can be used to check the performance of prepared media. Vol..0 buffer and. H2S reactions reactions affected depressed.1 4. J. When a container is opened for the first time the date should be noted on the container. penicillins). Barry.

Ensure a clear. Borosilicate glass is preferable to soda-glass because the latter may leach alkali into any solution contained in it. i. The total time above 50˚C is important because both nutrients and agar will be undergoing a denaturation process. The aim of effective sterilisation is to ensure all spores are destroyed with the minimum necessary heat input in order to ensure no ill effects are caused to the medium. Add the powder to the flask and slowly pour on the water with frequent swirling. a lethal rate table can be produced: (see facing page) x . However these processes must be carefully controlled to ensure the minimum possible heat input is used. From a chart recording of the sterilisation process (the recording must be from a thermocouple placed inside the medium during processing). As soon as the medium begins to boil it should be removed from the source of heat. 3. The higher the temperature the faster the denaturation. CAUTION: Agar media. Using this equation. Media should never be left at high temperatures for prolonged periods for example holding in a 56˚C water bath overnight will noticeably reduce a culture medium’s performance and gel strength. Heat Remember that heat denatures the nutrients and the agar in culture media. 30 minutes at 121˚C will noticeably affect the gel properties of most agars whilst the medium could be held at 50˚C for several hours before denaturation can be detected. Allow the medium to cool to 47˚C before dispensing with constant mixing into final containers for sterilisation. agars and broths so the reduction in holding time must be calculated for different volumes of both. media preparators. attempting to mix the water and powder too quickly can cause the formation of lumps which are difficult to disperse.pH out of tolerance If pH needs to be adjusted because manufacturer’s instructions cannot be followed. Pour the full amount of powder onto the full amount of water in a flask.121. Different types of media differ in heat penetration i. This boiling should be done with frequent agitation to ensure even heat distribution. The total time spent above 100˚C is important. well mixed. Heat is necessary for sterilisation and for dissolving the agar if the medium is to be distributed into tubes or bottles prior to sterilisation. then add remaining water. 1. but only for the same volume of medium. Heat input is less in equipment that allows rapid heating and rapid cooling. Figure 4 Schematic diagram of heat input during sterilisation The total heat input of a sterilisation cycle can be shown graphically. media processed and pH measured when cooled and appropriate calculations carried out.1) Z Where L = Equivalent time at 121˚C T = Actual temperature for 1 minute Z = Temperature coefficient (= 10˚C for spores) Log-1 = antilog of the number. or N/10 NaOH into aliquots of media should be made. the portion of the graph between 100˚C and 121˚C for both the heating and cooling stages is divided into 1 minute intervals. This can be done by using the lethal rate equation or tables (Stumbo 1973). Allow to stand for 10 minutes before swirling to mix. Thus the instruction to ‘autoclave at 121˚C for 15 minutes’ should be taken to mean ‘sterilise by an equivalent heat process to 121˚C for 15 minutes’.g. This ensures that the heating process is sufficient to kill the spores. In our experience the commonest cause of problems with culture media is the heat sterilisation. e. 2. a dynamic process which has many variables that must be considered. because not only is this the temperature at which spores are being killed. Once calculated the new holding time can be used for future cycles. add the powder slowly whilst constantly swirling. The time is added together and the total time removed from the holding time.e. Agar containing media need to be brought to the boil to take the agar into solution. particularly those with low agar content. Pour approximately 1/3 of the volume of water required into a flask. NOTE: The water should always be measured before adding to the flask. Never heat water before adding to the medium. titration of small additions of N/10 HCl. To prevent this agitate frequently and gently as the medium approaches boiling and the agar begins to dissolve. Dispensing prior to Sterilisation Most broth media are readily soluble at room temperature or with the aid of gentle heat. Total Heat Input Addition of Powder to Water There are a number of ways to do this. and using the equation or table. (shown below) can be converted into an equivalent time at 121˚C. may boil unexpectedly and overflow out of the flask. Total heat input should be controlled to protect the medium from denaturation. but it is the temperature with the potential to cause most damage to the medium’s performance. These are: QUALITY CONTROL USING MICROBIOLOGICAL PERFORMANCE CRITERIA ARE ESSENTIAL Glassware All glassware must be undamaged and clean having been rinsed with purified water before storage. but keeps damaging heat input to a minimum. Prolonged heating and cooling can lead to excessive heat input.e. solution is obtained before dispensing into final containers and sterilising. Lethal Rate Equation: L = Log-1 (T . Autoclaving culture media To achieve optimum performance from reconstituted culture media it is important to ensure sufficient heat input to kill all spores whilst protecting the medium from excessive heat input that would damage the nutrient and gelling properties of the medium.

“General purpose lab autoclaves are difficult to standardise and more consistent results are obtained by purpose-built media sterilisers. The only disadvantage of media preparators is that they can only handle one medium at a time with a cycle time of just under 1 hour.062 0. Media preparators are dedicated to sterilising media.123 0. the volume of medium in the bottles is directly related to the heat input.078 Temp 111 112 113 114 115 116 117 118 119 120 121 Time 0.this can prolong cooling times. This type of equipment will have built-in thermocouples and chart recorders capable of monitoring both chamber and load temperatures throughout the cycle. xi . based on holding time alone.g. Larger laboratory autoclaves now have separate steam generation facilities allowing rapid heating and some models evacuate the air by vacuum and then inject the steam under pressure ensuring there are no air pockets causing cold spots.490 0. Be aware that a sterilisation cycle that is optimum for 1 litre of medium will be excessive for l00ml of medium. It is therefore impossible to properly control a cycle if mixed volumes (e.025 0. Therefore.049 0. The poor heat conductivity of agar has significant effects when trying to sterilise large volumes of medium (e. Modern media preparators have this ability which gives them a significant advantage over general purpose autoclaves.” Temp = Temperature for 1 minute interval Time = The equivalent time at 121˚C Load Distribution / Composition Heat penetration in an overloaded autoclave will be hampered as will evacuation of air. Corry et al made the following comment. reducing the effectiveness of the equipment. If a thermocouple is used in a 500ml bottle of water to control the sterilisation of 500ml of agar medium. Pre-heating of the medium to get the agar into solution before autoclaving is recommended. Volume of Medium The heating and cooling periods of large volumes of media are longer than for small volumes of media. It is important to allow this type of equipment to free steam before closing the outlet valve. Brecker & Bridson showed that 500ml of agar in a thin-walled bottle took 6 minutes longer to reach 121˚C than did 500ml of water in an identical bottle.389 0. Brecker & Bridson found that: “In 4 litres of medium. although they all operate by using steam under pressure to attain temperatures above 100˚C. Agitation The continuous agitation of a medium during sterilisation greatly shortens the heating and cooling times.039 0. This type of equipment also incorporates a cooling jacket through which cold water is run after the completion of the holding time. We consider the lack of proper controls make it unsuitable for routine use. Mixed volume loads should therefore be avoided if at all possible.g. the difference in heat penetration must be taken into account.TABLE 2 – Lethal Rate Table: Temp 100 101 102 103 104 105 106 107 108 109 110 Time 0. The main advantage of this equipment is that they agitate the medium speeding up heating and cooling and ensuring thorough mixing. Simple bench top autoclaves are larger versions of pressure cookers but with the ability to measure the temperature and pressure of the chamber. both these factors affect the rate of heat penetration.015 0.000 Heat Conductance of Agar Agar is a very poor conductor of heat and heat penetration into agar containing media is significantly slower than into non-agar containing media.098 0.019 0.776 1. It is important to remember that the temperature of the medium will lag behind that of the chamber both in heating up and cooling down.309 0. Brecker & Bridson gave the following guide times for heat penetration at 121˚C in glass bottles: The equipment used for autoclaving varies from the very simple to very complex. The crudest autoclave is the domestic pressure cooker. Ensure sufficient space is left between items in a load to allow free passage of steam. Measurement Points Some autoclaves will measure chamber temperature.155 0.617 0. can be significantly greater for large volumes than for small volumes. 1 litre flasks and 20ml bottles) are put together in the same load. with a 5 minute hold at 100˚C. TABLE 3 – Heat Penetration Times Volume 500ml 1 litre 2 litre 5 litre Time to reach 121˚C 18 minutes 22 minutes 27 minutes 37 minutes The thickness of the glass and the shape of the container were not specified. the total heat treatment in cycles. As covered previously.031 0. They are well equipped with both time and temperature controls. In our experience. otherwise. some will measure the temperature of the medium (by use of a thermocouple).245 0.008 0.010 0. 2 litres or more). The cooling is brought about by use of a water jacket. For safety reasons the door will not open until the chamber has cooled to 80˚C . Adjustments have to be made to the fluid heat penetration time in the sterilisation cycle shown below. These times seem to be based on both autoclave and medium being heated from cold to 121˚C without the effective ‘holding’ at 100˚C that occurs with modern pulsing autoclaves or with traditional autoclaves before the outlet valve is closed.012 0. not all the air will be flushed out of the chamber. the temperature at the centre of the agar mass had not reached 121˚C 1 hour after the autoclave chamber had. Equipment Figure 5 Schematic diagram showing the lag of medium temperature compared to chamber temperature. some will measure both. with the agar settling undissolved to the bottom. a 1 litre Pyrex flask of medium will attain 121˚C within 5 minutes of the chamber reaching that temperature.” It is important that whatever equipment is used is properly maintained to ensure accuracy of gauges.195 0. This equipment is suitable for the sterilisation of small to medium sized volumes and the heating and cooling periods are short.

(1970) Design and Formulation of Microbial Culture Media. In most of the applications to which Bowie-Dick tape is put in the microbiology laboratory. New York. Browne’s Tubes The Browne’s tube is a chemical indicator comprising a heat sensitive solution within a glass tube. Failure to do this can result in traces of inhibitory substances being incorporated into non-selective agars. but again no information is given about the total heat input. In this respect it is a more reliable indicator of sterility than Bowie-Dick tape. and none can demonstrate overheating of the medium. media should be poured as soon as possible. The addition of extra agar to compensate for the reduction in gelling caused by overheating is not recommended as the overheating will have also damaged the nutrient properties. Single vent Petri dishes lose moisture more slowly than triple vent dishes but they are also more prone to condensation problems if not adequately dried. A.F. The amount of medium specified for poured plate techniques is generally less because the medium is not stored in vented Petri dishes. Most media will stand re-melting once in a boiling water bath. it is doubly important that holding at 47˚C in a water bath is minimised. This means it is necessary to use separate tubing for non-selective media and for inhibitory/antibiotic supplemented media. The only way to ensure sterilisation without overheating. Never re-melt an agar more than once. Methods in Microbiology Volume 39 edited by Norris. Immediately before pouring the medium should be swirled gently to ensure thorough mixing. Plates should contain at least 5mm. the tubes can deteriorate on storage which may lead to premature colour change and false information as to the performance of the sterilising equipment. It is therefore only an indicator of steam penetration. in a water bath. it was designed for use in the demonstration of adequate steam penetration. (1986) Introduction to Sterilisation and Disinfection. of medium if they are to be stored before use to minimise the effects of the drying that will occur during storage. and may deteriorate on storage giving false results. However the tube only indicates that the correct conditions have been achieved at that particular site in the steriliser. examples of which are given below: Molten Media Holding molten media in water baths at 47 ˚C or above for more than a few hours should be avoided. 2nd Edition. Spore Indicators The spores of Bacillus stearothermophilus are extremely resistant to heat and can thus be used as a biological indicator of sterilisation. All the above indicators are a compromise in the absence of accurate steriliser control. the autoclaving carefully controlled in an equivalent heat process. but only if they are used correctly and their limitations are fully understood. Again. Ideally. this type of indicator shows if the time and temperature requirements have been achieved but not if excessive heat has been used. and this was examined after sterilisation to make sure the diagonal stripes changed to black uniformly along the tape cross. C. M. Sterility indicators Sterility indicators can be a useful tool to highlight deficiencies in sterilising equipment. and Brecker. to 47˚C before pouring. spore strips only show ‘spot’ conditions. Three types of indicator are commonly used. (1973) Thermobacteriology in Food Processing. and Ribbon. Use of an inappropriate indicator for the product or equipment can lead to the mistaken belief that the process is being properly monitored. and Peel. After the cycle is complete they are removed aseptically and placed in a bottle of thioglycollate medium or cooked meat medium. Like Brown’s tubes. Academic Press. Drying Plates In order to achieve well isolated colonies on agar plates streaked for single colonies. which changes colour from red to green when the tube is subjected to a high temperature for the required length of time. Consequently they give no indication as to whether the medium has been subjected to excessive heat resulting in loss of performance of the sterilised product. linked to chart recorders. Pouring Plates Media should be cooled. Academic Press. are placed in various parts of the load. as the stripes will change colour before the full cycle is complete. When layered plates are used the total depth should not be less than 5 mm. during which they are examined for growth. J.References: Stumbo. D. Holding at 47˚C overnight will result in noticeable denaturation of agar gel. holding at 47˚C for 4 hours should be the maximum holding time any medium is subjected to. but give no information about the total heat input. and cooled to 47˚C before pouring. but caution must be taken to ensure over drying does not occur as this can be detrimental to the performance of the agar. Spore strips prepared in the laboratory or commercially produced strips. thereby reducing their ability to support the growth of fastidious organisms. New York.R. S. chemical and biological. Furthermore. and the sum of these can impair performance if not considered and kept to a minimum. physical. it will be a very unsatisfactory method for demonstrating the achievement of sterile conditions. autoclaving. The extra heat input will damage both the nutrient and gelling properties of the medium.Y. Re-melting Agar Adhesive Tape Bowie-Dick tape is classed as a physical indicator even though it relies upon a chemical change to produce the black diagonal stripes which indicate the tape has been subjected to heat. The major drawback of sterility indicators is that even when used carefully for the correct application. it is necessary to make sure the agar is free from surface moisture by drying. Bridson. Churchill Livingstone. as it shows both temperature and time are sufficient for sterility. A cross of the tape was placed on a sheet of steam permeable paper across a towel at the mid-point of a stack of towels. and in accordance with a previously determined equivalent heat process for the equipment and volume of media being sterilised. The use of multiple tubes throughout the load is more satisfactory. is to use thermocouples introduced into the medium being processed. they will show that the minimum conditions have been achieved. However. It is worth noting that numerous procedures in the stages of preparation of agar plates can result in loss of moisture. Wherever possible an appropriate equivalent heat process should be calculated for the load to be sterilised. and incubated at 55˚C for 7 days. the length of time needed to obtain a result means they are inconvenient for routine use. and pouring at temperatures above 50˚C can all result in moisture loss. E. whereas Bowie-Dick tape merely indicates a heating process has occurred.W. If a medium is likely to be allowed to set then re-melted. Boiling of the medium to dissolve agar before autoclaving. Again xii . Gardner. NB: -It is important to note that the tubing through which the medium is dispensed can retain inhibitory substances on the inner surfaces. the total heat input into the medium has to be considered.R. and so media should only be boiled before autoclaving if absolutely necessary. More commonly referred to as autoclave tape.M.

If a medium is stored as a gel and then is re-heated before use. inaccurate weighing. 2. Indeed. The following is a list of potential problems and their possible causes: FAULT Soft Gel POSSIBLE CAUSE Excess heat. Contaminated water or glassware. Any antibiotic supplement reconstituted but not used should be thrown away. To check for sterility. excess heat. Most sterile supplements such as blood.C. Plates containing antibiotics have a shelf life governed by the stability of the antibiotics. agar not dissolved. Excess heat. Sterile Supplements The addition of sterile supplements is performed after the medium has been sterilised and cooled to approximately 47˚C. deterioration of dehydrated medium.even deep frozen. Plates should be brought up to room temperature before use to avoid any ‘thermal’ shock to the bacteria. Plates showing visible signs of shrinkage (drying) should not be used. incorrect weighing and mixing. A plate with an original medium depth of 5mm will have its ingredients concentrated 20% by the time its gel has shrunk to a depth of 4 mm. deterioration of dehydrated medium. Serum products are best stored frozen then thawed. 6 Rapid drying of plates There are 2 commonly used methods of drying plates: 1. All media should be stored in the dark to prevent the formation of bacteriostatic and bactericidal substances (e. Any plates left on the bench for more than 8 hours should be discarded as unsuitable for use. Carefully place the plate in the incubator with the medium containing side up. repeat of a simple Q. the humidity of the surroundings and the amount of excess moisture present. The supplement itself should be warmed up to room temperature before adding to the medium and it should be added whilst mixing to prevent the formation of ‘cold spots’ and premature gelling of the agar. pH wrong. Antibiotics vary widely in their stability once reconstituted and can deteriorate rapidly . deterioration of dehydrated medium. procedure after 3 months. impure water or glassware.C. To produce ‘chocolate’ or heated blood plates add the sterile blood to the medium at 80˚C or add at a lower temperature and gently re-heat with frequent swirling until the medium ‘chocolates’.g. Excess heat. The supplemented medium should be thoroughly mixed by swirling before any plates are poured. the form in which it is stored and the conditions of storage. The effectiveness of this technique will depend on the venting of the Petri dishes. pH taken at wrong temperature. peroxides). deterioration of dehydrated medium. dirty glassware. (See specific section on pH of culture media) Impure water. Each time a batch of medium is prepared some form of performance quality control should be carried out. chemical contamination. inadequate mixing. serum and antibiotics are denatured at higher temperatures. Troubleshooting Guide The laboratory that routinely quality controls the media it produces will occasionally find it has produced a batch that is not up to standard. This allows excess moisture to evaporate whilst minimising the possibility of contaminating the agar. deterioration of dehydrated medium. deterioration of dehydrated medium. Contaminated glassware. Care must be taken to observe strict aseptic precautions whilst adding the supplement. Excess heat. pH equipment faulty or poorly standardised. Most plates stored medium side up at 4˚C in the dark will have a minimum life of 7 days. Excess heat. The practice of incubating plates overnight could lead to excessive moisture loss and reduced medium performance. Excess heat (scorching or burning). Bottled Media Any medium in an airtight capped container will have a longer shelf life than in a plate. deterioration of dehydrated medium. Plates are allowed to stand (with lids on) overnight on the bench. It is unwise to use media beyond their minimum shelf life without repeating the quality control and comparing it with the initial result. many can be stored for longer but we advise repeat of a simple Q. pH incorrect Storage of prepared media The shelf life of prepared media is dependent upon the composition of the medium. Contaminated water or glassware. procedure is recommended. Generally speaking it is unwise to extend the shelf life of an antibiotic containing medium beyond 7 days. impure water. pH too low causing acid hydrolysis. This can be extended up to 3-4 weeks for simple nutrient media by using some form of airtight packing. Bottled media containing antibiotics will have shelf lives governed by the activity of the antibiotic. Lift the base of the dish up and rest it on the lid as in the diagram. overheating. As a medium loses moisture the ingredients of the medium will be concentrated making selective media progressively more selective.Plates agar Lid Fig. incorrect weighing and mixing. incubate a representative sample at the temperature and time parameters used for performing the test for which the medium is employed. Many simple nutrient media can be stored at 1520˚C for 3 months in the dark. Abnormal colour Darkening Precipitation Toxicity Poor growth Poor selective or differential properties xiii . This chocolating of the blood destroys enzymes which would otherwise inactivate the nicotinamide adenosinedinucleotide (NAD or V factor) required for growth by Haemophilus spp. It is wise to aliquot serum on arrival into suitable volumes to be frozen until required.

Examples of less effective selection occurs with Salmonella media. It is important to note that the ecometric method is a simple technique by which laboratories can check the media they are producing. Charge a 1 microlitre loop with the incubated 4-hour culture and spread the test plate. it has to be performed with care. Keeping of records . Each medium used in the laboratory should have its own quality control protocol and the necessary organisms should be maintained. and record the segment and line. The ecometric technique of Mossel is simple and gives numerical readings that can form the basis of records suitable for trend analysis. The results can also be compared with results obtained using the same organisms on non-selective media. However. This can be overcome by using a spiral plater to inoculate the plates to ensure the plating method is identical for test and control agars. and CLAS etc. The results obtained can be compared with previous batches of the same medium or with batches of the same medium from different manufacturers. as simple errors such as holding the inoculation loop at a different angle may introduce errors. use that as the yardstick to measure the performance of new batches and to check that the efficiency of all media-making processes are being maintained.C. 2. use a suitable non-inhibitory plate. xiv . This is the end point and can be used to calculate the absolute growth index (AGI) and relative growth index (RGI) of a medium. without flaming or recharging the loop and finish at D5. For example.Dl . We recommend the following: A full quality control when a new batch of dehydrated medium is introduced into the laboratory. AGI = 75 Using the formula RGI = RGI = AGI Test AGI Control 75 85 88. e. and achieve suppression of bacteria without inhibiting the organism which is to be isolated. AGI = 85 Plate B—selective agar test —end point C4. For a batch of dehydrated media new to the laboratory. Inoculate 5ml of Brain Heart Infusion Broth (LAB 49) with a loopful of the chosen test organism (see individual products) and incubate for 4 hours. to be repeated every 3-4 months on opened containers. use a plate of the same medium from the last batch to check consistency.24 % x 100 x 100 The Ecometric Technique This plating technique is simple enough to use in both full and shortform quality controls. The performance of a selective agar can be thoroughly tested using an organism which it has been designed to isolate and one which it is designed to inhibit. Techniques The parameters of growth on culture media are: ● ● ● ● ● ● ● Lag time Organisms grown from known inoculum Organisms inhibited from known inoculum Comparative growth with standard inoculum Comparative inhibition with standard inoculum Colony size Colonial appearance TABLE 4 – Absolute Growth Index (AGI) Al = 5 A2 = 25 A3 = 45 A4 = 65 A5 = 85 Bl = 10 B2 = 30 B3 = 50 B4 = 70 B5 = 90 Cl = 15 C2 = 35 C3 = 55 C4 = 75 C5 = 95 Dl = 20 D2 = 40 D3 = 60 D4 = 80 D5 = 100 Figure 7 Schematic diagram of the ecometric method In practice absolute measurements of growth are time-consuming or require sophisticated equipment whilst colonial appearance is subjective and difficult to record. A short-form quality control whenever prepared media are used beyond their minimum shelf life or are re-heated more than once. on the test and control plates. For a new batch of plates from the laboratory stock of dehydrated medium. More recently it is a requirement of many laboratory accreditation schemes such as UKAS.g.g.24% as efficient as the non-selective control plate. 4. For example: Plate A—non-selective control—end point A5.both full and short-form quality controls . C4 or D5 etc. fall off in performance can be detected.g. Campylobacter Selective Agar (LAB 112) is a very good selective medium as it will give high RGI’s for Campylobacter spp. C and D as shown below: 3. e. The AGI is obtained by noting the end point in Table 4. Divide the plate to be tested into quarters designated A. The RGI is a comparison of the AGI of the test plate and that of the control plate.Cl .A2 . A short-form quality control when a new batch of medium is reconstituted from dehydrated media previously quality controlled. It is based on streaking an inoculum to extinction. This full Q. Repeat this process with a control plate. B. going from Al . e. Incubate both plates for 18 hours.B2 etc. Thus. Colony size is easily measured but is an insensitive indicator of performance. if the end point is C4 then the AGI is 75. where there is a close relationship between pathogens (Salmonella and Shigella) and the other enteric organisms.Quality Control of Culture Media The routine quality control of culture media is an essential ‘good laboratory practice’ necessary to maintain the standards and performance of any bacteriological culture technique. below and this gives the AGI.Bl . making it difficult to inhibit one without reducing the isolation rate of the other. Comparative methods are the most suitable ones for routine quality control of culture media they can be used for comparisons of growth and inhibition. 5. = Method 1. Having found a medium which is suitable. The onus is therefore on the end user to decide which medium is best for their use. Note the last point at which growth occurs. for this particular organism the test plate was 88.so that trends. Both absolute growth index (AGI) and relative growth index (RGI) can be obtained by this method. For the former the RGI should be as close to 100 as possible and the latter as close to 0 as possible. The very nature of selective agars and the bacteria which must be selected or repressed means that effectiveness of different media will vary quite considerably.

J. of colonies on test x dilution factor .A.R. From an overnight culture of the test organism. prepared in Tryptone Soy Broth (LAB 4). is calculated by counting the colonies on the test and control media: P. 3. Appl. 5. of colonies on control x dilution factor 2.R. prepared in Buffered Peptone Water (LAB 46). Starting with the highest dilution (10-8 in example below) place one drop of the dilutions on the relevant quadrant. Calculate the P. Moulds and Yeasts: An Attempt at Standardisation at the International Level.R. Spread each drop over the quadrant and incubate the plates at 37˚C for 18 hours. For example: Test = 20 colonies at 10 -3 dilution Control = 25 colonies at 10 -3 dilution P.R. which should be a nutritious agar such as Tryptone Soy Agar (LAB 11). et al (1983) Quality Assurance of Selective Culture Media for Bacteria. of a medium is using the Modified Miles-Misra technique: 1. The inoculum used must be the same for both media and the P. This method can be used in conjunction with the Miles-Misra technique to demonstrate recovery of known levels of CFU’s in broth media. xv . it is less prone to error than the ecometric technique and its accuracy can be further increased by using duplicate plates. 3. Examine the broths and note the highest dilution showing growth (turbidity of the broth).R.Reference: Mossel D. This technique is not as simple as the ecometric method and is probably more usefully used to quality control broth media. Repeat for control plate. = 0. A simple method to obtain the P. Count colonies at the lowest dilution they can be easily counted. However.A. = 20 25 P. Figure 8 Template for Miles-Misra plates for both test and control. 1. 2. Incubate at 37˚C for 18 hours. prepare a tenfold serial dilution in Maximum Recovery Diluent (LAB 103).8 Thus in this example the test medium is 80% as efficient as the control medium.) Determining the productivity ratio of a medium is another way to check its performance related to a control medium. = No.R. 54 313-327 Liquid Media Another simple technique can be used to quality control fluid media such as Fastidious Anaerobe Broth (LAB 71) and Fluid Thioglycollate Medium (LAB 25). Productivity Ratio (P.R. Divide the test plates into quadrants and mark each quadrant with the dilution to be used as shown below.R. Bacteriol. = 20 x 103 25 x 103 P. From an overnight culture of the test organism. prepare a tenfold serial dilution (to 10 12) in Maximum Recovery Diluent (LAB 103). Add 1ml of each dilution to 9ml of test and control broths. Repeat with the control plates. No. 4.

using a permanent marker pen. Replace on template after incubation to read result. going from Al-Bl-Cl-Dl-A2-B2-etc.Templates for the Ecometric Method Mark the bottom of the plate as shown below. place the plate face-up on the template below. Once the quadrants are marked. xvi . without flaming or recharging the loop. Charge a lµl loop with a 4-hour broth culture and spread the plate.

C. and incubated under identical conditions. xvii . All plates must be spread at the same time. Organism 1 2 3 4 End Point Test Control AG1 Test Control RG1 Q. Organism 1 2 3 4 End Point Test Control AG1 Test Control RG1 Mean RG1 N.B.C. The same inoculum should be used for all plates.Suggested Quality Control Record Sheet Quality Control Record (A) DATE: MEDIUM: LAB CODE: BATCH NO: WEIGHT OF AGAR: CONTROL MEDIUM USED: VOLUME OF WATER ADDED: CONDUCTIVITY OF WATER ADDED: METHOD OF STERILISATION: SUPPLEMENTS ADDED: EXPIRY DATE OF SUPPLEMENTS: Q.

Variation between individual figures will occur due to inevitable inaccuracies of the technique. xviii .Suggested Quality Control Record Sheet Quality Control Record (B) MEDIUM: BATCH NO: APPEARANCE OF POWDER WHEN OPENED: DATE RECEIVED: DATE OPENED: Date 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 RGI Appearance Comments N.B. Comparisons of relative growth index are only useful to show a trend in the condition of the medium.

Cap the tube.Preservation of Stock Cultures The following method has been used in our laboratory for several years. A commercially available form of the above system called Protect Beads is available from LAB M. which combines certified media control cultures and Protect storage beads. Carefully pipette the suspension on to sterile glass beads with a hole for threading. 5. remove the beads using forceps. then place them in a plastic freezer tube. 2. a domestic freezer at -20˚C will suffice. 6. Morganella morganii Neisseria gonorrhoeae Pediococcus damnosus Proteus mirabilis Pseudomonas aeruginosa Pseudomonas fluorescens Pseudomonas fragi Pseudomonas putida Salmonella dublin Salmonella enteritidis Salmonella gallinarum Salmonella saint paul Salmonella senftenberg NCIMB 9240 NCIMB 9365 NCIMB 9375 NCIMB 9376 Salmonella typhi Salmonella typhimurium ATCC 11775 NCTC 9001 10090 Other NCIMB 9483 9111 12900 10479 13337 19992 14869 11842 7469 14917 15521 12706 DSM 20241 19111 7973 NCIMB 50007 NCIMB 8707 NCIMB 8540 15307 8076h 19242 29258 25933 25668 10662 10038 10689 10936 9676 5188 9240 6022 10384 8394 13311 74 5742 NCIMB 9321 13880 29903 29930 25923 6538 CMCC 2656 19990 19435 1466 Kiel 27421 6681 10319 8198 19258 11348 11327 11344 9610 CCUG 11291 CCUG 4588 CCUG 4586 NCIMB 6681 637 10211 2665 NCIMB 10762 235 8375 NCIMB 235 8109 CMCC 2703 (non-toxigenic) Preservation of Bacteria using a Modified Glass Bead Technique 1. QC Assays. Replace the cap and tap the bottle to remove the bubbles from bead centres. TABLE – 5 Culture collection strains for the quality control of culture media Organism/Bacteria Acinetobacter calcoaceticus Aerococcus viridans Aeromonas hydrophila Bacillus cereus Bacillus coagulans Bacillus licheniformis Bacillus megaterium Bacillus subtilis Bacteroides fragilis Brocothrix thermosphacta Campylobacter coli Campylobacter jejuni Campylobacter laridis Citrobacter freundii Clostridium bifermentans Clostridium perfringens Clostridium perfringens Corynebacterium diptheriae Enterobacter aerogenes Enterobacter cloacae Enterococcus faecalis Enterococcus faecium 19409 13048 13047 8043 25285 11509 ATCC 15309 11563 7966 11778 7050 14580 14581 10334 10341 10342 5398 9343 10822 11366 11168 11352 6272 506 8237 8238 3984 10006 10005 775 DSM 2918 NCIMB 10018 CIP 7080 NCTC 5866 8251 8049 Other Salmonella virchow Serratia liquefaciens Serratia marcescens Shigella flexneri Shigella sonnei Staphylococcus aureus Staphylococcus aureus Staphylococcus aureus Staphylococcus epidermidis Streptococcus bovis Streptococcus lactis Streptococcus pneumoniae Streptococcus pyogenes Streptococcus thermophilus Vibrio cholerae Vibrio fluvialis Vibrio parahaemolyticus Yersinia enterocolitica Yersinia enterocolitica Yersinia enterocolitica xix . label and place in freezer. A block of aluminium or copper drilled out to take the plastic tubes will prevent thawing. It does not require ultra low temperature equipment. The block is kept in the freezer along with the beads. is also available as a convenient kit. 3. There are potential problems with this technique which can be avoided: (i) The tubes should be duplicated and then stored in separate pots . the other as a back-up in a separate freezer. Organism/Bacteria Escherichia coli Escherichia coli Escherichia coli Escherichia coli O111 Escherichia coli O157 Haemophilus influenzae Hafnia alvei Klebsiella oxytoca Lactobacillus acidophilus Lactobacillus brevis Lactobacillus bulgaricus Lactobacillus casei Lactobacillus plantarum Lactobacillus sake Lactobacillus viridescens Leuconostoc mesenteroides Listeria monocytogenes Microbacterium flavum Microbacterium lacticum Micrococcus luteus Moraxella sp. (ii) The frozen beads will soon defrost on the bench.one for routine access. 4. and removed wearing a glove. Grow the organism on an appropriate solid medium until a heavy growth is obtained. To recover the organisms. Pipette off the excess fluid into phenolic disinfectant in a discard jar. Use several plates with organisms forming small colonies. Remove growth from plate using a sterile cotton swab and emulsify in 10% w/v glycerol in Brain Heart Infusion Broth. Details of recommended QC strains of organisms are contained in the individual entries for media. roll on an appropriate medium and streak out to obtain single colonies. please call for further details.

Aberdeen. UK CIP Collection de l’Institut Pasteur. Colworth House. KIEL Federal Dairy Research Institute. NCIMB National Collections of Industrial and Marine Bacteria Ltd. USA. but narrowed depending upon the nature of testing carried out. personnel. CMI Commonwealth Mycological Institute. K. FRG. xx . those laboratories who will be monitoring food with regard to enforcement of the law. 12301 Parklawn Drive Rockville. 18804 45534 10620 61274 432 17364 19795 439 61269 79 1522 ATCC American Type Culture Collection.ICFMH Working party on Culture media. Kew. monitored and challenged within a continuously improving quality system. i. Further consideration might be given to possible future legislation. U. However. in the past. Food Research Institute. The impetus for laboratory accreditation has. There are a number of schemes which provide assessment and certification of laboratory performance. FRG. and methods of a testing laboratory are appropriate. D3400. stating that this must be to the EN 45000 Series of standards. Food Microbiol. Paris. Whilst these standards relate specifically to official food testing laboratories. U. Int. Ferry Lane. The impetus for this growth is essentially the Food Safety Act of 1990. as changes between systems will almost certainly involve extra cost if it becomes necessary. The choice of scheme should not be taken lightly. The above table is adapted from the original published by the IUMS . France. 1987 5 297 -299. In addition. Part of this sets out the requirements for accreditation. Central Public Health Laboratory. ISO9000. UKAS. and industry practice. Grisebachstrasse 8.e. University of Goteborg. GLP. and probably the greatest area of growth is the food industry. U. Gottingen. K. NCTC National Collection of Type Cultures. it will reflect the best practice to which all testing facilities should aspire. other schemes are available and the choice may continue to expand. a food laboratory would not use CPA (Clinical Pathology Accreditation scheme) and may also consider GLP (Good Laboratory Practice) not targeted to its specific needs. CPA. Accreditation is the means by which external assessment ensures that the facility. AB2 1RY. CCUG Culture Collection.Colney Lane. 5-413 46.g. Hermann Weigmanstrasse 1.Yeasts & Moulds Alternaria alternata Aspergillus amstelodami Aspergillus flavus Aspergillus niger Aurobasidium cladosporoides Candida albicans Cladosporum cladosporoides Debaryomyces kloeckeri Fusarium moniliforme Hansenula anomala Mucor racemosus Penicillium cyclopium Pichia burtonii Rhizopus stolonifer Saccharomyces cerevisiae Zygosaccharomyces rouxii Abbreviation key: ATTC NCYC CMI 89343 17455 91856 NCIMB 50097 45534 Laboratory Accreditation The accreditation of a microbial testing facility must be the goal of all involved in the production of goods or provision of services to consumers. differed by industry sector. Sharnbrook. DSM Deutsche Sammlung von Mikroorganismen. The choice of scheme is bewildering. the benefits of accreditation are being applied in diagnostic and food testing laboratories. It follows from this that ensuring the testing laboratory is operating to agreed standards could be interpreted as a reasonable precaution which should be taken.. Norwich NR4 7UA. and introducing the ‘Due Diligence Defence’ whereby the defendant must show that all reasonable precautions which could have been taken to avoid an incident were indeed in place. 23 St. Scotland. for example pharmaceutical laboratories have for many years been subjected to Good Laboratory Practice and regular inspection by bodies such as the FDA and MCA. K. CLAS. Guldhedsgaten lOA. Kiel. One driving force for any future regulations is likely to be The Official Control of Foodstuffs Directive (89/397) which is an EC Directive concerned with the establishment of the Single Market in Foodstuffs. and should take into consideration the demands of the laboratory customers (internal and external). and sets out the requirements for official food testing laboratories. Bedford MK44 1 LQ. Goteborg. more recently. London NW9 5HT. Colindale Avenue. Currently only UKAS in the UK meet the criteria for accreditation bodies laid down in European Standard EN 45003. J. MAFF. Maryland. e. (German Collection of Micro-organisms). Machar Drive. which laid down the requirements for food manufacturers with regard to the provision of safe food. [strains from this source will be deposited with NCIMB]. Surrey. NCYC National Collection of Yeast Cultures. Sweden. AURIS Business Centre. CMCC Colworth Microbiological Culture Collection.

However it may be useful for monitoring contamination levels of raw materials prior to use. to ensure that the control is in place. as well as physical examination of the whole procedure. engineering. Documentation of all monitoring results is essential. It is therefore advisable to seek professional help via research associations or expert consultants with many years experience of developing HACCP systems. failure to do so would pose a threat to the consumer. ATP) or visual (handling procedures. adjusted and verified in a time scale relevant to the production of the food. microbiology. Establish Control Criteria The control criteria must be a parameter or practice which can be monitored. As with verification. practice. equipment). Like all systems. This is a critical control point. if a product must reach 70˚C on cooking.Hazard Analysis Critical Control Points (HACCP) In the same way that European legislation is driving the issue of laboratory accreditation. or omission of important detail. This must include audit of the documentation and monitoring records. which allow positive feedback and rapid corrective action. Once hazards are agreed then the CCP’s can be identified. Documentation is again essential to show the ‘quality loop’ monitoring > non-conformance > action > monitoring > control. and hygiene. oven temperature can be set accurately determined limits. A poorly operated system could make things appear under control. HACCP is only effective if implemented properly with the co-operation of all employees involved. With the advent of rapid technology the hygienic status of surfaces can be measured in real time and re-cleaning performed if necessary. It does not replace end product testing. and a prime example of this is in the use of HACCP in the production of food. and the monitoring parameters are correct. For example. Auditing and Review The system must be reviewed and audited at regular intervals to ensure that nothing has changed which would affect the efficacy of the control process. and so the HACCP system used to control it also has to be designed specifically and reviewed regularly. Developing a HACCP system can be a time consuming and complex business. If a product fails verification when the monitoring process shows no problem. The team will then follow the principles of HACCP to implement a system which will ensure the safety of the product: Establish Monitoring Procedures The parameters which ensure that the critical control point is actually in control of the safety of the product. For example. it puts end product testing into the correct context as a validation that the control system is functioning correctly. Developed in the USA to ensure the absolute safety of food used in the NASA space programme (can you imagine suffering food poisoning in zero gravity?). xxi . location. monitored by probes and adjusted in real time. will allow the product to pose a threat to consumer of the product. Verification Procedures Further actions taken to verify the monitoring of CCP’s. as could measuring the level of microbial contamination of the manufacturing environment. temperature) chemical (pH. it also affects other areas where the expertise of the food microbiologist is essential. if not controlled. any areas of concern must be subject to the corrective action and demonstrate the ‘quality loop’. and the above representation is vastly over-simplified. Hazard Analysis This is the identification of the hazards associated with the production of the product and the ranking of the risk involved with each and involves the input of relevant members of the team with the necessary technical background. it indicates a failure in the system and a review is necessary to identify a missing CCP or other source of the problem. Microbiology is seldom an effective monitoring tool due to the time involved. Establish Corrective Action Plans Agreed actions necessary to regain control if monitoring of a CCP indicates that it is not controlling the safety of the product. biocide levels. The hazards inherent in any food production process are specific to that process in that company at that time. aw. End product testing falls into this category. Like Quality Systems HACCP can only work with the backing of the entire organisation starting from the top and requires a team approach to implement. These are generally physical (time. The European Directive on the Hygiene of Foodstuffs 93/43/EEC includes the requirement that all businesses shall develop a system in accordance with the HACCP principles to identify and control food hazards. or raw material which. The team must be multi-disciplinary and typically will include personnel from production. Critical Control Points This is any process. when the reality is that potentially dangerous products may be produced. it is a system which recognises the limitations of end-product testing and identifies the points in the production process critical to producing a safe product.

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plus one or two newer methods.4 x 103) This method can be adapted to perform an anaerobic count by incubating in anaerobic conditions. overlay with more of the same agar to cover surface | | Incubate at 37˚C for 24hr (some USA methods 35˚C.S. However incubation times may vary to those above and specific texts should be consulted. Manual of Microbiological Methods for the Food and Drink Industry. ISBN 0-935584-59-5. nor replace other sources of information. coliforms for dairy purposes 30˚C) | | Count all colonies >0. Methods for the Examination of Water and Associated Materials.Microbiology Methods We have set out some of the more common methods in use. Milton Keynes. M.. American Public Health Association.S. Micro organisms in foods . Select plates with between 15 and 150 colonies for enumeration.g.4 x 103) *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. Foodborne Pathogens An Illustrated Text (1991) Varnam. 2. cooled: Plate Count Agar APHA (LAB 10) or Plate Count Agar (LAB 149) or Milk Plate Count Agar (LAB 115) or Milk Agar (LAB 19) or Yeast Extract Agar (LAB 18) | | Incubate at the appropriate temperature to count the required bacterial population: 30˚C for 48hrs (aerobic mesotroph count) 21˚C for 5 days or 6. GL55 6LD. Remember that facultative organisms will give a count for both the aerobic and anaerobic counts. and Splittstoesser D. 43 2nd edition. MK14 6LE. C. Total Viable Count (Total Aerobic Count) Prepare an appropriate 1:10 dilution of the sample e.5mm diameter present on the plate. A.g. There are numerous sources of information regarding microbiology methods. Goucestershire.Their significance and methods of enumeration 2nd edition (1978) ICMSF edited by Thatcher. Numerous microbiological standards e. xxiii . just some of which are listed below: British Standards Institute. ISBN 0-7234-1521-8). Compendium of Methods for the Microbiological Examination of Foods (1992) Third edition.g. This is not intended to be a comprehensive list. Wolfe Publishing (now Times Mirror International). D. indicating the LAB M products required to perform them. M.G. D. Part 11. ISBN 0-8020-2293-6 Practical Food Microbiology. 2. and Evans. If the highest dilution has more than this use 300 as the figure to calculate the original count and express as ‘greater than…’ | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e. Hooper. Enumeration of Bacillus cereus. F. Linford Wood. Public Health Laboratory Service ISBN 0-901144-36-3 Environment Agency: The Microbiology of Drinking Water (2002). (1995) Campden & Chorleywood Food Research Association.g. Customer Services. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 1ml of appropriate dilutions into a sterile Petri dish and add aseptically 15ml of molten. Technical Manual No. Chipping Campden. and Greenwood. but as a simple guide only.H. and Clark. ISBN 0-87553-173-3 Food and Drug Administration. Methods for the Examination of Food for Micro organisms of Public Health Significance 2nd edition (1995) Edited by Roberts. Bacteriological Analytical Manual 8th edition (1995) AOAC International.F. Coliform/Enterobacteriaceae (Enumeration) Prepare an appropriate 1:10 dilution of the sample e.5˚C for 10 days (aerobic psycrotroph count) 55˚C for 48hrs (aerobic thermotroph count) | | Select plates with between 30 and 300 colonies for enumeration. W. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 1ml of appropriate dilutions into a sterile Petri dish and add aseptically 15ml of molten.g. BS5763 Methods for Microbiological examination of food and animal feeding stuffs. If the highest dilution has more than this use 150 as the figure to calculate the original count and express as ‘greater than…’ | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e. cooled: VRBA (LAB 31) (Coliform count) or VRBGA (LAB 88) (Enterobacteriaceae count) When set. Edited by Vanderzant. *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions.

Incubate plates at 37˚C for 24hrs. coli (Enumeration Using Membranes) Prepare an appropriate 1:10 dilution of the sample e. E.5˚C for 18-24hrs | | *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. | | Indole positive colonies will turn pink. other serovars of E. and Tryptone Water (LAB 129) | | Incubate at 44 ± 0. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate membrane on the surface of Minerals Modified Glutamate Agar (LAB 80A + LAB 80B) + 12g/litre of Agar No. E. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate the surface of Harlequin™ Tryptone Bile Glucuronide Agar (HAL 3) | | Incubate at 30˚C for 4hrs then transfer to 44 ± 0.1g of product. expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e.5˚C for 18-24hrs | | Put 2ml of Vracko-Sherris reagent into lid of dish.g.5˚C for 24hrs and examine for turbidity and gas production | | If negative incubate for a further 24hrs and examine again | | For all positive tubes add 2-4 drops of indole reagent to the matching Tryptone Water.1ml of positive broth into 10ml single strength Brilliant Green Bile Broth (LAB 51). coli and calculate the original count in the sample. Allow to soak in reagent for 5 min.g.g. Alternatively Harlequin™ SMAC-BCIG (HAL 6) can be used. coli O157:H7 (Presence or Absence) Add 25g sample to 225ml of Modified Tryptone Soy Broth (MTSB) (LAB 165) | | Homogenise for 2 mins | | Incubate at 42˚C for 6 and 24hrs | | Perform Captivate™ immunomagnetic separation at 6 and 24hrs | | Inoculate 50µl of Captivate™ beads onto SMAC (LAB 161) and CT SMAC (LAB 161 + X161). and examine again | | Subculture 0. Count all pink colonies as presumptive E. | | Examine plates for sorbitol negative colonies | | Confirm as E. coli can produce the verocytotoxin and therefore be pathogenic for humans. indicating a positive indole reaction | | Record result as presumptive presence of E.4 x 103 *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. 2.g. Furthermore.Escherichia coli (Presence or Absence) Prepare an appropriate 1:10 dilution of the sample e. and place membrane on top. | | Dry membrane in natural daylight or expose to UV for 5-30 min. inoculate 1ml of homogenate into 10ml of single strength MacConkey Broth Purple (LAB 5) or Brilliant Green Bile Broth (LAB 51) (with inverted Durham tubes).1g or 1g of product as appropriate *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions.2 (MC 6) | | Incubate at 37˚C for 4hrs then transfer membrane to a Tryptone Bile Agar plate (LAB 72) | | Incubate with membrane uppermost at 44 ± 0. xxiv . coli in 0. coli (Enumeration Without Membranes) Prepare an appropriate 1:10 dilution of the sample e. E. Wait one minute and examine for a red colour in the alcohol layer at the top of the broth. To detect presence in 1g of product. coli O157 by serology and biochemistry (commercial ID kits) It should be noted that not all O157 isolates produce the verocytotoxin which is the mode of pathogenesis of the organism. inoculate 10ml of double strength medium with 10ml of homogenate | | Incubate at 37˚C for 24hr and examine for acid and gas (LAB 5) or turbidity and gas (LAB 51) (30˚C is used in the dairy industry) | | If negative incubate for a further 24hrs. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | To detect presence in 0.

xxv . *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. (Enumeration) Prepare an appropriate 1:10 dilution of the sample e. although this is not straight forward. 2-4mm. and in particular damaged organisms may lose the ability to produce them.Yeasts and Moulds (Enumeration) Prepare an appropriate 1:10 dilution of the sample e. 2.g.5ml by spreading over the surface of Bacillus Cereus Medium (PREP.1ml or 0.g. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 0. as other Bacillus spp. and these may be differentiated by performing a wet preparation and examining under the microscope for typical cells. Pseudomonas species (Enumeration) Prepare an appropriate 1:10 dilution of the sample e. 2. with or without precipitate in surrounding medium). will grow on PREP agar (also known as MYP agar). Use reference schemes such as Bergey’s Determinative Bacteriology Volume 2 or Cowan and Steele. rounded.g. Alternatively a proprietary kit for detection of specific enterotoxin may be used. 2. *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. rough. or a proprietary kit. pink in colour. LAB 193) | | Incubate at 30˚C for 24hrs | | Examine the plate for colonies. count the dilution with less than 15 colonies on the plate if possible. or Baird-Parker medium with RPF supplement (X086) | | Incubate at 37˚C for 48hr | | Select plates with between 15 and 150 black colonies for enumeration. If the highest dilution has more than this use 150 as the figure to calculate the original count and express as ‘greater than…’ | | Confirm at least 5 colonies of each colony type present using slide or tube coagulase tests. aureus (and other staphylococci) on Baird-Parker Medium. LAB 73 or PEMBA. other staphylococci can mimic these reactions and so confirmation of all colony types is necessary to reduce the risk of reporting false counts. Examine microscopically to confirm large.g. Bacillus cereus Staphylococcus aureus (Enumeration) Prepare an appropriate 1:10 dilution of the sample e. or Oxytetracycline Glucose Yeast Extract Agar (LAB 89) | | Incubate at 25˚C for 5 days | | Select plates with between 15 and 150 colonies for enumeration. usually budding yeast cells as opposed to the much smaller bacterial rods and cocci). have been reported to produce enterotoxin. or Malt Extract Agar (LAB 37). taking into account the ratio of coagulase positive and negative colonies. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 0.1ml or 0. *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. Moreover.4 x 103).4 x 103) This is only a presumptive count.g. Due to the large size of the colonies. Confirmation can be done by biochemical methods. | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 1ml of appropriate dilutions onto the surface of : Rose Bengal Chloramphenicol Agar (LAB 36).1ml or 0.5ml of appropriate dilutions by spreading over the entire surface of Baird-Parker medium (LAB 85 and X085). | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e. Yeasts and moulds may grow on the medium. The advice to count all black colonies is a reflection of the variation of appearance of S.4 x 103) It is advisable to confirm growth on the plate as Pseudomonas spp by performing an oxidase test.5ml by spreading over the surface of Pseudomonas Agar (LAB 108) supplemented with CFC (X108) | | Incubate at 25˚C for 48hrs | | Select plates with between 15 and 150 colonies for enumeration. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 0.49 x 103) It is always prudent to perform a simple wet preparation of suspect yeast colonies by emulsifying a colony in a drop of saline on a slide and placing a coverslip on top. The production of the ‘typical’ lecithinase and lipase reactions is not a stable characteristic.g. If not clearly visible incubate for a further 24hrs.g. If the highest dilution has more than this use 150 as the figure to calculate the original count and express as ‘greater than…’ | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e.g. If the highest dilution has more than this use 150 as the figure to calculate the original count and express as ‘greater than…’ | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e. *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. | | Count typical colonies (large. and strains of other Bacillus spp. 2.

Perfringens Agar (OPSP. 2. similar to the use of different plating media for the isolation of Salmonella. and a blood agar plate to determine haemolytic activity. Incubate plates at 30˚C for 24 and 48hrs.4 x 103) *For liquid samples use 10ml to 90ml No homogenisation is required prior to preparing serial dilutions. and speciate using biochemical tests Colonies can be speciated using Listeriazym (T500). LAB 138) or Buffered LEB (LAB 139) | | Homogenise for 2 mins | | Incubate at 30˚C for 24 and 48hrs | | Streak a loopful (10 µl) onto Oxford Agar (LAB 122) or Palcam Agar (LAB 148).1ml of enriched UVM I into 10ml Fraser Broth (LAB 164) and incubate at 35˚C for 24 and 48hrs | | Streak a loopful of Fraser Broth (10 µl) onto Oxford Agar (LAB 122) or Palcam Agar (LAB 148). and a blood agar plate to determine haemolytic activity. Some workers have reported improved isolation by using both Oxford and Palcam agars together. similar to the use of different plating media for the isolation of Salmonella. | | Examine plates for typical colonies | | Confirm colonies as Listeria spp. Some workers have reported improved isolation by using both Oxford and Palcam agars together. whereas non-salmonellas will grow up to the disc. 10g* sample + 90ml MRD (LAB 103) | | Homogenise for 2 mins | | Prepare serial dilutions by adding 1ml of homogenate to 9ml of MRD (LAB 103) | | Inoculate 1ml of appropriate dilutions into a sterile Petri dish and add aseptically 15ml of molten. Place the disc mid way between the inoculation point and the edge of the dish before incubation. Isolation of Listeria species (the ‘modified USDA’ method) Add 25g sample to 225ml of UVM I (LAB 155) | | Homogenise for 2 mins | | Incubate at 30˚C for 24hrs | | Subculture 0.Clostridium perfringens (Enumeration) Prepare an appropriate 1:10 dilution of the sample e. Isolation of Salmonella species (Semi-solid technology) Isolation of Listeria species (the ‘FDA’ method) Add 25g sample to 225ml of Listeria Enrichment Broth (LEB. and speciate using biochemical tests Colonies can be speciated using Listeriazym (T500).g. If no growth incubate for a further 24hrs | | Subculture spreading growth to XLD (LAB 32) and Brilliant Green Agar (LAB 34) to obtain pure growth for confirmation by biochemical and serological methods Use of a filter paper disc soaked in polyvalent H Salmonella antiserum can increase the specificity of the media. Use of the disc with Diassalm enhances the H2S reaction on this medium. If the highest dilution has more than this use 150 as the figure to calculate the original count and express as ‘greater than…’ | | Calculate the original count in the sample expressed as CFU’s per gram or ml (The number should be given in standard scientific notation e. cooled. xxvi . Add 25g sample to 225ml of Buffered Peptone Water (LAB 46) | | Homogenise for 2 minutes and incubate at 37˚C for 24hrs | | Subculture three drops of enriched BPW centrally onto the surface of MSRV (LAB 150) or Diassalm (LAB 537) plates in duplicate. overlay with more of the same agar to cover surface | | Incubate anaerobically at 37˚C for 24hrs Examine plates for typical colonies| | Select plates with between 15 and 150 colonies for enumeration.When set. | | Examine plates for typical colonies | | Confirm colonies as Listeria spp. Incubate plates at 30˚C for 24 and 48hrs. LAB 109) or Perfringens Agar (TSC.g. | | Incubate with the lid uppermost at 37˚C or 42˚C for 24hrs | | Examine for a spreading zone of growth. The motility of salmonellas will be inhibited around the disc. LAB 194). The area where the motility is inhibited by the disc has a high concentration of Salmonella and gives a good H2S reaction indicated by blackening of the growth around the disc.

Incubate at 41. 1989. Both these formulations require the addition of chemicals not supplied by LAB M. Appl. Bact.1ml of enriched BPW into 10ml of Rappaport Vassiliadis (Soy) Broth (LAB 86). To ensure this lower temperatures are recommended: 41. 42 ± 0.5 ± 0. Isolation of Campylobacter species Add 25g sample to 75ml (or 1:4 w/v) of Campylobacter Enrichment Broth (LAB 135) | | Homogenise for 2 mins | | Incubate at 37˚C for 2-4 hrs followed by a further 14-44 hrs at 42˚C | | Streak a loopful (10µl) onto Campylobacter Blood Free Medium (LAB 112) and incubate at 37˚C for 40-48 hrs | | Examine plates for typical colonies | | Confirm colonies as Campylobacter spp. Typical colony descriptions for Brilliant Green Agar and XLD agar can be found under the individual listings in the main body of the catalogue.. such as Mueller Kauffman Tetrathionate Broth (LAB 42) or Tetrathionate Broth (LAB 97).5 ± 0. (Reference: Peterz M. Wiberg C.5˚C for incubators. and are also available as colony cards which show typical colonies in full colour. and Norberg P. The effect of incubation temperature and magnesium chloride concentration on growth of Salmonella in home-made and in commercially available dehydrated Rappaport-Vassiliadis broths. then an alternative medium may be substituted.1˚C for water baths.5˚C* for 24hrs | | Inoculate selective agars: Brilliant Green Agar (LAB 34) and XLD Agar (LAB 32) | | Incubate at 37˚C for 24 hrs and examine for typical colonies | | Confirm suspect colonies by serological and biochemical methods If Selenite cystine broth is not acceptable because of the use of sodium biselenite. using biochemical tests xxvii .Isolation of Salmonella species (conventional method) Add 25g sample to 225ml of Buffered Peptone Water (LAB 46) | | Homogenise for 2 minutes and incubate at 37˚C for 24hrs | | | | Subculture 1ml of enriched BPW into 9ml of Selenite Cystine Broth (LAB 55A + LAB 44B). *It is critical that the incubation temperature does not exceed 43˚C as this can inhibit some salmonellas. Incubate at 37˚C for 24 and 48hrs | | Inoculate selective agars: Brilliant Green Agar (LAB 34) and XLD Agar (LAB 32) | | Incubate at 37˚C for 24 hrs and examine for typical colonies | | Confirm suspect colonies by serological and biochemical methods Subculture 0.. J. 66 523-528).

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2 Columbia Agar Base Eugon Agar Fastidious Anaerobe Agar (F.B. 1 Bacteriological-High Clarity Agar No. Minimum Q.) Perfringens Agar (O. “Allow to soak times” are not critical. pH: at 20˚C. 1 Beef Extract Bile Salts No.A.A.P. first bring to the boil to dissolve the agar. Peptones and other culture media ingredients MC 7 MC 2 MC 6 MC 29 MC 24 MC 4 MC 19 MC 25 MC 12 MC 33 MC 15 MC 13 MC 402 MC 40 MC 20 MC 23 MC 22 MC 14 MC 18 MC 406 MC 9 MC 10 MC 11 MC 27 MC 17 MC 26 MC 16 MC 3 MC 35 MC 5 MC 8 MC 405 MC 32 MC 1 Acid Hydrolysed Casein Agar No. If agar media are to be dispensed prior to sterilising.C. of the medium. 2 Bacteriological-General Purpose Agar No.P. However it is occasionally necessary to make minor adjustments to meet performance criteria.S.B.S. Storage of Prepared Media – All prepared media should be stored in the dark.) Tryptone Soy Agar (U.A.) Fluid Thioglycollate Medium (U. pour a small quantity into a universal bottle.P. allow to set and plunge the probe into the medium. References A list of related publications and sources of information. For agars. The formulae in this manual and on the product label are adhered to wherever possible.P. Where an organism should show inhibition this could be complete or partial. This short form check should not be confused with a full Q. minor adjustments to the published formula may be made to meet performance criteria. 3 Columbia peptone FMV (Foot and Mouth Vaccine) peptone Gelatine Glucose (Dextrose) IPTG Lactalbumin Hydrolysate Lactose Malt Extract Powder Maltose monohydrate Mannitol Meat Peptone MUG Mycological Peptone Ox Bile Proteose Peptone A Skim Milk Powder Sodium Chloride Sodium Desoxycholate Sodium Thioglycollate Soy Peptone Special Tryptone Tryptone Tryptose X-gal Xylose Yeast Extract Powder PRODUCT CODE Description: A brief outline which may include any of the following information on the medium: ● ● ● ● ● HISTORY MECHANISMS APPLICATIONS RECOGNITION BY REGULATORY/ADVISORY BODIES ADVANTAGES Formula: The product composition in grams per litre.A. appropriate quality control should be performed to demonstrate that there has been no detectable fall off in performance.) Perfringens Agar (TSC) Reinforced Clostridial Agar Reinforced Clostridial Medium (Broth) Thioglycollate Medium (Brewer) Growth characteristics Abbreviation key for colonial descriptions: CV = convex F = flat E = entire P. Anaerobes LAB 160 LAB 24 LAB 90 LAB 71 LAB 25 LAB 109 LAB 194 LAB 23 LAB 22 LAB 64 Brazier's CCEY Agar Cooked Meat Granules Fastidious Anaerobe Agar (F. N. = pinpoint CR = crenated Rz = rhizoid G = glossy D = dull Biomolecular Productss HAL 4 HAL 5 LAB 168 LAB 174 LAB 169 LAB 173 LAB 191 LAB 182 LAB 181 LAB 177 LAB 178 LAB 183 LAB 175 LAB 176 LAB 180 LAB 179 Harlequin™ LB Agar Harlequin™ LB Top Agar LB Agar LB Agar (Lennox) LB Broth LB Broth (Lennox) Luria Bertani (Hi-Salt) Broth NZCYM Broth NZY Broth (NZYM) Superbroth Superbroth with Agar Terrific Broth YPD Broth YPD with Agar 2xYT Agar 2xYT Broth ( ) brackets are used to denote occasional variations.A. Blood Agar Bases LAB 28 LAB 15 LAB 1 LAB 525 LAB 90 LAB 11 Blood Agar Base Blood Agar Base No.S. Records should be kept of these results to help recognise changes in performance over a period of time. 4 Plant Tissue Culture Bacteriological Peptone Balanced Peptone No. Method for reconstitution Distilled water can be substituted for deionised water.Format and abbreviation guide Product Name (Alternative name or commonly used abbreviation) Dehydrated culture media selection guide Agars. organisms – for use every time a new batch of prepared medium is reconstituted.) xxix . If a medium is to be used beyond the suggested shelf life. Appearance: – of the finished cooled medium.P.) Fastidious Anaerobe Broth (F.C.

R.E.S.C.V.Improved) Campylobacter Enrinchment Broth Sorbitol MacConkey Agar (SMAC) D.S.S.R.B.L.S.P.V.R.H.A.C. Diluents/Isotonic solutions LAB103 Maximum Recovery Diluent LAB 100 Ringer’s Solution (1/4 Strength) LAB 100Z Ringer’s Solution (1/4 Strength) . LAB 155 U. Agar-examination of worts. LAB 79 W.S.M.C.A.C.P.R. Mauller Kauffman Tetrathionate Broth MLCB Agar O157 Broth (MTSB) Orange Serum Agar P.A.S.C. Agar-cultivation of Bacillus cereus Palcam Agar Palcam Broth PEMBA Agar . coli/Coliform Medium Harlequin™ mLGA Harlequin™ Tryptone Bile Glucuronide Agar (TBGA) Lactose Broth Lauryl Tryptose Broth MacConkey Agar No.) Tryptone Soy Broth (USP) Food Microbiology LAB 167 LAB 85 LAB 34 LAB 46 LAB 112 LAB 135 LAB 105 LAB 537 LAB 136 LAB 137 LAB 171 LAB 16 LAB 164 HAL 8 HAL 2 HAL 6 HAL 3 LAB 196 LAB 158 LAB 138 LAB 139 LAB 122 LAB 172 LAB 93 LAB 94 LAB 150 LAB 42 LAB 116 LAB 165 LAB 147 LAB 73 LAB 148 LAB 144 LAB 193 LAB 194 LAB 109 LAB 149 LAB 10 LAB 98 LAB 108 LAB 86 LAB 23 LAB 55 LAB 161 LAB 87 Aeromonas Agar Baird-Parker Medium Brilliant Green Agar (Modified) Buffered Peptone Water Campylobacter (Blood Free .B. enrichment LAB 35 T.Bacillus Cereus Medium Perfringens Agar (TSC) Perfringens Agar (O. Cholera Medium Tetrathionate Broth Base XLD Agar Yersinia CIN Agar Identification Media LAB 59 LAB 126 LAB 54 LAB 104 LAB 69 LAB 53 LAB 129 LAB 130 LAB 131 Kligler Iron Agar Lactose Broth Lysine Iron Agar Peptone Water Simmons Citrate Agar Triple Sugar Iron Agar Tryptone Water Urea Agar Urea Broth Neutralising Media LAB 188 LAB 187 LAB 186 LAB 185 LAB 184 LAB 189 D/E Neutralising Agar D/E Neutralising Broth D/E Neutralising Broth Base Letheen Agar (AOAC) Letheen Broth (AOAC) Microbial Content Test Agar Enterococci Test Media LAB 106 Kanamycin Aesculin Azide Agar LAB 107 Kanamycin Aesculin Azide Broth LAB 166 Slanetz and Bartley (m Enterococcus Medium) xxx .B.E. Agar M.B. Agar Base HAL 7 Harlequin™ C.D. Potato Dextrose Agar Pseudomonas Agar Rappaport Vassiliadis Medium (Broth) Reinforced Clostridial Agar-enumeration of anaerobes Selenite Cystine Broth Sorbitol MacConkey Agar Sugar Free Agar-enumeration of organisms in butter and similar products LAB 97 Tetrathionate Broth Base A. MacConkey Agar (with salt) MacConkey Agar (without salt) Mueller Kauffman Tetrathionate Broth O157 Broth (MTSB) Rappapport Vassiliadis Medium S.E. (Bevis)-double indicator LAB 41 C. E. beers and yeast cultures LAB 38 Wort Agar-yeasts in dairy and sugar products LAB 99 Wort Broth LAB 32 XLD Agar Coliform/Enterobacteriaceae Media LAB 51 LAB 91 LAB 60 LAB 61 HAL 8 HAL 9 HAL 3 LAB 126 LAB 196 LAB 45 LAB 5 LAB 72 LAB 573 LAB 31 LAB 88 Brilliant Green Bile 2% Broth E. LAB 27 Hoyle's Medium LAB 123 Kirschner’s-T. with MUG Violet Red Bile Agar (VRBA) Violet Red Bile Glucose Agar (VRBGA) Diagnostic Medical Microbiology LAB 121 Bromocresol Purple Lactose Agar LAB 6 C.A.D.H.V.R.Tablets LAB 101 Ringer’s Solution (Calgon) LAB 102 Ringer’s Solution (Thiosulphate) Enteric Pathogens LAB 167 LAB 13 LAB 34 LAB 46 LAB 112 LAB 135 LAB 161 LAB 3 LAB 29 LAB 65 LAB 537 HAL 1 HAL 6 LAB 110 LAB 116 LAB 150 LAB 30 LAB 2 LAB 42 LAB 165 LAB 86 LAB 52 LAB 44 LAB 55 LAB 96 LAB 97 LAB 32 LAB 120 Aeromonas Agar Bismuth Sulphite Agar Brilliant Green Agar Buffered Peptone Water .P. with MUG LAB 88 V.B. Broth LAB 31 V.pre-enrichment broth Campylobacter Agar (Blood Free .G. Broth M.R.Blood Culture Media LAB 49 LAB 71 LAB 4 Brain Heart Infusion Broth Fastidious Anaerobe Broth (F.B. (Mackey & Sandys)-single indicator LAB 67 G.L.P.Pseudomonas spp. LAB 573 V.coli/Coliform Medium Harlequin™ Listeria Medium Harlequin™ Sorbitol MacConkey Agar (SMAC-BCIG) Harlequin™ Tryptone Bile Glucuronide Agar (TBGA) Lauryl Tryptose Broth Liquid Baird-Parker Medium Listeria Enrichment Broth Listeria Enrichment Broth (Buffered) Listeria Isolation Medium (Oxford) Listeria Monocytogenes Blood Agar (LMBA) M.S. Fraser Broth Harlequin™ E. Agar (Salmonella Shigella Agar) Selenite Broth Selenite Cystine Broth T. 3 MacConkey Broth (Purple) Tryptone Bile Agar V. Desoxycholate Citrate Agar (DCA) Desoxycholate Citrate Agar (Hynes) Diassalm Harlequin™ Salmonella ABC Harlequin™ Sorbitol MacConkey Agar (SMAC-BCIG) Hektoen Enteric Agar M.S.L.B.R. Agar M.L.L.) Plate Count Agar Plate Count Agar A. Medium-investigation of dental caries Refer to other sections for our full range of Medical Products.A.R. coli Medium) Flourescence Agar .L. Broth (Enterobacteriacae Enrichment Broth) Endo Agar Eosin Methylene Blue Agar Harlequin™ E.B.Improved) Campylobacter Enrichment Broth China Blue Lactose Agar Diassalm Easter and Gibson Pre-enrichment Broth Easter and Gibson Salmonella Medium EC Medium (E.R.A.E.P.A.D.Y.

-standard methods Plate Count Agar-for spiral platers or pour plates R2A Medium Tryptone Glucose Extract Agar A.S.S. Transport Medium LAB 124 Amies with charcoal LAB 125 Amies without charcoal LAB 505 Carey-Blair Medium Total Viable Counts LAB 19 LAB 115 LAB 10 LAB 149 LAB 163 LAB 63 LAB 11 LAB 197 LAB 18 Milk Agar Milk Plate Count Agar Plate Count Agar A.E. Malt Extract Broth Nutrient Broth No.Nutrient Media for general use LAB 48 LAB 49 LAB 525 LAB 526 LAB 8 LAB 68 LAB 8 LAB 62 LAB 18 Brain Heart Infusion Agar Brain Heart Infusion Broth Eugon Agar Eugon Broth Nutrient Agar Nutrient Broth 'E' Nutrient Broth No. Sabouraud Liquid Medium U.H. 2 B.Y.P.P.S. Sensitivity Test Media LAB 39 LAB 114 LAB 74 LAB 12 LAB 170 Mueller Hinton Agar (II) Mueller Hinton Broth (II) Nu-sens Agar Sensitivity Test Agar Susceptibility Test 'ISO' Agar Yeasts and Moulds LAB 117 LAB 37 LAB 159 LAB 89 LAB 98 LAB 36 LAB 9 LAB 33 LAB 111 LAB 38 LAB 99 LAB 119 Dermatophyte Test medium Malt Extract Agar Malt Extract Broth O.H.P. Tryptone Soy Broth U.G.S. aureus Streptococci Test Media LAB 523 LAB 92 LAB 35 LAB 75 Azide Blood Agar Base M17 Agar T.P.Y. Medium Todd Hewitt Broth Sterility Test Media LAB 25 LAB 159 LAB 14 LAB 33 LAB 11 LAB 4 Fluid Thioglycollate U.P.A.Oxacillin Resistant Staphylococci Isolation Medium LAB 113Z Salt Meat Broth (Tablets) LAB 84 Single Step Staph Selective Agar (4S)-rapid method for S. Tryptone Soy Agar Water Plate Count Agar (ISO) Yeast Extract Agar xxxi .P.C. 2 Tryptose Phosphate Broth Yeast Extract Agar Water Testing HAL 9 Harlequin™ mLGA LAB 126 Lactose Broth LAB 196 Lauryl Tryptose Broth LAB 5 MacConkey Broth (Purple) LAB 82 Membrame Lauryl Sulphate Broth LAB 80 Minerals Modified Glutamate Broth LAB 163 R2A Medium LAB 197 Water Plate Count Agar (ISO) Refer to other sections for our full range of Water Test Media.A. Agar Potato Dextrose Agar Rose Bengal Chloramphenicol Agar Sabouraud Dextrose Agar Sabouraud Liquid Medium Sabouraud Maltose Agar Wort Agar Wort Broth Yeast Extract Dextrose Chloramphenicol Agar Staphylococci Media LAB 85 LAB 95 LAB 158 LAB 7 LAB 192 Baird-Parker Medium DN'ase Test Agar Liquid Baird-Parker Medium Mannitol Salt Agar ORSIM . Tryptone Soy Agar U.P.

xxxii .

Samples requiring enrichment: Inoculate alkaline peptone water and incubate at 37˚C for 18-24 hr. Aeromonas spp. will give a positive oxidase reaction and demonstrate both oxidative and fermentative metabolism. will remain unchanged in both the butt and slant.0 0.0 0.1.5-3.1 . spreading for single colonies.1 0.025 11. surface spreading for single colonies.5-1. To fully identify colonies as Aeromonas spp. pH: 7. Pseudomonas spp. Calcium and magnesium salts are added to control the permeability of the bacterial cell wall and thus prolong their survival. Screw caps down after autoclaving.0 No growth No growth Shape CV. gonorrhoeae S. this medium will not inhibit those strains of Aeromonas sensitive to ampicillin used in other media.5-3.G Colour Translucent pink Translucent pink LAB 167 Description Aeromonas Agar is a highly selective medium for the isolation of Aeromonas spp.G CV. Amies Transport Medium with Charcoal LAB 124 Description Amies introduced his modification of Stuarts transport medium to overcome a number of problems.5 *The selective nature of the medium may mean occasional strains do not grow.E. Allow to soak for 10 minutes. or grow poorly.3 Sodium thiosulphate Irgasan Brilliant green Neutral red Agar Appearance: Purple. Storage: Poured plates: 7 days at 2-8˚C in the dark (may be extended if moisture tight packaging used). Constantly mix whilst distributing. swirl to mix and sterilise by bringing to the boil. Appearance: Soft translucent gel.C.1 0.0 10. swirl to mix then bring to boil to dissolve agar.5 5. Based on the selective agents.C. mix well and dispense into Petri dishes. Allow to soak for 10 minutes.0mm diameter) should be confirmed as presumptive Aeromonas spp.0 Method for reconstitution Weigh 45. Method for reconstitution Weigh 9. brilliant green and irgasan. 1.44 0. will also be oxidase positive.2 g/litre 5. Formula Sodium chloride Potassium chloride Disodium hydrogen phosphate Sodium thioglycollate Calcium chloride Magnesium chloride Potassium phosphate Agar No.0 8. but do not possess fermentative metabolism.0 5. the above tests should be supported using a proprietary kit such as API 2ONE or Microbact 24E (other products may be available).75 grams of powder.aureus E. Media Range Aeromonas Agar Bile Salt Irgasan Brilliant Green Agar Interpretation Organism Aeromonas spp. Sterilise by autoclaving at 121˚C for 15 minutes. An alternative method is to inoculate triple sugar iron tubes. from food. Inoculation: Faecal specimens: Inoculate surface of medium directly.E.* Pseudomonas spp. pyogenes Storage of Prepared Medium: Capped containers up to 6 months at 15-20˚C in the dark.0 ± 0. organisms: N. by performing an oxidase test and inoculating into Hugh & Leifsons O/F medium. Formulation Beef Extract Meat Peptone Xylose Bile Salts No. The addition of charcoal to the medium extended the survival time of Neisseria gonorrhoeae from 24 to 72 hours. Examine for typical colonies and confirm as Aeromonas spp.2 4.005 0.005 0.2 Minimum Q.1 ± 0. S.0 0. Distribute into bijou bottles filling to shoulder. clinical and environmental samples. Incubation: Incubate plates aerobically at 37˚C for 18-24 hr. coli Size 0. Amies replaced the problem component with an inorganic phosphate buffer system. Inoculation: As LAB 124. coli NCIMB 50034 (inhibited) Confirmation Typical colonies (translucent pink colonies 0. It is used when microscopic examination of a film is an important part of the procedure and the charcoal may interfere with interpretation. Minimum Q.5 grams of powder and disperse in 1 litre of deionised water. Stuarts transport medium suffered from overgrowth by coliforms that were capable of utilising sodium glycerophosphate. 1 g/litre 3. Aeromonas will typically produce an acid butt (yellow) and an alkaline or unchanged slant (red) whilst Pseudomonas spp. disperse in 1 litre of deionised water. organisms: Aeromonas hydrophila NCIMB 9240 E.15 1.2 1. Cool to 47˚C. Amies Transport Medium without Charcoal LAB 125 Description This medium is as LAB 124 without the charcoal. Clear gel pH: 7. Subculture onto Aeromonas Agar.

Inoculation: Heavily inoculate a small area of the plate with a 1.2 4. Colour changes due to production of acidity will develop in a few seconds and should be viewed against a white background. and phenol red to indicate growth of organisms producing acid. References Phillips K. Constantly mix whilst distributing. pH: 7. disperse in 1 litre of deionised water. 58: 296-300. J. Up to 4 organisms per plate can be tested.2 Minimum Q.2 + 0. It is suitable for sterilisation by filtration.0 loopful of a fresh culture of the test organism. Sterilise by autoclaving at 121˚C for 15 minutes.5 grams of powder and disperse in 1 litre of deionised water. a plug taken from an area well away from any growth can be used as a negative control. Mix well and pour.0 0. pH: 7. Capped Container – up to 3 months at 15-20˚C in the dark. 41: 325-328. clear liquid.75 grams of powder.2 Inoculation: Push the swab containing the sample into the gel to approximately one third of the gels depth. Cap tightly and keep cool during transport to the laboratory. A Simple and sensitive technique for determining the fermentation reactions of non-sporing anaerobes.2 ± 0. Cover the plug with bromothymol blue indicator (0. Aseptic Commissioning Broth LAB 157 Description A general growth medium specifically designed for the commissioning of aseptic filling machines in the pharmaceutical and allied industries. Method for reconstitution Weigh 19. (turbidity.1 0.0 5. Bact. 2 g/litre 4.0 15. swirl to mix then bring to boil to dissolve agar. no acid or gas) Inoculation: Dispense medium through filling line into the final packaging. as appropriate.2 Minimum Q. Sterilise at 121˚C for 15 minutes. by filtration. Formula Tryptone Yeast Extract Sucrose Sodium chloride Phenol red g/litre 5. pyogenes Storage of Prepared Medium: Capped containers – up to 6 months at 15-20˚C in the dark. (1976). Appl. Can. organisms: B. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. Anaerobe Identification Medium Base LAB 66 Description A medium introduced by Phillips in 1976 for testing the fermentation capabilities of non-sporing anaerobes. gonorrhoeae S. Comparison with controls is useful.04%). Formula Beef Extract Peptone mixture Sodium chloride Agar No. Health. References Amies. and reducing blockage. Appearance: Soft gel with heavy concentration of evenly suspended charcoal.. Gas production and turbidity also indicate growth and are indicated by bubbles in Durham’s tubes or distortion of plastic packaging.0 0. Typically 20-25˚C for up to 14 days. Appearance: Red. J. fragilis Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Pub. Incubation: As per routine sterility testing laid down in Pharmacopoeias or in house methodology.0 2. organisms: Escherichia coli (acid and gas production) NCIMB 50034 Proteus spp.0 5. swirl to mix and heat gently to dissolve. Allow to soak for 10 minutes. It is a simple formulation incorporating peptone.0 16.2 ± 0. organisms: N. Allow to soak for 10 minutes.C. yeast extract and sucrose as energy sources. Allow to soak for 10 minutes. Screw caps down after autoclaving. Method for reconstitution Weigh 40 grams of powder. 1 g/litre 10.2 .D.R. pH: 7.0 0. disperse in 1 litre of deionised water. C. The base is carbohydrate free and nutritious. Cut off the unwanted swab stick then screw on the cap pushing the swab further down into the gel.C.5 5.Formula Charcoal-activated Sodium chloride Potassium chloride Disodium hydrogen phosphate Sodium thioglycollate Calcium chloride Magnesium chloride Potassium phosphate Agar No.2 1. A modified formula for the preparation of Stuarts Transport Medium. or alternatively. Cool to 47˚C and aseptically add 50-70ml of sterile defibrinated horse blood.15 1. (1967).0 3.1 0.0 Method for reconstitution Weigh 17. Incubation: 37˚C anaerobically for 24-48 hours.C. Before use.005 Minimum Q. Distribute into bijou bottles filling to shoulder. Appearance: Blood agar plate. or suitable container. flood the surface with 1ml of a sterile solution of the substrate under test. Recognition of fermentation: Remove a small plug of agar from below the growth. passing quickly through the filters.

Polymyxin is added to suppress coliforms but some Proteus spp and Gram positive cocci may grow through. (2) S.0 12. licheniformis Proteus spp. B. cereus B. F. and Jongerius. E.G.A.CR.0 10.E.S.2 Minimum Q. aureus isolates on traditional Baird-Parker Medium have to be confirmed with a separate coagulase reaction.0 0. Yellow (white halo) shape & surface F. 1 D-Mannitol Sodium chloride Phenol red Agar No. swirl to mix then sterilise by autoclaving at 121°C for 15 minutes. IV.D. disperse in 1 litre of deionised water.0 LAB 73 Description Introduced by Mossel and his co-workers in 1967 for the enumeration of Bacillus cereus in foods.E. Incubation: 30˚C aerobically for 24-48 hours. Appearance: Cream/pale fawn.E. (1978) Micro-organisms in foods. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: – up to 7 days at 2-8˚C in the dark. The typical reactions of S. coagulans B. cereus from other members of the Bacillus group. Mitteilung: Fleischwritsch. F.2 ± 0.5 grams of powder.0 20. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes.Rz. pH: 7. cereus is mannitol negative and produces red colonies. coli S. 51: 1629-1632 (1971).0 1.0 5.0 10. organisms: B.. Colonies suspected of being S. Formula Beef Extract Balanced Peptone No. D. epidermidis NCIMB 50082 E. aureus colony size (mm) 3. Formula Tryptone Beef Extract Yeast Extract Lithium chloride Glycine Sodium pyruvate Agar No. Alternatively the base medium can be used with the RPF (Rabbit Plasma Fibrinogen) supplement (X086). Allow to soak for 10 minutes. 52: 1160-1162 (1972). Incubation: 37˚C aerobically for 48 hours. aureus should be confirmed by the coagulase test or by a latex agglutination kit.A. Mossel. faecalis E.J.0 7. The medium is highly selective due to potassium tellurite and lithium chloride. 1 g/litre 1.3 . disperse in 900ml of deionised water.025 15. Inoculation: Surface.5 no growth 1.8 ± 0. aureus NCIMB 50080 S.0 2. F.) Mannitol Egg Yolk Polymyxin Agar Baird-Parker Medium Base LAB 85 Description Baird-Parker introduced this complex medium in 1962 to overcome the problems of recovering damaged Staphylococcus aureus from foodstuffs. Mannitol fermentation on this medium produces a yellow colour. pH: 6. opaque with X085. This supplement is a more specific alternative to Egg Yolk Tellurite Emulsion (EYT) for the direct detection of coagulase positive S. 2 g/litre 10. spreading or streaking for single colonies. Cool to 47˚C and aseptically add 100ml of X073 egg yolk emulsion and 2 vials of X074 Polymyxin. Thatcher. 15: 650-653.: Vergleictiende Untersuchungen über die Selektivmedien zum qualitativen und quantitativen Nachweis von Vacillus cereus in Lebensmitteln. : Fleischwritsch. Sulphamethazine may be added to inhibit Proteus spp. T.CR. Translucent. aureus are detected by the egg yolk emulsion: (1) lecithinase production .0 CV. Microbiol. E. The tellurite inhibits most coliforms and is also reduced by S.G. Mix well and dispense into Petri dishes. cereus is indicated by a white precipitate around the colonies. Cool to 47°C and add 50ml of X085 sterile egg yolk tellurite emulsion.0 2. Clarke. white halo Yellow Yellow Yellow Pink (swarms) Yellow Minimum Q. organisms: S..0 Method for reconstitution Weigh 46 grams of powder. For Baird-Parker RPF medium Weigh 5. Glycine and sodium pyruvate are both utilised by staphylococci as growth factors.9 grams of powder and disperse in to 90ml of deionised water.0 2.0 10. pyruvate also neutralises toxic peroxides that may be formed in the medium. Koopman.G. cereus NCIMB 50014 E.D.R. aureus does not always give typical egg yolk reactions. opaque gel. D.D.S.2 Growth Characteristics organism B. The use of the RPF supplement overcomes several disadvantages of EYT: (1) S. subtilis B. ISO7932:(1993) 3/100 1. Enumeration of Bacillus cereus in foods. BS5763 Part 1L:1994. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 3 days at 2-8˚C in the dark. Allow the medium to soak for 10 minutes. M. Cool to 47°C and add 1 vial of X086 RPF supplement. aureus to telluride giving typical black colonies. F. swirl to mix then sterilise by autoclaving at 121°C for 15 minutes. Mitteilung I. colour Pink.0-3. (1967).P. pale straw with X086. The lecithinase production of B. Two reactions on this medium differentiate B.Bacillus Cereus Medium Phenol Red Egg Yolk Polymyxin Agar (P. CV. References Inal. University of Toronto. these are mannitol fermentation and lecithinase production. Appl.E. CV.0 0.0-4.a zone of clearing outside the opaque zone. (2) lipase production . this formula was shown to be the most effective for this purpose by Inal in 1972.D. Inoculation: Surface spread.5 1.CR. Appearance: Pink.C. aureus. Mix well and pour into Petri dishes. Volume 1 second edition. Allow the medium to soak for 10 minutes.C.an opaque zone round the colony.0 10. Method for reconstitution Weigh 65.

0-2.5-1. . A. The presence of bismuth sulphite and brilliant green make this medium highly selective. colour other Agar Base ‘A’: Weigh 36. (H2S-ve strains green) Metallic sheen especially in heavy growth. Green Green Green Green/ Brown (black centre) (black centre) Klebsiella spp.0-3.C.0 0.0 0.E. Bring to boil over a tripod and gauze. 29: 310-311.0 0.0 CV. M’V (1926).5-1.5-2. W. An improved diagnostic and selective medium for isolating coagulase positive staphylococci.L.. E.E. A.5 LAB 13A + LAB 13B Description A modification of Wilson and Blair’s original medium for the isolation of Salmonella typhi and other Salmonella from clinical samples.E.5-2. Chemical Mixture ‘B’: Suspend 18 grams of powder in 100ml of deionised water.0 Bismuth Sulphite Agar (Wilson and Blair Medium) Citrobacter spp. Appl. A combination of bismuth and sodium sulphites affording an enrichment and selective medium for the typhoid-paratyphoid groups of bacteria. Appl.0 10. 1 Ferric citrate BPC Brilliant Green Agar No. coli P. Appearance: Pale green. 2nd edition Univ of Toronto Press. (1986) J. I. egg yolk nonclearing staphylococci. Mix well and pour thin plates. 1.S. (1984) 2nd edition. aureus after heat treatment and after storage of frozen or dried cells.Rz.D. Appl.Rz. Appl Bact.J.M. Bact 27(1): 78-82. References Baird-Parker.-1. Beckers N J.G. 30: 470-474. (1978) Micro organisms in Foods I. organisms: Salmonella sp. typhi colony size shape & (mm) surface 1.E.0 no growth F. Black Black 3.0 5. pH: 7. Speck M. streak out to single colonies. Enterobacteriaceae colony size shape & (mm) surface 1.0 5. Bacteriol.01 20. 1.0 CV.G F. A. Sawhney D. Washington. Baird-Parker.G.0 0. References Wilson.0 5.D.4 0.G. J.G.4 . .0 S.4 grams of powder and mix with 1 litre of deionised water.Rz. Enterobacteriaceae 0.0-2. Ten Broeke. R. on Baird-Parker’s isolation medium for Staphylococcus aureus. P.0 CV. The effect of Recovery medium on the isolation of S. J.E.5-2.G. (1962).C.G F.0 CV. 25(1): 12-19. J.P. Canad.0 CV. CV. Antonie van Leeuwenhoek 33: 220-236.G. Growth characteristics organism S. As the medium contains neither lactose nor sucrose it can be used to detect lactose and sucrose fermenting Salmonella.Growth characteristics(with X085) organism S. single colonies may give rabbit eye appearance Other Salmonella spp.0 no growth Minimum Q. Inoculation: Surface. Bact.5 CV.5 Proteus spp. and Davenport. Bacillus spp. Their significance and enumeration. et al (1984).E. 1.0 0. colour Black other Metallic sheen black deposit in medium. The occurrence of coagulase-positive.2 White Narrow opaque Grey zone of fibrin Black precipitation White Grey Black Brown Black Brown (poor growth) (no growth) (no growth) 0. Cool to 50˚C approx. Bacillus spp. 61:149-155. and add 100ml of Chemical Mixture ‘B’.E. B.C.Rz. aureus colony size shape & (mm) surface 1.C. CV. Store at 4˚C for 3 days to mature. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – store 3 days before use.5-2. opaque gel. Bact 28: 390-402. Add to 1 litre of Agar Base ‘A’ prepared as above.E.0-2. Use within 7 days.G.C.0-3. sewage and other materials. Brown Black Brown (no growth) (no growth) Sodium sulphite Disodium phosphate Glucose Method for reconstitution Growth characteristics(with X086) organism Coagulase positive S. colour Black other Narrow opaque margin surrounded by a 2-5 mm zone of clearing (poor growth) (no growth) Formula Bismuth Sulphite Agar Base ‘A’ LAB 13a Beef Extract Balanced Peptone No. Pathol.G.G. 3/102 1. The use of sulphamethazine for inhibiting Proteus spp. Black/ Green E.E.E. and Blair. J.P-2. CV. F.F. Store at 2-8˚C in the dark.5-1. (1976). Incubation: 37˚C for 24 hours aerobically.E. 2 Bismuth Chemical Mixture ‘B’ LAB 13b Bismuth ammonium citrate g/litre 6.G. before use. Compendium of methods for microbiological examination of foods. International Journal of Food Microbiology (1987) 5:3:200-202.G. E. The Staphylococcus medium of Baird-Parker in practical use. NCIMB 50076 E. . Sterilise for 15 minutes at 121˚C. American Public Health Association. (1964).A.6 ± 0. (1965). and cool quickly in cold water. Smith. saprophyticus 0. Microbiol. CV.0 Other Coagulase negative staphylococci Proteus spp. and Baird-Parker. J. aureus Coagulase negative staphylococci Proteus spp. CV.M.5-2.

CR. streaking out to single colonies.E.G.C. Cool to 47˚C then aseptically add 5-7% sterile.0 5.0 5. – Colistin/Oxolinic acid (XO11) C. Grey Grey Grey Grey Ps. fragilis P.0-3. Weigh 39. perfringens – Neomycin (XO15) (XO16) Staphylococci/streptococci – Colistin/Naladixic acid (XO12) Formula g/litre 15. When the blood is ‘chocolated’ the medium gives good recovery of Haemophilus spp.0 Blood Agar Base No. F. Inoculation: Surface.P. (1972). 2 Method for reconstitution Minimum Q. CV. haemolysis patterns and pigment production of diagnostic value. disperse in 1 litre of deionised water.G.G.0 F. with the addition of 5% sterile blood. anaerobically for 24 and 48 hours. pH: 7.5 P. Allow to soak for 10 minutes.5 grams of powder.0 CV. London 1.P. CV.D. colour White/ Golden other (haemolytic) S. Soak for 10 minutes.4 ± 0. Appearance: Dependent upon blood additive.G.5-1.2 Tryptose Soy Peptone Yeast Extract Sodium chloride Agar No.0 N.2 Minimum Q.0-3. Livingstone. aureus S.5-1.5 . pyogenes ATCC 19615 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. R.5-1.G.5-1.G. pyogenes colony size shape & (mm) surface 1.E.5-2.0 2. aureus NCIMB 50080 S. CV.0-1. C. aeruginosa 1. CV.G. Incubation: 37˚C aerobically or microaerobically for 24 hours.E.5-3. pneumoniae 0.0-1. organisms: S. 11th edn. pH: 7.G. CV. can be used to cultivate a wide range of micro organisms of clinical significance.E. Typical haemolysis patterns are obtained with this medium.-1.5 CV. F.0 F. pneumoniae P.0 1. NCIMB 50080 S. defibrinated horse or sheep blood. anaerobius F.-(E)G B. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C. Inoculation: Surface.E. The medium gives colonial appearances.5 E.G.0 CV. with the addition of blood.0 C. CV.5-2. Grey White non haemolytic 1.Blood Agar base LAB 28 Description An inexpensive general purpose agar base which.5 0. aeruginosa 0. colour WhiteGolden Grey other haemolytic beta haemolytic alpha haemolytic non-haemolytic Growth characteristics organism S.CR.E. streaking to single colonies. disperse in 1 litre of deionised water.CR.G. anaerobically or microaerobically for 24 hours.0-2.E. 1 Sodium chloride Agar No. coli 1.5-1.5 0. aureus S.C.E. fragilis P.5-1.D. is capable of growing delicate clinical pathogens.E. organisms: S.5 P.0 12. CV. pyogenes ATCC 19615 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.P.0 CV. Mix by swirling the flask and pour into Petri dishes.G. Cool to 47˚C and add 5-7% sterile defibrinated blood.5 5.G. perfringens References Cruikshank.0 12. CV. The medium can be made selective for various groups by the addition of appropriate antibiotic mixtures eg: Streptococci – Colistin/Oxolinic acid (XO13) Gardnerella spp. perfringens B. necrophorum 0. coli 2.-1.E.5 Ps. swirl to mix then sterilise for 15 minutes at 121˚C.5 CV.E. Grey Target haemolysis non-haemolytic Grey GreyWhite Transparent haemolytic mucoid N.0 Method for reconstitution Weigh 37 grams of powder.G. 2 LAB 15 Description A very rich agar base which. 2 g/litre 10. Growth characteristics organism S. Medical Microbiology. Formula Beef Extract Balanced Peptone No.P. CV. pyogenes colony size shape & (mm) surface 0.0 E.E.-1. Incubation: 37˚C aerobically. anaerobius 1.CR. meningitidis P.G.E. Grey alpha haemolytic draughtsman Grey Grey Grey mucoid haemolytic many colonial forms green pigment Grey beta haemolytic) (alpha or non haemolytic) Grey (draughtsman (alpha haemolytic) (mucoid) (require CO2) (May require CO2) (haemolytic) (green pigment (haemolytic) “Target”haemolysis (non-haemolytic) non haemolytic S. Mix well before pouring.E.G.E.5 P. aureus. CV.-0.G. Appearance: Dependent upon blood additive. meningitidis 0.4 ± 0.0 10.P.

5-1. clear liquid.5-1. The use of porcine material in this product ensures there are no Specified Risk Materials (SRM’S) with respect to Transmissible Spongeform Encephalopathies (TSE’S).5 10.difficile.G. Finally the addition of lysed horse blood optimises the recognition of colony fluorescence when cultures are examined using UV light.0 5. Growth characteristics (with horse blood) organism S.2 Method for reconstitution Weigh 49 grams of powder. Pbl.g. Formula Brain-Heart Infusion solids (porcine) Tryptose Glucose Sodium chloride Disodium hydrogen phosphate g/litre 17. Allow to stand for 10 minutes then dissolve with gentle heat before dispensing into tubes or bottles.difficile Selectivity is achieved by addition of supplement X093 (cefoxitin cycloserine) and egg yolk emulsion X073 is added to help differentiate C.0 Brain Heart Infusion Broth LAB 49 Description A rich isotonic infusion medium with tryptose (a mixture of meat and milk peptones) providing a wide range of substrates.C.E. pH: 7. Hlth.D. it incorporates additional ingredients to improve the isolation and differentiation of C. faecalis S. A. 3/108 1. Based upon the market leading anaerobe medium. Rep.6 .0-1. Use in conjunction with an anaerobic culture medium e.5 shape & surface CV. The medium is suitable for use as a blood culture medium or as an enrichment broth for fastidious organisms. E.difficile from lecithinase positive clostridia. the phosphate buffer will help neutralise the acids produced from the utilisation of glucose and thus maintain viability.0-4.G.G. 74: 131-149. streaking out to single colonies. (White) Grey/ Green Grey/ Green Grey Grey References Rosenow. Incubation: Time and temperature to suit specimen/organisms. 63: 173-178. (1919). (1944).P. Allow to stand for 10 minutes then swirl to mix.difficile from clinical specimens. The medium is lightly buffered to prevent the early death of some species due to acid production. Inoculation: Surface. J. Sterilise at 121˚C for 15 minutes.25 0. CV. pneumoniae E.5 Method for reconstitution Weigh 37 grams of powder then disperse in 1 litre of deionised water. inf. aureus NCIMB 50080 E.5 12.0 2. subculture after 1. and Clarke.0 2.5 S.5 10. cultivation and pathogenicity of Actinomyces israeli recovered from the human mouth and from actinomycosis in man. focal infection with special reference to oral sepsis. Cholic acid is present to promote spore germination following alcohol shock treatment. T. aureus colony size (mm) 1.E. F.R. Res. clear gel.E. The mixture of brain and heart infusions is particularly useful in the isolation of Actinomyces israeli and Histoplasma capsulatum. 7 and 15 days or immediately on showing signs of growth.4 ± 0. coli NCIMB 50034 Storage of Prepared Medium: Capped container – up to 3 months at 15-20˚C in the dark. disperse in 1 litre of deionised water. Inoculation: (as a blood culture medium).E.CR.0 Brazier’s CCEY Agar LAB 160 Description Brazier’s CCEY agar is the formulation currently used by the Anaerobe Reference Unit for the isolation of C.0 2. and p-hydroxyphenylacetic acid to enhance the production of p-cresol. aureus NCIMB 50080 E. 1:205-267. other Staphylococci 0. milleri E.. J. Dent.E. CV. Minimum Q.0 2. Howell. This medium was first used for the isolation of dental pathogens. Capped container – up to 3 months at 15-20˚C in the dark. Interpretation: Observe daily.Brain Heart Infusion Agar LAB 48 Description A general purpose nutritious agar base. Organisms which produce significant amounts of acid may well overwhelm the buffering system and autosterilise.2 Minimum Q.0 5.C. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.0-3. CV. Cool to 47˚C then pour into Petri dishes. The medium is not recommended for the determination of haemolytic reactions because of the glucose content. Appearance: Pale Straw colour.0 P.0 2. Dis. 2. pH: 7. Incubation: 37˚C aerobically for 7 to 15 days. CV.G.4 ± 0.G. References Roseburg. A low concentration of glucose is used to stimulate early growth. Appearance: Straw colour. a distinctive metabolite of C..C.0-1.5-1. 3. Fastidious Anaerobe Broth LAB 71. Overheating will cause caramelisation and darkening of the medium. organisms: S.0 2. Epps. coli Pseudomonas aeruginosa 0. L. (1948) Efficiency of methods of isolation of Histoplasma capsulatum. A study of the isolation. F.E. Studies on selective localisation.G.E. Formula Brain-Heart Infusion Solids (porcine) Tryptose Glucose Sodium chloride Disodium phosphate Agar No. and completed by Jon Brazier. colour White/ Golden White/ Yellow White Transp.1 1. organisms: S. Fastidious Anaerobe Agar. CV.G.J. Using a minimum volume of 50ml of medium add the blood to a dilution of from 1:10 to 1:20. Sterilise by autoclaving at 121˚C for 15 minutes. With the addition of 7% defibrinated blood the medium will support the growth of a wide range of fastidious organisms. resulting from work initiated by Ken Phillips and Paul Levett.-0. pyogenes S. The use of porcine material in this product ensures there are no Specified Risk Materials (SRM’S) with respect to Transmissible Spongeform Encephalopathies (TSE’S). E. 2 g/litre 17.

Phenolic odour due to the production of p-cresol. and Kamplemacher. fimbriate edge. generally circular but tending to elongate in the direction of spreading. Org. 2-3mm diameter.C. Salmonella infections in nine European laboratories using a standard technique. difficile E. Formula Beef Extract Balanced Peptone No.0 1. The modification was to increase the selectivity of the medium by increasing the dye concentration.5 mm red) LAB 34 Description First introduced by Kristensen et al in 1925 as a selective medium for the isolation of salmonellae (except S. Food Microbiol. Utrecht. The medium is suitable for subcultures from selective enrichment media. (1968).Formula Peptone Mix Sodium chloride Soluble Starch Agar Sodium bicarbonate Glucose Sodium pyruvate Cysteine HCl Haemin Vitamin K L-arginine Soluble pyrophosphate Sodium succinate Cholic acid p-Hydroxyphenylacetic acid g/litre 23. and sterilise by autoclaving at 121˚C for 15 minutes.5-1.C. Incubation: 37˚C for 24-48hrs under anaerobic conditions Characteristics of C.E. (1969). 39: 487-491.H. raised elevation. 2nd end. coli (inhibition) NCIMB 50034 Storage of prepared medium: Plates – up to 7 days at 2-8˚C in the dark.0 yellow colony) (crenated small red colonies) CV. Washington D. American Public Health Association (1966). Incubation longer than 48hrs may result in a lighter gray or white centre to the colony. Pharmacopoeia of culture media for food microbiology. Recommended Methods for the Microbiological Examination of Foods.9 ± 0. the American Public Health Association and the Association of Official Analytical Chemists. typhi References Edel.0 12.D. However because this medium is highly selective. Clinical Infectious Diseases 16 (4) 228-33.0 and Shigella spp.G. BPLS) Enterococcus spp. Appearance: Tan opaque gel. sonnei no growth (0. Confirm by latex agglutination. This medium is definitely not recommended for S.0 no growth no growth Pink/Red (may not grow) References Brazier J.G. Int.P. coli Proteus spp 1.5 S.G. pH: 6. small numbers of salmonellae may be missed. (ed.H.4 1. Hlth. Green (yellow) (NG) colonies Salmonella spp. (1987). (1993) Rôle of the Laboratory in Investigations of Clostridium difficile Diarrhoea. Appearance: Tan.H.0 Method for reconstitution Weigh 52 grams of powder and disperse in 1 litre of deionised water.difficile: Gray opaque flat colonies.0 0. 1 Yeast Extract Disodium hydrogen phosphate Sodium dihydrogen phosphate Lactose Sucrose Phenol red Brilliant green Agar No. CV.S.2 Minimum Q. typhi E. Ps aeruginosa no growth Klebsiella spp. Pour plates and dry the surface before inoculation. Growth characteristics organism colony size shape & (mm) surface CV. Hlth. DO NOT remelt or autoclave: overheating causes precipitation of the medium.C. colour Pink colonies other (red zone in medium) (0. organisms: C. swirl to mix. clear gel.0 + 0. Lecithinase negative.5 0. 2 g/litre 5.7 .0 3. Cool to 47˚C and aseptically add the following: 2 vials of X093.L. Org. Comparative studies on Salmonella isolation in eight European laboratories.2 Method for reconstitution Weigh 48 grams of powder and add to 1 litre of deionised water. Minimum Q. 1-1. E.25 0. organisms: Salmonella sp. Mix well and pour into Petri dishes.E. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. NCIMB 50076 E. W.5 Brilliant Green Agar (modified) (Phenol Red Brilliant Green Agar. Bull. Bull Wld. Sharf) A. 1-1. E. This formulation is quoted by the International Standards Organisation. Inoculation: Surface streaking for single colonies. Washington. 40ml of Egg Yolk Emulsion X073 and 10ml lysed horse blood.0 1. J.0 5. Less inhibitory media such as X. and Hektoen Enteric Agar will be useful in detecting salmonellae and shigellae inhibited by Brilliant Green Agar. Allow to soak for 10 minutes. pH: 7. 5th edn.0 0.01 0. standard European Community Methods. 3/112 1.0 1.5 1. ground glass appearance and a rough. Store plates away from light. Inoculation: Surface streak untreated or alcohol shocked specimens for single colonies.001 1.0 10.0 0. Colonies fluoresce yellowgreen under UV light. a heavy inoculum can be used. J.0047 12.A. Incubation: 37˚C for 18-24 hours aerobically.6 10.M.E.0 10. and Kamplemacher. 513: 245-247.0 1.09 0. Edel. 41: 297-306. Association of Official Analytical Chemists (AOAC) (1978) Bacteriological Analytical Manual. Wld. The medium was modified by the Netherlands Institute for Public Health..0 0. W. typhi).0 0. no growth S. Allow to soak for ten minutes and then bring to the boil with frequent swirling to dissolve the solids and cool to 47˚C in a water bath.

pH: 6.0 Method for reconstitution Weigh 40 grams of powder.0 20. Shane.E. Bacteriological Analytical Manual.4 ± 0. Confirmation by indole production in Tryptone Water LAB 129 (44˚C for E.2 Method for reconstitution Weigh 28 grams of powder and disperse in 1 litre of deionised water. CV. Interpretation: Turbidity.B. Mesopholic coliforms 32˚C for 24-48 hours aerobically. plating either over entire surface for colony count or streak out to single colonies. 1 g/litre 7.C.5-2. coli and thermotrophs 44˚C for 18 hours aerobically. Incubation: 37˚C aerobically for 18-24 hours.E.5 Proteus spp. Int.5 0.L. 5:3:206-207. Washington. American Water Works Association and Water Pollution Control Federation.G. faecalis References Pharmacopoeia of Culture Media for Food Microbiology (1987).C. Infekt. CV.5 0. 5th ed. coli).) Yellow other (mucoid) Yellow (N.C. D.G.Brilliant Green Bile 2% Broth LAB 51 Description A modification of MacConkey’s medium. and comparison studies on Lauryl Sulfate Tryptose Broth as presumptive medium.0 Citrobacter spp. Association of Official Analytical Chemists.E. Washington D.C. pH: 7. coli colony size shape & (mm) surface 1. formulated in 1926 by Dunham and Schoenlein.0-2..0 S. Hyg. (1902). Ensure the Durham tube is free from gas bubbles before commencing inoculation. 4. 2. M.S.. 1. Dispense into tubes or bottles with inverted Durham tubes. Standard Methods for the Examination of Dairy Products. 1.G.8 . C. NCIMB 50076 E. Formula Balanced Peptone No. coli. Inoculation: Surface. American Public Health Association. Formula Peptone mixture Lactose Bromocresol purple Agar No. water and food products.F.G. Allow to soak for 10 minutes. Allow to cool to 47˚C then pour into Petri dishes.G. Hausler. 337. (1947). coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 1 month at 2-8˚C in the dark. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. for the recovery of coliform bacteria in foodstuffs and water.G. Standard Methods for the Examination of Water and Wastewater.C. Water Works Assoc.E. References Drigalski.. colour Yellow Yellow Purple Purple Cream (purple if N.8 ± 0. Uber ein Verfahren zum Nachweis der Typhusbacillen.. Studies on false confirmed test using B. Am.0 CV. Growth Characteristics organism E.G. (ED) (1972).0-1.0-6. The brilliant green and bile inhibit most Gram positive organisms thus overcoming the problem of some Clostridium spp.2 Minimum Q. aureus E.025 12. Z. CV. J.0 10. 13th ed. Lactose fermenting organisms produce yellow colonies. Psychrotrophic coliforms 4˚C for 10 days aerobically. NCIMB 50034 S. J. 39: (4). Allow to soak for 10 minutes. 1978. disperse in 1 litre of deionised water.5 0. non lactose fermenters produce purple colonies. CV. Appearance: Green.G. aureus NCIMB 50080 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.5 Salmonella spp. Sterilise by autoclaving at 115˚C for 15 minutes. Washington. organisms: Salmonella sp.L. broth can be used at double strength if required but cannot be sterilised by autoclaving.E.0133 Bromocresol Purple Lactose Agar (Drigalski agar) LAB 121 Description A non selective differential medium for the isolation and enumeration of Enterobacteriaceae from urine. American Public Health Association. colour changes (to yellow or yellowish green) and production of gas are all presumptive evidence of the growth of organisms of the coli-aerogenes group. 14th ed. organisms: E. CV. Minimum Q.B. (1975). Inoculation: Serial 1:10 dilutions of homogenised sample are inoculated into the broth in the proportion of 1ml sample to 9ml broth. B.0 0. Appearance: Purple. D. clear. fermenting lactose and giving false positive results.G. W. 1.F.0-2.E. Incubation: E. pasteurisation must be used instead. 39:283-300. 1 Lactose Ox Bile Brilliant green g/litre 10.E. CV. clear agar.4 8. J. Food Microbiol. – purple) Klebsiella spp. Association of Official Analytical Chemists (AOAC). swirl to mix then warm to dissolve.

1ml into 10ml Rappaport Vassiliadis Medium LAB 86. Incubation: 30˚C aerobically for up to 48 hours. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 14 days at 2-8˚C in the dark. then sterilise by autoclaving at 121˚C for 15 minutes. coli (inhibition) NCIMB 50034 Candida albicans (inhibition) NCIMB 50010 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Buffered Peptone Water LAB 46 Description A pre-enrichment medium design to help sublethally damaged salmonellae recover before introducing them into a selective medium. Subculture: 10ml aliquots in 100ml of Selenite Cystine Broth LAB 55 and 0.0 1.0 3.4 ± 0. Formula Peptone Sodium chloride Disodium hydrogen phosphate Potassium dihydrogen phosphate g/litre 10. and Silliker J.7 1. Sublethal injury to salmonellae occurs in many food processes and this preenrichment step greatly increases recovery of these organisms. Bull.C.0 4. jejuni E. (1976) Salmonella in Compendium of Methods for microbiological examination of foods. The selective cocktail X112 (X212) makes the medium selective for C. clear liquid. Inoculation: Add 25 grams of sample to 225ml of Buffered Peptone Water and homogenise. coli and C.6 Minimum Q. coli. Washington. This nutrient medium is free from inhibitors and is well buffered to maintain the pH at 7..0 1. 48: 167-174.H. Sterilise by autoclaving at 121˚C for 15 minutes. laridis when incubated at 37˚C. Wld Hlth Org.2 ± 0. C. Appearance: Black agar. pH: 7. Health Ass. 2 Method for reconstitution Weigh 45.5 2.0 5. Inoculation: Add 25 grams of sample to 225mls of Buffered Listeria Enrichment Broth and homogenise. With this product incubation at 42˚C is no longer necessary and higher recovery rates have been reported at 37˚C than at 42˚C. Continuously mix whilst pouring to prevent the charcoal settling.K. on this medium are distinctive. Poemla P. Campylobacter Blood Free Selective Medium (Modified CCDA-Improved) Method for reconstitution Weigh 47 grams of powder and add to 1 litre of deionised water. Incubation: 37˚C for 48 hours in an atmosphere of 5% oxygen.2 for the incubation period.C. laridis surface streaking to single colonies. pH: 7.0 5. fennelliae require up to 7 days. monocytogenes NCIMB 50007 E. Formula Peptone blend Bacteriological Charcoal Sodium chloride Sodium desoxycholate Ferrous sulphate Sodium pyruvate g/litre 25. organisms: E. Appearance: Yellow.0 2.0 3. C. Allow to soak for 10 minutes then swirl to mix and sterilise by autoclaving at 121˚C for 15 minutes. Formula Tryptone Soy peptone Sodium chloride Dipotassium hydrogen phosphate Glucose Yeast Extract Potassium dihydrogen phosphate Disodium hydrogen phosphate g/litre 17. Cool to 47˚C and add 2 vials of X138 (X139/X539 can be used as an alternative) reconstituted in 50% alcohol. Subculture: After 24 and 48 hours onto Listeria Isolation Medium – LAB 122. Incubation: Aerobically at 37˚C for 18-24 hours.2 Minimum Q. LAB 139 is a buffered version of the ‘FDA’ broth LAB 138. Cool to 47˚C then add 2 vials of X112 supplement. Butzler and Blazer-Wang all of which contain antibiotics shown to be inhibitors to C. The extra buffering capacity maintains the pH of the enrichment culture during incubation.25 12. pH: 7. clear. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. The colonial morphologies of Campylobacter spp. cinaedi and C. organisms: L. coli. and Kampelmacher E.2 LAB 112 Description A blood free medium which will support the growth of most enteric campylobacters. The supplement X112 (X212) consists of cefoperazone and amphotericin and is superior to the selective cocktails of Skirrow. C. (1973). Pub.Buffered Listeria Enrichment Broth LAB 139 Description A medium for the selective enrichment of food and environmental samples for Listeria spp. References Edel W. jejuni. jejuni. Mix to dissolve then distribute into tubes or bottles.25 0.2 ± 0.5 Agar No.2 1. organisms: C. Aseptically dispense into sterile tubes or bottles. Method for reconstitution Weigh 20 grams of powder and disperse in 1 litre of deionised water. C.35 9. 10% carbon dioxide and 85% nitrogen.5 6. mix well and pour into Petri dishes. disperse in 1 litre of deionised water and allow to soak for 10 minutes.9 . Inoculation: C. ensuring optimum conditions for the recovery of Listeria spp. Am. Swirl to mix.H.C.0 Minimum Q.0 3. Appearance: Pale straw.0 0.5 grams of powder.

environmental samples and faeces.. Hodge D. Hunt J.G.E. disperse in 1 litre of deionised water and allow to soak for 10 minutes. Incubation: 37˚C in 10% CO2 for 2-3 days.G.0 0. Herbert G.0mg 0. Charcoal-Based. coli C. Appearance: Chocolated Blood Agar.. Incubation: Aerobically at 37˚C for 2-4 hours. Clin Micro.L. and Tran T. disperse in 1 litre of deionised water.V.E. C. equigenitalis E.G. J. J.E.3 0. Smith. laridis C. colour Grey/ White Creamy Grey Grey Pale Grey other Efflorescent (spreading moist) Moist (C. Appearance: Translucent.0 CV.2 Minimum Q. Goosens H.0 10. Clin.0 F. References Bolton..J..Clin. Modified Selective Medium for Isolation of Campylobacter spp from Feces: Comparison with Preston Medium. mix well and dispense into sterile containers.E.0 5. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers: 7 days at 2-8˚C in the dark.G. (1987).E. C.5 0. Dis. Parker G. Further cool to 47˚C before adding antibiotic selective agents.A. Thornley J..A.C. Butzler J. This broth has been shown to give appreciably better results than Preston Broth. or if small numbers of campylobacters are present in heavily contaminated specimens.E. Clin.2 C.3 ± 0..G. 1.0-3.Growth Characteristics organism C.. Micro. (1988) 7 p 155-160. Fleming P. wine-red with a fine black suspension..5 1. The medium is a sugar free base with a mixture of high grade casein and soy peptones as nutrients and with L-cystine and sodium sulphite as supplements and reducing agents. Eur.. The medium is made selective with the addition of amphotericin (5 mg/L) and trimethoprim (10 mg/L). (1987) 40 p 702-703. Cool to 47˚C..N.0 5.6 Tryptone Soy Peptone Sodium chloride Agar No. 25 p 2274-2277. (1998) Chapter 7 Campylobacter in FDA Bacteriological Analytical Manual 8th Edition. De. Hutchinson D.H. Allow to soak for 10 minutes. Inoculation: Surface. Abeyta C. Vlaes L.0 5. The use of a selective enrichment broth enhances the recovery of sub-lethally damaged organisms due to processing of foods. Simor A.10 . Agar Base (Contagious Equine Metritis Organism) LAB 78 Description This medium is a selective isolation medium for Taylorella equigenitalis the causative organism of contagious equine metritis. followed by a further 16-44 hours at 42˚C.O. Coignau H.4 ± 0. Personal Communication..0 5. Minimum Q.5 cm. moist) requires 7 days in absence of X112 requires 7 days in absence of X112 Method for reconstitution Weigh 27. Boeck M. from food.5 1. Infects. and a Filtration System. organisms: Campylobacter jejuni E. (1986) 24 p 840-843... coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. S. For faeces 1ml of a 10% suspension in Buffered Peptone Water LAB 46 is added to 5ml of broth.S. Formula g/litre 15.E. streaking out for single colonies. References Bolton F.0-3.0 5.2 Campylobacter Enrichment Broth (Bolton Formulation) LAB 135 Description A selective enrichment broth for the isolation of Campylobacter spp.6 grams of powder. Lane J. Subculture: Onto Campylobacter Blood Free Selective Medium LAB 112.. Microbiol.Clin..0 2. organisms: T. Reassessment of Selective Agars and Filtration Techniques for Isolation of Campylobacter Species from Feces. Allow to cool to 80˚C. Van Den Borre C. Hutchinson D. Weaver R. Pale Grey Other Enterobacteriacae – No growth if sensitive to cefoperazone.. F. Parker G.J.J.J. F. add 2 vials of selective supplement X131 ( X132 can be used as an alternative) reconstituted with 5ml of 50% alcohol and 50ml of saponin lysed horse blood. Roscoe M. Karmali M. Bolton F. Isolation of Campylobacter: What are we missing? J. cinaedi 1. (1986) 23 p 456-459.5 grams of powder. Rennie R. a Blood Free Medium. jejuni colony size shape & (mm) surface 2.. equigenitalis have been described. Formula Meat Peptone Lactalbumin Hydrolsates Yeast Extract Sodium chloride Haemin Sodium pyruvate α – ketoglutaric acid Sodium metabisulphite Sodium carbonate g/litre 10. Lynch J. (1988) Personal Communication..0-3. Micro.0 F. F. Swirl to mix and autoclave at 121˚C for 15 minutes. Bolton F.P.C..N.M. add 50ml of sterile horse blood and allow to ‘chocolate’. Hollis D.P. Streptomycin (200 mg/L) can also be used but sensitive variants of T.Path..E. Laboratory and Clinical Evaluation of Isolation Media for Campylobacter jejuni J.M. Selective Medium for the Isolation of Campylobacter organisms from Faeces. pH: 7. Richardson H.5-3. Gun-Munro J. fenneliae 2. Inoculation: Food homogenate is added to broth in a ratio of 1:4 (w/v) in screw cap containers leaving a head space of 1. J. Mix well and pour into Petri dishes. Evaluation of a BloodFree.0 0.C. pH: 7.0-2.0 12.G. 2 L-Cystine Sodium sulphite Method for reconstitution Weigh 37. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes.

0 10. epidermidis Growth Characteristics colony size shape & (mm) surface 1.L. spread inoculum evenly over entire surface. 0. Bacteriological techniques in the diagnosis of equine genital infections.G. Allow to soak for 10 minutes.E.S.0 1. 752-756. aureus S.0 0. organisms: P. Salmonella spp.5-1. CV. 1470.G. The formulation is the same as that described by the Methodenkomission für Milchivirtschaft. S.G. Formula g/litre 5. aureus Proteus spp. organism in mixed cultures. M. organism E.R. pale yellow agar. 52-55. N. Appearance: Opalescent.P.G. References Methodenbuch Band VI. coli Proteus spp. fluorescens E.1-1. equigenitalis 0. Rec. 18..2 Minimum Q.5-1. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.5-2.G. CV. Add 10ml of glycerol. Interpretation: Count all colonies Method for reconstitution Weigh 45. J. P.0 5. pH: 7.11 .0 0. Clin. F.5-3.C. Inhibition of the C. 1985.0 ± 0. 2 Method for reconstitution Weigh 35 grams of powder. G.P.2 Minimum Q. aeruginosa to develop typical colonies which will fluoresce in ultraviolet light and produce green pigment.E. pH: 7. aeruginosa NCIMB 50067 E. Tribe. W.D.I.E. allow to soak for 10 minutes then swirl to mix. coli NCIMB 50034 Staph. Rec.0 References Atherton. (1981).0-2.D. Brown V.G. Vet. Incubation: 30-35˚C aerobically for 24-48 hours. Cetrimide inhibits the growth of many micro organisms whilst allowing P. Use of an improved Cetrimide Agar Medium and other culture methods for Pseudomonas aeruginosa.5-1. aeruginosa colony size (mm) 0.3 13. J. LAB 133 Description A medium recommended by the United States Pharmacopoeia for the isolation of Pseudomonas aeruginosa from pharmacological preparations. Atherton. Vet.0 0. coli S. Fleming. 108.4 10. Formula Pancreatic Digest of Gelatin Magnesium chloride Potassium sulphate Cetyl trimethylammonium bromide (cetrimide) Agar g/litre 20.0 2. 432.G. Inoculation: Subculture from enrichment broth. J. CV. (1965). disperse in 1 litre of deionised water.0 0.E. 101. 1.0 0.325 12.M.G.G.5 N. CV.5-2. (1978).6 Balanced Peptone No.G. making it a useful medium for the detection of streptococci and staphylococci as well as coliform organisms. Appearance: Blue clear agar.G. Shigella spp.3 grams of powder.C. N. Inoculation: Surface. disperse in 1 litre of deionised water. organisms: E. Mackintosh.G.E.5 Growth Characteristics organism P. (1977). Sterilise at 121˚C for 10 minutes.E.0 1. Lactose fermenters in milk form blue colonies.2 ± 0. The medium is non-inhibitory to the growth of cocci. 1 Beef Extract Lactose Sodium chloride Aniline blue Agar No. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. streak out for single colonies.0 shape & surface F.Growth Characteristics organism colony size shape & (mm) surface CV. Pathol.0 3.E. Lowbury E. aureus NCIMB 50080 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. T. colour green pigment (non pigment) green/yellow fluorescence green/yellow fluorescence Pseudomonas spp.5 2. colour Cream other colony size variation is common References United States Pharmacopoeia XXI. Rec.E. Personal Communication.G. Subculture is carried out onto the medium after enrichment in LAB 4 Tryptone Soy Broth.CR. China Blue Lactose Agar LAB 105 Description China Blue Lactose Agar is a non-inhibitory medium for the differentiation and enumeration of bacteria in milk. Cool to 47˚C and pour into Petri dishes. M.E.G. CV. Incubation: 37˚C aerobically for 18-24 hours. F.E.0 CV. Vergand Deutscher Landwirtschaftlicher Untersuchungs – und Forschugsanstallen.CR.J. Vet. 2. colour Blue colourless colourless colourless colourless Pale Blue Blue other (mucoid) (spreading) Blue (colourless if NLF) Klebsiella spp.0 Cetrimide Agar U.Rz. CV.

with a description of a new medium suitable for use in routine laboratory practice. Medium (Mackey and Sandys) (Cystine Lactose Electrolyte Deficient-Single Indicator) LAB 41 Description A medium for urine culture first described by Mackey and Sandys in 1960. J.02 15. colour Yellow/ Orange Yellow/ Orange Blue Blue Blue Blue Yellow/ Orange (Yellow-orange if lactose +ve) (Blue if non-lactose fermenting) (Yellow if lactose fermenting) (Green pigment & odour) other (Blue if non-lactose fermenter) mucoid Minimum Q. 1 Beef Extract Tryptone Lactose L-Cystine Bromothymol blue indicator Andrade’s indicator Agar No.G. (Bevis modification) (Cystine Lactose Electrolyte Deficient – Double Indicator) References Bevis.C.0 Method for reconstitution Weigh 36 grams of powder. The absence of electrolytes inhibits the swarming of Proteus spp. Sandys. Appearance: Green/blue clear gel. J.E. pH: 7.E. LAB 6 Description Bevis modified Mackey and Sandys original medium by introducing a double indicator to improve the differentiation of lactose and non lactose fermenting coliforms.G.P. coli colony size shape & (mm) surface 2.5 ± 0.2 Growth Characteristics organism E. CV. Allow to soak for 10 minutes.CR. organisms: E. G.0 3.L.0 3. disperse in 1 litre of deionised water. A modified electrolyte-deficient culture medium. G and F. or halophilic organisms. Diagnosis of urinary infections.H.0-1. C. Method for reconstitution Weigh 36 grams of powder.E. Cool to 47˚C mix and distribute into Petri dishes. 2.02 0.5-1. 3.G.0-4.. This medium may not grow Pasteurella spp.5 CV.0 CV. Tech. F. Inoculation method: Surface inoculation. Allow to soak for 10 minutes. and Sandys. either streaking for single colonies or spread evenly over entire surface for colony counts.D. Differentiation of lactose and non lactose fermenters is achieved using bromothymol blue as pH indicator. clear gel.E.C. (1960). Incubation: 37˚C for 24 hours aerobically. aerogenes Proteus spp. Cystine is added for the benefit of those organisms which have a specific cystine requirement.G. (1968).0 Shigella spp.D.0-3. G.0 0. Ps.D. K.G.0 S.D. Cool to 47˚C and mix before pouring.Med. B.E. T. CV. J. CV.0 4.E.0-4. Lab. The swarming of Proteus spp.0 10. disperse in 1 litre of deionised water. Blue-White CV.E. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C.0 4. Inoculation: Surface inoculation either spreading for single colonies or spread evenly over entire surface for colony counts. will grow many of the more demanding streptococci of Lancefield groups A. 17: 224-233.5-2. 1 g/litre 4. Tech.0 C. CV.128 0. organisms: E.G. pH: 7. J.0-3. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C.E. Incubation: 37˚C aerobically for 18-24 hours.0 2..0 10. Med.5 Other staphylococci 0. Appearance: Green/blue.0-3. 0. aureus NCIMB 50080 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. aeruginosa 1. (1966). Brit.08 15.0 CV.128 0. Med.E..12 .C.G. Formula Balanced Peptone No. 1. 1: 1173. YellowOrange Enterococcus spp. coli NCIMB 50034 S.5 Salmonella spp.E. LAB M C. is inhibited.G.Lab. This medium supports the growth of Streptococcus pyogenes and most other fastidious organisms that do not require blood. 1 Beef Extract Tryptone Lactose L-Cystine Bromothymol blue indicator Agar No.L.L. aureus NCIMB 50080 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. A new medium for preventing swarming of Proteus spp.D.E. 25: 38-41.0 0.3 ± 0. coli NCIMB 50034 S.2 Minimum Q.5 1. staphylococci and streptococci. 1 g/litre 4. Mackey. aureus 1. Formula Balanced Peptone No.H.

R. aeruginosa 0. Clin Pathol. CV. P.CR. 1. coli K.0 5.0 1.E. Salmonella spp.0-3. Columbia agar can be used for the isolation of most clinically significant pathogens. 11th Edn. perfringens – Neomycin (XO15) (X016) Campylobacters . Livingstone. Brit. Storage of Prepared Medium: – up to 6 months at 15-20˚C in the dark.E.L. (1966). Hlth. pyogenes ATCC 19615 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. CV. B.. Anaerobically for 24 and 48 hours.0-4.G. Allow to soak for 10 minutes. 1. E.D. Inoculation: Surface plating.G. 1972.5-2. aureus S.5-1. Clin.E. R.0-3. The medium gives good growth of most organisms and is especially useful in the recovery of fastidious anaerobes.E.G.5 Enterococcus spp.0 Ellner. Blue-white lactose fermenting) CV. defibrinated horse or sheep blood.. Yellow Ps.E.Med. 81:559-562.0-1.D.A. The blood can be ‘chocolated’ if required. and Martin.J. D and Naylor. C.13 .-0.(X214) Staphylococci/streptococci – Colistin/Naladixic acid (X012) Formula Columbia Peptone Mixture Corn Starch Sodium chloride Agar No.E. J.P.5 F.E. Goldberg. G. Dip-slide: an aid to quantitative urine culture in general practice.G.D. CV.0 CV. An improved medium for the cultivation of N.G. (1967).E. aeruginosa 1. 2 g/litre 23. H.5-4. anaerobius 1.0 Shigella spp. β-haemolytic dependent on strain Grey greenish discolouration in medium. Incubation: 37˚C aerobically or microaerobically for 24 hours.5-1. CV. Report.E. Microbiol.0 0. CV. Comparison of isolation of Haemophilus vaginalis (Corynebacterium vaginale) from Peptone-Starch-Dextrose Agar and Columbia Colistin-Nalidixic Acid Agar. colour Yellow Yellow Blue Blue Blue Blue Yellow (yellow if lactose +ve) (blue if nonlactose fermenting) Neisseria meningitidis E.5 P.0 2. J. References Cruickshank. Guttman. Publ. Thayer..G.CR.E.E.G. Method for reconstitution Weigh 41 grams of powder.P. The medium can be made selective for various groups by the addition of appropriate antibiotic mixtures eg: Streptococci – Colistin/Oxolinic acid (X013) Gardnerella spp. F. 2.0-4. meningitidis.5 S. aureus NCIMB 50080 S.G. J. (1976). CV.0-1. E and Vasi.E.0 CV.Med.0-2.G. pneumoniae 0. aureus 1. Medical Microbiology. F.5 CV.5-2. J.CR.(D) colour WhiteYellow other Haemolytic White α.0-3. mucoid in H2/C02 (mucoid) (haemolytic) Ps.2 Method for reconstitution To make Cooked Meat Medium the granules should be added to Nutrient Broth (LAB 14) or Fastidious Anaerobe Broth (LAB 71) in the ratio of 15 grams of granules to 200ml of broth.G.G.G. Drakeford.E. London. Grey White/ Grey References Columbia Agar Base LAB 1 Description A general purpose nutritious agar base formulated by Ellner et al. (1966). 3: 343-345. R.0 12.5-1. Mix well before pouring.0 2. Brit.0 CV. Medium prepared with Nutrient Broth LAB 14 should be re-steamed when used after a period of storage. The medium is also suitable for prolonged storage of cultures. coli (green pigment & odour) other (blue if non lactose fermenters) (mucoid) organism S. aerogenes Proteus spp. organisms: S. (1966).G. pyogenes Growth Characteristics colony size shape & (mm) surface 1. D.H. When further enriched by the addition of sterile blood. perfringens 1. colony size shape & (mm) surface 2. trans/ Grey Opaque/ Grey other Staphylococcus spp. 4:245-247. Amer. (yellow if CV. 45:502-504. Medium made with Fastidious Anaerobe Broth LAB 71 will not require re-steaming after storage. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C. and Sandys. CV. Minimum Q. streaking out for single colonies. Cooked Meat Granules (for Cooked Meat Medium) LAB 24 Description Dried minced ox heart which has been trimmed of excess fat and prepared according to the Martin and Lekpers modification of Robertsons original formulation. Diagnosis of urinary infections.E. Appearance: Cherry red if blood is fresh and well oxygenated.G. Cool to 48˚C and add 5-7% sterile. CV. G.E. F.5-2.. Stoessel.0-3. disperse in 1 litre of deionised water. fragilis P.E.Growth Characteristics organism E. 1: 1173.0 Staph. J. – Colistin/Nalidixic acid (X011) C. 0.J. gonorrhoeae and N.3 ± 0.G. C.0 CV. To sterilise autoclave at 121˚C for 15 minutes in capped tubes which should be tightened after autoclaving to prevent re-oxygenation.C.G.0 3. pH: 7.D.5 1..0 0.5 Opaque many colonial Grey forms (green pigment) (haemolytic) (mucoid) Grey usually target haemolysis (non haemolytic) (mucoid) References Mackey. Cooked Meat Medium should be dispensed into containers allowing at least 2025mm depth of meat particles and a broth supernatant of at least 1015mm.. A new culture medium for medical bacteriology. and Washington.

P. 40: 581-589. disperse in 1 litre of deionised water. The medium uses sodium citrate and sodium desoxycholate as inhibitors. Incubation: 37˚C for mesophiles. New culture media based on Sodium desoxycholate for the isolation of intestinal pathogens and for the enumeration of colon bacilli in milk and water. NCIMB 50076 E. Sterilise at 121˚C for 10 minutes. (1942). Allow to soak for 15 minutes then sterilise by autoclaving at 121˚C for 15 minutes. flexneri group 1.-1.5.G. 193-207. 1.2 Method for reconstitution Use a calibrated scoop to distribute 0. E.G.5 0. streaking for single colonies. 20 327-349. P. (G) Yellow/ (green pigment) Pink References Hynes.0 5.0 10. Interpretation: Reddening of meat – saccharolytic organism – Blackening and digestion – proteolytic organism.0 2. (uninhibited) (G) CV. Appearance: Pale pink. 1. M.E. Sodium thiosulphate is the substrate for the enzyme thiosulphate reductase being broken down to form sulphite and hydrogen sulphide. S. coli 0.E.0 Formula Beef Extract Balanced Peptone No.C. When the medium boils up into the neck of the flask. (1935).0 P.5-1. Cooked Meat Medium Tablets LAB 24Z Description As Cooked Meat Granules (LAB 24) with Nutrient Broth (LAB 14) incorporated and presented in tablet form. Use proportionately more granules and water if greater depths of medium are required. translucent.0 1. Method for reconstitution Add 2 tablets to 10ml of deionised water in a narrow container. from faeces and environmental samples.5 CV.0 5.0-1.0 Pseudomonas spp. Inoculation: Surface. & Bact.G. dysenteriae E. Allow to soak for 10 minutes. Bact. typhi Growth Characteristics colony size shape & (mm) surface 0.D.0 ± 0. Appearance: Granules covered in slightly opalescent pale yellow liquid. Incubation: 37˚C for 18-24 hours aerobically. Add 10ml deionised water.0 Proteus spp. Path.G. LAB 29 Description This is Leifson’s original formulation of this selective medium for the isolation of Salmonella spp.P. before pouring plates.G.0 10.A. (1916) J.5 grams of powder. The isolation of intestinal pathogens by selective media.E. Path. Remove at once and cool to 47˚C approx. appropriate temperature for thermophiles.5-2. Formulation Cooked meat granules Beef Extract Balanced Peptone No. a fine precipitate of desoxycholate may be present which may clear if the pH is increased by the growth of organisms. (G) CV.0 10. pH: 7. Dry the surface before inoculation.5 D.D.-2. 2 g/litre 5. J. Citrobacter spp.E.0 5.5-2.0 CV.14 . Bact. 1 Sodium chloride g/litre 75. pH: 7. and Shigella spp. quickly remove from the source of heat and allow the froth to subside.D.025 12.5-1.9 gram amounts of granules into tubes or bottles. organisms: Salmonella sp.0-2. It has approximately half the quantity of inhibitors used in the Hynes modification. M. colour other transp (black centre) Yellow transp (black centre) (Opaque) (clearing Yellow around colony) transp (more opaque (pinkish) centre) transp(pinkish) transp Red/Pink ppt in medium Red/Pink ppt in medium (black centre) Yellow (black/grey centre) fishy odour (clearing around colony) Other Salmonella spp. 54. 1 Lactose Sodium citrate Sodium thiosulphate Ferric citrate Sodium desoxycholate Neutral Red Agar No. Path.E.5 CV. Return to the heat and allow the foam to boil up into the neck of the flask once more. swirl to mix. S. Liefson.0 5. organism S.0 CV.5 CV. sonnei 1.E. J. 1. 1. 0. Minimum Q.G. CV. The hydrogen sulphide reacts with the ferric ions to produce a black precipitate of ferrous sulphide.E.. This gives a typical black centre to the colonies of most species of Salmonella.2 Inoculation: Samples or swabs directly into the medium.C. References Robertson. DO NOT REMELT OR AUTOCLAVE THIS MEDIUM.Cooked Meat Medium LAB 127 Description This is a complete version of Robertson’s Cooked Meat Broth which was designed to grow all anaerobes found in battlefield wounds during the First World War. CV.0 ± 0.E. then bring to the boil with frequent stirring.E. The medium is based on LAB M Cooked Meat Granules – LAB 24 and Nutrient Broth – LAB 14.0 Method for reconstitution Weigh 45. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. (Desoxycholate Citrate Agar) S.

clear.G.0 0. DO NOT REMELT OR AUTOCLAVE THIS MEDIUM.5-2. streaking out for single colonies. Growth Characteristics organism other S.-1. 1.0-2.5 CV.0 3.2 Method for reconstitution Weigh 52 grams of powder. bile aggregates may appear on the surface on refrigeration.G.A.E.0 5. aerogenes Proteus spp P. flexneri S.G.2 ± 0.0 CV. Yellow (Green pigment) Pink 1. CV.0 5.E.0 CV.E. coli K. sonnei S.CR. Cool to 47˚C then distribute 20ml into 90mm Petri dishes.0 5.4 ± 0.G. Salmonella spp.0 10. Agar (Desoxycholate Citrate Lactose Sucrose Agar) LAB 65 Description This modification of Leifson’s D. P.D.0-4. colourless Slight cloudy colourless Trans. colour Trans.P.G.D. Pseudomonas spp.0 0.E.C.E. To keep the desoxycholate in solution the pH also had to be increased.0 0. The isolation of intestinal pathogens by selective media.L. Incubation: 37˚C aerobically for 24 hours. 1 Beef Extract Lactose Sucrose Sodium citrate Sodium thiosulphate Sodium desoxycholate Agar No.C.G.E.D.G.-2.0 5. Formula Beef Extract Balanced Peptone No. swirling frequently to prevent burning.G. NCIMB 50076 E. Colourless Colourless (black centre) Colourless (Black/grey centre) Red Pink Colourless (No growth) (mucoid) (Yellow) fishy odour (green) Other Salmonella spp. 54: 193-207 Citrobacter spp.(D) Colourless . disperse in 1 litre of deionised water in a two litre flask.0 CV. (1942). The medium was designed to be more inhibitory to commensal flora whilst allowing for adequate growth of Salmonella spp and Shigella spp. sonnei S. typhi colony size shape & (mm) surface 0.P.0 2.0 colour CV. coli 1. The citrate and desoxycholate levels are significantly increased.5-1.0 5.4 8.D. medium which incorporates sucrose as an additional fermentable substrate to differentiate lactose negative sucrose positive coliforms from Salmonella spp. aeruginosa colony size shape & (mm) surface 1. which are sucrose positive.5 12. This medium is unsuitable for the isolation of Yersinia spp. Path.E.(G) (inhibited) Proteus spp.2 Minimum Q. Cool to 47˚C before pouring plates. Simmer for 30 seconds to dissolve. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.CR. CV.5 P. 1. Formula Balanced Peptone No.5-2. 1. J. colourless Red Red Yellow (ppt around colonies) (ppt around colonies) (Fishy odour) other Growth Characteristics organism S.0-2.S.E.5-1. CV. organisms: Salmonella sp.G. CV. CV.0 0.E.5-1. Bact.C.0-2.C.5-2.A. Inoculation: Surface plating.-1.0 10.0 LAB 3 Description A modification of Leifson’s D. 2 g/litre 5.C. Colourless P.pale pink CV.0 CV. Bring to the boil over a gauze.E. Simmer for 1 minute to complete dissolution of the solids. dysenteriae E.G. typhi E. Incubation: 37˚C aerobically for 24 hours. CV. Pinkish Trans.P.0 0. clear.03 Method for reconstitution Weigh 50 grams of powder.C. Inoculation: Surface.E.5 0. CV.5-1.E. Hynes (Desoxycholate Citrate Agar -Hyne’s modification) D.02 12. Appearance: Pale Pink. before use.G.5 1.0 1.0-2.5-1. CV. flexneri S. 2 Neutral Red g/litre 7.0 0. DO NOT REMELT OR AUTOCLAVE THIS MEDIUM.G. pH: 7. Allow to soak for 10 minutes then heat gently with frequent mixing and bring to the boil.E.0 0. Appearance: Pink. Dry the surface by partial exposure. Minimum Q. M. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.15 . organisms: Salmonella sp. CV. (More opaque Pinkish centre) Trans.E. streaking out to single colonies. pH: 7. disperse in 1 litre of deionised water.5 5.E.0 S.A.5 1. medium was introduced in 1942.G. NCIMB 50076 E.D.5 CV. Dry the surface before inoculation.(D) (inhibited) References Hynes. The medium still uses lactose fermentation and hydrogen sulphide production as differential indicators. 1 Lactose Sodium thiosulphate Sodium citrate Ferric citrate Sodium desoxycholate Neutral red Agar No.

Non acid producing colonies are grey to colourless. the medium is reddened.0 12. O. Enterococci Proteus spp. (3): 217-221. 1 Glucose Agar No. pH: 6.0 0. F. and Hersom. colour varies with species.0 References Taplin.2 Method for reconstitution Weigh 27 grams of powder. Yeasts appear as white creamy colonies.. The medium also detects the ‘flat sour’ organism Bacillus stearothermophilus in sugar and other sweetening agents used in the preparation of frozen dairy foods. RzD colour Yellow zone mauve centre Mauve (Yellow halo) Yellow Yellow Yellow (mucoid) Yellow Yellow (spreads) B.E. Formula Tryptone Glucose Bromocresol purple Agar No. count yellow colonies for differential acid producer count.G. New culture media based on sodium desoxycholate for the isolation of colon bacilli in milk and water. mix well and distribute into tubes or universal containers. stearothermophilus Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.References Hynes.5-2. Canned Foods.A. Fungi other than dermatophytes cause the medium to become yellow due to acid production. A.0-3.. H.. 2 Phenol Red g/litre 10. Isolation and recognition of dermatophytes on a new medium. (1942). D. J. 13th Edn. organisms: Aspergillus sp.G.M.E. though these are rarely encountered in lesions associated with ring worm. Standard Methods for the Examination of Dairy Products.P. Allow to soak for 10 minutes. Sterilise at 121˚C for 15 minutes. G.5 2.5 ± 0. For Mesophiles – Aerobically for 48-72 hours at 30-32˚C. (1963).0-1.D CV. Zaias. Washington. (1946).G. Sterilise by autoclaving at 121˚C for 15 minutes. 2 g/litre 10. CV. Incubation: For Thermophiles – Aerobically for 48 hours at 55˚C. Tanner. coli Klebsiella spp.5 0. Blank. 40: 581-589. Food Research 1. Bact. (1956). Incubation: 22-25˚C aerobically for 10-14 days. (1972). J. If incubation is prolonged the medium may become reddened. disperse in 1 litre of deionised water. The medium was designed to detect the thermophilic bacteria causing ‘flat sour’ spoilage of canned foods.0 40. 4th Edn. Note: Do not exceed the times stated for sterilisation.E. Rebell.C.16 . Champners. Minimum Q. This medium helps in the differentiation between saprophytic and environmental fungi.) LAB 117 Description A modification of the formulation of Taplin. (1935).9 ± 0.G. Path. Bact. overheated acidified agar loses gel strength and the sugars are caramelised.H. pH: 5. Inoculation: Pour plate technique.E.0 5. Capped container – up to 3 months at 15-20˚C in the dark. 1.C. allow to cool in the sloped position.G. The Microbiology of Food 2nd edn. London. pre-heat sample by steaming for 20 minutes if a spore count is required. Appearance: Purple clear agar. Dissolve 2 vials of Chloramphenicol X009 (X209) in ethanol and add these to the agar. The isolation of intestinal pathogens by selective media.0 0. 54: 193-207. cereals and other food products. (DTM) Arch. Growth Characteristics organism colony size (mm) shape & surface Rz.5 1. clear gel.C. Dermatol. Formula Balanced Peptone No. stearothermophilus 2. Tryptone Medium for the Detection of Flat Sour Spores. N. Interpretation: Count all colonies for total counts.0 Bacillus spp. swirl to mix then bring to the boil to dissolve agar before dispensing in 20ml amounts for poured plate technique.2 Method for reconstitution Weigh 62 grams of powder. Interpretation: Dermatophytes appear as fluffy colonies. References Williams. American Public Health Association. 1. (1969). Appearance: Orange. Garrard Press. 99: 203-209.5-3. organisms: B. Hausler A. J. CV.2 Minimum Q. Churchill.5-1. S. Dextrose Tryptone Agar LAB 20 Description A medium for the enumeration of thermophilic spore bearers in foods. Storage of Prepared Medium: Slopes – up to 1 month at 2-8˚C in the dark. Ed. Zaias. Baumgartner.5 1. Path. disperse in 1 litre of deionised water.D Rz.0 Dermatophyte Test Medium (D. M. Rebell and Blank for the detection of dermatophytic fungi. aureus E. Inoculation: Surface plating or stab inoculation. W.0 0. E.04 12. NCIMB 50097 Trichophyton sp. Blastomyces..J. Leifson.B.W.T. CV. Histoplasma and Coccidiodes may also turn the medium red. Allow to soak for 10 minutes then bring to the boil with frequent stirring.

H.M. Diagnostic semi-solid media based on Rappaport-Vassiliadis Broth for the detection of Salmonella spp. saccharose combined with bromocresol purple.0 7. The efficiency of Diassalm is due to the ability of salmonellae to move through the highly selective mobility medium in a Petri dish. D. epidermidis NCIMB 50082 Storage of Prepared Medium: Plates – up to 7 days: at 2-8˚C in the dark. as developed by Van Netten et al (1991). and Van der Moosdijk.17 . and S.. Food Protection 50.0 grams of powder. Allow to cool to 47˚C then pour into Petri dishes. from food and water. Keep the lid uppermost at all times. Appearance: Pale cream.3 ± 0. Four or more organisms can be tested on one 90mm Petri dish.. Proceedings of the 10th Symposium on the quality of poultry meat.08 0. disperse in 1 litre of deionised water.Green transparent. Diassalm can be seeded after preenrichment or after 8hr enrichment in selective broth (De Smedt and Bolderdijk 1987). J. non-motile salmonellae may be present. Food Protection 52. Evaluation of semi-solid Rappaport medium for detection of Salmonellae in meat products. P. It is an improved modification of MSRV (De Smedt and Bolderdijk 1988) and SR (Perales and Audicana 1989) with regard to the composition of the basal medium. Letters in Applied Microbiology Van Netten. Gram positive. Appearance: . DN’ase producing organisms will be surrounded by a clear area where the DNA has been broken down into fractions which are not precipitated by the Hydrochloric acid.. D. Minimum Q.0 2. H. enteritidis in foods. DN’ases produced by the organisms hydrolyse the DNA molecule to a mixture of smaller mono and poly nucleotides. (1992). Agar No. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. 688 De Smedt J.1 g/litre 20. I and Audicana. Proceedings of the International Symposium of Salmonella and Salmonellosis. When the mobility zone is absent. 16. Perales. whilst the double diagnostic system allows visualisation of motile and non-motile suspected salmonellae due to blacking zones against the turquoise background. catalase positive cocci that 1.. (1968) Appl. bring quickly to the boil. Capped container – up to 1 month at 4˚C in the dark. A. (1992).1ml) of 8 to 20hr. A. LAB 537 Description Diassalm. Allow to cool to 47˚C and add 1 vial of Novobiocin supplement – X150 (10mg/vial). aureus strains from clinical specimens. The use of diagnostic selective semi-solid medium for the isolation of Salmonella enteritidis from poultry. and Van Netten. H. (1991).037 11.. Mix well.0 6. clear.0 12.C. Van der Zee. is a semi-solid differential medium for the isolation of Salmonella spp. and Mossel. Mix well and pour plates. LAB M have substituted their raw materials into Blazevic’s formula to create a richer base for Diassalm.. magnesium chloride and novobiocin.’ase AGAR LAB 95 Description DN’ase agar provides a convenient means of identifying potentially pathogenic staphylococci. Van de Moosdijk. Van Netten. Berlin.N.A. Nitrofurantoin may be used instead of Novobiocin to improve the isolation of S. and ferro-iron in combination with thiosulphate. DiSalvo observed perfect correlation between coagulase activity and DN’ase production using S.0 Method for reconstitution Weigh 53. Proceedings Third World Congress Foodborne infections and intoxications. Inoculation: Use a heavy inoculum on a small area. I. This will precipitate the DNA in the medium..0 5. P.8 References Blazevics.Diagnostic Semi-Solid Salmonella Agar (Diassalm) According to Van Netten and Van der Zee et al inoculum (or the blackened centre if non-motile) is sub-cultured onto brilliant green agar and XLD agar.5 0.. organisms: S. A loopfull of the motile zone which is the farthest from the sample Interpretation: Having obtained good growth flood the plate with 1N hydrochloric acid. aureus NCIMB 50080 S. Incubation: 37˚C aerobically for 18-24 hours. Detection of Salmonella spp. Microbiol. Futher biochemical and serological identification are performed according to recognised procedure.2 pH: 5. Van der Zee. soft gel Sodium chloride Agar No. Formula Tryptone Deoxyribonucleic acid (DNA) g/litre 20. 2 Method for reconstitution Weigh 39 grams of powder. D. enteritidis. selective system and the introduction of a differential system. Incubation: At 42 ± 0. Allow to soak for 10 minutes. Spelderholt Beckbergen.5 ± 0. The original basal medium was a commercially available sulphide mobility-indole medium (SIM BBL) (Blazevic 1968). Formula Tryptone Meat Peptone Ferrous ammonium sulphate Sodium thiosulphate Sucrose Lactose Bromocresol purple Malachite green oxalate Magnesium chloride anhyd. with the use of a standard method. Van der Zee. The diagnostic properties of Diassalm are based on the use of two indicator systems. diagnostic semi-solid agars and immunocapture kit. 658 Perales. et al 1987 J. P. Direct latex agglutination may also be carried out from the edge of the mobility zone. Interpretation After incubation the plates are examined for a mobility zone with a purple/black colour change.2 Minimum QC organisms: Salmonella typhimurium NCIMB 50076 Proteus mirabilis (Inhibition) Storage of Prepared Medium: Plates – up to 7 days: at 2-8˚C in the dark. Other publications have also reported a close correlation. disperse in 1 litre of deionised water.5 0.1 0.5 or 37˚C for 18-24 hours.A. pp. Selectivity is achieved by the use of malachite green oxalate. pH: 7. Inoculation: 3 drops (0.2 5. based on the ability of coagulase-positive species to split DNA.0 2.J. but the centre is blackened. incubated preenrichment broth are inoculated in one spot in the centre of one plate of Diassalm. 59-67.

M.).M. Gibson D. W. Cann P.S. J. The deoxryibonuclease test as applied to certain gram-negative bacteria. Gen. Fed. Sub-culture: Into LAB 137 Easter and Gibson Salmonella Medium.. 1970. Some modifications to the media for rapid. B. Messinova. organisms: E. disperse in 1 litre of deionised water. 245-262. J.C. pH: 7. The Streptococcus. Zierdt. Inoculation: From LAB 136 Easter and Gibson Pre-Enrichment Broth.2 Minimum Q.A.5 2. 363-387. to T. These media are both manufactured with the same peptones. J. disperse in 100ml of deionised water. G. Yusupova. and McKechnie.M. W. H. DO NOT AUTOCLAVE. Med. Method for reconstitution Weigh 25 grams of powder.C.. (1987). A. Incubation: 37˚C aerobically connected to monitoring equipment. J and Ewing. Hyg. Salmonella reduce T. J. 22.produce DN’ase can be provisionally classified as S. so that organisms “switched on” to utilising the substrates in LAB 136 can carry on metabolising the same substrates in LAB 137. Streptococcal deoxryribonuclease. V. to produce a significant change in the medium that can be detected by the conductance/impedance equipment. 616618. Pseudomonas aeruginosa. Appl. T1033. NCIMB 50076 E. Heat gently to dissolve supplement. J. 1964. References References Baird-Parker. Ogden I. Streitfeld. Weckman.5 2.2 Minimum Q. This broth is developed from LAB 46 Buffered Peptone Water and will enhance the recovery of sub-lethally damaged bacteria.O. 20(1). (trimethylamine-N-oxide).O. J. Uhr (ed.O.0 5. 54-57. 77. Black.5 grams of powder. C. Deoxyribonuclease activity of Corynebacterium and its relation to virulence. 1958 Deoxyribunuclease and coagulase activity of micrococci. Keep refrigerated. heat to boiling to effect sterilisation. and Shamsutdinov. J. Appl.C. then sterilise by autoclaving at 121˚C for 15 minutes. 191.0 5.A. 94.2 ± 0.M. and Janklow. pp. Gibson D. This medium is based on LAB 55 Selenite Cystine Broth and is supplemented with dulcitol and T. (trimethylamine) and. 13. W. Appearance: Clear reddish/orange solution.C.M. Some corynebacteria and streptococci may also produce DN’ase.M.D. Bacteriol.A. 1967. Baltimore: Williams & Williams. Incubation: 37˚C for 18 hours aerobically. Bacteriol. Proc. O. DiSalvo. Rheumatic Fever.O. S. D. Easter and Gibson Salmonella Medium LAB 137 Description A medium for the rapid detection of Salmonella spp by conductance/impedance techniques. When cool add 1ml of stock solution of L-cystine. Add 1 vial of X137 T. and Catlin. 140-165. M.1gm L-cystine dissolved in 15ml of normal NaOH – add to 100ml sterile distilled water. Microbiol.56 1. D.A. Discard after 1 month.A. Bacteriol. Stock solution of L-cystine 0. H. 1957 Deoxryribonuclease activity of micrococci from clinical sources. DN. A. W. Deoxryribonuclease-positive Staphylococcus epidermidis strains.M. 1. 84. increase the conductivity of the medium which can be detected and measured by monitoring equipment such as Malthus. Easter and Gibson Pre-enrichment Broth LAB 136 Description A pre-enrichment broth for use in detection procedures utilising conductance/impedance techniques. (1987). Tech. dispense into appropriate containers. coli NCIMB 50034 Storage of Prepared Medium: Capped container – up to 1 month at 15-20˚C in dark. W. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: capped container – up to 1 month at 15-20˚C in dark. 9. Bull.0 Method for reconstitution Weigh 2. These media are both manufactured with the same peptones. J. 1965. H. Glomerulophritis. Salmonella reduce T. organisms: Salmonella sp. 1963.’ase is also produced by some Gram negative bacilli such as Serratia marcescens. Wannamaker. W. Distribute into sterile conductance tubes or bottles. Bacteriol.0 Formula Meat Peptone Tryptone Sodium dihydrogen phosphate Dulcitol Sodium carbonate g/litre 2. and Golde.18 .M. in so doing. U. Microbiol. Martin. Can. 38. 63. B. A slight precipitate may form. Best results will be obtained by those customers using PreEnrichment Broth (LAB 136) and Salmonella Medium (LAB 137) bought from LAB M.5 5.. A modified conductance medium for the detection of Salmonella spp..2 ± 0. The classification of staphylococci and micrococci from world-wide sources. Evaluation of extracellular deoxyribonuclease activity in Pseudomonas. 1971. pH: 7. Armed Forces Med.5 10. L.5 5. 459-464. so that organisms ‘switched on’ to utilising the substrates in LAB 136 can carry on metabolising the same substrates in LAB 137 significantly shortening the lag time.0 3. M. J. Rapid and automated detection of Salmonella by electrical measurements. N. aureus. W. automated detection of salmonellas by conductance. 63. R. M. W. (1985). Appl. Swirl to mix. Microbiol. significantly shortening the lag time. Appearance: Very pale yellow clear solution. Best results will be obtained by those customers using PreEnrichment Broth (LAB 136) and Salmonella Medium (LAB 137) bought from LAB M. 747-753. Hoffmann. Formula Tryptone Meat Peptone Sodium chloride Disodium hydrogen phosphate Potassium di-hydrogen phosphate Mannitol g/litre 7. E. V. Easter M. 73. Inoculation: Homogenised food samples. A./Selenite supplement. C. Hodgson.. 1962. J. 299-304. and confirmed by tube coagluase or thermostable DN’ase tests.

4˚C for 10 days for psychrotrophs. clear.References Easter M.0 1. coli K. Inc.5 15.0 0. 63. OVERHEATING THIS MEDIUM WILL ADVERSELY AFFECT ITS PERFORMANCE.0 20.C.0 6. E. A. Interpretation: Turbidity and a colour change to yellow-green is presumptive evidence of Enterobacteriaceae.0 CV. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Add 4ml of a 10% w/v alcoholic solution of basic fuchsins (95% ethyl alcohol). Appearance: Green. J. 1. 32˚C for 24-48 hours for mesotrophs. G. Harrewijn.. and Cornelissen. 63. J.E.C.0 5. Standard Methods for the Examination of Water and Wastewater.A. In: Quality assurance and quality control of microbiological culture media. The precipitate typically associated with this medium should be dispersed by gentle swirling prior to pouring the plates. D. Broth uses glucose instead of lactose to make the medium a test for all enterobacteria including non lactose fermenting organisms.5-1.. disperse in 1 litre of deionised water. F.B. American Public Health Association. Standard Methods for the Examination of Dairy Products. E. 1974. 51-57. Gibson D. Inc. whilst Gram positive organisms are mostly inhibited.. Washington. 1975. Mossel. Inoculation: Surface. edited by J.0 Shigella spp 0. Some modifications to the media for rapid automated detection of salmonellas by conductance. 444-452. organisms: E.2 Method for reconstitution Weigh 43. American Public Health Association. M. Sci. 245-262.G. preferably under refrigeration. Monitoring the quality of selective liquid media by the official French dilution technique used for the bacteriological examination of foods. It is recommended by the American Public Health Association as a standard medium for the enumeration of coliforms in water and dairy products. LAB 88 V.E.45 2. 1987. 1963. G. Bacteriol.Appl..5 Gram positive no growth.G.D..0-1. Basic Fuchsin is a potential Carcinogen and care should be taken when handling it to avoid inhalation of the powdered dye and contamination of the skin. A modified conductance medium for the detection of Salmonella spp. Cann P. L. 26.-Verlag Darmstadt.M. with an improved buffering capacity to encourage early growth and prevent autosterilization. 1 Lactose Dipotassium phosphate Sodium sulphite Agar No.E.0 10. 11. 299-304.0 3. Gibson D. 260-267. 1987. Orig. Subculture onto confirmatory media e. Cool to 47˚C in a water bath before pouring. warm gently if necessary. streaking out for single colonies. 359-464. 1 Method for reconstitution Weigh 41 grams of powder. 1972.5 2. Washington D. 35: 109.I. A. Richard. aeruginosa 0. Health Lab.0-3. Swirl to dissolve. CV. subtilis (inhibition) Storage of Prepared Medium: capped containers – up to 3 months at 15-20˚C in the dark. R. M. 14th Edn.5 ± 0. Minimum Q. C. Inoculation: Add 1 part of sample suspension or dilution to 10 parts of medium. then distribute into bottles or tubes and heat at 100˚C for 30 minutes only. The examination of foods for Enterobacteriaceae using a test of the type generally adopted for the detection of salmonellae.D. 13th End.5-1. Appearance: Pale pink/orange pH: 7. Most enteric Gram negative organisms will grow well. CV. Centr.Appl.19 . Endo Agar LAB 60 Description This medium was developed in 1914 for the isolation of Salmonella typhi. American Public Health Association.2 Minimum Q.C. Formula Balanced Peptone No. Appl.2 ± 0. organisms References Mossel. CV. J. Sterilise by autoclaving at 121˚C for 15 minutes. Incubation: 44˚C for 18 hours for thermotrophs.T. Bacteriol. A.E.0-2. American Public Health Association. Standardisation of the selective inhibitory effect of surface active compounds used in media for the detection of Enterobacteriaceae in food and water.0135 Balanced Peptone No. C. D. pH: 7. aerogenes Proteus spp colony size shape & (mm) surface 1. 1914. coli NCIMB 50034 B. Broth is recommended as an enrichment medium when examining food and feedstuffs for Enterobacteriaceae. A. Corry. A. References Endo. Rapid and automated detection of Salmonella by electrical measurements. 1985.5 grams of powder and add to 1 litre of deionised water. colour Red Pale Pink colourless Pale Pink Pale Pink other (mucoid) Deep Red (Metallic sheen) Ps.0 Salmonella spp 1. Bacteriol.0-2. must be carried out. Formula g/litre 10. pp. It is a modification of LAB 51 Brilliant Green Bile Broth. This medium is light sensitive and should therefore be stored in the dark.. Growth Characteristics organism E. 1 Dextrose Disodium hydrogen phosphate Potassium dihydrogen phosphate Bile Salts Brilliant green g/litre 10. organisms: E.C. Bakt. Bring to the boil with frequent swirling to dissolve the solids. F.0 E.E.g.E.G. D.E. M. 1982. Hyg.G. Abt 1. but Endo agar has a role as a coliform medium. Visser. and Nesselrooy-van Zadelhoff. other media have since proved superior for this purpose. Incubation: 37˚C for 18-48 hours aerobically. E. M. In this medium acetaldehyde is produced by coliforms and then fixed by the sulphite to produce a metallic sheen with the basic fuchsin dye.CR. Ogden I. N. Broth (Enterobacteriaceae Enrichment Broth) LAB 91 Description E. 94.5 2. The medium will become dark red in colour if exposed to light.G.C. J.R.. Cool rapidly.

Eosin Methylene Blue Agar (Levine) LAB 61 Description This medium was introduced in 1916 by Holt-Harris and Teague to differentiate Escherichia spp. Inhibitory effect of light on growth supporting properties of Eosin Methylene Blue Agar. pH: 7. disperse in 1 litre of deionised water.. asaccharolyticus.G.. soluble pyrophosphate for Porph.E. (1976).0 shape & surface colour other (Metallic sheen) (mucoid) CV. J. 1. Gram negative anaerobes Non-sporing anaerobes Actinomyces spp. CV. Washington.0 Fastidious Anaerobe Agar (F. Appearance: Blue/purple with a light precipitate.0 0.0 12. 14th Edn.0 Salmonella spp. Pyruvate helps neutralise hydrogen peroxide and is also utilised by Veillionella spp.20 .2 ± 0. D. American Public Health Association. arginine for Eubacterium spp.2 Minimum Q.5 P. Incubation: 37˚C anaerobically with 10% CO2 for 48 hours to 5 days.0 1. organisms: B.0 0.M. American Public Health Association.7 1. Haesler. fragilis P.4 0.2 Agar No.G. 31:1 141-142. pH: 6. Appearance: Red due to addition of blood. Infect.1% glucose. Blue Black CV.0 1. J.G.3 0.E. streaking out to single colonies.C. Allow to soak for 10 minutes.C.065 15. streaking for single colonies. The low level of glucose prevents the production of high levels of acids and alcohols which would inhibit colonial development.0 0. and Stamm. Vitamin K and sodium succinate provide essential growth factors for some anaerobes as does the 0. organisms: E.G.0 5.0-3. Developed by LAB M.0-4. Girolami. aureus E.G. 23: 43-47. Propionibacterium acne and Bacteriodes fragilis. other coliforms do not produce enough acid to cause this reaction. References American Public Health Association. 1 Lactose Dipotassium phosphate Monopotassium phosphate Eosin Y Methylene Blue Agar No. Microbiol. STORE IN THE DARK.P.25 0. Formula Peptone mix Sodium chloride Soluble starch g/litre 23.A. Cool to 47˚C then aseptically add 5-10% of sterile defibrinated horse blood..P. coli and B. gingivalis and Porph.0 0. Standard Methods for the Examination of Water and Wastewater. Cool to 50˚C and agitate gently to ensure uniform distribution of the flocculant precipitate which is a feature of this medium before pouring into Petri dishes. Dis. coli on this medium is due to acid production resulting in an amide bonding between the eosin and methylene blue. (1918). M. CV.) LAB 90 Description A primary isolation medium capable of growing most clinically significant anaerobes. Eosin inhibits most Gram positive organisms.A.0 0. 2. Inoculation: Surface.C. (D) CV.4 1. S.E. P. 3. anaerobious Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. This medium can be made selective for various species of anaerobes by the addition of appropriate selective cocktails e.. Formula Balanced Peptone No. mix well and pour into Petri dishes.8 ± 0. (1975). CV. The peptones included have been chosen for maximum growth stimulation.5 0. Brown Blue Colourless Colourless White Colourless Colourless Klebsiella spp. Washington.. American Water Works Association and Water Pollution Control Federation. Standard Methods for the Examination of Dairy Products. comparisons have shown this medium to be superior to other formulations as a primary isolation medium for fastidious organisms. 2 g/litre 10. The prepared medium is sensitive to light. Appl.C.01 0. J. Incubation: 37˚C aerobically for 24 hours. Inoculation: Surface plating. Specific growth promoting agents are Cysteine for Fusobacterium necrophorum. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. R.Rz.g. 13th edn. Levine. Starch and sodium bicarbonate act as de-toxification agents whilst haemin encourages pigment production in Porphyromonas melaninogenicus.E. 2 Sodium bicarbonate Glucose Sodium pyruvate Cysteine HCl monohydrate Haemin Vitamin K L-Arginine Minimum Q. D. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Environ. X291 X092 X093 Growth Characteristics organism E. aerogenes on a simplified Eosin-Methylene Blue agar. The distinctive metallic sheen produced by E. W.E.L. The blood will darken (reduce) because of the presence of reducing agents. coli colony size (mm) 2. Clostridium difficile X090.0-3.001 1.G. Soluble pyrophosphate Sodium succinate Method for reconstitution Weigh 46 grams of powder and add to 1 litre of deionised water. Candida spp. Differentiation of E. as an energy source. CV. and Aerobacter spp. (ed) (1972).0 10.5 Method for reconstitution Weigh 37.5 grams of powder.G. faecalis 1. Allow to soak for 10 minutes. X290 X091. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes.0-2.E. It was modified by Levine in 1918 who removed sucrose from the formula and increased the lactose content.0 Shigella spp.5-1.

P.005 0. 8: 83-85. Price. Dent. Tillotson. Pathol. References Brazier. Formula Peptone mixture Yeast Extract Sodium thioglycollate Sodium chloride Agar No.0 A.5 0.G. G. J.A.C. 34: 930-934. Brazier. Fastidious Anaerobe Broth (F. Res. Comparison of a home made blood culture broth containing a papain digest of liver.0-2. May have a narrow band of red/purple at the surface due to action of oxygen on the resazurin. J.L. Microbiol.21 . (clearing) pitting Minimum Q.5 0. K. 38: 1146-1149.2 ± 0. (1982). Pathol. Tillotson. More Rapid identification of bacteraemia by manual rather than radiometric methods. M. fragilis C. colour Grey Grey ‘Target’ haemolysis (non haemolytic) (grey) (haemolytic) other Method for reconstitution Weigh 29. Clin. The medium is very rich in nutrients from the specially selected peptone mixture. and Wade W. Wade W. Selective and differential medium for isolation of Bacteriodes ureolyticus from clinical specimens.. Eley.H. O’Hare. J. J. 60: 121-126. Grey CV..I.E. Comparison of Media for cultivation of subgingival bacteria. Allow to soak for 10 minutes.75 0. viscous. israeli 0. Boil to dissolve the agar then dispense into screw cap containers.E. Heginbotham M. 17: 311-315. anaerobius 0. (1987). with four commercially available media. fragilis Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.B. Blood culture – current state and future prospects. Yellow fluorescence of Fusobacteria Letters in Applied Microbiol. clear. ureolyticus Prop. References Gould. B.G.2 F. Fraser.0005 Fluid Thioglycollate Medium (U.A.5-1..A.0 2.S. Hyde. CV. Keaney.. Comparison of solid media for the culture of anaerobes.S. Pathol. B.J.) LAB 25 Description A medium for sterility tests. Sterilise by autoclaving at 121˚C for 15 minutes.B.5 0. Appl.D.E. organisms: B. J..5 2. W.. Bennett. Clin. J. Ganguli. (1985). the medium should be reheated to deoxygenate.0-2. 2: 124-126. Inoculation: If used as a blood culture medium a minimum dilution of 1:10 should be used. Aerobic and anaerobic organisms grow well in this medium even from small inocula.0 10. (1982). G.7 grams of powder. Pathol.G. Turton. 43: 253-256. J. (1984). (1990). The medium was designed to give optimum growth of fastidious anaerobes and has found applications as a blood culture medium and an enrichment broth for the isolation of anaerobes.0 1. Clin.E. 4 abstract 334.B.0 shape & surface CV.75 1. Pept. Vitamin K. A. The thioglycollate will also serve to inactivate any mercurial compounds used as preservatives. L-cysteine together with sodium thioglycollate reduce the Eh of the medium and the agar content inhibits absorption of oxygen and convection currents. J. Clin. (1981).H. 35: 458-461.G. Clin.S.Growth Characteristics (48 hours) organism B. T. Resazurin is a redox indicator. The low level of agar reduces oxygen diffusion into the medium.A.5 5. L. W.5 5. Path.(D) transparent Grey/ Brown translucent White White/ White Porphyromonas asaccharolyticus 1..G.H. Clarry.A. (1986). Clin.E. Infectious Diseases. haemin and L-cysteine are all growth factors required by some anaerobes. necrophorum 1. Junt.0-2.W. Ganguli. B. swirl to mix. prepared according to the specification of the United States Pharmacopeia.A.G. J. If the medium is reddish this indicates too much oxygen has been absorbed. Formula Tryptone L-Cystine Glucose Yeast Extract Sodium chloride Sodium thioglycollate Resazurin Agar g/litre 15. European Journal of Clinical Microbiology.5 F.0 (‘molar tooth’) (smooth) Incubation: 37˚C for 24-72 hours.G. for the isolation of anaerobes from simulated paediatoic blood cultures.0 0.001 0.) LAB 71 Description F.. CV. 66: no. Evaluation of ten commercial blood culture systems to isolate a pryridoxal dependent streptococcus. to be the liquid medium of choice for fastidious anaerobes. M. A note on ultra violet red fluorescence of anaerobic bacteria in vitro.0 0. G.4 0. disperse in 1 litre of deionised water. was developed by LAB M working in conjunction with the microbiology department of a University of Manchester teaching hospital. Bact. perfringens colony size (mm) 1.. 35: 1142-1149.5 0.5-2.0 CV. 1 L-Cysteine HCl Resazurin Sodium bicarbonate Haemin Vitamin K g/litre 15.A. W.G.. In appropriate tubes or bottles the thioglycollate ensures adequate anaerobic conditions. Growth indicators: The broth may become turbid or individual colonies may form suspended in the medium. Do not reheat more than once..J.. Evaluation of Fastidious Anaerobe Broth as a blood culture medium..G. Duerden. E. pH: 7.S. Fitzgerald T.0-2.001 0.5 0. Ganguli.0 CV. Appearance: Pale straw. Keep the container airtight. (1989). acne 0.A. Griffiths.S. (1983). J.C. J.E. (1986). 36: 963-977. Pathol. J. Several published evaluations show F.E. Tighten the caps as soon as possible after autoclaving. G. Hyde. CV. L. L. Rapid Detection of Bacteraemia by early subculture. Med.E.

The pyocyanin pigment produced by most strains of Pseudomonas aeruginosa is also produced. Formula Balanced Peptone No. Inoculation: Surface spreading.2 Minimum Q. and is similar to Palcam broth in that it contains aesculin to indicate the presence of a potential Listeria isolate.Method for reconstitution Weigh 29. disperse in 1 litre of deionised water containing 10ml Glycerin B. McClain D.Comm. sporogenes S.CR. Growth indicators: Turbidity.2 Method for reconstitution Weigh 35 grams of powder. 21st End.35 20.I. E. Fraser broth is a secondary enrichment broth for the isolation of Listeria spp. Appearance: Straw coloured. Store at ambient temperature in the dark. Slant over a generous butt if required. Minimum Q. M. Fraser – 35˚C aerobically for 24hrs and 48hrs. organisms: Ps. 2 simple media for demonstration of pyocyanin and fluorescein.. D.D. Allow to soak for 10 minutes. All broths should be subcultured before discarding.C.22 . Inoculation: 1/2 Fraser – Add 25g sample to 225ml of 1/2 Fraser broth and homogenise Fraser – Subculture 0. (1988) Rapid detection of Listeria spp in food and environmental samples by esculin hydrolysis. Surface may be pink/blue due to oxidation of Resazurin.0-2. Formula g/litre 15. Ward and Raney’s medium formulated for the demonstration of the fluorescein pigment produced by many strains of Pseudomonas.1ml of primary enrichment broth (UVM I or 1/2 Fraser) into 10ml of Fraser broth. J. organisms: Listeria sp NCIMB 50007 e. swirl to mix. aeruginosa NCIMB 50067 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.0 Minimum Q. Use of an improved cetrimide agar medium and other culture methods for Pseudomonas aeruginosa.. but not in the refrigerator.H. Div. swirl to mix and sterilise at 121˚C for 15 minutes. 18: 752-756. aeruginosa 0. Clin. Brown. Allow to soak for 10 minutes. Appearance: Straw opalescent broth with precipitate (clears on storage). mix well and aseptically dispense into sterile tubes or bottles. US Dept of Agric. irrespective of colour change.5 Grey (colony size varies Greenwith strain) fluorescent-(non pigmented) pigment (mucoid) Grey fluorescent pigment Ps.2 + 0.CR. Lab. Microbiol.0 3. Fraser broth may also be used as a primary enrichment medium by incorporating 1/2 strength supplement into the broth base (X164 or X564). If more than 30% of the medium turns pink (oxidised) the Eh may be restored (once only) by heating in a boiling water bath or by free-steaming. Sterilise by autoclaving for 15 minutes at 121˚C. and Raney. Lab.5 F. 1.C. and Sperber W. Revised May 24. J.. J. Incubation: 30-35˚C aerobically for 14 days.75 grams. (1954). References The Pharmacopeia of the United States of America.1 ± 0. aureus NCIMB 50080 Storage of Prepared Medium: Capped container – up to 3 months at 15-20˚C in the dark.. Fluorescence Agar LAB 16 Description This medium is a modification of King.57.0 9. swirl to mix then sterilise for 15 minutes at 121˚C. colonies in medium.0 1. Subculture onto selective agars at 24 and 48hrs. Path.. 1 Dipotassium phosphate Magnesium sulphate Agar No. disperse in 1 litre of deionised water. fluorescens 1.O. Can be made selective by the addition of selective agents such as X108 Cetrimide Fucidin Cephaloridine.5-2.monocytogenes from processed meat and poultry products. then bring to the boil to dissolve and dispense 15ml into 6mm x 150mm tubes.No. pH: 7. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Bottles – up to 14 days at 2-8˚C. organisms: C.6 1. Appearance: Pale straw colour.0 Peptone mixture Yeast extract Aesculin Disodium hydrogen phosphate Potassium dihydrogen phosphate Sodium chloride Lithium chloride Method for reconstitution Weigh 55 grams of power and add to 1 litre of deionised water (add to 900ml if preparing 1/2 Fraser). and Lowbury.E. E.0 5.C.0 1. Ward.. Cool to 47˚C and add 2 vials of Fraser supplement X165 (or 2 vials of 1/2 Fraser supplement X164). Mix well before pouring.H. pH 7. (1989) FSIS method for isolation of L.2 References King.5 12. 44: 301-307.A.5 1.2 ± 0. (1989). It also contains lithium chloride in an attempt to suppress the growth of enterococci in the medium (as does Palcam).D. (1965). Ps. Incubation: 30-37˚C for 24 and 48 hours aerobically.FSIS. clear gel. and Lee W.Food Protection 51 (10) 762-765.L.K. Interpretation Blackening of the broth indicates the presence of a potential Listeria and should be subcultured onto Listeria isolation medium (Oxford) LAB122 or Palcam agar LAB148. Growth Characteristics organism colony size shape & (mm) surface F. Clin Med. Incubation: 1/2 Fraser – 30˚C aerobically for 24hrs. V. clear. pH: 7. Fraser Broth LAB 164 Description Developed as a modification of UVM II medium.J. 2 g/litre 20. colour other References Fraser J.P. Soak for 10 minutes. (1985).

G.C. Agar Base
LAB 67
Description
A nutritious agar base described by Thayer and Martin for the isolation of Neisseria gonorrhoeae. The rich peptone mixture is enhanced by the use of corn starch to absorb toxic metabolites and a buffering system is used to maintain neutral pH. The medium is made selective by the use of various antibiotic cocktails. Thayer and Martin originally recommended the use of vancomycin, colistin and nystatin V.C.N. but the addition of trimethoprim (X068, X268 V.C.N.T.) is useful in preventing the swarming of proteus. More recently the emergence of vancomycin sensitive gonococci has made the New York City selective agents (lincomycin, colistin, amphotericin, trimethoprim X070, X270 LCAT) the combination of choice. Enrichment of the base is usually by the addition of lysed blood. Alternatively chocolated blood or haemoglobin powder and Thayer and Martin’s mixture of vitamins, amino acids and coenzymes can be used. The supplement X069, X269 can be added as this is without Amphotericin and this permits the growth of yeasts. The growth supplement X271 can be added to this medium to aid in the isolation of Neisseria spp. Formula Special Peptone Corn Starch Sodium chloride Dipotassium hydrogen phosphate Potassium dihydrogen phosphate Agar No. 2 g/litre 15.0 1.0 5.0 4.0 1.0 10.0

Hektoen Enteric Agar
LAB 110
Description
A medium developed at the Hektoen Institute in Chicago for the enhanced recovery of shigellae from clinical specimens. This medium has high levels of peptones and sugar which counteract some of the toxic effects of bile salts used to make the medium selective. This allows the shigellae to grow as well as the salmonellae. Salicin is fermented by many coliforms including those that do not ferment lactose and sucrose. The medium employs a double indicator system similar to that used in LAB 6 C.L.E.D., (Bevis) and an H2S indicator system similar to that used in LAB 32 XLD. Although intended primarily for clinical use this medium is quoted in B.S. 4285 as suitable for the examination of dairy products for salmonellae. Formula Meat Peptone Yeast Extract Lactose Sucrose Salicin Bile Salts No. 3 Sodium desoxycholate Sodium chloride Sodium thiosulphate Ammonium ferric citrate Acid fuchsin g/litre 12.0 3.0 12.0 12.0 2.0 7.0 2.4 5.0 5.0 1.5 0.1 0.065 14.0

Method for reconstitution
Weigh 36 grams of powder, disperse in 1 litre of deionised water. Allow to soak for 10 minutes, swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. Cool to 48˚C and add 50-70ml of lysed blood and 2 vials of X070 selective agent. Mix well and pour into Petri dishes. Appearance: Dependent on blood supplement used. pH: 7.2 ± 0.2

Bromothymol blue Agar No. 1

Method for reconstitution
Weigh 76 grams of powder, disperse in 1 litre of deionised water. Allow to soak for 10 minutes, swirl to mix then heat gently and bring to the boil. Cool to 47˚C and pour plates. DO NOT AUTOCLAVE OR OVERHEAT THIS MEDIUM. Appearance: Green, clear. pH: 7.5 ± 0.2

Minimum Q.C. organisms: N. gonorrhoeae ATCC CDC98 E. coli (inhibition) NCIMB 50034
Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Inoculation: Surface, streaking out for single colonies. Incubation: 37˚C microaerobically for 24-48 hours.

Minimum Q.C. organisms: Salmonella sp. NCIMB 50076 Shigella sp. E. coli (some inhibition) NCIMB 50034
Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Inoculation: Surface plating, streak out to single colonies.

Growth Characteristics
organism colony size shape & (mm) surface CV.E.G. CV.E.G. CV.E.G. CV.E.G. colour other

Incubation: 37˚C aerobically for 18-24 hours.

N. gonorrhoeae 1.0-2.0 N. lactamica B. catarrhalis 1.0-2.0 2.0-3.0

Transparent variations in colony size Grey Grey Cream

N. meningitidis 2.0-3.0

Other organisms should not grow with the exception of antibiotic resistant variants.

References
Young, H. 1978. Cultural diagnoses of gonorrhoea with modified New York City (MNYC) medium. Brit. Journ. Ven. Dis. 54: 36-40: Thayer, J. D. and Martin, J. E. 1966. Improved medium selective for the cultivation of N. gonorrhoeae and N. Meningitidis: Public Health rep. 81: 559-562.

1.23

Growth Characteristics
organism H2S +ve Salmonella H2S -ve Salmonella S. sonnei S. flexneri S. dysenteriae E. coli colony size shape & (mm) surface 2-3 2-3 2-2.5 1.0-2.5 1-2 0.5-2 CV.E.G. CV.E.G. CV.E.G. CV.E.G. CV.E.G. colour Green+ Black Green Green Green Green (Rough) other

Method for reconstitution
Weigh 45.0 grams of powder and disperse in 1 litre of deionised water. Soak for 10 minutes, swirl to mix and sterilise at 121˚C for 15 minutes. Cool to 47˚C and add 100ml of Horse Serum, and 2 vials of VCA supplement X040. Mix well and pour, continuing to mix whilst pouring to keep the charcoal in suspension. Appearance: Black Agar pH: 7.4 ± 0.2

Minimum Q.C. organisms: Helicobacter pylori S.aureus (inhibition) NCIMB 50080
Storage of prepared medium: Plates – up to 7 days at 2-8˚C the dark. Inoculation: Surface streaking for single colonies. Incubation: 37 ˚C for 72 hours.

CV.E.G. Salmon ppt. (Rough) around (No growth) colonies CV.E.G. CV.E.G. CV.E.G. Salmon Salmon (Rough) (Mucoid)

Citrobacter spp. 1.0-2.0 Klebsiella spp. 0.5-2.0 Proteus spp. 1.0-2.0

Green/ (No growth Black brownish centre) centre Green (No growth)

Growth Characteristics
Organisms H. pylori B. catarrhalis Colony Size (mm) 1.0-1.5 1.0-2.0 Shape and Surface CVEG CVEG Colour Grey White/Cream

Pseudomonas spp.

0.5-1.5

F.Rz.D.

References
King, S. and Metzger, W.I. (1967). A new medium for the isolation of Salmonella and Shigella species. Bact. Proc. Am. Soc. Microbiol. 77. King, S. and Metzger, W.I. (1968). A new plating medium for the isolation of enteric pathogens. Hektoen Enteric Agar, Appl. Microbiol., 16(4), 577. King, S. and Metzger, W.I. (1968). A new plating medium for the isolation of enteric pathogens. II. Comparison of Hektoen Agar with SS and EMB agar. Appl. Microbiol., 16(4), 579. Speck, M.L. (ed.). (1976). Compendium of Methods for the Microbiological Examination of Food. Washington, D.C.: American Public Health Association.

References
Bolton et al. Public Health Laboratory, Preston. Personal communication.

Hoyle’s Medium
LAB 27
Description
A highly selective culture medium for the isolation and differentiation of Corynebacterium diphtheriae types gravis, mitis and intermedius. Hoyle’s medium gives rapid growth of all types of C. diphtheriae, which results in most specimens giving adequate growth with overnight incubation. Formula Beef Extract g/litre 10.0 10.0 5.0 12.0

Helicobacter Pylori Medium
LAB 140
Description
A selective medium for the isolation of Helicobacter pylori, the causative agent of chronic gastritis. This campylobacter-like organism was described in 1983 colonising the gastric mucosa, a site previously thought to be sterile due to the low PH. The high level of Urease production by this organism appears to be the major pathogenicity factor enabling it to withstand the strongly acidic environment. Helicobacter Pylori Medium is a modification of CCDA Medium for the isolation of Campylobacter spp. Formulated by Bolton et al at Preston Public Health Laboratory, it incorporates a rich agar base supplemented with horse serum to promote optimum growth, and Vancomycin, Cefsulodin, and Amphotericin as selective agents. Formula Beef Extract Meat Peptone Tryptone Sodium chloride Charcoal Acid Hydrolysed Casein Ferrous sulphate Sodium pyruvate Sodium carbonate Agar no.2 g/litre 10.0 5.0 5.0 5.0 4.0 3.0 0.25 0.25 0.4 12.0

Peptone Sodium chloride Agar

Method of Reconstitution
Weigh 37 grams of powder and disperse in 1 litre of deionised water. Allow to soak for 10 minutes, and sterilise by autoclaving at 121˚C for 15 minutes. Cool to 47˚C, add 50ml of lysed horse or sheep blood and 10ml of X027 potassium tellurite solution. Mix well before pouring.

Appearance: Dark Red, clear gel.
pH: 7.8 ± 0.2

Inoculation
Spread the entire surface with the swab or sample under investigation. Hoyle’s medium is very selective and spreading for single colonies using a wire loop is not necessary. Use of a nonselective blood agar alongside Hoyle’s is recommended. Incubation: 37˚C for 18-48 hrs, aerobically

1.24

Interpretation
organism C. diphtheriae var gravis C. diphtheriae var mitis colony size shape & (mm) surface 1.5-2.5 0.5-2.0 CV.CR.D (daisy head) CV.E.G. CV.E.G. CV.E.G. colour other

Minimum Q.C. organisms: E. faecalis NCIMB 50030 E. coli (inhibition) NCIMB 50034
Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Inoculation: Surface, spread 0.1ml to 0.5ml over entire surface of plate. Incubation: 37˚C or 42˚C aerobically for 18-24 hours. Interpretation: Count all white/grey colonies, approx 2mm diameter, surrounded by a black halo to give presumptive enterococcus/faecal streptococcus count.

Grey Colonies difficult to emulsify

Grey Easily (dark centre) emulsified Grey (dark centre) Grey Streptococcal (dark centre) appearance in Gram stain

C. diphtheriae 0.5-1.0 var intermedius C. Ulcerans 1.0-1.5

References
Mossel, D.A.A., Bijken, P.H.G., Eelderink, I. and van Spreekens, K.A. (1978). Streptococci, edited by Skinner, F. A. and Quesnel, L.B. SAB Symposium Series No. 7 Academic Press, London.

C. hoffmanii C. xerosis Streptococcus spp. H. influenzae

0.5-1.0 0.5-2.5 pp - 1.5 pp - 1.5

CV.E.G. Black White/grey (confluent growth)colonies CV.E.G. CV.E.G. Black Black Enterococci may be larger

CV.E.G. Grey/black Some strains no growth

Kanamycin Aesculin Azide Broth
(K.A.A. Broth)

Storage: Plates – up to 7 days at 2-8˚C

Minimum Q.C. Organisms C. diphtheriae var mitis (non-toxigenic) E. coli NCTC 10418 (inhibition) Reference:
Hoyle L. (1941) A Tellurite Blood Agar Medium for the Rapid Diagnosis of Diphtheria. Lancet 1 175-176 176. Elek S.D. (1948) The Recognition of Toxigenic Bacterial Strains in vitro. Brit. Med. J. 1 493-496.

LAB 107
Description
An enrichment and isolation medium for enterococci. The medium can be used with the M.P.N. technique to enumerate enterococci in food. This broth is identical to LAB 106 K.A.A. agar with the omission of the agar. Formula Tryptone Yeast Extract g/litre 20.0 5.0 5.0 1.0 1.0 0.5 0.15 0.02

Kanamycin Aesculin Azide Agar
(K.A.A. Agar)

Sodium chloride Sodium citrate Aesculin

LAB 106
Description
A selective isolation and enumeration medium for enterococci (Lancefield group D streptococci) in food. Sodium azide and kanamycin provide the selective inhibition required whilst aesculin and iron salts form an indicator system for the presumptive identification of enterococci. Incubation at 42˚C will increase the medium’s selectivity. Formula Tryptone Yeast Extract Sodium chloride Sodium citrate Aesculin Ferric ammonium citrate Sodium azide Kanamycin sulphate Agar No. 1 g/litre 20.0 5.0 5.0 1.0 1.0 0.5 0.15 0.02 10.0

Ferric ammonium citrate Sodium azide Kanamycin sulphate

Method for reconstitution
Weigh 33 grams of powder, disperse in 1 litre of deionised water. Allow to soak for 10 minutes, warm gently to dissolve completely then disperse into tubes or bottles. Sterilise by autoclaving at 121˚C for 15 minutes. Appearance: Light straw, clear. pH: 7.0 ± 0.2

Minimum Q.C. organisms: E. faecalis NCIMB 50030 E. coli (inhibition) NCIMB 50034
Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. Inoculation: Inoculate tubes with decimal dilutions of food suspension. Incubation: 37˚C or 42˚C aerobically for 18-24 hours. Interpretation: Blackening of the medium suggests the presence of enterococci/faecal streptococci.

Method for reconstitution
Weigh 43 grams of powder, disperse in 1 litre of deionised water. Allow to soak for 10 minutes, swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. Cool to 47˚C, then dispense into Petri dishes. Appearance: Pale straw, clear. pH: 7.0 ± 0.2

References
Mossel, D.A.A., Bijken, P.H.G., Eelderink, I. and. van Spreekens. K.A. (1978). Streptococci, edited by Skinner, F.A. and Quesnel, L.B. SAB Symposium Series No. 7 Academic Press, London.

1.25

Fermentation of lactose as well as glucose. kansasiiwhite floccular ‘snowflake’ colonies ‘ropey’ deposit fine granular deposit. Pus with no other bacteria Tissue Concentration and acid or alkali decontamination.kansasii and M. J. Medium LAB 123 Description The cultivation of non-sputum specimens for mycobacteria requires special attention. it is recommended that specimens should also be inoculated onto Lowenstein Jensens slopes for the following reasons: . organisms: M.012 Growth Characteristics organism M. scrofulaceumM. produces acidity in both slope and butt (yellow). Fermentation of glucose only.025 12.5 5. if required. 2 g/litre 20.0 0. D. tuberculosis M. Urine & Faeces C. Incubation: 37˚C aerobically for 18-24 hours.2 Minimum Q. Erfahrungen bei diagnosticher Verwendung der Tiefenkulter. scrofulaceum may be inhibited by the antibiotic cocktail.F. Minimum Q. turning phenol red indicator yellow.0 5. (inhibition) NCIMB 50010 Storage of Prepared Medium: up to 3 months at 2-8˚C in the dark. Inoculation: Kirchner’s Medium is suitable for the cultivation of mycobacteria from the following specimens: Pre-treatment Required Sputum. Abt 1. which is made selective by the addition of an antibiotic cocktail. Tubercle (1972).. O.F. Liberation of sulphide results in the formation of iron sulphide (blackening of either slope or butt).Clin. (1932).A.. M .Growth suitable for further characterisation may be obtained more rapidly .0 0.4±0.Kirchner’s T. 36: 1357-1361. Although Kirchner’s Medium has been shown to give higher isolation rates than solid media. Formula Sodium dihydrogen phosphate Potassium dihydrogen phosphate Magnesium sulphate Trisodium citrate L-Asparagine Phenol red g/litre 7. B.0 1. can be obtained from the use of liquid culture media. References Kirchner.C. Appearance: Pale red.4 grams disperse in 1 litre deionised water. The pH is then adjusted using the appropriate normal solution observing colour or indicator. None. tissue or body fluids obtained by surgical procedures are not easily repeated and may contain small numbers of mycobacteria.0 ± 0. pH: 7. 409-412. Historically. Allow the mixture to soak for 10 minutes. 53: 31-34. Sbl fur Bact. Allen.S. Selective Kirchner’s Medium is a liquid culture media. such specimens were inoculated into guinea pigs. coli (inhibition) NCIMB 50034 Candida albicans. Marks. that as large a portion of specimen as possible be inoculated into suitable growth media. Bring to the boil with frequent stirring to dissolve completely.. Dispense into tubes and sterilise for 15 minutes at 121˚C. however improvements in culture methods have led to a situation where comparable results. but not in the anaerobic conditions of the butt. Appearance: Reddish brown agar.R. Originale 124. Disintegration followed by acid or alkali decontamination. It is important.54 2. if necessary. Incubation: 37˚C aerobically for up to six weeks.0 0. Path. Specimens such as C.0 10.S. which remains acid (yellow). J. Aseptically dispense in 9ml volumes into sterile Macartney bottles. Kligler Iron Agar LAB 59 Description A differential medium for the recognition of enteric pathogens by their ability to ferment glucose and/or lactose. and has been shown to be sufficient in the recovery of mycobacteria from non-sputum specimens. Cool in a slanted position such that slopes are formed over deep butts approx.B. pH: 7. clear fluid.5 0.Some Mycobacterium species including M.26 . add 20ml Glycerol A. Mitchison. D. therefore. 3cm in depth. tuberculosis (H37RV strain) E. and liberate sulphides.W.C. is followed by reversion in pH on the slope. Organisms Salmonella typhimurium NCIMB 50076 Pseudomonas aeruginosa NCIMB 50067 Inoculation Subcultures for further identification are picked from the centre of isolated colonies on selective media and streaked across the slant and stabbed deep into the butt of tubes of Kligler Iron Agar.2 The acidified or alkaline de-contaminated material may be added directly to the Kirchner medium.. 1.34 2. Manickavasagar. Method of reconstitution Weigh 49 grams of powder and mix with 1 litre of distilled water. Selective Kirchner’s medium in the culture of specimens other than sputum for mycobacteria. Ending the routine guinea pig test. Fermentation liberates acid. from initial acidity to final alkalinity (red colour). swirl to mix and sterilise by autoclaving at 121°C for 10 minutes. (1972).0 Method for reconstitution Weigh 17. intracellulare. (1983). with or without gas. Allow the medium to cool and supplement with 1ml sterile heat in activated horse serum and 2 vials of X124.3 0. Formula Peptone Lactose Glucose Sodium chloride Ferric ammonium citrate Sodium thiosulphate Phenol red Agar No.

flexneri Butt Acid Acid Acid/gas Acid/gas Acid/gas Acid Acid Slope Alkaline Alkaline Alkaline Acid Alkaline Alkaline Alkaline Sulphide + + + - Liquid Baird-Parker Medium LAB 158 Description Developed by Van Doorne et al in 1981.0 5. and Lacey.J.2.0 5. R.M. A modification of the Kligler Lead Acetate Medium. this medium is essentially Baird-Parker Agar LAB 085 without the agar and egg yolk components. Hendriksz. pH: 6. 1985. Method for reconstitution Weigh 13 grams of powder. (1981) Liquid Modification of Baird-Parker’s Medium for the Selective Enrichment of Staphylococcus aureus. but are not ideal due to potential inhibition of S. H. Antonie Van Leeuwenhoek 47 267-278. D. pH: 6. Even Giolitti Cantoni Medium may be inhibitory to some strains of S. Incubation: 37˚C anaerobically for 48 hours.. (1917). References: Kligler.Interpretation Organism Salmonella typhi S. (1927). Non selective enrichments have been suggested. (1975).1ml of 1% potassium tellurite solution to each tube.M. coli (inhibition) NCIMB 50034 Storage of prepared medium: Up to 1 month at 2-8˚C (without tellurite added). Bact. When cool. This medium was chosen for development to overcome the problems of other selective enrichments and non-selective enrichments. mix to dissolve and dispense 10ml amounts in tubes.F. United States Pharmocopeia. Sterilise at 121˚C for 15 minutes. 13:182-189. Interpretation: Culture tubes showing growth (blackening of the medium and turbidity) onto Baird-Parker Agar LAB 085... 1.0 1.0 5.Paratyphoid Group. H. Formula Beef Extract Gelatin Peptone Lactose g/litre 3. Minimum Q. Inoculation: See methods for standard techniques. cool and add 0.P. Washington. Van der Kreek. References American Public Health Association. Lactose Broth LAB 126 Description A medium for the performance and confirmation of the Presumptive Test for members of the coliform group in water and dairy products.2ml of 1% potassium tellurite solution to each tube.R. in the isolation of Staphlococcus aureus. Single strength. Formulation Meat Peptone Tryptone g/litre 8. Single Strength Medium: Weigh 43 grams of powder and proceed as above. J.0 12. Soak for 10 minutes. paratyhi A + B Other Salmonella E. Before use.2 Minimum Q.. I. S. 10ml of sample homogenate to each tube. organisms: S.0 2. disperse in 1 litre of deionised water. Appearance: Clear straw broth. Pauwels. 1ml of sample homogenate to each tube. aureus NCIMB 50080 E. Standard Methods for the examination of water and waste water. Sterilise by autoclaving at 121˚C for 15 minutes. heat to dissolve then distribute into bottles with Durham tubes.0 Storage: Tightly capped containers . D. Hlth.9 ± 0. Appearance: Straw coloured. Bailey. Other selective broth media suffer from the potential of inhibiting sub lethally damaged S. XXI. aureus.27 . Incubation: 35˚C aerobically for 48 hours. References Van Doorne. Liquid Baird-Parker Medium is the ideal solution for detecting low numbers (<20/g) of S.S. J. organisms: E. clear.P.0 Beef Extract Yeast Extract Sodium pyruvate Glycine Lithium chloride Method for Reconstitution Double Strength Medium: Weigh 86 grams of powder and disperse in 1 litre of deionised water. coli Proteus spp Shigella sonnei S. A Simple Medium for the Differentiation of Members of the Typhoid .up to 3 months at 15-20˚C in the dark.0 5. aureus by microbial antagonism in a mixed bacterial population. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. aureus as it has been shown to give acceptable selectivity whilst being non-inhibitory to injured cells. Inoculation: Double strength.T.C. add 0. Publ. Am. aureus cells where the salt content is greater than 40g/l. recommended by the U.C. 892. heat to 100˚C in a water bath or steamer for 15 minutes to remove oxygen.0 10.8 ± 0. 7:1042-1044. Baird. Anaerobic conditions can be achieved using an anerobic jar (container caps must be loose fitting) or by overlaying the surface of the broth with 1cm of molten agar or liquid parafin. Interpretation: Coliforms are presumptively identified by their ability to ferment lactose and produce gas within 48 hours at 35˚C. G.

swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes.0 Method for reconstitution Weigh 36 grams of powder and add to 1 litre of deionised water.3 ± 0. 689-695. Can. monocytogenes NCIMB 50007 E. Minimum Q. J.W.C. streak out to single colonies. Incidence and Pathogenicity. A.D. 1. Inoculation: Surface. mix well and pour plates. fosfomycin. clear. Method for flow cytometric detection of Listeria monocytogenes in milk. Journ. Food Protect.0 Listeria Isolation Medium (Oxford Formulation) LAB 122 Description A selective identification medium for the isolation of Listeria monocytogenes from food and clinical material.F. Listeria monocytogenes dans le lait pasteurise.W.0 1. Add to 1 litre of deionised water. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 14 days at 2-8˚C in the dark. 50. Listeria monocytogenes in raw milk: Detection.Oct.5 grams of powder.0 5. Boland.K. J.L.. monocytogenes colony size shape & (mm) surface 0. Cool to 47˚C and add 2 vials of X138 (X139/X539 can be used as an alternative) reconstituted in 50% alcohol. Formula Tryptone Soy Peptone Sodium chloride Dipotassium hydrogen phosphate Glucose Yeast Extract g/litre 17.2 Method for reconstitution Weigh 57. P. Cancelo. Lithium chloride is used to inhibit enterococci and acriflavine to inhibit some Gram negative and Gram positive species.Listeria Enrichment Broth LAB 138 Description A medium for the selective enrichment of food and environmental samples for Listeria spp. Columbia agar is the nutrient base to which selective inhibitors have been added.G.A. Lovett. Personal Communication Public Health Laboratory. Appearance: Pale yellow.G. these are colistin.A. Black Usually no growth References Garayzabal. Van Netten. Gregory J. (1986). J.. J. The medium offers more rapid enrichment than the low temperature enrichment techniques.E. This medium is now recommended by the Commission of European Communities and the International Dairy Federation for the examination of soft cheeses for Listeria monocytogenes. organisms: L. The advisability of using this medium at two levels of selectivity has been recognised.C. Perales. I. pH: 7. P.. a heavy inoculum can be used. colour other Grey/ Black/brown Green around colonies diffusion Gram-ve Bacilli Enterococci No growth p. Francis. D.M. Further selective agents may be added after autoclaving to increase the selectivity. Aesculin is included in the formula as a differential indicator. J. Microbiol.B. first described in 1987 by J. Appearance: Yellow.. Hunt.. A selective and diagnostic medium for use in enumeration of Listeria spp. Hunt.A. Lovett. No.5 15. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. slightly opaque gel. D.28 . J. Bolton.F.2 ± 0.0 CV. The medium is incubated at 30˚C and utilises acriflavine.M.M. Preston U.5 CV. C. L. pH: 7.J.L. Microbiol. Appl. Personal communication. Bolton. Allow to cool to 47˚C. International Journal of Food Microbiology 6:187-198. 3: 188-192. Growth Characteristics organism L. in foods. Allow to soak for 10 minutes then swirl to mix and sterilise by autoclaving at 121˚C for 15 minutes. F. Mossel.0 2. L. Fernandez.. This medium is highly selective.S. (1988)..2 Minimum Q. Inoculation: Add 25 grams of sample to 225mls of Listeria Enrichment Broth and homogenise. 32: 149-150. G. Vol. add 2 vials of selective supplement X122 (X123 can be used as an alternative). C.F. Donnelly. 0. Preston P.5-1. (1987). J.E.W. Subculture: After 24 and 48 hours onto Listeria Isolation Medium – LAB 122.5 6. cefotetan and cyclohexamide.0 3. J.0 0.p.V. nalidixic acid and cyclohexamide as selective agents. LAB M’s formulation has been used to successfully isolate Listeria from such diverse products as chicken giblets and dairy cheeses. monocytogenes NCIMB 50007 E. References Lovett. Frances. Aseptically dispense into sterile tubes or bottles. monocytogenes will hydrolyse aesculin to aesculutin which reacts with the iron salt to give a black precipitate around the colonies. & Environ. Van de Van. Incubation: 30˚C aerobically for 24-48 hours. Formula Columbia Agar Base Aesculin Ferric ammonium citrate Lithium chloride g/litre 41.5 2. Baigent (1986). Food Protect. Incubation: 30˚C aerobically for up to 48 hours. Rodriguez.. organisms: L. Allow to soak for 10 minutes. 50: 188-192. J. J.

2 g/litre 20.H. will spread on this medium making interpretation difficult. Formula Peptone Lactose Bile Salts Sodium chloride Neutral red Agar No.E.E. (pink) Transp. Interpretation: Count all colonies.E.0 0. 1.25 0. Streptococcus diacetylactis) in dairy products.G.0 5.G. CV.O. Allow to soak for 10 minutes. Minimum Q. (spreading) CV. Improved medium for lactic Streptococci and their Bacteriophages.0 Proteus spp.0 MacConkey Agar (With Salt) LAB 30 Description A selective medium for the isolation of bile tolerant organisms from faeces. for this reason LAB 2 MacConkey Agar (without salt) may be preferred as it is less prone to this phenomenon.CR.0 5. P.0-3. CV.-0. Sandine. Streptococci form colonies of 1-2mm in diameter.0 10. B. aerogenes Growth Characteristics approx. organisms: E.5-2.5 15.G. (1975).E. organism E.5-2. disperse in 1 litre of deionised water. The medium can also be used to investigate the bacteriophage susceptibilities of these organisms. Red (ppt around colony) Citrobacter spp. Sterilise by autoclaving at 115˚C for 20 minutes.0 F.0-3. Formula Balanced Peptone Soy Peptone Yeast Extract Beef Extract Lactose Sodium glycerophosphate Magnesium sulphate Ascorbic acid Agar No. swirl to mix then bring to boil to dissolve agar before dispensing in 15ml aliquots.E.0 5. organisms: S.G.2 Minimum Q. Microbiol. Streptococcus cremoris.G. Another application is for the enumeration of Streptococcus thermophilus in yoghurts. Appearance: Pink/red.4 ± 0. This formula is recommended by W.P.0 0.0-4. 1. aureus 0.G. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. Allow to soak for 10 minutes.5 5. Methods for the Examination of Water and Associated Materials.0 19. Incubation: 30˚C for 48-72 hours for mesophilic streptococci. 29 No. coli K.E.29 .0 S.0 (dependent on lactose fermentation Orange and pigment Opaque production) Pink/Deep Red Opaque Transp.C.E.D.5 CV. CV.G. References Environment Agency: The Microbiology of Drinking Water (2002). and other bodies for the examination of water and milk.C.G.0 2. Incubation: 37˚C aerobically for 24 hours.0-2.5 Salmonella spp.2 Weigh 52 grams of powder. CV. pH: 7. colour Pink/Red Pink/Red Yellow Colourless Colourless other (mucoid) (ppt around colony) References Terzaghi.M17 Agar LAB 92 Description A medium for the enumeration of lactic streptococci (Streptococcus lactis. 1. size shape & (mm) surface 2.0 2.0 5.5 Shigella spp. Pinkish (colonialvariation) White/ Pink Enterococcus spp. CV. translucent agar. Bile-tolerant Gram positive organisms as well as Gram negative organisms will grow on this medium. Appl. Inoculation: Pour plate technique. 2. Appearance: Pale straw.E. Some strains of Proteus spp.E. 1.1 ± 0. 37˚C for 48 hours for Streptococcus thermophilus. lactis Storage of Prepared Medium: Capped containers – up to 1 month at 15-20˚C in the dark. W. Inoculation: Surface plating..0 CV. P. Cool to 47˚C and mix well before pouring into Petri dishes.05 12. aeruginosa 1. urine. sewage and foodstuffs.0-4. clear pH: 7. disperse in 1 litre of deionised water.5-2. 2 g/litre 5. 6 pp 807-813. streaking out to single colonies.E.2 grams of powder.0 Method for reconstitution Method for reconstitution Weigh 57.

(1958). Appearance: Pink/red. The medium has been made more selective than MacConkey’s original formula by the use of crystal violet as well as bile salts.T.G. 2. R. water and dairy bacteriology.4 ± 0.C. T.5 grams of powder. International Standards for Drinking Water.E.E.0-5. Formula Mixed Peptones Lactose Bile Neutral red Agar No. CV. clear. Incubation: 37˚C aerobically for 18-24 hours. coli NCIMB 50034 Ent. Cool to 47˚C and mix well before pouring plates.G. Medical Microbiology. and the American Public Health Association for the isolation of Enterobacteriaceae from waters and sewage. swirl to mix then sterilise for 15 minutes at 121˚C.G.0 CV. (Camb). 3.5 Shigella spp..). London.0 Staph.E. aerogenes Proteus spp.D. F.G. Geneva.0 2.0 Shigella spp. faecalis (inhibition) NCIMB 50030 Storage of prepared medium: Plates – up to 7 days at 2-8˚C in the dark.G. coli NCIMB 50034 S.G. disperse in 1 litre of deionised water. Allow to soak for 10 minutes.G. 3. MacConkey Agar (without salt) MacConkey Agar No. No growth Enterococcus spp. aureus NCIMB 50080 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.G. pH: 7. No growth Enterococcus spp. CV.1 ± 0.0 CV.G. 12th Edn. (D) CV.characteristic Yellow.5-1.E. Cruikshank. coli colony size shape & (mm) surface 3. aerogenes 4. CV.W. 5: 333-379.E. A Guide to the Laboratory Diagnosis and Control of Infection. MacConkey.0 10. W. colour Red other (non lactose fermenting yellow) (mucoid) fishy odour Growth Characteristics organism E. (1973).03 0. A. London.001 15. 3 Sodium chloride Neutral red Crystal violet g/litre 20.0-4. streaking for single colonies.0-3.0 0.0 CV. Inoculation: Surface. aureus No growth Other Staphylococcus spp. (D) colour Red Pink-red Paleyellow Yellowgreen Paleyellow (pink) other (red ppt around colony) (mucoid) (fishy odour) K. 8: 322-341. dry the surface of the agar by partial exposure at 37˚C.0-3.E. Taylor.0-3. E. aeruginosa 2.30 . aureus 1.0 Ps. streaking for single colonies.0 10.2 Agar No.CR. Methods for the Examination of Water and Associated Materials.0-3.0 Proteus spp. (1908) Bile salt media and their advantages in some bacteriological examinations.2 Minimum Q. Prior to inoculation. Churchill. Pink-Red Yellow Kleb. The Examination of Water Supplies. Dry the surface before inoculation. (1958).H. Salmonella spp.H. (Camb.O. 7th Edn. Growth Characteristics organism E. organisms: E. CV.0 0. Formula Peptone Lactose Bile Salts No.. 7th Edn.5 5.0 Description A medium first introduced by MacConkey in 1905 for the isolation and differentiation of lactose and non lactose fermenting enteric bacteria.(lactose-negative) Orange PinkDeep Red Ps.0-4.E. J. CV.H. Gram positive organisms will not grow on this medium.D. it is used for culturing a wide range of clinical material and has applications in food. S. 0.0-3.O.0 5.0 1. 2 g/litre 20. W.5 Method for reconstitution Weigh 48.0-4. F.0 CV. Churchill.odour if green (Green) Yellow Yellow Pink. Churchill.E. Minimum Q.CR. 2 Method for reconstitution Weigh 51. 3rd Edn.C. The Examination of Water and Water Supplies. E.0 Pink.W. 3 LAB 2 LAB 45 Description A modification recommended by the W.E.O.E. Incubation: 37˚C aerobically for 24 hours.0-4. organisms: E. Geneva. pH: 7. 2.G. Appearance: Pale red slight violet tinge. A.Hyg. 3rd Edn. (1905) Lactose-fermenting bacteria in faeces. CV.World Health Organisation (1971).Hyg. J.5 grams of powder and add to 1 litre of deionised water.05 13. aeruginosa 1. International Standards for Drinking Water.5 References MacConkey. Inoculation: Surface inoculation.G.0 0. Taylor. World Health Organisation (1971).0-1.E. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C. Environment Agency: The Microbiology of Drinking Water (2002). coli colony size shape & (mm) surface 2. The medium has since been modified to improve the recovery of staphylococci and enterococci. Cool to 47˚C and pour into Petri dishes. 1. Allow to soak for 10 minutes.

.P.0 15. 44˚C aerobically for E. It should be noted that excess heating of this medium together with its low pH can easily result in hydrolysis of the agar gel producing soft plates. Formula Malt Extract Mycological Peptone Agar No. 2 g/litre 30.References American Public Health Association (1950). Diagnostic Procedures and Reagents. Selectivity can be increased by further lowering the pH with the addition.H. Leonard Hill. 4th Edn. London. H. subtilis (inhibition) Storage of Prepared Medium: Capped containers – up to 1 month at 15-20˚C in the dark. H.H.S. New York. Lactose-fermenting organisms cause a colour change from purple to yellow. Incubation: 37˚C aerobically for coliforms.4 ± 0. Applied Mycology and Bacteriology.. coli.P. (1952). coli. Thom and Church.C. L. R.2 Minimum Q.. Prepare single strength broth (35g/l) if 1ml or 0. The Aspergilli. clear.C.D. Public Health Laboratory Service Water Committee (1969). and Burgess.0 0.0) Method for reconstitution Weigh 35 grams of powder. Mix well and dispense into tubes or bottles with inverted Durham tubes. International Standards for Drinking Water. Formula Peptone Lactose Bile Salts Bromocresol purple g/litre 20. A. A. swirl to mix then sterilise at 115˚C for 10 minutes.0. It was first described by Thom and Church in 1926 in a study of Aspergillus spp. London. Appearance: Purple. If the addition of XO37 Lactic Acid is required this should be done after sterilisation. One 5ml vial of XO37 will lower the pH of 250ml of medium to 3. Aspergillus niger 25 References Galloway. Sterilise by autoclaving for 15 minutes at 121˚C.. 9th edn. disperse in 1 litre of deionised water. NCIMB 50010 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. organism Candida krusei Penicillium notatum Growth Characteristics colony size shape & (mm) surface CV.2 (if XO37 is added pH 3.A.0 10. World Health Organisation (1971). Prepare double strength broth (70g/l) if 50ml or 10ml amounts of inoculum are to be added to equal volumes of broth.CR. 3rd edn.1ml amounts of inoculum are to be added to 10ml of broth.O.D. Growth Indicators: Turbidity. 3rd Edn. of XO37 Lactic Acid. Malt Extract Agar LAB 37 Description An acidic medium which will support the growth of most yeasts and moulds whilst inhibiting most bacteria.0 MacConkey Broth (Purple) LAB 5 Description This medium is used in the detection and enumeration of faecal coliforms (37˚C) and E. Minimum Q.H. coli (44˚C).O. The replacement of neutral red used in the original formulation by bromocresol purple makes the colour change caused by acid producing organisms easier to read. Minister of Health.A.3 ± 0. organisms: E.S. Use Durham tubes to detect gas production for E.O. American Public Health Association (1946). Standard Methods for the examination of Water and Sewage. Capped container – up to 1 month at 15-20˚C in the dark.. New York.M. W.31 . claiming the high carbohydrate content ensured rapid growth. Inoculation: Pour plate technique or surface streaking for single colonies. allow to soak for 10 minutes.0 5.D. disperse in 1 litre of deionised water. London. Cool to 47˚C before making additions and pouring plates. Bacteriological Tests for Graded Milk. Incubation: 25˚C aerobically for 5 days. pH: 7. 10 25 colour White White Green (white/yellow velvet strain dependent) White border. report No. 71. clear. The Bacteriological Examination of Water Supplies. black centre (yellow/black centre) other Candida albicans 4 References Ministry of Health (1937). gas production.5-4. 1.E. 1926.M. organisms: Candida spp. pH: 5. coli NCIMB 50034 B. Geneva.5-4. F.01 Method for reconstitution Weigh 50 grams of powder. after sterilisation.0 5. Memo 139/Foods. Appearance: Pale brown/straw.

Davis. clear gel. Formula Peptone Sodium chloride g/litre 1. Incubation: 25˚C (or 37˚C) for up to 7 days aerobically.4 ± 0. This medium can be made selective for methicillin resistant S.0 3.G. swirl to dissolve and dispense into final containers.G.2 1. colour Bright Yellow White or (some ferment Yellow mannitol) other Method for reconstitution Weigh 20g of powder and disperse in 1 litre of deionised water.5-2. 5: 21-25.0-1.0 12. Method for reconstitution Weigh 108 grams of powder. (1959). (1983). 1/4 strength Ringer’s LAB 100.0 CV.5-4. L. Milk Testing 2nd edn. clear broth.5 CV. Appearance: Red.C. 1 Sodium chloride D-Mannitol Agar No. The majority of Staphylococcus aureus ferment mannitol producing yellow colonies. R.P. Growth Characteristics organism S. J.Malt Extract Broth LAB 159 Description A liquid medium of low pH for the growth of yeasts and moulds. aureus colonies by other means e. Incubation: 37˚C for 48 hours aerobically. London pp 54 & 57. depending upon protocol used. BS5763 Part 6. Enterobacteriaceae no growth Vibrio spp.0 various usually Pink (yellow if mannitol +ve) Inoculation: Inoculate samples direct into tubes of broth according to the particular method being employed.E. Sterilise by autoclaving at 121˚C for 15 minutes.0 75.. coagulase. Preparation of dilutions. Washington. Allow to soak for 10 minutes.0 Minimum Q.0 0. mix by swirling. 2nd Edn. aureus by the addition of X207 (methicillin supplement).L. protein A. The presence of a low level of peptone lessens the physiological shock normally experienced by bacterial cells when they are introduced to a diluent such as Ringer’s Solution. Appearance: Clear fluid. The high carbohydrate content of the medium ensures rapid growth of yeasts and moulds. disperse in 1 litre of deionised water. It is necessary to confirm the identity of presumptive S.M. aureus Other Staphylococci colony size shape & (mm) surface 1. DN’ase. & other halophiles 1. pH: 7.5 grams of powder in 1 litre of deionised water.D. Recommended Methods for Microbiological Examination of Foods.G.5 Method for reconstitution Dissolve 9. Formula Beef Extract Balanced Peptone No. Sterilise by autoclaving at 115˚C for 10 minutes. Sharf) A. London.0 8. J. This formula is recommended by ISO 6887: BS5763. coli (inhibition) Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. R. ISO 6887.4 + 0. Minimum QC organism: Candida albicans NCIMB 50010 Aspergillus niger NCIMB 50097 References Galloway. Cool to 47˚C before pouring into Petri dishes. and Stokes.0 10. (1957). Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. organisms: S.2 1.025 Description An osmotically controlled solution which is an alternative to. (1952) Applied Mycology and Bacteriology. aureus E. typically employed as part of sterility testing protocols for various products. J. Appl.E. Appearance: Pale brown/straw. References American Public Health Association (1966). occasional strains of coagulase negative staphylococci may also ferment mannitol. Maximum Recovery Diluent (Peptone/Saline diluent) LAB 103 Mannitol Salt Agar LAB 7 Description A medium for staphylococci which is selective because the high sodium chloride level inhibits most other species with the exception of halophilic vibrios.g. (ed.32 . References Straka.0 10. 3rd ed. Dairy Industries. heat gently to dissolve then distribute into final containers. Inoculation Method: Surface plating. and Burgess.H. and a replacement for. 2 Phenol Red g/litre 1. Allow to soak for 10 minutes. pH 5. Subculture turbid tubes onto solid media for identification of growth.A. Microbiology-General guidance for the preparation of dilutions for microbiological examination. Rapid destruction of bacteria in commonly used diluents and its elimination. Formula Malt Extract Mycological Peptone g/litre 17. streak out for single colonies. The level of peptone is such that multiplication of the organisms is not possible in the time in which the sample will be present in the diluent (1-2 hours). Leonard Hill. Microbiol.P. thermonuclease or latex agglutination. Sterilise by autoclaving for 15 minutes at 121˚C.

.0 30. rinse waters and dairy products. 40th Ann Rep. Incubation: E.P.0 Method for reconstitution Weigh 76.return to incubation for a full period. With the addition of a further 5 g/l Agar No. LAB 129 to confirm gas and indole production respectively.2 ± 0. After 48 hours incubation at 37˚C a result should be obtained regarding the production of gas. coli NCIMB 50034 S. Met.O. a sample of 100ml should be filtered. coli 4 hours at 30˚C 14 hours at 44˚C. Soc.4 ± 0. The volumes should be chosen so as the number of colonies on the membrane lies between 10 and 100. 16:40.8). 1. clear solution. Appearance: Red. disperse in 1 litre of deionised water.Membrane Lauryl Sulphate Broth LAB 82 Description This medium superseded Membrane Enriched Teepol broth when Shell Chemicals withdrew Teepol 610 from sale. pH: 7.0 1. Full details of the methodology can be found in The Bacteriological Examination of Water Supplies 71.2 1. Formula Yeast Extract Peptone Antibiotic Free Skim Milk Powder Agar No. Div.2 Minimum Q. Incubation: Aerobically at 30˚C for 72 hours. 1969.C. disperse in 1 litre of deionised water. Inoculation: Pour plate technique. Memo 139/Foods H. after 24 hours incubation at 44˚C. Interpretation: No colonies:. References Ministry of Health (1937). References Burnham. after filtration. Proc. These membranes should be incubated in a container which does not allow evaporation to occur. Water Exam. E. swirl to mix then sterilise for 15 minutes at 121˚C.0 0. Minimum Q. Coliforms 4 hours at 30˚C 14 hours at 35˚C. Formula Peptone Yeast Extract Lactose Phenol red Sodium lauryl sulphate g/litre 39. coli and coliform counts should be made on separate samples of water.2 grams of powder.33 . With waters expected to contain less than 1 coliform per ml. Appearance: White. X219) for accelerated shelf-life determination of dairy products.0 6. (1967). opalescent gel.2 Method for reconstitution Weigh 24 grams of powder. organisms: E. British Standard 4285: Methods of Microbiological Examination for Diary Purposes. Wat. pH: 7. aureus NCIMB 50080 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.assume a nil count. Inoculation: E. Sodium lauryl sulphate was found to be an adequate reproducible substitute and this medium is recommended for the enumeration of coliform and organisms in water and sewage. Also see Milk Plate Count Agar LAB 115.0 15. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. Milk Agar can also be used with the P-INC supplement (X019. organisms: E. Allow to soak for 15 minutes. Environment Agency: The Microbiology of Drinking Water (2002). Water tight metal containers placed in an accurate water bath are required for incubation of membranes at 44˚C.0 5. N. Distribute into screw cap containers and sterilise by autoclaving at 115˚C for 10 minutes. Water Board London pp 18-22. E.0 Milk Agar LAB 19 Description An approved formulation for the enumeration of micro-organisms in milk.M. Bacteriological Tests for Graded Milk. Treat. Coliform organisms: Yellow colonies from membranes incubated at 35˚C or 37˚C should be subcultured into Lactose Broth LAB 126. 1 the medium is suitable for the preparation of RollTubes using established mechanical equipment. coli: Yellow-coloured colonies from membranes incubated at 44˚C should be subcultured to Lactose Broth LAB 126 and Tryptone Water.C. Exam. Windle Taylor. (see page 5.S. Cool to 45˚C before mixing with sample dilutions. 1 g/litre 3. London. (1961) Glutamic acid medium. Methods for the Examination of Water and Associated Materials. The membrane filter should be placed face upwards on a pad soaked in Membrane Lauryl Sulphate Broth. Small colonies of an intermediate colour:.

2 Method for reconstitution Weigh 19. Formula LAB 80a Lactose Sodium formate L-Cystine L(-) Aspartic acid L(+) Arginine Thiamine Nicotinic acid Pantothenic acid Magnesium sulphate (MgSO4. 10192. disperse in 1 litre of deionised water. The production at 44˚C of gas from lactose and the formation of indole are evidence of E. (1964).35 grams of base medium (LAB 80a) together with 6. Single strength: Dissolve in 11.002 0. Incubation: 37˚C for 18-24 hours aerobically. (1968).C. Sterilise by autoclaving at 121˚C for 15 minutes. 62: 495-508.04 0. Formula Tryptone Yeast Extract Dextrose Antibiotic Free Skim Milk Powder Agar No. Methods for the Examination of Water and Associated Materials. 56: 377-388.9 ± 0.5 0. A tube of Tryptone Water LAB 129 should also be inoculated and incubated at 44˚C for 24 hours for the production of indole. Hyg. Calculate back to determine viable organisms per ml References British Standards Institute. 1 g/litre 5. alternatively heat to 100˚C for 30 minutes on three successive days. J. A comparison between MacConkey broth and Glutamic acid media for the detection of coliform organisms in water.I. aureus NCIMB 50080 Storage of Prepared Medium: Capped container – up to 3 months at 15-20˚C in the dark. Incubation: 30˚C aerobically for 72 hours.0 10.7 grams of base medium (LAB 80a) together with 12. An improved formate lactose glutamate medium for the detection of Escherichia coli and other coliform organisms in water. D. coli NCIMB 50034 S. Hyg. organisms: E.Milk Plate Count Agar LAB 115 Description A medium recommended by the British Standards Institute and the International Organisation for Standardisation for the enumeration of viable bacteria in milk and other dairy products.7 ± 0. (1984). PHLS Water Sub-Committee. Interpretation Tubes showing the production of acid (medium turns yellow) and gas in the Durham’s tube are considered presumptive positive.C. (1958). Minimum Q. clear solution.N. Interpretation: Count all colonies.2 Minimum Q. BS 4285 Section 1. organisms: E. Inoculation: Use the Most Probable Number technique.0 2.34 .02 12.7 Method for reconstitution Double strength: Dissolve 22. opalescent gel. The product is supplied in two parts because it has been shown that separating the sodium glutamate from the base improves its stability.2H2O) Dipotassium hydrogen phosphate Bromocresol purple LAB 80b Glutamic acid (sodium salt) 0. pH: 6.048 0.5 1. Camb.0 1. Camb. 1. pH: 6.7 grams of sodium glutamate (LAB 80b) in 1 litre of deionised water containing 5 grams of ammonium chloride eg BDH cat no.P.0 Calcium chloride (CaCl2.35 grams of sodium glutamate (LAB 80b) in 1 litre of distilled water containing 2. swirl to mix then bring to the boil to dissolve the agar.2.02 1.04 0. 27149. Teepol Broth and Glutamic Acid Media for the enumeration of Coliform organisms in water.002 0. Appearance: Pale cream. coli. International Organisation for Standardisation Draft International Standard. Dispense 10ml and 50ml volumes into tubes with inverted Durham tube. PHLS Standing Committee on Bacteriological Examination of Water Supplies.2 0.02 Gray.7H20) Ferric ammonium citrate g/litre (double strength) 20. R. Comparison of MacConkey Broth. Hyg. With 10ml and 50ml of sample add to equal volumes of double strength medium. References Minerals Modified Glutamate Medium LAB 80A & LAB 80B Description This medium was developed for use with the Most Probable Numbers Technique (M. Appearance: Purple. Dispense 5ml volumes into tubes with inverted Durham tubes. J.5 grams ammonium chloride. Each presumptive positive tube should be subcultured to Brilliant Green Bile Broth LAB 51 with Durham tube and incubated at 44˚C for 24 hours and examined for gas production.8 0.0 0. Inoculation: Pour plate technique. With 1ml volumes of sample add to 5ml of single strength medium.5 grams of powder. Sterilise by autoclaving for 10 minutes at 115˚C.) for the enumeration of coliforms in water supplies. Camb.D. Allow to soak for 10 minutes. 66: 67-87. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. The medium is an improved version of the chemically defined glutamic acid medium described by Gray in 1964. (1982) ISO/DIS 6610. Ensure the Durham tube is free of bubbles.N. J. Allow to cool to 47˚C and dispense into suitable containers. Environment Agency: The Microbiology of Drinking Water (2002).002 0.

clear.0 2. DO NOT AUTOCLAVE OR OVERHEAT. Heat to dissolve and sterilise by autoclaving at 121˚C for 15 minutes.01 15.0 2.E.0-3. Citrobacter spp.0 20. Some problems may occur with H2S negative strains. M. Allow to soak for 10 minutes. Formula Yeast Extract Tryptone Meat Peptones Sodium chloride Mannitol L-Lysine HCL Sodium thiosulphate Ferric ammonium citrate Brilliant green Crystal violet Agar No. NCIMB 50076 E.R.0 3. Cool to 47˚C and adjust pH if the acidified medium is required. 66th meeting of the Japanese Vet. sendai and S.R.4 ± 0. pH: 6.0 10. typhi and S. Allow to soak for 10 minutes. swirl to mix then bring to the boil with frequent agitation to completely dissolve the powder. CV. Agar can be used to enumerate Lactobacillus bulgaricus in yoghurts. berta. (H2S negative) Proteus spp.0 1.0 5. Appearance: Light amber. organisms: Lactobacillus acidophilus 1. this medium will detect lactose and sucrose fermenting strains. eg S. 2 g/litre 5. senftenberg.4 M.0 0. Agar (Mannitol Lysine Crystal Violet Brilliant Green Agar) organism Salmonella spp. Salmonella spp. sodium acetate. Appearance: Pale purple. Inoculation: Surface plating.0 0.35 . swirl to mix. Mainly inhibited pale may have black centre LAB 116 Description A medium for the selective isolation of Salmonella spp.0 4.0 2. as these strains are more susceptible to the brilliant green dye. Method for reconstitution Weigh 70 grams of powder and add 1 litre of deionised water.G.G. Agar (de Man. Shigella spp. pH: 6.E. typhi and S.C.S.5 CV. or from enrichment broths. translucent gel. Incubation: 37˚C aerobically for 24 hours.0 4. streaking for single colonies.2 Minimum Q.08 15.E. Because of the low selectivity of this medium the inoculum should not be heavy. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.0 E.012 0. disperse in 1 litre of deionised water.5-2. The medium is suitable for most lactic acid bacteria.0 7.0-3. and it is recommended that this medium should be used in conjunction with other more selective media.B.0 5. Formula Mixed Peptones Yeast Extract Beef Extract Glucose Potassium phosphate Sodium acetate Ammonium citrate Magnesium sulphate Manganese sulphate Tween® 80 Agar No. Tween® 80.0-2.C.S.2 0. This medium should not be used to detect S.G.05 1. Salmonella colonies are recognised by distinctive colonial appearance and H2S production and like the Bismuth Sulphite Agar of Wilson & Blair. 1 g/litre 10. Growth Characteristics colony size (mm) 2. from various sources and is intended as a substitute for Tomato Juice Agar. S.2 Minimum Q.0 1. paratyphi A) from food and faeces. (with the exception of S.0 Method for reconstitution Weigh 49 grams of powder. CV. paratyphi A.M. Rogosa and Sharpe Agar) LAB 93 Description This medium was developed for the cultivation and enumeration of Lactobacillus spp.L. organisms: Salmonella spp. coli Klebsiella spp Gram positive organisms mainly inhibited mainly inhibited mainly inhibited References Inove et al. S. When acidified to pH 5. Cool to 47˚C and pour plates.8 ± 0. magnesium and manganese sulphates act as growth stimulants. pullorum. Medicine Society.0 5.0 shape & surface CV.E. Inoculation can be carried out directly.G.0 5. colour Black Pale Grey brown mainly inhibited 0.C.

130-135. Motile organisms spread from the centre in the semi-solid agar. 23. clear. clear. M. but non-salmonellas are inhibited by the selective agents. coli (inhibition) NCIMB 50034 Storage of prepared medium: Plates – up to 7 days at 4˚C. Incubation: 25˚C microaerobically for 2-5 days.3 References de Man. After overnight incubation the use of polyvalent salmonella antisera or a latex kit can confirm the presence of a Salmonella.9 0. and Sharpe. G.5˚C. Interpretation: Count all colonies exhibiting typical morphology.0 0.F. Keep lid uppermost at all times. this product has the same formulation as LAB 93 M. Formula Mixed Peptones Yeast Extract Beef Extract Glucose Potassium phosphate Sodium acetate Magnesium sulphate Manganese sulphate Tween® 80 Ammonium citrate g/litre 10.05 1. R. is inoculated directly from the preenrichment medium. Bolderdijk and Rappold as a rapid means of Salmonella detection. A medium for the cultivation of lactobacilli.S. for 18-24 hours. Soak for 10 minutes. and Bolderdijk.5 grams of powder and disperse in 1 litre of deionised water.3 1.5 10. and will signal the presence of a Salmonella by inhibiting the mobility of the organism around the disc. This should be confirmed by subculturing from the edge of the mobility zone onto XLD and brilliant green agar and performing biochemical and serological tests. in the centre of the plate. Capped containers – up to 1 month at 15-20˚C in the dark. Inoculation: Surface. Rogosa. Appl. CV.. Interpretation: For enumeration purposes count tubes showing signs of growth as positive. M. based upon Rappaport Vassiliadis broth. organisms: Lactobacillus acidophilus Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. Direct latex agglutination may be carried out from the edge of the mobility zone.5-1.5-2. substantiated if a disc with polyvalent H antiserum has been added and is inhibiting the zone.G.5 0. Brixia Academic Press.E. Inoculation: Either with suspect colonies from M.S. soft gel. CV. A medium for the cultivation of lactobacilli.0 2. pH: 6. Alternatively.E.0-1.C.G. Growth Characteristics organism Lactobacillus acidophilus Lactobacillus sake Lactobacillus bulgaricus colony size (mm) 0. Interpretation: A spreading growth indicates a Salmonella may be present.08 2. or use pour plate technique.Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.4 ± 0.S. M. Appearance: Turquoise/blue. 23: 130-135.0 Formula Tryptone Meat Peptone Acid Hydrolysed Casein Sodium chloride Potassium dihydrogen phosphate Magnesium chloride Malachite green Agar No.3 2.E. Appl.C. or 42 ±0.2 0. agar with the omission of agar. (1960). Inoculation: From pre-enrichment broth (6-24hrs) adding 0. Incubation: 37˚C. Mix well before dispensing. J.E. F.. spread to cover surface. References De Smedt.7 7. pH: 5. References de Man.0 shape & surface F.S. J.R.36 . Bacteriol. Minimum Q. Appearance: Light amber. a paper disc wetted with polyvalent H antiserum can be placed 1/3 of the way from the edge of the dish.0 5. agar.R. Fifth International Symposium on Rapid Methods and Automation in Microbiology and Immunology. M and Sharpe.2 Minimum Q. Dispense into suitable tubes or bottles then sterilise by autoclaving at 121˚C for 15 minutes.5 Method for reconstitution Weigh 31. agar or with serial dilutions of test material.2 0. MSRV colour White Rough White White White (Semi-solid Rappaport Medium) LAB 150 Description MSRV was developed in 1986 by De Smedt. (1960). The medium can be used for confirmatory tests on organisms isolated on M. (1987): ‘One Day Detection of Salmonella from Foods and Environmental Samples by Mobility Enrichment’.0 5. J. Bacteriol. Cool to 47˚C.R.E. 1. The medium can also be used for enumeration by the Miles and Misra technique.. Rogosa and Sharpe Broth) LAB 94 Description A medium for the cultivation and enumeration of Lactobacillus spp. J.M. Florence (1987).5-1. Using this medium De Smedt and Bolderdijk have reported the possibility of detecting Salmonella in 24hrs (1987) Streptococcus lactis 0.1ml to the centre of the plate. swirl to mix and bring to the boil.G.R.5 1.2 Method for reconstitution Weigh 55 grams of powder. 1 g/litre 2. Incubation: 25˚C microaerobically for 2-5 days. disperse in 1 litre of deionised water.C.E. The medium.3 4. Rogosa. Allow to soak for 10 minutes. swirl to mix then warm to completely dissolve solids. M. and add 2 vials of X150 novobiocin supplement (10mg/vial).0 20. Broth (de Man.C organisms: Salmonella typhimurium NCIMB 50076 E.037 2. J.0 10.

Incubation: As recommended by methodology for particular organisms and antibiotics by NCCLS. aureus (antibiotic sensitivity zones) ATCC 25923 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. 3rd edn.3 ± 0. References Mueller.0 17. (1941). Janssens. This medium. M. controlled to ensure correct zone sizes with aminoglycoside and tetracyline antibiotics. H. J.37 ..4 ± 0.A. (1987).S. 48: 330-333. De Smedt. L. values. A.C. and Schwab. American Public Health Association.C.F. R. NCCLS..P.0 17.. (1943). and Butzler. R. R. J. New York.S. Collaborative Study of the International Office of Cocoa.L. Proc. J. Baltimore.M. (1985). has been adopted by the National Committee for Clinical Laboratory Standards (NCCLS) in the USA as the definitive method for susceptibility testing. organisms: E. (1990). Clin.I. Wauters. standards for antimicrobial susceptibility tests on bacteria which grow aerobically.H.F. Journal of Food Protection 50 658-661. Semi-solid selective mobility enrichment medium for isolation of Salmonella from faecal specimens J.. M.M. J. Media for isolation cultivation. disperse in 1 litre distilled water. and Lautenschlaeger.. swirl to mix then sterilise at 121˚C for 15 minutes. Formula Beef Extract Acid Hydrolysed Casein Starch Agar No.C. (1986). (1985). and Bolderdijk. coli ATCC 25922 S. Method for reconstitution Weigh 38 grams of powder. Allow to soak for 10 minutes.5 17. J.F. Protein-free medium for primary isolation of gonococcus and meningococcus. Rapid Salmonella Detection in Foods in Mobility Enrichment on a Modified Semi-Solid RappaportVassiliadis Medium.. pH: 7.C.1 Minimum Q. It is an antagonist free medium for use in the tube dilution technique for the determination of antibiotic M. De Smedt. and sterilise at 121˚C for 10 minutes. Appearance: Pale straw.C.C.0 50-100mg/litre 20-35mg/litre Magnesium ions Method for reconstitution Weigh 21 grams powder. Med.S. Gould. Mueller Hinton Broth II LAB 114 Description This medium is the broth version of Mueller Hinton Agar.L. values) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.I. aureus ATCC 25923 E. Microbiol 19 940-941.. 1. Incubation: As recommended by methodology for particular organisms and antibiotics by NCCLS. Appearance: Straw coloured. Goodale. swirl to mix then heat gently to dissolve.H.. Williams & Williams. pH: 7. N. used in the technique of Bauer and Kirby. Soc. organisms: S. Methods for dilution antimicrobiol susceptibility tests for bacteria that grow aerobically. Exp. Rappold H. making it suitable for trimethoprim and sulphonamide testing.5 50 mg/litre 20 mg/litre Mueller Hinton Agar II LAB 39 Description A medium for antimicrobial sensitivity testing by the disc diffusion method. coli ATCC 25922 (M.C.I. J.. and Bolderdijk. clear gel. 1 Calcium ions Magnesium ions g/litre 2.Am. (1986). Journal of Food Protection 49 510-514. J. Ass. D. The medium is carefully standardised to meet N. J. mix well and pour plates. Formula Beef Extract Acid Hydrolysed Casein Starch Calcium ions g/litre 2.. disperse in 1 litre of deionised water. Inoculation: Standard inocula are required. W. Cool to 47˚C. inoculum as described by N. Laboratory Identification of sulphonamide resistant gonococcic infection. The medium has a very low thymine and thymidine content. Distribute into tubes or bottles. G. Diagnostic Procedures and Reagents. Inoculation: Surface. and Hinton. Goossens. 123: 547-549. (1984).L.P.De Smedt. Journal of Food Protection 53 659-664.C. Biol. G. (1950). Dynamics of Salmonella Isolation with Modified Semi-Solid Rappaport-Vassiliadis Medium.C.5 1.M. and Bolderdijk. identification maintenance of medical bacteria. De Boeck. References MacFaddin.5 1. Approved Standard.2 Minimum Q. Performance standards for antimicrobial susceptibility testing – second informational supplement. The medium was originally formulated as a heat labile protein free medium for the isolation of pathogenic Neisseriaceae. and Med.-M7-A. clear. Chocolate and Sugar Confectionery on the Use of Mobility Enrichment for Salmonella Detection in Cocoa and Chocolate.C. As described by NCCLS. Allow to soak for 10 minutes.

Hyg. However some salmonellas will be missed in this medium either because they are sensitive to Brilliant Green or cannot utilise tetrathionate (included in this group is S. and Kampelmacher. Bring to the boil and cool to below 45˚C.C. Appearance: Transparent.5 grams of powder. pH: 7. D.C. P. W. Formula Tryptone Soy Peptone Sodium chloride Calcium carbonate Sodium thiosulphate anhydrous Ox Bile g/litre 7. ISO 3565 (E).0 7. Suppl. The standard Salmonellae isolation method.R. Appearance: Green.L. 217: 1-90.0 15. XLD LAB 32. J. 41: 297-306. References Ericsson. 117: 26-32.P.. Wld.A. Z. Incubation: 37˚C aerobically for 18-24 hours. L. organisms: Salmonella sp.C.4 ± 0.M.H. aureus NCTC 6571 E. Formula Acid Hydrolysed Casein Peptone Beef Extract Sodium chloride Glucose Agar No. Weitere Erfahrungen mit dem kombinierten Anreicherungsverfahren fur Salmonellabacillen. Pathol. LAB 34. ISO 6579(E).G. disperse in 1 litre of deionised water.8 ± 0. Milk and milk products. or breakpoint technique. LAB 116.2 Weigh 36. H.) This medium is used in the standard European Community Salmonella Isolation Procedure and in International Standards Organisation (ISO) Methods. and Waterworth. International Organisation for Standardisation. (1923). Detection of salmonellae (Reference method). and Skinner.J.1985. Effect of medium composition on the apparent sensitivity of Pseudomonas aeruginosa to gentamicin. Incubation: 43˚C aerobically for 24-48 hours. Microbiol. typhi. This medium will grow all organisms not having a specific need for blood. edited by Corry.0 40. Roberts. In: Isolation and identification methods for food poisoning organisms. International Organisation for Standardisation Microbiology-1981 General guidance on methods for the detection of Salmonella. Hlth. pH: 7. F. No. 17 35-49. Kauffman. Scand. F.0 2. Report of an international collaborative study. (1971). Subculture: Subculture onto selective agars. (1982). potassium iodine 25g. International Organisation for Standardisation. (1969). SAB Technical Series. J. Antibiotic sensitivity testing. Academic Press. Biol (Paris) 89: 434. Minimum Q. van Leusden. Pathol. Organisms which can utilise tetrathionate.75 Nu Sens Agar LAB 74 Description This is a sensitivity test medium developed by LAB M in response to requests for a nutritionally rich sensitivity medium. 1. London.0 Method for reconstitution Method for reconstitution Weigh 82 grams of powder. Edel.M.38 . distilled water 100ml) and 9.. Soc. turbid solution which precipitates on standing. and Beckers. disperse in 1 litre of deionised water.E.1% w/v solution.7 4. solidified with a very pure agar and is free from antagonists. References Mueller. organisms: S. 1 g/litre 2.L. (1935). coli NCTC 10418 (antibiotic sensitivity zones) Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. B. flourish. Comparative studies on Salmonella isolations in eight European laboratories.Mueller Kauffman Tetrathionate Broth LAB 42 Description A selective enrichment broth for salmonellae first described by Mueller in 1923 then modified by Kauffman in 1935 with the addition of Brilliant Green and Ox Bile increasing its selectivity. almost colourless. J.2 Minimum Q. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped container – up to 7 days at 2-8˚C in the dark. Inoculation: 1 part sample to 9 parts medium or 1 part preenrichment medium to 10 parts medium. Mix well and dispense into sterile containers.C. giving reproducible zone sizes.A. Inoculation method: Joan Stokes method.5ml of brilliant green 0.g. Allow to soak for 10 minutes. M. It is composed of specially selected peptones with a small amount of glucose. Clin.5 5. Prior to use. NCIMB 50076 E.0 5. Un nouveau milieu d’enrichissment pour la recherche du bacille typhique et des paratyphiques C. (1975). M.M. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. Org. Sherris. e. E. keeping the chalk in suspension. Garrod. F. (1968). add 30ml 1N iodine solution (or 19ml of a solution containing iodine 20g... Meat and meat products. Acta. such as most Salmonella. ISO 6785(E).3 25. surface inoculum for semi confluent growth.B. Detection of Salmonella.0 2. H. Cool to 47˚C and pour plates. 22: 534-538. Bull.3 2. van Schothorst.

Medical Microbiology.0 Nutrient Broth No. Strep. Bacillus spp. Sterilise by autoclaving at 121˚C for 15 minutes.0 spp. water.39 . pH: 7. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C. Growth Characteristics organism S. 1 Sodium chloride g/litre 10. organisms: S. pyogenes P.E.E. aureus NCIMB 50080 E.G.C. Appearance: Straw coloured.5-2.0 5. Minimum Q.0 F.0 2. Formula Beef Extract Yeast Extract Peptone Sodium chloride g/litre 1. 11th edn. References British Pharmacopoeia. London.5 spreading 2.3 ± 0. disperse in 1 litre of deionised water. aureus NCIMB 50080 E. Allow to soak for 10 minutes. (1972). Incubation: 37˚C for 18-24 hours aerobically.5-2. pH: 7. swirl to mix then dispense into tubes or bottles.2 Method for reconstitution Weigh 28 grams of powder.P. Heat to dissolve then dispense into bottles or tubes. aureus NCIMB 50080 E.S. Appearance: Buff. Grey-Green Method for reconstitution Weigh 13 grams of powder. Inoculation and incubation: To suit chosen organism. it is isotonic and can be enriched with biological fluids such as sterile blood and egg yolk. Livingstone.M.E.0 12.0-4. and sterilise for 15 minutes at 121˚C..0 CV.0-4. Cool to 47˚C.4 ± 0. Incubation: Temperature and time to suit organisms.E. Formula Peptone Beef Extract Sodium chloride Agar No.E.0 Method for reconstitution Weigh 25 grams of powder.0 3.3 ± 0. disperse in 1 litre of deionised water.G.0 8. coli Proteus spp.0-6. Cruikshank. mix well then pour plates.0-2. Nutrient Agar is suitable for teaching and demonstration purposes. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. organisms: S. CV.P. Ideal for teaching purposes.2 Minimum Q. Usually aerobic. Formula Beef Extract Balanced Peptone No.0 5. clear. Growth indicator: Turbidity. CV. Capped containers – up to 3 months at 15-20˚C in the dark.CR.G. aureus colony size shape & (mm) surface 1. organisms: S.0 other Staphylococcus 0.D. 2. Inoculation: Surface streaking for single colonies. Allow to soak for 10 minutes. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. CV.2 Minimum Q.Nutrient Agar LAB 8 Description A general purpose medium for the cultivation of organisms that are not demanding in their nutritional requirements e. colour WhiteYellow WhiteYellow Transp. R.C. 1. add to1 litre of deionised water. This broth can also be used as the suspending medium for cooked meat granules for the cultivation of anaerobic organisms. 2 g/litre 5. Grey Grey Grey Grey fishy odour mucoid may spread odour if pigmented other Nutrient Broth “E” LAB 68 Description An inexpensive broth for the growth of nutritionally non-demanding organisms. London.G. H.-0. 2 B. pH: 7.0 10.5 E. various Klebsiella spp.O. opalescent gel. dust etc.0 CV. LAB 14 Description A general purpose broth which can be used for sterility testing for aerobic organisms as recommended by the British Pharmacopoeia.0 5.G. aeruginosa 2. (1973). organisms that can be isolated from air.0 Ps.g. 1.C. clear. Appearance: Pale straw.

(1993) Examination of raw beef products for the presence of verocytotoxin producing Escherichia coli.0 Description Modified tryptone soy broth has emerged as the medium of choice for the enrichment of E. Formulation Tryptone Sodium chloride K2HPO4 Dextrose Soy Peptone Bile Salts No. 1.0 5.0 4. Crozier. Reilly. 2) Willshaw...Microbiol 40 3-9. Smith. Symptoms start with severe stomach cramps and watery.0 7. and the increase in foodborne infection3. T. coli O157:H7 (non-toxigenic) NCTC 12900 E.0 2. MTSB was the most productive and selective for the isolation of E. Streptococcus and Clostridium.Bacteriol. E..M. G. Proc.. 3) Sharp. V. Rowe. and the addition of novobiocin supplement (X150).I. References 1) Bolton.0 4.S. cultivation and identification of organisms which have caused spoilage in frozen concentrated orange juice. Allow to soak for 10 minutes. Roberts.2 Method for reconstitution Weigh 33 grams of powder and add to 1 litre of deionised water. By having a low pH and incorporating orange extract. cool to 47˚C. coli O157:H7 in red meats1.0 3. Food Tech. 6 p181. (1952) Evaluation of plating media for citrus concentrates.Food Microbiol. Food Tech 6 p386. Capped containers – up to 3 months at 15-20˚C. D. Some workers1 recommend the use of an immunomagnetic separation step after 6hrs incubation.. Orange Serum Agar is the ideal isolation medium.F.2. G.Appl. L. (1994) Escherichia coli O157 infections in Scotland.2 Minimum QC organisms: E. Bring to the boil swirling frequently. MTSB is made selective for O157:H7 by including bile salts in the dehydrated medium..U. in the dark.0 3. pH: 5. Murdock. (1995) Optimisation of methods for the isolation of Escherichia coli O157 from beefburgers. J. Subculture onto CT-SMAC (LAB161 plus X161) or SMAC-BCIG (HAL 6) and examine for non-sorbitol fermenting colonies and/or glucuronidase negative organisms. J.. and Troy. for 15 minutes. 5 days at 30˚C for yeasts and moulds.J. for bacteria. so too has the need to optimise methods for its efficient isolation. and a percentage of individuals infected will develop Haemolytic Uraemic Syndrome (HUS) leading to acute renal failure4. Leuconostoc. 4) Doyle. Appearance: Amber.0 1.F.. acidophilus Pen. pH: 7.’s) per ml of the sample allowing for dilution factors.5 Method for reconstitution Weigh 42 grams of powder.K. M. Thirlwell. J. 75 420-426..J.Med. W. Int. Inoculation: Pour plate technique.R. Formula Tryptone Yeast Extract Orange Extract Glucose Di-potassium phosphate Agar No.P. disperse in 1 litre of deionised water. J..2 g/litre 10.5 ± 0. Interpretation: Count bacterial colonies and yeast/moulds separately. All broths should be subcultured to selective media whether turbid or not. J. J. slightly opalescent gel. Soc. and dispense into containers.O157 Broth MTSB (Modified Tryptone Soy Broth) Orange Serum Agar LAB 147 LAB 165 Description A medium developed for the investigation of organisms involved in the spoilage of citrus products including fruit juices and citrus concentrates. References Hays.40 . G. organisms L. roquefortii Storage of Prepared Medium: Plates – up to 7 days at 4˚C. D. Incubation: 3 days at 30˚C. J. J.E. and Reister. H. As concern regarding this organism has grown due to the severity of the disease syndromes caused. coli O157:H7. Florida State Hort. D. Sterilise by autoclaving at 115˚C. particularly those of serogroup O157.5 3. Coia. Appearance: Clear straw broth Minimum Q. (1991) Escherichia coli O157:H7 and its significancein foods. Cool to 47˚C and add 2 vials of Novobiocin supplement X150. (1951) The isolation. Incubation: 42˚C aerobically for 24hrs. Allow to soak for 10 minutes. Mix well and distribute aseptically into sterile containers.2 ± 0. PHLS Microbiology Digest 12 (2) 67-70. in the dark. The low pH of these products restricts the growth of organisms to those capable of tolerating an acid environment such as yeasts and moulds and bacteria belonging to the genera Bacillus. Cheasty.A. Curnow.C.J. Calculate the colony forming units (C. Williamson.. Lactobacillus. Hays. coli NCIMB 50034 (inhibition) Inoculation: Add 25g sample to 225ml of supplemented MTSB and homogenise for 2 minutes. swirl to mix and autoclave at 121˚C for 15 minutes..3 g/litre 17. B. 12 289-302.W.L. In a comparison of 4 different selective broth media.L. (1952) The control of ‘off-odour’ spoilage in frozen concentrated orange juice.C.0 5. bloody diarrhoea. Folinazzo. Interpretation: Turbidity in the broth indicates growth.

.001 12. Bact.E.CR. Acriflavine. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 4˚C in the dark. cerevisiae Pen. introduced by Mossel in 1970. Cool to 47˚C and add 2 vials of P. D. (1987). Inoculation: 0.0 20.41 . 35: 454-457.G.0 Formula Columbia Peptone Mix Sodium chloride Corn Starch Yeast Extract LAB 148 Description Palcam Agar was developed by Van Netten et al in 1989 as an improved selective differential medium for the isolation of Listeria monocytogenes from food. has a neutral pH and it has been shown to give better recovery rates than those media with a low pH.0 confluent shape & surface CV.5-2.D.08 12.E. Grey/green Black halo. Appearance: Pale yellow. Formula Yeast Extract Dextrose Biotin Agar No. CV. Board. (Small white/ yellow colonies with yellow/ green halo) - Enterococci Inhibited - Staphylococci Inhibited - - Bacillus spp.Y. clinical and environmental specimens. whilst the double indicator system of aesculin hydrolysis and mannitol fermentation aids differentiation of Listeria spp from enterococci and staphylococci which can be confused with Listeria spp on other types of culture media. swirl to mix then sterilise by autoclaving at 115˚C for 10 minutes.0 1. et al.5-2.0-5. Some factors influencing the recovery of yeasts and moulds from chilled foods.0-8.0 0. (draughtsman colonies). Mix thoroughly and pour into Petri dishes. for Selective Enumeration of Moulds and Yeast in Foods and Clinical Material.G. J.0-4. Improved selectivity is achieved by the combination of antibiotic supplements and microaerobic incubation. monocytogenes NCIMB 50007 E. Rose Bengal Chloramphenicol LAB 36 is recommended in these instances. (Small yellow colonies with yellow/green halo). 2 g/litre 5.0 ± 0. Method for reconstitution Weigh 71 grams of powder and disperse in 1 litre of deionised water.) Palcam Agar Base (Polymyxin.0 1.A. R.E. Soak for 10 minutes. O. Banks.C.0 3.E. notatum Pen.1ml of sample selectively enriched in Palcam Broth (or other enrichment medium) spread over surface of plate.5 10.Y.2 Growth Characteristics organism Candida spp. Aesculin. Inoculation: Surface spreading or pour plate.A. colour other Tetracycline resistant bacteria may grow.8 15. colour Cream White White Green/blue centre Yellow centre Yellow with black centre Aerial hyphae with black conidia Minimum Q. organisms L. Lithium chloride. g/litre 23.E. L. F. 2 Minimum Q.0-4. translucent pH: 7.0 Method for reconstitution Weigh 37 grams of powder. Mix thoroughly and pour into Petri dishes.D. Cool to 47˚C and aseptically add two vials of X089 Oxytetracycline selective supplement. organisms: Aspergillus sp.0 3. Grey/green Black halo.G.A.C. 4: 197-206. monocytogenes 1.0 3.D.2 ± 0. Appl.5 0.0 7. Food Microbiol. colony size (mm) 3.D. Incubation: 25˚C aerobically for 5 days.0 5. Allow to soak for 10 minutes. Inhibited - - 1.0mm F.2 Glucose Mannitol Aesculin Lithium chloride Ferric ammonium citrate Phenol red Agar No.D.E. J. Growth Characteristics organism colony size shape & (mm) surface F. Appearance: Red. J. disperse in 1 litre of deionised water. (1970).0 0.0mm References Mossel.5-2. Oxytetracycline is used to inhibit bacteria. (draughtsman colonies). pH: 7.0 0. Int. Ceftazidine. certain high protein foods may reduce the effectiveness of this antibiotic as a selective agent. Incubation: 30˚C aerobically or microaerobically for 24-48 hours.G.E. 0. clear. Other Listeria spp. Mannitol) LAB 89 Description A selective medium for the enumeration of yeasts and moulds in food.0 0. NCIMB 50097 Saccharomyces cerevisiae Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.5 1.Y. Capped containers – up to 1 month at 4˚C in the dark. colonial morphology dependent upon species. Supplement – X144.C. Unlike many selective media for yeasts O. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes.G. flavescens Aspergillus niger Rhizopus spp.Oxytetracycline-Glucose-Yeast Extract Agar Base (O. Candida krusei S.

Formula Tryptone Yeast Extract Soy Peptone Liver Extract Ferric ammonium citrate Sodium metabisulphite Tris buffer Agar No.2 ± 0.0 5. Subculture: Onto Palcam Agar – LAB 148.08 10.A. I.0 5. monocytogenes NCIMB 50007 E. The medium is buffered and utilises sodium metabisulphite and liver extract as sources of H2S with ferric ammonium citrate as the indicator. Incubation: 30˚C for 24 hours and 48 hours. coli (inhibition) NCIMB 50034 Storage of prepared medium: Capped containers – up to 7 days at 4˚C.8 0. Food Micro. LAB 109 Description This medium was developed by Handford in 1974 to overcome some of the problems associated with enumerating Clostridium perfringens in foods. pH: 7. 2nd edn. Inoculation: A light inoculum from a pure culture. organisms: E. Mannitol) LAB 144 Description Developed by Van Netten et al (1989) L-Palcam is a selective differential medium for the enrichment of Listeria spp. Mossel. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. Curtis. Sterilise by autoclaving at 121˚C for 15 minutes. Lithium chloride. the volume of water for reconstitution must be reduced accordingly.0 5.D. Method for Reconstitution Weigh 54.42 . D. 2 g/litre 15. monocytogenes Int.5 10..0 Palcam Broth (Liquid.0 References Van Netten.0 1. Williams and Wilkins.. the result being the indication of a potential problem up to 48 hours before growth on plating media can be observed.0 7. Polymyxin. Perales.P. in food. (1984). Aesculin. Peptone Water LAB 104 Description A general purpose growth medium that can be used as a base for carbohydrate fermentation studies.. monocytogenes Int. Food Micro. P. Baltimore/London. The medium is made selective with the addition of X109 Sulphadiazine and X110 Oleandomycin/Polymyxin supplements.A. Mix well and dispense into sterile tubes or bottles. swirl to mix and autoclave at 121˚C for 15 minutes.0 0. Formula Columbia Peptone Mix Yeast Extract Peptonised Milk Sodium chloride Mannitol Aesculin Ferric ammonium citrate Phenol red Lithium chloride g/litre 23. Baltimore/London.0 5. J.References Van Netten.A.A. J.2 ± 0. If low numbers of Listeria are present the medium may not produce the brown black colour. A pH indicator may be added to detect acid production from carbohydrate utilisation. Ceftazidime. 8 (4) 299-316.4 makes the medium an enrichment broth for Vibrio spp. Inoculation: Sample or pre-enriched sample added to the broth in the ratio 1 : 10.0 5. G. Soak for 10 minutes.0 5.4 grams of powder and disperse in 1 litre of deionised water. All tubes should be subcultured onto selective agar before a sample is scored as negative. colourless. Williams and Wilkins. Curtis. Acriflavine.. MacFadden.. environmental and clinical samples.C.5 0. (1989) Liquid and solid selective differential media for the enumeration of L.S.C. Biochemical Tests for the Identification of Medical Bacteria. (1983). Vol. 1. P.0 5.0 Sodium chloride Method for reconstitution Weigh 15 grams of powder. D.F. I.D.. Appearance: Clear red broth pH: 7.0 1.) Minimum Q. 1.: carbohydrates. If sterile additions are to be made to this medium e.W.2 Minimum Q. References Bergey’s Manual of Systematic Bacteriology. Perales.W. Formula Peptone Tryptone g/litre 5. Cool to 47˚C and add two vials of X144. The medium has a high level of tryptone making it suitable for use in the indole test. and disperse in 1 litre of deionised water. Allow to dissolve then distribute into final containers. G. 8 (4) 299-316.0 1.0 5. J. Mossel.g.P.2 Perfringens Agar (O. Appearance: Clear. Incubation: According to organism. It is unique amongst Listeria enrichment media in that it contains an indicator system (aesculin) which will signal the presence of a possible Listeria by a browning/blackening of the broth. organisms: L. (1989) Liquid and solid selective differential media for the enumeration of L. Adjustment of the pH to 8.

P.0 12.A. 6˚C aerobically for 10 days for aerobic psychrotroph count.H.0 ± 0. 6˚C aerobically for 10 days for aerobic psychrotroph count.A. Inoculation: Pour plates. organisms: C. clear. Interpretation: Count all colonies and calculate the number of organisms (or ‘colony forming units’ c. Cool to 44-46˚C for not more than 3 hours prior to use. Microbiol. A rapid technique for the enumeration of C. Minimum Q.A) A. 13th edn. Washington. perfringens in foods. The product uses agar of very high gel strength in order that it can be used in pour-plate as well as surface inoculation techniques. Appearance: Pale straw. E. Formula Tryptone Yeast Extract Glucose Agar No. (1987).u.0 ± 0.H. 1 prior to reconstitution of the medium. App & Env. 1985 p.R. Method for reconstitution Weigh 20. or pour plate. 2 g/litre 5.S.P.. 1 g/litre 5.5 grams of powder.P. Interpretation: Count large black colonies. disperse in 1 litre of deionised water. 37: 559-570. Plate Count Agar and is suitable for the determination of total viable counts in food products by surface count and pour plate methods. perfringens in food. P.2 References Reasoner.Method for reconstitution Weigh 45. Inoculation method: Pour plate technique or surface inoculation. 13: 559. Appl. Washington.F. perfringens.) per ml of sample allowing for dilution factors. pH: 7. disperse in 1 litre of deionised water. pH: 7. Food Microbiol.) (Standard Methods Agar.0 15. Washington D. Microbiol.2 Minimum Q. Geldreich. D.0 Method for reconstitution Weigh 23.H.f. and McClung.2 References American Public Health Association (1972). epidermidis NCIMB 50082 E. 14th edn. (1985) A New Medium for the Enumeration and Subculture of Bacteria from potable water. A new quantitative and confirmatory medium for C. Washington. American Public Health Association (1976).C. 16th Edition. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. 1.C. ROLL-TUBES. Allow to soak for 10 minutes. Plate Count Agar (A. S.0 2. Dispense into tubes and sterilise by autoclaving at 121˚C for 15 minutes.5 grams of powder. J. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. The product can be remelted prior to use although it should not be held for a prolonged period in the molten stage. 2nd edn.5 1. J.A. Add an additional 10g/litre Agar No. (1965). and Ferguson. Inoculation: Surface.A. Incubation: 37˚C anaerobically for 24 hours. Appearance: Pale straw colour.C. pH: 7. dairy and water bacteriology. disperse in 1 litre of deionised water.. epidermidis NCIMB 50082 E. Capped containers – up to 3 months at 15-20˚C in the dark. perfringens. specifications.M. Allow to soak for 10 minutes then sterilise by autoclaving at 121˚C for 15 minutes. perfringens NCIMB 50027 E. 5: 3: 240-241. Cool to 47˚C then pour into Petri dishes. (ed. presumptively identified as C.. A new medium for the detection and enumeration of C.A.A.) A. Appl. clear gel.43 .A.5 1. M. American Public Health Association Inc.1-7. The formula is equivalent to A. Standard Methods for the Examination of Dairy Products. Jan.0 2. Appl..P. Bact. 55˚C aerobically for 48 hours for aerobic thermotroph count. (1974). (ed. Environment Agency: The Microbiology of Drinking Water (2002). Methods for the Examination of Water and Associated Materials. Bring to the boil with frequent stirring to dissolve. organisms: S. Plate Count Agar LAB 149 Description A medium designed for use with the spiral plating system and other surface inoculation techniques. Incubation: 30˚C aerobically for 48 hours for aerobic mesotroph count. R.C. Franson. Hausler.M. Recommended Methods for the Microbiological Examination of Foods. 1ml sample plus 9ml medium.. When set overlay with sterile medium. Pharmacopoeia of culture media for food microbiology. Standard Methods for the Examination of Water and Waste Water.J. Tryptone Glucose Yeast Agar) LAB 10 Description Formulated to A. Incubation: 30˚C aerobically for 48 hours for aerobic mesotroph count. American Public Health Association (1985) Standard Methods for the Enumeration of Water and Wastewater.. Allow to cool to 47˚C before adding 2 vials each of selective supplements X109 and X110. 55˚C aerobically for 48 hours for aerobic thermotroph count. (1971). Shahidi. (ed. Sharf.J. J.P.5 grams of powder. Int. L. Microbiol.H.P. Minimum Q. A. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.H. organisms: S. Steenberger.E. W.) A. Appearance: Pale straw coloured. 21: 500-506. Marshall. clear.0 References Handford. J. this medium is used for establishing total viable counts for aerobes in food. American Public Health Association (1966).3 ± 0. coli NCIMB 50034 (inhibition) Storage of prepared medium: Use on day of preparation. Formula Tryptone Yeast Extract Glucose Agar No.S.H. Interpretation: Count all colonies or use spiral plating colony count equipment.

Appearance: Translucent white agar. and Adams. and Enomoto. M.0 4. colour Yellow Grey Grey Grey fluorescence yes yes no no no Ps. supplement. J. Jap.J.O.Cr.N. supplement or X108 C. Lab. Fluffy White Fluffy White C. Allow the medium to cool to 47˚C then add the contents of 2 vials of either X107 C. organisms: P.0 Pseudomonas Agar Base (C. 2 Method for reconstitution Weigh 48.Cr. soft drinks. Washington D.R. References Burton. maltophila P. Add 10ml of glycerol.. (1978). B.C.0 Tryc. Tryc. Inoculation: Surface. 5th ed.N. This addition must be after autoclaving and cooling to 47˚C. (ed.1 to 0. Formula Potato Extract Dextrose Agar No. disperse in 1 litre of deionised water.D.E. E. Mix well before pouring into sterile Petri dishes. Sterilise by autoclaving at 121˚C for 15 minutes.0 if X037 is added) Gelatin Peptone Potassium sulphate Magnesium chloride Agar No. The mineral requirement for pyocyanin production. supplement. putida colony size shape & (mm) surface 2.5-4.F. J.0 15. res. 26:15. notatum 4. dried and frozen foods and other types of product. 14: 65-72.0 3.N.0-3.0 References Association of Official Analytical Chemists (AOAC). Green/Blue Pen. Standard Methods for the Examination of Dairy Prod.C. glabrata Asp. Alternatively the medium can be made selective for Pseudomonas species generally by the addition of X108 C.Potato Dextrose Agar LAB 98 Description Potato Dextrose Agar is recommended by the American Public Health Association for the enumeration of yeasts and moulds in examination of dairy products.C. mentagrophytes 4.D. Appearance: Pale straw. Mix well and pour into Petri dishes. This can be done by adding 10ml of sterile 10% Lactic Acid X037.A.D. B.G. (1977). 1. Depending on whether the medium is to be used as a selective or non-selective agar it can be used with or without acidification. aeruginosa.O. Hausler. NCIMB 50097 Saccharomyces cerevisiae Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. organisms: Aspergillus sp. Formula Acid Hydrolysed Casein g/litre 10. fragi P. (1970). M.4 grams of powder and disperse in 1 litre of deionised water.J.F. Black spores centre White surround Green spores centre White surround colour White Grey/White Yellow obverse Yellow obverse White Yellow obverse Green obverse Incubation: 25-30˚C aerobically for 48 hours. In certain cases it may be desirable to lower the pH of the medium to 3.G. Bacteriological Analytical Manual. Campbell.C.C. Sci.0 Growth Characteristics organism P.1 ± 0. S. This medium can be made selective for the isolation of Burkholderia cepacia by the addition of X140.0-3.K. Sci.. Br.0 10. aeruginosa 2. G. CV.V. and Raney.6 ± 0. Sect.C. and Eagles. Poult. Incubation: 21˚C aerobically for 5 days. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.0 2.E. pH: 5. aeruginosa NCIMB 50067 E.Cr.0 20. 44: 301. S. (1976). Clin.V. supplement.Rz.0-3. CV.0 2. J. Growth Characteristics organism Candida spp.5 in order to suppress bacterial growth. W.F.Cr.Cr.D.0 2.0 16.0-3.D.2 Minimum Q.E.). 14th edn. J.0 1.0 CV.4 11. D. 1 g/litre 4. Interpretation: Count all colonies as Pseudomonas species. Mead. Ward./blue light).0 CV. then sterilise at 121˚C for 15 minutes. spread 0. (1948). Med. fluorescens P. Two simple media for the demonstration of pyocyanin and fluorescein. King. Capped containers – up to 1 month at 15-20˚C in the dark. Washington D. The medium is made selective for Pseudomonas aeruginosa by the addition of X107 C. Candida krusei colony size (mm) 2. Can./C. Agar) LAB 108 Description The base medium is a modification of King’s medium A which uses magnesium and potassium salts to enhance production of the pigments pyocyanin (green) and fluorescein (detected by U.W.. Once the pH has been lowered the medium may not be heated again without resultant loss of gel strength caused by agar hydrolysis. Goto. C.44 . Bot. Minimum Q.D. opaque. pH: 7.2 (3.C. verrucusum Toro. Inoculum: Pour plate technique. Microbiol. (1954).0 shape & surface C.V. to one litre of Potato Dextrose Agar LAB 98. niger 5.5ml of sample over entire surface.0-3.: American Public Health Association. 18: 661-667. F. F. Colonies that exhibit the pyocyanin and fluorescein pigments count as P.0 1.C.0 Method for reconstitution Weigh 39 grams of powder.

Pour a further 10ml as an overlay and again allow to set. C. 1985 p. Methods for the Examination of Waters and Associated Materials. Agar or other salmonella selective agars. Environment Agency: The Microbiology of Drinking Water (2002).L. incubate at 22˚C for 5-7 days or 30˚C for 3 days. P. and Standard Methods for the Enumeration of Water and Wastewater.18 13. Preston. Incubation: When plates have set. when dissolved dispense in 10ml volumes in screw capped bottles and sterilise by autoclaving at 115˚C for 15 minutes.5 0. Microbiol. (1985). M. and has been shown to give significantly higher counts than plate count agar (PCA) or similar high-nutrient media.2 + 0. and Norberg. (If required.) Single Component Enrichment Broth LAB 86 Introduction Rappaport Vassiliadis Broth (R10 modification) was born out of a long series of experiments carried out to determine the correct levels of malachite green and magnesium chloride that would allow Salmonella to multiply freely yet still inhibit the other enteric organisms.1˚C for 24hrs (water bath). and Chronas. Bacteriol.C. D.E. American Public Health Association (1985) Standard Methods for the Enumeration of Water and Wastewater. 1-7. 59 143-145. typhimurium NCIMB 50076 Storage of Prepared Medium: Capped container – 6 months at 2-8˚C Inoculation: From pre-enrichment broth in the proportions of 1-part inoculum to 99 parts R. there will be a population of heterotrophic bacteria which cannot grow at all under the conditions of the SPC method or may grow so slowly that the colonies fail to reach a size detectable to the eye in the 48-h incubation period. M. The standard plate count (SPC) method using PCA provides an enumeration of bacteria which grow best at.. a medium of low nutritional content (R2A) and extended incubation times are required. organisms: E. Interpretation: Count all colonies and report the number of bacteria in the original sample as the heterotrophic plate count.5 0.0 Rappaport Vassiliadis (R.. Incubation: 41. App & Env.5 0. Personal communication. It can also be used with membrane filters if required. Minimum Q. In order to enumerate this section of the bacterial population in water.2 pH: 7. This formulation is very hygroscopic and will produce a slight exothermic reaction when mixed with water. J. However.2 ± 0. Geldreich. A note on the stability of Rappaport-Vassiliadis Enrichment Medium J. Jan..R2A Medium LAB 163 Description R2A medium was developed to determine the bacterial count in potable waters during treatment and distribution.J.H. R2A medium is recommended by the Environment Agency. This formulation has been shown to be superior to Mueller Kauffmann and Selenite Broth for the isolation of Salmonella from meat products.2 1.2 Method for reconstitution Weigh 18 grams of powder and disperse in 1 litre of deionised water.V.58 0... Formulation Yeast Extract Meat Peptone Casamino acids Glucose Starch Dipotassium hydrogen Phosphate Magnesium sulphate Sodium pyruvate Agar No. American Public Health Association Inc. Washington DC. Swirl to mix and sterilise at 121˚C for 15 minutes. Broth. Appl. bring to the boil to dissolve the agar.V. body temperature and this estimation at best may correlate to the coliforms present in the sample. Mavromatti. P.3 0. Aeromonas hydrophila References Reasoner. (1983) The Rappaport Vassiliadis (R.1ml onto the plate and spreading over the entire surface of the medium. Inoculation: Pour 15ml into a Petri dish containing 1ml of sample. Cool to 44-46˚C for not more than 3 hours before use.. Bacteriol. 16th Edition. F.) Enrichment Medium for the Isolation of salmonellas: An overview J. Food Micro. Appearance: Clear.C.5 7. green fluid. Int. CH.5 ± 0.B. pH: 5. P.5 0. References Vassiliadis.L. Minimum QC organisms: Pseudomonas fluorescens. Sub-culture onto either XLD Agar. Wiberg. Formula Soy Peptone Sodium chloride Potassium dihydrogen phosphate Dipotassium hydrogen phosphate Magnesium chloride anhydrous Malachite green g/litre 4.5 0. mix well and allow to set.3 15. (1985) A New Medium for the Enumeration and Subculture of Bacteria from potable water. Other incubation temperatures between 20˚C and 28˚C may be used. G.45 . disperse in 1 litre of deionised water. P. 1. E. Alternatively it may be used as a spread plate. 56 69-76. coli (inhibited) NCIMB 50034 S.G. The development work carried out on the formulation shows that it is extremely efficient in detecting small numbers of Salmonella in heavily contaminated products. inoculating 0.2 Appearance: Clear opalescent gel g/litre 0.5˚C for 24 hours (incubator) or 42 ± 0. Vassiliadis. 1989. Efstratiou.05 0. Methods for the Examination of Water and Associated Materials.V. Peterz. The effect of incubation temperature and magnesium chloride concentration on growth of Salmonella in home-made and in commercially available dehydrated Rappaport-Vassiliadis broths. M.26 0. or near. Pharmacopoeia of culture media for Food Microbiology. Bolton.033 Method for reconstitution Weigh 26. Appl. swirl to mix.8 grams powder. and pour into smaller volumes before sterilising).

J. Appearance: Straw coloured.0 5.Reinforced Clostridial Agar LAB 23 Description This is a solidified version of R. Sterilise for 15 minutes at 121˚C. Food Res. (LAB 22) and can be used for the enumeration of anaerobes by pour plate. (1939). is rich.M.0 3.M. Cool to 47˚C. Growth Indicators Turbidity. Inoculation: Homogenised food sample to a ratio of 1:10 with R.0 Reinforced Clostridial Medium (R. The small amount of agar reduces diffusion of oxygen through the fluid. R. Incubation: 30˚C for up to 72 hours. Garrett. Appearance: Pale straw. Formula Yeast Extract Beef Extract Peptone Soluble Starch Glucose L-Cysteine hydrochloride Sodium chloride Sodium acetate g/litre 3. N. Barnes. disperse in 1 litre of deionised water. Dairy Res. disperse in 1 litre of deionised water. Interpretation: Count all colonies as presumptive clostridia. Anaerobic technique – a modified deep agar shake.. Ingram. perfringens NCIMB 50027 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. A. Inoculation: Pour plate technique or tube culture. J.0 5.2 Agar No.0 1.0 0. 5: 145.5 5.C. References Miller. translucent gel.W.5 12. Methods for the growth and enumeration of anaerobic spore-formers from cheese. When solidified in tubes or bottles with minimal head space it can be used for anaerobic culture without the need for anaerobic atmosphere.M. clear. organisms: C.0 10.J.8 ± 0.0 3.) LAB 22 Description This medium was formulated by Hirsch and Grinstead to recover small numbers of Clostridium spp. shake tube or membrane filtration methods. O.S. and Prickett. Various workers have reported its efficiency with many products and specimens. Lab. (1956). 4: 447-451. 25: 94-106.C. Appl.75 Method for reconstitution Weigh 49.8 ± 0. 1 Method for reconstitution Weigh 38 grams of powder. Anaerobic conditions for pour plate.M. (1954). pH: 6. E.C.0 0. swirl to mix then sterilise for 15 minutes at 121˚C. E. P. swirl to mix then bring to the boil to dissolve.M. organisms: C. and Barnes. H. non-selective and uses cysteine hydrochloride and glucose as reducing agents. perfringens NCIMB 50027 Storage of Prepared Medium: Capped container – up to 3 months at 15-20˚C in the dark.2 Minimum Q. and Grinstead. and distribute into sterile dishes or tubes containing decimal dilutions of the sample under test. The isolation of anaerobic Gram-negative bacteria from poultry. Minimum Q. and Goldberg.C. gas production.S. M. A simple modification of the deep shake tube for counting anaerobic bacteria. (1962). Pract. Formula Yeast Extract Beef Extract Peptone Glucose Soluble Starch Sodium chloride Sodium acetate L-Cysteine hydrochloride Agar No.0 1.C.0 5. E. colonies in medium.0 10. 1. Bact. 21: 101-110. 2 g/litre 3.0 0.0 10. Count as early as possible as prolonged incubation may result in the medium being disrupted due to gas production. References Hirsch. pH: 6. from a variety of sources.5 grams of powder.46 . allow to soak for 10 minutes.C. Allow to soak for 10 minutes. Distribute 25ml into 1oz Universal containers. Incubation: 30˚C for up to 72 hours.0 10.M.

this has been substituted by chloramphenicol because of superior selectivity. Ringer’s Solution (Thiosulphate) LAB 102 organism Growth Characteristics colony size shape & (mm) surface 13. Sacchromyces spp.H. Chloramphenicol (or 2 vials X089 oxytetracycline). NCIMB 50097 Sacchromyces cerevisiae E. Staphylococcus spp.0 0.M.5 grams of powder in 1 litre of deionised water. The solution can also be used in the sampling of food production apparatus by the rinse and swab method.105 0.5 grams of powder. Higgins.12 0. disperse in 1 litre of deionised water. Formula Mycological Peptone g/litre 5. Incubation: 25˚C aerobically for 24 hours to 5 days. The original formulation of Jarvis (1973) used chlortetracycline. London.S. of Health.5 1.2 Method for reconstitution Dissolve 12.0 10.2 ± 0. London.5 Rhizopus spp.. Allow to cool to 47˚C then add 2 vials of X009 (X209). LAB 100Z – Dissolve 1 tablet in 500ml of deionised water. (1937). Formula Sodium chloride Potassium chloride Calcium chloride Sodium bicarbonate ‘Calgon’ (Sodium hexametaphosphate) g/litre 2. Capped Container – up to 1 month at 2-8˚C in the dark.25 0. Formula Sodium chloride Potassium chloride Calcium chloride Sodium thiosulphate g/litre 2.G. Bacteriological Tests for Graded Milk.O. (1956). Bull. H.0 Method for reconstitution LAB 100 – Dissolve 2. M. Mix well. 100ml of LAB M Thiosulphate Ringer Solution will neutralise 7mg of chlorine. pH: 7. (1950). Min.12 0.15 0.25 0. faecalis B.L. Minimum Q. Rose Bengal Chloramphenicol Agar Base LAB 36 Description A selective medium for the enumeration of moulds and yeasts in foods. Davis. 2 Method for reconstitution Weigh 28. Formula Sodium chloride Potassium chloride Calcium chloride Sodium bicarbonate g/litre 2. This ensures total release of all the organisms taken up by the swab to increase the accuracy of a quantitative bacterial count.05 10.12 0.0 Dextrose Dipotassium phosphate Magnesium sulphate Rose bengal Agar No. Proceed as for LAB 100. Dispense 10ml amounts into screw cap containers and sterilise by autoclaving at 121˚C for 15 minutes. Fusarium spp. swirl to mix then sterilise for 15 minutes at 121˚C. CV.C. organisms: Aspergillus sp. no growth 1. then pour into Petri dishes. Dispense into containers as required then sterilise by autoclaving at 121˚C for 15 minutes. This medium should be protected from light.G. Appearance: Deep purple/red. Rose Bengal becomes increasingly toxic on exposure to light so it is important to store plates in the dark.E.Ringer’s Solution (1/4 strength) LAB 100 LAB 100Z (TABLETS) Description An osmotically controlled solution for the preparation of suspensions of food samples and for use as a diluent in dilution techniques for bacterial enumeration. coli Ent. Aspergillus flavus 8 Candida spp. when completely dissolved dispense into containers as required and sterilise by autoclaving at 121˚C for 15 minutes.075 0. 9: 50-51. Inoculation: Surface spreading.105 0.0 1. Health. The Rose Bengal dye is taken up by the growing colonies making them easier to see and inhibiting their spreading. E. Memo 139/Foods. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. allow to soak for 10 minutes.5 0.S.47 .G. clear. Comparison of recovery rate of organisms from cotton wool and calcium alginate wool swabs. Dairy Industries.05 References Min.0 no growth no growth no growth Method for reconstitution Dissolve 2. Chloramphenicol may be added before autoclaving. Mucor spp. The sodium hexametaphosphate in the formula will enable the alginate swabs to completely dissolve. Ringer’s Solution (Calgon) LAB 101 Description For use with calcium alginate swabs.5 12. J. Laboratory Control of Dairy Plant.05 12.E. subtilis 1. colour White White White Yellow/ Green White White hyphae other Description An isotonic rinse for use in hygiene studies on plant and equipment that have been treated with chlorine disinfectants.5 grams of powder in 1 litre of deionised water. P. Mon.8 grams of powder in 1 litre of deionised water.5 10 Fluffy Fluffy Fluffy Flat & hyphae CV.

G.D. Allow to soak for 10 minutes. Appearance: Cream/yellow.S. Minimum Q.W. Comparison of an improvised Rose-Bengal Chlortetracycline Agar with other media for the selective isolation and enumeration of moulds and yeasts in food. Formula Balanced Peptone Maltose Agar No. swirl to mix. B.G. rubrum colony size shape & (mm) surface 0. J. Pagano.0 25 CV.0 40. organisms: Candida sp.References Jarvis. J. (1910). Diagnostic medium for the differentiation of species of Candida. albicans C. Incubation: Aerobically. such as sporing structures and pigmentation are well developed on this medium.2 T. Because of its low pH this medium is very sensitive to overheating which will soften the agar and caramelise the carbohydrate. disperse in 1 litre of deionised water. Whitefluffy Whitefluffy Whitecentreyellow radial Whitefluffy colour White GreyWhite Reverseshades of red Reverseyelloworange reverseyellowother Yeasty smell Yeasty smell Method for reconstitution Weigh 62 grams of powder. Sabouraud Liquid Medium U. disperse in 1 litre of deionised water. floccosum 25 Reversetan 1.0 1.0 5.2 Minimum Q. The medium can be made more selective by the addition of chloramphenicol supplement (X009) (X209). Appearance: Buff opalescent gel. Appl. mentagraphytes 25 M.0 20. (1957-8).0-3.0 Method for reconstitution Weigh 30 grams of powder.CR. pH: 5.Milk and Fd. Sabouraud Maltose Agar LAB 111 Description This is a modification of Sabouraud Dextrose Agar. This medium may be made selective by the addition of 2 ampoules X009 Chloramphenicol selective supplement which may be added either before or after autoclaving. An aureomycin rosebengal agar for the enumeration of yeast and mould in cottage cheese.C. R. Diagnostic features. Appearance: Pale straw clear. NOTE: The gel strength of the medium may diminish if recommended sterilising time or temperature is exceeded. It can also be used as a growth medium for the determination of fungistatic activity in pharmaceutical products. Heat to dissolve. J. (Fluid Sabouraud Medium) LAB 33 Description A liquid sterility test medium for the detection of yeasts.0 12.L. Levin.D. Overcast. 4: 197-206. J. 2 g/litre 10. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C. The acidic pH (5. moulds and acidophilic bacteria in pharmaceutical products. then dispense into final containers before sterilising at 121˚C for 15 minutes.P.5-2. Do not overheat or the agar gel will be softened and the carbohydrate will be caramelised. J. Formula Pancreatic digest of casein Peptic digest of fresh meat Glucose g/litre 5. 32: 442-445. yeasts 37˚C for 48 hours. canis 25 E. This medium conforms with the United States Pharmacopeia.0 12. Bact. 137-143.C.0 Growth Characteristics organism C. NCIMB 50010 E. F. (1969). translucent. Capped container – up to 3 months at 15-20˚C in the dark.S.2 Method for reconstitution Weigh 62 grams of powder. J. References The Pharmacopeia of the United States of America. Int. mix well then pour plates.E. swirl to mix then autoclave at 121˚C for 15 minutes.Tech.0 40. (1987). W. 36: 723-727. Les Teignes Paris. krusei T.6) of this medium inhibits many species of bacteria. D. Incubation: 22-25˚C aerobically for 10 days..48 . coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. allow to soak for 10 minutes.P. Board. substituting maltose for dextrose. Formula Balanced Peptone No. disperse in 1 litre of deionised water. Food Microbiol. allow to soak for 10 minutes. 1 Dextrose Agar No.6 ± 0.6 ± 0. XVII (1965). recommended by the American Public Health Association. NCIMB 50010 E. pH: 5. R.7 ± 0. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. (1973). Antibiotics Annual. and Weakley. Inoculation: As recommended in the U.D. W. and Trejo. Banks. Cool to 47˚C and mix well before pouring plates. References Sabouraud. Some factors influencing the recovery of yeasts and moulds from chilled foods. Inoculation method: Surface streaking for single colonies or stab method. Cool to 47˚C. pH: 5.0 Sabouraud Dextrose Agar LAB 9 Description Introduced by Sabouraud in 1910 as a selective medium for fungi and yeasts. organisms: Candida sp. 2 g/litre 10. fungi 25-30˚C for up to 3 weeks.

J.mentagraphytes 25mm M.0 1. and Trejo. Williams and Wilkins. Subculture at 24 and 48 hours.6 ± 0. Add 19 grams of Selenite Broth Base and warm to dissolve. 137-143. Subculture: On to Baird-Parker Medium LAB 85.1 ± 0.0 g/litre Growth Characteristics organism C. E. Subculture: Onto two or more selective agars. Appearance: Pale orange/red with slight precipitate (overheating will cause excessive precipitate and loss of selectivity). food and sewage. Levin. J. References Leifson. Antibiotics Annual. NCIMB 50010 E. Causes severe burns.Minimum Q. Danger of cumulative effects. Incubation: Aerobic. canis 25mm Method for reconstitution Dissolve 4 grams of sodium biselenite in 1 litre of cold deionised waer. (1936). 1.0 30. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. WARNING: SODIUM BISELENITE Toxic by inhalation and ingestion. R.E. F. Subcultures should be performed after no more than 24 hours incubation as there is an increasing loss of selectivity if incubation is prolonged. Capped containers – up to 3 months at 15-20˚C in the dark. Inoculation: Approximately 0.0 Incubation: Up to 24 hours aerobically at 35-43˚C. D. Allow to soak for 15 minutes then sterilise by autoclaving at 121˚C for 15 minutes.0 5. Amer. First described by Leifson in 1936 the medium is a peptone lactose broth.. J. organisms: Salmonella sp. or by free steaming. rubrum colony size shape & (mm) surface 0. This medium can be incubated at various temperatures from 35-43˚C to vary the selectivity. cultivation. albicans C. (1985). Inoculation: Surface streaking or stab inoculum.5-2. Media for the isolation. New selenite enrichment media for isolation of typhoid and paratyphoid (Salmonella) bacilli.0 100. Distribute into tubes or bottles and sterilise for 5-10 minutes in a boiling water bath. moderately buffered. which utilises sodium biselenite as a selective agent. organisms: S. aureus NCIMB 50080 E. After contact with skin wash with water immediately. Baltimore. Formula Beef Extract Meat Peptone Tryptone Cooked Meat Granules Sodium Chloride g/litre 10.CR.C. Inoculation: Up to 1g of sample to 10ml of medium. It can also be used for the isolation of halophilic micrococci which contaminate hides and raw salt. Les Teignes. DO NOT AUTOCLAVE. Incubation: 37˚C aerobically. pH: 7. identification of Medical Bacteria Vol 1. Appearance: Straw broth over meat particles pH: 7.0 4. 24: 423-432. Minimum Q.D.2 (complete medium) E.D. Paris. Pagano.F. other fungi 25˚C for up to 3 weeks. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.C. floccosum 25mm References Sabouraud. MacFaddin. NCIMB 50076 E.C. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Whitefluffy Whitefluffy Whitecentre yellow radial Whitefluffy colour White GreyWhite Reverseshades of Red Reverseyelloworange Reverseyelloworange Reversetan other Yeasty smell Yeasty smell T. krusii T. yeasts 37˚C for 48 hours. Formula Selenite Broth Base LAB 44A: Peptone Lactose Sodium phosphate Sodium biselenite LAB 44B: Sodium hydrogen selenite 4. (1910).0 10. If you feel unwell seek medical advice. Method for reconstitution Add 2 tablets to 10ml of deionised water in a narrow container. Selenite Broth LAB 44A & LAB 44B Description A medium for the selective enrichment of salmonellae from faeces. (1957-8) Diagnostic medium for the differentiation of species of Candida. W.5-1 gram of sample per 10ml tube.Hyg. Mannitol Salt Agar LAB 7 or 4S agar LAB 84.0 5. J.49 .0 25mm CV. Salt Meat Broth Tablets LAB 113Z Description An enrichment medium for the isolation of staphylococci from heavily contaminated samples.0-3. organisms: Candida sp.0 5.2 Minimum Q.

1 12.A.0 5. Incubation: 37˚C. Milk and milk products (1985)..A. (1936). It is necessary to add lysed or ‘chocolated’ blood for some organisms. M. e. Appearance: Pale straw colour. The efficiency of selenite broth of different compositions in the isolation of Salmonella.H. 130. 2nd edn. but all antibiotics should give adequate zone sizes when compared to controls using standard organisms.C. Detection of Salmonella. coli NCTC 10418 (antibiotic sensitivity zones) Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. (1978). (1968). 15: 31-34. Minimum Q. G.M. M. 24. 6579-1981. Compendium of methods for the microbiological examination of foods. Antibiot. Appl.T. (1958). atmosphere to suit organisms metabollic requirements. 134. NCIMB 50076 E. Ont. organisms: Salmonella sp. Am. American Public Health Association.2 Minimum Q. New selenite enrichment media for the isolation of typhoid and paratyphoid (Salmonella) bacilli. Pathol.G. E.. Arnold.C.01 0. Inoculation method: Surface. aeruginosa NCTC 10662. 8: 599-606. Appl.01 0. E. R. Inoculation: Add sample to broth in the ratio of 1:10. Bartram. University of Toronto Press. Use a preenrichment broth if damaged organisms are to be recovered. (A brick red precipitate indicates overheating). according to technique. J.. If you feel unwell seek medical advice. Appearance: Dependent upon the blood additive. To prepare blood agar cool to 45˚C and add 7% lysed blood or 6% defibrinated blood according to preference. disperse in 1 litre of deionised water. 2 Method for reconstitution Weigh 40 grams of powder. Hanus. organisms: S. and. 1 Lactose Sodium phosphate L-Cystine Sodium biselenite LAB 44B Sodium hydrogen selenite 4. W.2 Aneurine hydrochloride Uridine Agar No. Their significance and methods of enumeration. ISO 6785-2985 (E). (1953).0 4. Report on the Antibiotic Sensitivity test trial organised by the bacteriology committee of the Association of Clinical Pathologists. Formula Peptone-Infusion Solids Starch Sodium chloride Disodium citrate Adenine sulphate Guanine hydrochloride Uracil Xanthine g/litre 21. Micro-organisms in foods. 2nd edn.01 0.0 ± 0.R.01 0. A new agar for in vitro antimicrobial sensitivity testing. Microbiol. Sands.5 0.P.Selenite Cystine Broth LAB 55A & 44B Description This formulation is a result of the investigation of North and Bartram in 1953.4 ± 0. International Organization for Standardization. E. Formula Selenite Cystine Broth Base LAB 55A Balanced Peptone No. and sterilise for 10 minutes in a boiling water bath. (1965). Mix well then pour plates.50 . aureus NCTC 6571. Distribute into tubes or bottles. F. North. 18: 1-5. DO NOT AUTOCLAVE THIS MEDIUM. General guidance on methods for the detection of Salmonella. Speck.0 0. ISO. S.T. 1. Microbiology (1981).0 1.0 10.T. ICMSF.J. S. Ps.. and Scherr. Causes severe burns. It was found that the addition of 10 micrograms/ml of cystine to Leifson’s selenite broth enhanced recovery of salmonellae. 1. coli NCTC 10418.O.L. pH: 7. 1. Add 19 grams of Selenite Cystine Broth Base and heat to dissolve.Clin. Interpretation: There are no defined zone sizes as in Mueller Hinton. Bechtle.C. J. Hyg. After contact with skin wash with water immediately. swirl to mix then sterilise by autoclaving for 15 minutes at 121˚C. London.) LAB 12 Description A medium formulated for antibiotic susceptibility testing by the Joan Stokes technique.6 5.. J.J. WARNING: SODIUM BISELENITE Toxic by inhalation and ingestion. Antibiotic activity in the presence of agar. allow to soak for 10 minutes. Toronto. and Bennett. (1984). Subculture onto Salmonella selective media. aureus NCTC 6571 E. Clinical Bacteriology 3rd edn. They examined the effect of varying concentrations of cystine and phosphate on the recovery of salmonellae in egg products using selenite broth. very rich and includes various nucleotides to enable fastidious organisms to be tested. E. Incubation: 37˚C for 24-48 hours aerobically.01 0. Chemother. Danger of cumulative effects.01 g/litre Sensitivity Test Agar (S.0 Method for reconstitution Dissolve 4 grams of Sodium biselenite (LAB 44B) in 1 litre of deionised water. is inhibitor-free. or steamer. clear with slight precipitate. Committee of the A. References International Organisation for Standardization. (1967).g. pH: 7. Leifson.0 0. References Stokes. 423-432. Microbiol. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.

0 0.colon aerogenes groups and for isolation of certain fungi. Proteus mirabilis P. Mix well then pour plates. Appearance: Green. Dis. opalescent.5-1. (1926).2 Growth Characteristics organism (most) Salmonella spp S. CV. Allow to soak for 10 minutes. freundii NCTC 9750 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. Incubation: 37˚C aerobically for 24-48 hours. MacFaddin. J. Incubation: 42˚C aerobically for 48 hours. pH: 6. CV. J.2 1. Koser. Food Protect. Certain Enterobacteriacae have the ability to utilize citrate as the sole source of carbon and utilize inorganic ammonium salts as the sole source of nitrogen resulting in an increase in alkalinity. providenciae E. Klebsiella spp.0 1.5% potassium tellurite solution.S. paratyphi A Serratia spp. coli C. Single Step Staphyloccocus Selective Agar (4-S medium) LAB 84 Description This staphylococcal isolation and identification medium is based on the ability of S.0 3. 8: 493-520. L.E. faecalis colony size shape & (mm) surface 0.Inf.5 P.2 Minimum Q. pH: 6. aureus to grow and produce lecithinase in the presence of a high sodium chloride concentration and potassium tellurite at 42˚C. J. epidermidis S. (1983). Appearance: Pale cream.Simmons Citrate Agar LAB 69 Description A medium devised by Simmons in 1926 to help in the differentiation of enteric bacteria and in the isolation of fungi.0 Method for reconstitution Weigh 24 grams of powder. The high temperature inhibits the production of lipase (which causes clearing of egg protein) allowing the effect of lecithinase (which causes opacity of egg protein) to be more easily detected. aureus NCIMB 50080 S. (1923). and Katzenelson.A. coli NCIMB 50034 C. 2 g/litre 4. organisms: S. Yersinia spp.E. Bact. with loose caps to allow gaseous exchange.0 13.P. 1. opaque. aureus S. Allow to set as slopes.C. A Culture medium for differentiating organisms of typhoid .9 ± 0. Cool to 48˚C and add aseptically 30ml X073 egg yolk emulsion and 1ml X027 3. epidermidis NCIMB 50082 Storage of Prepared Medium: Plates.08 15. J. J.0 References Simmons.G. Williams and Wilkins.0 10.C. Bromothymol Blue is used as a pH indicator. swirl to mix then heat to dissolve the agar and solids. Allow to soak for 10 minutes. colour other Grey (Opaque zone of ppt around colony) White/ grey Grey (No ppt zone) (Usually no growth) Most other organisms are inhibited by this medium at 42˚C.E. Biochemical Tests for Identification of Medical Bacteria. References Mintzer-Morgenstern.0 2.7 ± 0. Inoculation: Streak on surface and stab into the butt.51 . disperse in 1 litre of deionised water. 39: 209-215. Dispense into tubes or bottles then sterilise by autoclaving at 121˚C for 15 minutes. Growth Characteristics organism S. vulgaris Providencia rettgeri P. Interpretation: Utilisation of citrate and ammonium salts results in growth and a change in colour of the medium from green to blue. Shigella spp. CV. organisms: E. Inoculation: Surface spreading or streak out to single colonies. 45:3:218-22. (1982).0 0. Formula Tryptone Yeast Extract Dextrose Sodium chloride Agar No. 2 g/litre 0. Method for reconstitution Weigh 80 grams of powder. disperse in 1 litre of deionised water. A Simple Medium for Isolation of Coagulase-Positive Staphylococci in a Single Step. Utilisation of the salts of organic Acids by the Colon-aerogenes group.up to 7 days at 2-8°C in the dark.0 50. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. S. typhi S. For the recovery of heat stressed staphylococci a 3 hour preincubation in Brain Heart Infusion Broth LAB 49 at 37˚C is recommended. Formula Magnesium sulphate Ammonium dihydrogen phosphate Dipotassium phosphate Sodium citrate Sodium chloride Bromothymol blue Agar No.0 5. gallinarum S.G.G.F. cholerasuis S. E. freundii growth yes no no no no yes no no yes yes yes no no no yes colour of medium blue green green green green blue green green blue blue blue green green green blue Minimum Q.

2 ± 0.L. Acheson. and allow to soak into the agar. Storage in bottles is not recommended as re-melting the medium will cause damage. 74 591-595. J.W. C.D. coli NCIMB 50034 (Pink/red) Ent.. Riley. but it may also be used as a spread plate for the examination of other sample types.5 – 4.2 Crystal violet Agar Method for reconstitution Weigh 48.. DO NOT AUTOCLAVE. Incubation: 37˚C aerobically for 18-24 hr. coli strains are sorbitol positive and so produce pink/red colonies. L.001 12. Interpretation Count all red and maroon colonies as presumptive enterococci.Bact.. Pathogenicity of the organism is linked to the production of verocytotoxins (VT1 and VT2).0 2.1 + 0.0 Shape CV.. (1957) J. Allow to soak for 10 minutes.Slanetz and Bartley Medium (Membrane Enterococcus Agar) Sorbitol MacConkey Agar (SMAC. D.W.5 grams of powder and add to 1 litre of de-ionised water. Minimum QC organisms: E.03 0. Rev. (1991) Ann.A. Dry the surface prior to inoculation.D. O157:H7 is sorbitol negative and produces translucent colonies whereas most other E. and Todd. D. J. Cool to 47˚C and pour into sterile Petri dishes. Selectivity of the medium can be increased by adding Cefixime-Tellurite (C. Methods for the Examination of Water and Associated Materials. OR LEAVE FOR GREATER THAN 4hr AT 47˚C. (1991) Foodborne Pathogens an Illustrated Text. 1. Published by Wolfe Publishing Ltd. O157:H7 has been associated epidemiologically with food poisoning outbreaks involving beefburgers and cold cooked meats.D. This reaction is not exclusive to enterococci. pH: 7. I.. 3 with the substitution of the fermentable carbohydrate from lactose to sorbitol.5 – 5.0 1.E.g. and place this on the surface of a properly dried Slanetz and Bartley plate. Inoculation Water: Filter 100ml of the water through a suitable membrane. Ganguli. coli O157:H7 (non-toxigenic) NCTC 12900 (translucent) E. R.0 4.5ml over the surface of the plate using a spreader. At 37˚C for 4hr then 44˚C for 44hr if testing untreated waters or raw materials.24.3 Sodium chloride Neutral red LAB 161 Description This is a selective differential medium for the isolation of Escherichia coli 0157:H7. O26.G CV. (1989) Epid. CT-SMAC) LAB 166 Description This medium was originally described by Slanetz and Bartley for the enumeration of enterococci from water samples using a membrane filtration technique.0 5. Farrel. 36 198-202.G. OVERHEAT. Environment Agency: The Microbiology of Drinking Water (2002). pH: 7.P.5 grams of powder and mix with 1 litre of deionised water.52 . and the count at this stage should be considered presumptive. Inoculation: Surface streak for single colonies. O145)..0 10. Enterococci reduce tetrazolium chloride to the insoluble red dye formazan. Med 308 681-685.5 – 4. E. O113.G Any Colour Translucent Pink/red Pink/red E.A. et al (1983) New Eng. Evans. Salmon. Colonies may be confirmed as enterococci by demonstrating aesculin hydrolysis using Kanamycin Aesculin Azide Agar LAB 106 Formula Tryptose Yeast Extract Glucose Dipotassium hydrogen phosphate Sodium azide 2. coli Sorbitol +ve organisms References Law..0 Formula Peptone Sorbitol Bile salts no.) supplement X161. Varnam. coli O157:H7 Other E. L.E. and Bartley.K. g/litre 20. producing colonies which are dark red or maroon on the surface of the membrane or agar. A. Riley.H. 103 249-254.2 Mininmum QC organisms: Enterococcus faecalis NCIMB 50030 Escherichia coli NCIMB 50034 (inhibition). Inf. O111..0 Method for reconstitution Weigh 43. Appearance: Rose coloured gel. L.H. A. slight violet tinge.W. Hutchinson.C. Cool to 47˚C.1 12.4 0.0 2. Published by American Public Health Association. Micro. 41 383-408. Donohue-Rolfe.W. Appearance: Pale red. Other samples: Dilute as necessary and spread 0. the primary serovar associated with haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS).0 0. Med. P. Confirmation of isolates can be achieved by demonstration of a positive aesculin reaction on KAAA LAB106. Incubation Water: at 37˚C for 48hr if testing potable waters or processed foods.0 0. Reference: Slanetz. Hitchins. A. (1992) J.G.. Growth Characteristics: Organism Size (mm) 2. Bring to the boil with frequent stirring to dissolve completely.5 5. and pour plates. Storage: Plates – upto 7 days at 2-8˚C. but it should be noted that not all strains of O157:H7 produce verocytotoxins. M.3. add 2 vials of CT supplement X161. Hartman. swirl to mix and sterilise by autoclaving at 121˚C for 15 minutes.4 Tetrazolium chloride Agar g/litre 20. Micro..0 2. and that strains from other serovars can be toxin producers (e. (1992) in “Compendium of methods for the microbiological examination of foods” Ch.T. L. The medium is a modification of MacConkey Agar No. faecalis NCIMB 50030 (inhibition) Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.

H. Am. B. K. P. 269-296. (1965) Isolation of shigellae. 1.E. 476-479. 32. clear. There is also the extra thiosulphate giving good H2S production which reduces the ferrous ammonium sulphide giving black centred colonies with H2S positive organisms.S. 656-659. organisms: Salmonella sp. 44: 4. Microbiol. Clin. then bring to the boil. International Standard FIL-IDF30. Krol.I. Appl. References International Dairy Federation (1964). Formula Beef Extract Balanced Peptone No.5 8.E.E. disperse in 1 litre of deionised water. S. coli colony size shape & (mm) surface 0..1-2. 2 g/litre 5.. whilst some enterococci are inhibited by this formula. pH: 7.5 8.0 Method for reconstitution Weigh 34 grams of powder and disperse in 1 litre of deionised water. Alimenta 5(2). and allow to cool to 47˚C. D.0 10. (1969).2 Method for reconstitution Weigh 60 grams of powder. boil to dissolve and disperse into tubes or flasks. Sterilise by autoclaving at 121˚C for 15 minutes. Pink-Red (No growth) Yellow Ps. (1969). Interpretation: Count colonies.6 ± 0.0 Pink (Green pigment) Yellow Yellow PinkYellow (black centre) Gram positive organisms – no growth. Isolation of salmonellae and shigellae from an artificial mixture of fecal bacteria. J. Bacteriol. Pathol. colour Red K. 1 Lactose Bile Salts No. J. (1966).. Agar (Salmonella Shigella Agar) LAB 52 Description This medium is a modification of Leifson’s DCA Medium first described in 1941 by Mayfield and Goeber shortly before Hynes described his modification of DCA. Formula Gelatin Peptone Tryptone Sodium chloride Agar No. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 4˚C in the dark. Appearance: Pale Pink. Leifson.1-1.0 1. (1972).0 CV. B. Incubation: 30˚C for 2 days then 20˚C for a further two days – aerobically. Mossel. New culture media based on sodium desoxycholate for the isolation of intestinal pathogens and for the enumeration of colon bacilli in milk and water. organisms: E.025 13. Swirl to mix.). pH: 7. 40: 581-589. International standard count of contaminating organisms in butter. streaking for single colonies.2 Minimum Q. aerogenes Proteus spp. 18: 4.. DO NOT AUTOCLAVE THIS MEDIUM. S.C. Kominos.1-3. Mix well then pour plates.G. Allow to soak for 10 minutes.D.0-3. Incubation: 37˚C aerobically for 18-24 hours. P. (1935). W.G.0 ± 0. clear.F. Inoculation: 0. Taylor. The medium conforms to the formulation of the International Dairy Federation (I. Pathol. 1 g/litre 7. Inoculation method: Surface plating. and Moerman.0 CV. 51-60.E. Comparison of plating media and enrichment broths. 0. M. H. other Red ppt around colonies (No growth) (grey centre) (fishy odour) Ritter.S. Alimenta 11(2). sodium citrate and the addition of brilliant green dye. The selectivity of this medium can be such that it was suggested by Taylor et al in 1965 to be unsuitable for the isolation of Shigella species. Thomas.0 5. CV.. 3 Sodium citrate Sodium thiosulphate Ferric citrate Brilliant Green Neutral Red Agar No.D. Growth Characteristics organism E. The Gram negative rods are able to deaminate proteins as a carbon source. Dry the surface before incubation. E. 0. J.E.0 14.0 0.5 7. aeruginosa 0.00033 0.CR. Appl.0 Shigella spp.A.C. NCIMB 50076 E. Minimum Q. Appearance: Light straw. 2. CV.5 References Isenberg.G. Harris.D. Sugar Free Agar LAB 87 Description A formula described by the International Dairy Federation for the enumeration of psychrotrophic and mesophilic Gram negative rods in butter and other dairy products.0 8.0 Salmonella spp.5 1.D.C.. Greater understanding of the selection mechanisms involved enable us to adjust the reaction and allow the more delicate Shigella to grow without unduly impairing the medium’s selective properties. Bact.53 . and Siegel.2ml of butter fat in a pour plate technique.A. and Eschmann. B. coli NCIMB 50034 Storage of Prepared Medium: Capped container – up to 3 months at 15-20˚C in the dark. CV.G. II.0-2. 43-45. CV.5 5. The selectivity of the medium was increased by the addition of extra bile salts.5-2. and allow to soak for 10 minutes.

E. Growth Characteristics organism Vibrio. vulnificus V. Formula Balanced Peptone No. Inoculation: Add 1 part of sample suspension or inoculated pre-enrichment medium to 9 parts of Tetrathionate Broth.J. from faeces. and Kuwahara.C.0 9.4 ± 0.04 0. It conforms to the formulation recommended by the American Public Health Association for use in the examination of dairy products and foods for salmonellae.0 Method for reconstitution Weigh 46 grams of powder and add to 1 litre of deionised water. DO NOT AUTOCLAVE OR OVERHEAT THIS MEDIUM. Yellow CV.0 10.G. Jap. (1953). Incubation: 12-24 hours at 37˚C. 1 g/litre 5.0 17. Furniss. Organisms which reduce tetrathionate. Iodine solution: Dissolve 5g of potassium iodide and 6g of iodine crystals in 20ml of distilled water.. T.V.0 10.0-3. Yellow CV. whilst most enteric organisms are inhibited. NCIMB 50076 E.0 0. pH: 8. Blue or Green CV. Subculture: Onto LAB 34 Brilliant Green Agar and either LAB 32 XLD or LAB 110 Hektoen Enteric or other Salmonella selective media. parahaemolyticus 3. Inoculation: Surface plating with a heavy inoculum. Therefore heavy inoculation of the medium is possible.0-4. To overcome this. cholerae.P. S.0 1. shigelloides 3. Ples.0 1. proliferate in the medium. PHLS Monograph No. Mix well before dispensing into bottles and continue swirling whilst dispensing to avoid the calcium carbonate sedimenting. The Vibrios.0 V.0 P. pH: 8. 387-391. APHA.T. Sakazaki.0-3. Yellow CV.04 15.H.G.6 ± 0. T.E.G.0 10. and Donovan. is designed for the selective isolation of Vibrio species.0 1.0 2. Enomoto.2 Method of reconstitution Weigh 88 grams of powder and add to 1 litre of deionised water.S. 11. S.G. Bring to the boil with frequent swirling to fully dissolve the medium. Lee J.54 .0 10. Yellow CV.0-3. For the best results the medium should be used the same day as prepared. such as salmonellae.G.G.0 2.2 Minimum Q. organisms: Salmonella sp. References Standard methods for the Examination of Dairy products. cholerae colony size (mm) 2.0-3. Yellow CV. 1 Bile Salts Calcium carbonate Sodium thiosulphate g/litre 5. foods etc.E.0 Description T. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 4˚C in the dark.A.. The formulation was developed by Kobayashi.S.B. Enomoto. Appearance: Dark green clear agar.E. Sakazaki and Kuwahara and inhibits most of the Enterobacteriaceae for at least 24 hours.. Yellow F.E.G.0 2.P. (1963). R.E. Minimum Q.0-5.T. Cool to 45˚C and add 20ml of iodine solution prepared as indicated below. Formula Yeast Extract Peptone Mix Sodium thiosulphate Sodium citrate Bile salts Sucrose Sodium chloride Ferric citrate Bromothymol blue Thymol blue Agar No. LAB 97 LAB 96 Description A selective enrichment broth for the growth of Salmonella typhi and other Salmonella spp.0-5. Incubation: 37˚C aerobically for 18-24 hours. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 7 days at 4˚C in the dark (without iodine solution).E. swirl to mix then bring to the boil.C. Appearance: Turbid white. Bacteriol 18: 10-11. mimicus Enterococci Proteus spp.C. alginolyticus V. streak out to single colonies. V. particularly V.0 30. Gram-positive organisms are inhibited by the inclusion of bile salts.B. Novobiocin may be added to the medium at a level of 40 microgram/ml before addition of the iodine.G. Certain members of the Proteus group will also reduce tetrathionate thereby impairing the performance of the medium in some cases. References Kobayashi. cholerae (type F) E.0 2. Cholera Medium (Thiosulphate Citrate Bile Salts Sucrose Agar) Tetrathionate Broth Base A.E. Allow to soak for 10 minutes. Green/ Yellow Green other may revert to green at R. 10th Edition. organisms: V.L. A.5 10.CR.0 1. Cool to 47˚C and pour into Petri dishes. fluvialis V. metschnikovii V. 1. Capped containers – up to 1 month at 15-20˚C in the dark. (1978). Green CV.C.G.0 shape & surface colour CV.

0 2.4grams of powder and disperse in 1 litre of deionised water.0 5. Allow to soak for 10 minutes.1 0.. pH: 7. The use of a fermentable sugar in the formulation leads to the production of acid which would normally inactivate the haemolysin. aerobically. Appearance: Pale straw. Med. 115: 598-600.L. 35 (1) 973-974. Organisms Streptococcus pyogenes ATCC 19615. Path. Updyke.55 .0 5.I.Thioglycollate Medium (Brewer) LAB 64 Description This is the original formula introduced by Brewer in 1940 as a clear medium for the cultivation of anaerobes.0 1. J. 1. The agar makes the medium viscous slowing down the permeation of oxygen and any convection currents.W. Formula Beef Extract Yeast Extract Balanced Peptone No. This is prevented by the inclusion of buffers to maintain the pH of the medium thus preserving the haemolysin. Reference: Todd. and Hewitt. (1932) A New Culture Medium for the Production of Antigenic Streptococcal Haemolysin. Todd Hewitt Broth is also used to cultivate streptococci prior to serological grouping. Bact. E. Inoculation: Ensure adequate dispersal of the inoculum in the broth. E. (1954) A Dehydrated Medium for the Preparation of Type Specific Extracts of Group A Streptococci.0 5. Sodium thioglycollate acts as a reducing agent and also neutralises the bacteriostatic properties of mercurial compounds. L. swirl to mix and warm to dissolve.C.F.0 20. 1 g/litre 1. Storage: Capped containers .2 Inoculation: Pick a well isolated colony for subculture into Todd Hewitt Broth.. It has found applications as a sterility test medium and as a blood culture medium although it has been superseded by Fluid Thioglycollate LAB 25 and Fastidious Anaerobe Broth LAB 71 for these purposes. (1940). Method for reconstitution Weigh 20 grams of powder.. Formula Infusion from fat-free minced meat Tryptone Dextrose Sodium bicarbonate Sodium chloride Disodium phosphate anhydrous g/litre 10. References Brewer. Minimum Q. Growth Indicators: A diffuse turbidity or individual colonies. pH: 7. viscous liquid which may have a green surface due to contact with oxygen. Appearance: Straw coloured.H. Allow to soak for 10 minutes then bring to the boil with gentle agitation to dissolve the solids.002 1.0 0.up to 3 months at 15-20oC in the dark. 2 117-118.0 2. clear broth. J. Do not reheat more than once.. perfringens NCIMB 50027 Storage of Prepared Medium: Capped container – up to 3 months at 15-20˚C in the dark.0 Todd Hewitt Broth LAB 75 Description A nutritious broth medium formulated by Todd and Hewitt for the production of antigenic streptococcal haemolysin. Distribute into screw top containers leaving minimal headspace. Microbiol. translucent. Appl. Ass.0 2. Tighten caps as soon as possible after autoclaving. as well as promoting the growth of pneumococci. Dispense into 10ml volumes in screw capped containers and sterilise by autoclaving at 121˚C for 15 minutes. 1 Dextrose Sodium chloride Sodium thioglycollate Methylene blue Agar No. If the medium has a diffuse green tinge it should not be used until the oxygen has been driven off by holding in a boiling water bath for 5 minutes.C. Incubation: 37˚C for 24-72 hours. organisms: C. disperse in 1 litre of deionised water. M.4 Method for Reconstitution Weight 36. Amer.0 2.2 ± 0. Incubation: 37˚C for 18-48hrs.8 ± 0. and Nickle. Methylene blue is a redox indicator which is colourless at low Eh but turns green on exposure to oxygen. Clear liquid mediums for the culture of anaerobes.2 Minimum Q. Sterilise by autoclaving at 121˚C for 15 minutes. J.

Identification of Enterobacteriaceae.A.3 0. 2 g/litre 3. swirl to mix then bring to the boil with frequent swirling to dissolve the solids. and Ewing. coli Enterobacter aerogenes E.H.. paratyphi S.C. Inoculation: A heavy inoculum is streaked over the surface of the slope and stabbed into the butt. lactose. Recommended Methods for the Microbiological Examination of Foods..0 10.P. Minneapolis.56 . disperse in 1 litre of deionised water.Triple Sugar Iron Agar LAB 53 Description This is a modification of the Krumwiede and Kohn medium of 1917 which differentiates some of the Enterobacteriaceae on the basis of four reactions. A. fermentation of lactose.0 0.0 5. flexneri Salmonella typhi S. Edwards. Incubation: 37˚C aerobically for 24 hours. when screening for Salmonella spp.0 1. coli NCIMB 50034 Ps. cloacae Proteus mirabilis Butt NC NC Acid + + - Method for reconstitution Weigh 65 grams of powder. enteritidis S. Minimum Q. (1962). W. This medium should be used in conjunction with a urease test to eliminate Proteus spp. New York. Appearance: Reddish-orange gel. Burgess Publishing Co. Diagnostic Procedures and Reagents. cholerasuis S. glucose and sucrose and H2S production. organisms: E.025 12. pH: 7. American Public Health Association (1966).2 Acid NC + - Providencia rettgeri Acid NC = No Change References American Public Health Association (1963). New York. Allow to set as a slope ensuring that the slant is over a butt approximately 3cm deep. Formula Beef Extract Yeast Extract Balanced Peptone No.0 Interpretation Slant/butt Alkaline/acid Acid/acid Alkaline/alkaline Colour Red/yellow Yellow/yellow Red/Red Utilisation Glucose only fermented Peptones utilised Glucose fermented Lactose + or sucrose fermented Neither glucose. pullorum S..0 3.0 20. A. NC NC + Sulphide production Organism Shigella dysenteriae S. gallinarum E. 4th edn.P. Allow to soak for 10 minutes. typhimurium S.0 10. 1 Sodium chloride Lactose Sucrose Glucose Ferric citrate Sodium thiosulphate Phenol red Agar No.R. sonnei S.3 0.H.H. nor sucrose fermented Peptones utilised Slant NC Acid Acid Acid Gas Acid Gas Acid Acid Gas Acid Gas or Alk. P.A. Distribute into tubes and sterilise at 121˚C for 15 minutes. 1. 2nd edn.. aeruginosa NCIMB 50067 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark..4 ± 0.

0 Method for reconstitution Weigh 36. Capped container – up to 1 month at 2-8˚C in the dark. clear gel. Delaney.0 Tryptone Bile Glucuronide Agar LAB 162 Refer to Harlequin™ HAL 3 Tryptone Glucose Extract Agar LAB 63 Description A plate count agar suggested by the American Public Health Association (A.P.D. 14th edn. McCarthy and Grasso in 1962 as a method for detecting faecal coliforms in water supplies based on the production of indole on a bile medium at 44˚C.0-3. pH: 7. 276-277. coli colony size (mm) 1.: Association of Official Analytical Chemists. Inoculation: 1ml of a 1:10 dilution of homogenised sample onto a membrane. Cool to 47˚C and pour into Petri dishes.C. M.E. R. swirl to mix then boil to dissolve before distributing into tubes or bottles.E.2 ± 0. (Ed. The recommended membranes are 85mm in diameter with a 0. Formula Tryptone Bile Salts No. The membrane is then transferred to a Tryptone Bile Agar plate and incubated at 44˚C: after incubation the membrane is flooded with indole reagent.57 . 55˚C aerobically for 48 hours for aerobic thermophile count.J. Formula Beef Extract Tryptone Glucose Agar No.M.C. Bacteriological Analytical Manual.0 15.G. Sterilise at 121˚C for 15 minutes. 109.45 micron pore size manufactured from cellulose esters. Baird. This medium is also recommended by the Association of Official Analytical Chemists (A. J.). J. 39: 111-117. Standard Methods for the Examination of Dairy Products. Allow to stand for 5 minutes for reaction to develop. Measurement of E. Appearance: Pale straw colour. Bact.O. Indole positive colonies produce a red colour on the membrane and are easily counted. Indole reaction: Pipette 1-2ml of reagent into Petri dish lid. Speck. coli type I by the membrane filter technique Wat. References Association of Official Analytical Chemists (AOAC).J.) (1976)..2 Method for reconstitution Weigh 24 grams of powder. disperse in 1 litre of deionised water.J.5 15. disperse in 1 litre of deionised water. Food Microbiol.G. (1988). References Anderson.C. no growth Pseudomonas spp.. & Grasso.. organisms: S. 6˚C aerobically for 10 days for aerobic psychrotroph count. (1962). Appl. Minimum Q.0 1. R.0 1. A rapid and direct method for enumerating Eschericia coli biotype I in food. The idea was applied to foodstuffs by Anderson and Baird-Parker in 1975.A. J.: American Public Health Association.C. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 4 days at 2-8˚C in the dark. Pharmacopoeia of culture media for food microbiology. coli NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. pH: 7.L. Capped containers – up to 3 months at 15-20˚C in the dark. indole reaction on membrane positive – pink colour negative – no colour other Enterobacteriaceae 0. (1975). Growth characteristics organism E. 289. Hausler.M. G.0 shape & surface CV. Compendium of Methods for the Microbiological Examination of Foods.2 Minimum Q. Int. epidermidis NCIMB 50082 E.E.. Incubation: 4 hours at 37˚C on Nutrient Agar LAB 8 or Minerals Modified Glutamate Medium LAB 80A and 80B plus agar – then 1824 hours on Tryptone Bile Agar at 44˚C. remove membrane with forceps and place on reagent. Sewage Wks. W. Allow to soak for 10 minutes. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. organisms: E. 3 Agar No. Bacillus spp.5-2. Corry. Allow to soak for 10 minutes. (1995). A. McCarthy. J.U. J.A. Baird-Parker. J. Appearance: Straw coloured. 1.) for estimation of total viable counts in food and dairy products. clear.0 Klebsiella spp.H. no growth no growth no growth CV.E. coli. Staphylococcus spp.5 grams of powder. Inoculation: Pour plate technique.0 ± 0. Count all pink colonies as E. 2 g/litre 20.Tryptone Bile Agar LAB 72 Description First introduced by Delaney.0 5. Curtis. Incubation: 30˚C aerobically for 48 hours for aerobic mesophile count. (Ed. 8th ed. Indole reagent: 5% p-dimethylaminobenzaldehyde in N-HC1 (Vracko & Sherris 1963). The inoculum is placed onto the membrane on a resuscitation agar and incubated at 37˚C for 4 hours.: American Public Health Association. 1 g/litre 3. then dry in sunlight to ‘fix’ the colour.) (1976).A.

G.) LAB 11 Description A general purpose agar which will support the growth of a wide range of micro organisms.5-1. The medium is recommended by the United States Pharmacopeia for the sterility testing of a wide range of pharmaceutical products. J. coli NCIMB 50034 S.C. organisms: E.5 CV. pneumoniae Ent. GreyWhite White GreyGreen mucoid (marked strain variation) (CO2) other References United States Pharmacopeia 21st edition (1985).G.) (Soybean Casein Digest Medium U. 1.P. 2 g/litre 15.E. g/litre 17.P.0 5.0-1.5 S.P. New York.S. M.G. 93: 1-13.0 5. Capped containers – up to 3 months at 15-20˚C in the dark.E. F.S.0-1.E. colour WhiteYellow White. Strep. 37˚C aerobically for 14 days for blood cultures.E.0 5. Appearance: Pale straw coloured. Do not exceed temperature. aeruginosa 0.G. Dispense into tubes or flasks and sterilise at 121˚C for 15 minutes. Transp. The bacteriophage typing of staphylococci.C. The medium can be used for phage typing. Minimum Q. allow to soak for 10 minutes.5 CV. A. pyogenes PP-0. epidermidis NCIMB 50082 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. clear. Inoculation: Surface plating.E.P. Infect. pH: 7. disperse in 1 litre of deionised water.5 2. usually aerobic. organisms: E. Cool to 47˚C. 1. Swirl to mix and warm if necessary to dissolve.P.0 References United States Pharmacopeia 21st edition. Incubation: Time and temperature to suit organisms. Dis.G.G. CV. CV. (1953). Examination of Dairy Products. CV.) (Soybean Casein Digest Medium U. swirl to mix then sterilise for 15 minutes at 121˚C. faecalis PP 0.A.E.S. The formula conforms with that laid down by the United States Pharmacopeia for sterility testing.0 Klebsiella spp. colicine typing and for testing the X and V factor requirements of Haemophilus spp.) Tryptone Soy Broth (U. pH: 7.E. Growth Characteristics organism S.0 3. clear gel.0 Ps.0-3.D.2 Dextrose Method for reconstitution Weigh 30 grams of powder. for sterility tests.Tryptone Soy Agar (U. coli NCIMB 50034 S. J. Growth indicators: Turbidity or precipitate.3 ± 0.2 Minimum Q.CR. aureus NCIMB 50080 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark. and Carr.H.strain-dependent Yellow Transp.5 Method for reconstitution Weigh 37 grams of powder. The medium is also widely used for blood cultures although the high carbohydrate level may cause rapid growth and subsequent death of acid-producing organisms. Incubation: 20-25˚C aerobically for 14 days.0 12.3 ± 0. aureus Other staphylococci colony size shape & (mm) surface 1.. disperse in 1 litre of deionised water. 2. (1985).0 Formula Tryptone (Casein Digest USP) Soy Peptone Sodium chloride Dipotassium phosphate LAB 4 Description A general purpose nutritious broth capable of growing a wide range of bacteria and fungi. Formula Tryptone (Casein Digest USP) Soy Peptone Sodium chloride Agar No.5-3.58 . Appearance: Straw coloured.0 2. mix well and then pour plates. Blair.S. CV.

Allow to soak for 10 minutes.C. 3rd edn. organisms: E. The medium is a general purpose broth that has been used as a blood culture medium. Medium (Tryptone Yeast Cystine) Minimum Q. then sterilise for 15 minutes at 121˚C. Interpretation: Indole positive organisms will give a distinct colour change when either Kovac’s or Ehrlich’s indole reagent is added. J.3 ± 0.2 units per ml of Bacitracin. Method for reconstitution Weigh 29. New York.0 0.5 ± 0. coli NCIMB 50034 S. mix and pour plates. pH: 7. A. swirl to mix. organisms: E. (1983). 2nd edn. Inoculation: Surface. coli NCIMB 50034 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.5 grams of powder. clear.C. A.H. References American Public Health Association (1948). disperse in 1 litre of deionised water.8 2. (1950).H. Allow to soak for 10 minutes. pH: 7. 10th edn.0 5. aureus NCIMB 50080 Storage of Prepared Medium: Capped containers – up to 3 months at 15-20˚C in the dark.. Tryptose Phosphate Broth LAB 62 Description This is a versatile nutritionally rich buffered glucose broth. and with the addition of sodium azide 0.2 Minimum Q.2 0. Sterilise by autoclaving at 121˚C for 15 minutes. Method for reconstitution Weigh 15 grams of powder.A. Incubation: Dependent on application. Inoculation: For blood culture work dilute sample at least 1:10 in broth. Standard Method for the Examination of Dairy Products.025% as a selective medium for streptococci in dairy products. 10% CO2.0 Minimum Q. Diagnostic Procedures and Reagents. New York. D.59 .0 0. American Public Health Association. Sterilise by autoclaving at 121˚C for 15 minutes. American Water Works Association 10th edn.5 Disodium phosphate anhydrous Sodium bicarbonate Sodium acetate anhydrous Sucrose Agar No. Incubation: 37˚C for 24-48 hours. Appearance: White. Biochemical tests for the identification of medical bacteria.1 1. de Stoppelaar in 1967 to differentiate Streptococcus sanguis (frequently found in dental plaque) from Streptococcus mutans (implicated in dental caries). It can be made selective by the addition of 0.2 1. streaking out for single colonies.. Appearance: Very pale straw.P. Formula Tryptose Dextrose Sodium chloride Disodium phosphate g/litre 20. Appearance: Colourless. This product is preferable to peptone water LAB 104 because it has a higher content of tryptophan.0 12.0 2..3 ± 0.0 12. 2 Method for reconstitution Weigh 98 grams of powder. pH: 7.Tryptone Water LAB 129 Description A substrate for the testing of an organism’s ability to produce indole from tryptophan.A. mutans ATCC 25175 Storage of Prepared Medium: Plates – up to 7 days at 4˚C in the dark.0 5. a supplement for animal cell culture. clear. The indole test is frequently used in the classification of coliform organisms.Y.0 5.C. Williams & Wilkins. organisms: S. mutans thus forming distinctive colonies. Baltimore. The medium uses a high sucrose content to promote the formation of specific glucans by S. disperse in 1 litre of deionised water.2 T.P. translucent gel.0 References American Public Health Association. Heat to dissolve then distribute into screw cap bottles. LAB 35 Description A medium designed by J. Formula Tryptone Sodium chloride g/litre 10. Incubation: 37˚C for 4-5 days in an atmosphere of 90% H2. Cool to 47˚C. 391-392.0 2. heat to dissolve solids then distribute into final containers. MacFaddin. (1955). disperse in 1 litre of deionised water. Formula Tryptone Yeast Extract L-Cystine Sodium sulphite Sodium chloride g/litre 15.C. Inoculation: From pure culture.0 50.

.2 0. (1973). Jordon. but this will be delayed.0 As Type A Grey white soft consistency. J.. Walker. Formula Peptone Glucose Sodium chloride Disodium phosphate Potassium dihydrogen phosphate Phenol red Agar No.012 12. C.. Citrobacter spp. Inoculation: Pure culture using straight wire for stab/streak technique. aerobically. The indicator for ammonia production is phenol red. Bratthall. Distribute into sterile screw cap bijou bottles. Sandham. Urea Broth Base (Christensen) LAB 131 Description This is a liquid version of Christensen’s medium (LAB 130) introduced by Maslen in 1952. J. Alldred. clear. pH: 6. (1946). translucent. coli NCIMB 50034 Storage of Prepared Medium: Capped container – up to 1 month at 2-8˚C in the dark.2 Minimum Q. Emilson. Interpretation: Production of red colour in under 6 hours is positive for rapid urease production. (1972). organisms: Proteus spp. and it is easier to detect contamination in a fluid rather than in a slope. Microbiol.2 1. E.G.0-3. then sterilise at 121˚C for 15 minutes. J. Maslen also claimed that it is easier to detect positive results. Thesis. disperse in 95ml of deionised water. This delay is achieved by the incorporation of glucose and the introduction of a buffering system into the medium.B.J.G. 52: 461-466. aerobically. de Stoppelaar. The presence of dextran forming bacteria resembling Streptococcus bovis and Streptococcus sanguis in dental plaque. Incubation: 37˚C for 4-6 hours or overnight. Archives of Oral Biology 19: 1357-1364.G. Inoculation: Fluid culture by pasteur pipette or straight wire from pure growth. A selective medium for Streptococcus mutans. allow to set in the sloped position.9 grams of powder. M. (1971). Other enterobacteria will split the urea. Ikeda. Wade. add aseptically 5ml sterile urea solution X130/X135. Method for reconstitution Weigh 2. pH: 6. Staphylococcus spp.0 0.C. Incubation: 37˚C for 4-6 hours – preferably in a water bath for most rapid growth. H. Utrecht.0-3. This modification allows inoculation by Pasteur pipette. An improved medium for isolation of Streptococcus mutans.8 0. O.0 ‘heaped’ colony granular surface irregular edge colour other Minimum Q.0 1.0 1. J. Streptococcus mutans.1 grams of powder. E.V. and de Moor.C. van de Houte.60 . organisms: Proteus sp.. W. 12: 1199-1201. Journal of Clinical Microbiology 4: 95-98. Yellow Crumbles when touched with wire Type B 1. Interpretation: The production of a red colour in under six hours is a positive result for rapid urease. Streptococcus sanguis and dental caries. Distribute into sterile bottles and slopes. Swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. Archives of Oral Biol. D.8 ± 0.Growth Characteristics organism Strep. Allow to cool to 47˚C. (1967). (1986). J. Appearance: Yellow.8 5.0 5.0 1.8 ± 0. Van Houte. Growth of Streptococcus mutans on various selective media. Formula Peptone g/litre 1. (1976). mutans Type A colony size shape & (mm) surface 1.2 0. 4: 781-783. Helicobacter pylori Red colour Red colour Red colour Red colour Red colour 4-6 hours 18-24 hours 18-24 hours 24-48 Hours 30 minutes S.004 Urea Agar Base (Christensen) LAB 130 Description This is a modification of Christensen’s urea base for the detection of rapid urease production by Proteus spp.0 1. C. Oral Biol.. 1 g/litre 1. Klebsiella spp.D.M. Urea decomposition as a means of differentiating Proteus and paracolon cultures from each other and from Salmonella and Shigella types. add to 95ml of deionised water. Appearance: Yellow/pale pink. 22: 319-323.0 Glucose Disodium phosphate Potassium dihydrogen phosphate Sodium chloride Phenol red Method for reconstitution Weigh 0. Med. coli NCIMB 50034 Storage of Prepared Medium: Capped container – up to 1 month at 2-8˚C in the dark.0-3. sanguis 1.0 Convex glossy crenated White References de Stoppelaar. Arch. References Christensen. D.. Bacteriol. white precip in agar (glistening drop) very rubbery (glistening drop) Organism Growth Characteristics Proteus spp. swirl to mix. Allow to soak for 10 minutes. H. J. J.D. Allow to cool to 47˚C then add aseptically 5ml of X130/X135 sterile urea solution. W. Rijksuniversiteit.E. T. Gold. A high-sucrose medium for the identification of Streptococcus mutans.

66-79. Appl. Recommended Methods for the Microbiological Examination of Foods. Influence of carbon source. Hyg.61 . Microbiol. (1986).5 10. A.0 9.03 0.P. Inoculation: UVM I – Add 25g sample to 225ml of UVM I and homogenise.2 Appearance: Straw opalescent broth g/litre 5. Routine use of liquid urea medium for identifying Salmonella and Shigella organisms. Subculture onto selective agars at 24 and 48 hrs. Eelderink.H. Zbl. Koopmans.) LAB 31 Description A medium for the enumeration of coliform organisms in food and dairy products. Incubation: 37˚C for 18-24 hours for ‘coliforms’.. The selectivity of the medium is due to the presence of bile salts and crystal violet.4 ± 0.0 20. ulcerans Some strains Citrobacter Klebsiella Escherichia Yersinia Staphylococcus Pasteurella multocida Red colour Red colour Red colour Red colour . Mossel.A. J. (1977). Calculate the number of coliforms in original sample. Cool to 45˚C and distribute into bottles or tubes. and van Rossem. Method for reconstitution Weigh 52 grams of powder and add to 1 litre of deionised water. 13th edn. 1989. 32˚C for 24-48 hours for mesotrophs. D.G. Selectivity can be increased by incubation at 42-44˚C. Food Protec. I. 4˚C for 10 days for psychrotrophs..0 1. Corynebacterium hoffmani C. D.. bile salts and incubation temperature on the recovery of Enterobacteriaceae from foods using MacConkey type agars. The original method has been modified to replace the second stage broth (UVM II) with Fraser broth LAB164 (McClain & Lee 1989). P.4 ± 0.C.5 grams of powder. (1952). Incubate selective agars for 24 and 48 hrs. use within 3 hours.P.P.monocytogenes from processed meat and poultry products. Warburton D.0 5. Development and use of single..M. and forms the basis of the USDA method. J. D. J. I. NCIMB 50007 E.0 7. Capped containers – up to 1 month at 15-20˚C in the dark. 3 UVM Base LAB 155 Description UVM (University of Vermont Medium) Base is a two stage selective enrichment broth for the isolation of Listeria from meat products and environmental swabs. and Lee W..57.C. Appearance: Light purple-violet. Allow to soak for 10 minutes.1ml of enriched UVM I into 10ml UVM II. A 278. (1979). 1 Sodium chloride Bile Salts No. London. I. Washington. clear gel. Davis.0 References Maslen L. Revised May 24. 2: 545-546. coli (inhibition) NCIMB50034 References McClain D..No. . A.A.. Formula Tryptone Meat Peptone Beef Extract Yeast Extract Sodium chloride Disodium hydrogen phosphate Potassium dihydrogen phosphate Aesculin pH: 7. 18-24 hours Violet Red Bile Agar (V. 289-295. J. organisms: E. ‘polytropic’ diagnostic tubes for the approximate taxonomic grouping of bacteria. 2nd edn. Dissolve by bringing to the boil with frequent swirling of the flask to prevent overheating. UVM II – 30˚C for 24 and 48 hrs.A.A.A. Lab. isolated from foods. et al (1991) A Canadian comparative study of modified versions of the FDA and USDA methods for the detection of L. and Sutherland. . Bacteriol. Formula g/litre 3.H. Washington. . Hausler). J. water and medicinal preparations.Comm... US Dept of Agric. American Public Health Association (1966).G.Food Protection 54 (9) 669-676.. coli NCIMB 50034 S.H. (1951). If held molten in a water bath. Mix well and distribute into sterile tubes or bottles.A. Brit. Div..5mm in diameter. Mossel. Bakt. Med. F.H.0 5.W.6 1.0 Lactose Neutral red Crystal violet Agar No. Orig. (ed. van der Zee.. swirl to mix and sterilise at 121˚C for 15 minutes..A. Mossel. 60. epidermidis (inhibition) NCIMB 50082 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. 1..0 0. M. J. disperse in 1 litre of deionised water.. Milk Testing Dairy Industries.A.002 12. H.35 1.2 Minimum Q. Interpretation: Count all red/purple colonies > 0. Standard Methods for the Examination of Dairy Products. Incubation: UVM I – 30˚C aerobically for 24 hrs. . Cool to 47˚C and add 2 vials of UVM I supplement (X155/X555) or UVM II supplement (X156) as required.Organism Growth Characteristics Proteus spp.FSIS. (ed.B.R.. Eelderink. 42. W. Hardon. Inoculation: Pour plate (with or without overlay) or surface spread. 470-475. .A. References American Public Health Association (1972).. Minimum QC organism: Listeria sp. Subculture onto selective agars and into UVM II.0 5. Yeast Extract Balanced Peptone No. Red colour 4-6 hours 18-24 hours 18-24 hours 18-24 hours .0 5.monocytogenes.. pH: 7. 2 Method for reconstitution Weigh 38. . and van Netten. Non lactose fermenters produce pale colonies.P. Lactose fermenters produce red/purple colonies often surrounded by a halo of the same colour. UVM II – Subculture 0. J. The enumeration of thermotropic types amongst the Enterobacteriaceae colonizing perishable foods. (1989) FSIS method for isolation of L. 42˚C for 18 hours for thermotrophs. Autoclaving is not necessary. Sharf) A.

Glucose is fermented by all members of the Enterobacteriaceae thus V. cerevisiae. 77: 513-516.5 grams of powder. Wallerstein Lab. P. V.004 g/litre of Actidione to suppress the yeasts. and Scholts. Mengerink. Interpretation: Count all colonies. Inoculation: Pour plate method with overlay. Bacteriol.C. References Pharmacopoeia of Culture Medium for Food Microbiology (1987). J. D.2 Minimum Q. Lactobacillus and Pediococcus spp. coli NCIMB 50034 S..A. mix well and dispense into tubes or bottles. 1 Sodium chloride Bile Salts No. Further sterilisation is not required.F. disperse in 1 litre of deionised water. LAB 31 contains lactose which is fermented by members of the coli/aerogenes group. J. Incubation: 30˚C aerobically for 48 hours (bacteria).425 0. The overlay procedure ensures anaerobic conditions and suppresses the growth of non-fermentative Gram negative bacteria. Inst. gives a presumptive Enterobacteriaceae count. 1.C. Int. pH: 5.G.B. Calculate organisms per ml in original sample. Inoculation: Surface plating or pour plate. If a process involving bakers or distillers yeast is under examination the pH should be adjusted to 6. disperse in 1 litre of deionised water. (1950). Use of a modified MacConkey agar medium for the selective growth and enumeration of Enterobacteriacaea. 2 g/litre 3. S.B. 2 g/litre 4.0 1. 13. Nutrient Agar (Wallerstein Laboratory) LAB 88 Description A modification of Violet Red Bile Agar LAB 31 introduced by Mossel in 1978.Violet Red Bile Glucose Agar (V.5mm in diameter. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. 3 Glucose Neutral red Crystal violet Agar No.R.0 5.0 7. The medium may be adapted to detect bacteria only by the addition of 0. Brewing. Bring to the boil with frequent swirling to prevent overheating.R.A.P. use within 3 hours.022 15.002 12. (1962).5 10.125 0.0 0.J.R.B. 5: 3: 280-81.5 ± 0. organisms: S. Allow to soak for 10 minutes.0025 0. Formula Yeast Extract Tryptone Dextrose Potassium dihydrogen phosphate Potassium chloride Calcium chloride Magnesium sulphate Ferric chloride Manganese sulphate Bromocresol green Agar No.) W.0 LAB 79 Description This medium was developed by Green and Gray in 1950 for the isolation and enumeration of yeasts. Calculate the number of Enterobacteriaceae in original sample. H. Incubation: 37˚C aerobically for 18-24 hours. Capped containers – up to 1 month at 15-20˚C in the dark. The medium has a pH of 5. Flavobacterium.125 0.A.G. Comm. 20˚C aerobically for 48 hours (yeasts).0 50.4 ± 0.55 0.B. If held molten in a water bath. 84: 381. Bile salts and crystal violet are used to inhibit Gram positive and non-enteric organisms. If adjustment of pH to 6.A. clear. W. J.0 Method for reconstitution Weigh 38.03 0.0 5. Food Microbiol. (1971). L.H.H. Detection of Wild Yeasts in the Brewery. epidermidis (inhibition) NCIMB 50082 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Minimum Q. Mossel. moulds and bacteria in the brewing process. Formula Yeast Extract Balanced Peptone No.357. Cool to 45˚C. Interpretation: Count all red/purple colonies > 0. organisms: E.0025 0. Differential Procedure Applicable to Investigation in Brewing. as well as yeasts and moulds.A.R. References Green.5. Hall. this medium gives a ‘coliform’ count. clear.0 0. Capped container – up to 1 month at 4˚C in the dark.2 Method for reconstitution Weigh 75 grams of powder.5 is required used 1% sodium bicarbonate.62 . V. Allow to soak for 10 minutes then swirl to mix. Appearance: Light purple-violet. Storage of Prepared Medium: Plates – up to 7 days at 4˚C in the dark. pH: 7. LAB 88 has substituted lactose with glucose. J.A.5 which is optimum for Brewers yeast and will allow the growth of a wide range of organisms including Enterobacteriaceae. and Gray. Appearance: Green.G.R.

E. Storage of Prepared Medium: Capped container – up to 1 month at 15-20˚C in the dark. organisms: S. pH: 4. translucent. 2 g/litre 15. Dairy Sci.s. Formula Malt Extract Peptone Maltose Dextrin Dipotassium phosphate Ammonium chloride g/litre 15. swirl to mix then sterilise by autoclaving at 121˚C for 15 minutes. (1959). Incubation: 25˚C aerobically for 5 days. Sci. Allow to soak for 10 minutes.78 12. cerevisiae colony size shape & (mm) surface 4.0-3.Wort Agar LAB 38 Description A medium for the enumeration of yeasts and moulds in butter. cerevisiae. 1.78 12. Add 2. swirl to mix then sterilise for 15 minutes at 121°C.) for 20 minutes. Formula Malt Extract Peptone Maltose Dextrin Dipotassium phosphate Ammonium chloride Agar No. Minimum Q.0 15. E.0 ± 0.0 Varies with strain Cream CV.63 .i. pH: 5. Fd. 10: 678-681. M.0 1. organisms: S.G colour White other References Parfitt.C. Inoculation: Pour plate or surface spread.i. Use 60 grams per litre if required for inoculation by plate streaking with a wire loop. Agric.35ml of glycerol. If osmophilic modification is required add 48.3 grams of powder and disperse in 1 litre of deionised water. 16: 141-147. Incubation: 25˚C aerobically for 5 days.75 2. (1933).0 Wort Broth LAB 99 Description A broth version of the medium LAB 38 Wort Agar developed by Parfitt for the enumeration of yeasts and moulds. cerevisiae. J. translucent. Appearance: Light Brown. Selective media used in the microbiological examination of sugar products. Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Capped container – up to 1 month at 15-20˚C in the dark.0 0.C.) for 20 minutes.H.0 Varies with species 2.35ml of glycerol. Allow to soak for 10 minutes.2 Method for reconstitution Weigh 33. add 2.8 ± 0. Fungi S.0 Method for reconstitution Weigh 48.0 1. Scarr. The medium can be modified to enable it to isolate osmophilic yeasts from soft drinks and sugar products by the addition of high concentrations of sucrose and glucose.2 Minimum Q. If osmophilic version is required disperse 33.75 1.s. The medium can be modified for the isolation of osmophilic yeasts from soft drinks and sugar products by the addition of high concentrations of sucrose and glucose. Growth Characteristics organism Candida spp.75 2.0 0. in butter. Appearance: Light Brown. The influence of media upon the yeast and mould count of butter. developed by Parfitt in 1933.75 1.3 grams of powder to 1 litre of a solution containing 35% w/v sucrose and 10% w/v glucose then sterilise at 108˚C (5 p. J.3 grams of powder and disperse in 1 litre of deionised water.3 grams of powder in 1 litre of a solution of 35% w/v sucrose and 10% w/v glucose then sterilise at 108˚C (5 p. Do not exceed time or temperature of sterilisation.P.

E.G. (1967).5 Proteus spp 1. epidermidis NCIMB 50082 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. (1965). J. organisms: E. (1967).G.0 0. Capped container – up to 3 months at 15-20˚C in the dark.64 . Isolation of shigellae.2 Minimum Q.C. 1 g/litre 3. Clin. II.(D) Citrobacter spp. (1965). W. 48: 350-355.E. coli 1. Bring rapidly to the boil with frequent stirring. Pathol.0-2. CV.I.. To prepare Yeastrel Milk Agar add 10ml of fresh milk before autoclaving. Pink fishy odour (black centre) 1.0 15. will ferment xylose to produce acid. Comparison of plating media with stools. III. and dispense into containers.E.2 Method for reconstitution Weigh 53. swirl to mix. 44: 471-475.5 grams of powder. B.5 CV. 48: 356-362. Inoculation: Pour plate technique or surface spreading.5-1. Am.08 13. Pathol.0 5. disperse in 1 litre of deionised water. 1 Agar No.. (clearing of acid black ppt of coliforms) centre Pink Pink Pink Yellow Yellow inhibited (ppt around colony) (black centre) Shigella sonnei 1. disperse in 1 litre of deionised water. IV. However the salmonellae will also decarboxylate the lysine to keep the pH neutral. Isolation of shigellae.8 0. Formula Xylose L-Lysine Lactose Sucrose Sodium chloride Yeast Extract Phenol red Agar No. 2 Sodium desoxycholate Sodium thiosulphate Ferric ammonium citrate g/litre 3.E.G.I. Isolation of shigellae.G. Am.I. Citrobacter spp. Agar (Xylose Lysine Decarboxylase Agar) References Taylor. 1. CV. Methods for the Examination of Water and Associated Materials. Pour into plates as soon as the medium has cooled. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.5 0.0 0.C. Sterilise for 15 minutes at 121˚C.G. Allow to soak for 10 minutes. 55˚C aerobically for 48 hours for aerobic thermophile count.. 6˚C aerobically for 10 days for aerobic psychrotroph count.0 6. Mix well.0-1. Yeast Extract Agar (Yeastrel Milk Agar) LAB 18 Description A nutrient agar corresponding to the Standard Formulation for the plate count of micro-organisms in water and dairy products. colour other References Environment Agency: The Microbiology of Drinking Water (2002). NCIMB 50076 E. J. Protracted boiling or prolonged holding at elevated temperature induces precipitation.5-1.D.G.4 ± 0.5 CV.L. Salmonella spp. and transfer immediately to a 47˚C water bath. Most enteric organisms except Shigella. pH: 7. pH: 7. W. Free steam or boil to dissolve. B. Formula Yeast Extract Balanced Peptone No. Clin. Comparison of new and traditional media with stool specimens. clear gel.0 7. Appearance: Pale straw.2 ± 0. Clin. Minimum Q. and Harris. Incubation: 37˚C for 18-24 hours aerobically. Appearance: Light rose. D. Xylose Lysine Agars: New media for the isolation of enteric pathogens. Incubation: 30˚C aerobically for 48 hours for aerobic mesophile count.0 3. 44(4). The indicator system is novel and complex. 1.5-2. Taylor.E. and Harris. Am. The medium is low in nutrients and relies on a small amount of sodium desoxycholate for selectivity. CV.0-2. Isolation of shigellae.0 1. W. Comparison of plating media and enrichment broths. LAB 32 Description This medium was introduced by Taylor in 1965 to improve the recovery and recognition of Shigella spp. At near neutral pH Salmononella can produce H2S from the reduction of thiosulphate producing black or black centred colonies. J. clear gel. Growth Characteristics organism colony size shape & (mm) surface CV...G.I. Taylor. J.. W. coli NCIMB 50034 S. and has proved to be an excellent medium for Salmonella spp.5 Trans. organisms: Salmonella sp. I. British Standard 4285: Methods of Microbiological Examination for Dairy Purposes. 476-479. streaking out for single colonies. Inoculation: Surface. Pathol.X. the acid produced by fermentation of both lactose and sucrose will keep the pH too acid for H2S to be produced. Pathol.5 5. Clin.E. dysenteriae E. Am.5 S.E. This medium is also useful for teaching and demonstration purposes using non-fastidious organisms.8 Taylor.(D) CV.0 Method for reconstitution Weigh 23 grams of powder..0-2. however.75 5.5 7. flexneri S. Trans. and Schelhart. can also decarboxylate lysine.

pH: 6. The selective components are sodium desoxycholate. Development of a two step enrichment procedure for recovery of Yersinia enterocolitica from food.N. The ratio of transparent border to red centre varies with serotype and environmental strains may appear rough with an irregular edge. THIS MEDIUM MUST NOT BE RE-AUTOCLAVED. Appearance: Pale yellow. Synthesis of a selective agar medium for Yersinia enterocolitica. Cefsulodin Irgasan Novobiocin (C. if they grow.A. Interpretation: Count all colonies. organisms: Aspergillus sp.N. Microbiol.A. Can. Milchwiss.I. Formula Yeast Extract Dextrose Agar No. clear. J. International Milchwirtschaftsverband: Milch und Milchprodukten – Zählung von Hefen und Schimmelpilzen (Kolonieählung bel 25 C). (1979). Agar) LAB 120 Description This medium is based on the work of Schiemann. – International IMV Standard 94: (1980) in Milchwiss. D.6: (1986).N. Microbiol. Red centre Citrobacter spp.G. Minimum Q.0-2. Formula Peptone Mixture Mannitol g/litre 22.01 2.S. Incubation: 30˚C aerobically for 24 hours. (1982). 2 Minimum Q. Appl. crystal violet. Inoculation: Surface.A. Allow to cool to 45˚C before using with poured plate technique. G. Incubation: 25˚C for 5 days. Schiemann. organisms: Y.65 . 4285. disperse in 1 litre of deionised water. 37: 727-730. 2. Bestimmung der Anzahl von Hefen und Schimmelpilzen. the International Organisation for Standardisation (I.5 0. swirl to mix then bring to boil.A. cerevisiae E. 43: 14-27. Allow to soak for 10 minutes. J. B. produce a larger colony with a diffuse pinkish centre and opaque outer zone. Pale pink colony Gram +ve organisms no growth References Schiemann.5 CV. 36: 220-222. Mikrobiologische Milchuntersuchung. then bring to the boil for 1 minute only.5-3. The medium is said to have superior storage properties to O. International Organisation for Standardization (ISO): Milk and milk products – enumeration of yeasts and moulds – colony count technique at 25˚C – standard method ISO/DIS 6611. 1 g/litre 5. localised.C. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark.6 ± 0.E.2 References Engel.V.Yeast Extract Dextrose Chloramphenicol Agar LAB 119 Description A selective medium for the enumeration of yeasts and moulds in milk and other dairy products. 5: 208. may be rough & irregular (may not grow) Y. Method for reconstitution Weigh 58 grams of powder.03 0.5 20.0 CV. Eniviron.0 1.001 12. Mix well.0 0. (1982).0 15. Produkte in DIN Deutsches Institut für Normung e.E.O. NCIMB 50097 S.I. 209.I. D.). Growth Characteristics organism colony size shape & (mm) surface colour other Colony varies with strain. Inoculation: Pour plate technique. It is used for the isolation and enumeration of Yersinia spp.0 20.G. Food. Allow to soak for 10 minutes.I. Verglich verschieden Nährböden zum quantitativen Nachweis von Hefen und Schimmelpilzen in Milch und Milchprodukten. disperse in 1 litre of deionised water. Yersiniae ferment mannitol with an intense. Section 3. pH: 7.Y. cefsulodin. from clinical samples and from food. (1987). enterocolitica 1.2 Sodium chloride Magnesium sulphate Sodium pyruvate Sodium desoxycholate Neutral red Crystal violet Agar No. pour plates. Mossel.0 0.0 Yersinia Selective Agar (Schiemann’s C. 25: 1298-1304. D. Add 2 vials of X009 (X209) which have been dissolved in ethanol and autoclave at 121˚C for 10 minutes. irgasan and novobiocin. aerobically.) and the British Standards Institute (B. Appearance: Red. clear.4 ± 0.) agar. Normenausschufl Lebensmittel und landwirtshaft. enterocolitica E. DO NOT AUTOCLAVE.S. and also has the advantage of incorporating an autoclavable supplement. Microbiol. Allow to cool to 47˚C add 2 ampoules C. Int. Reference method DIN 10186. The formulation meets the requirements of the International Milk Union (1980).S.E.0 Method for reconstitution Weigh 40 grams of powder.G. British Standards Institute.C. streaking out for single colonies. 1. supplement X120. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates – up to 7 days at 2-8˚C in the dark. Most other enteric bacteria. acid production in the centre of the colony which produces a red ‘bull’s eye’ appearance.

1.66 .

(1965) Amer. I. Mossel. The low nutrient content of the medium and the inclusion of phosphate buffer prevents bacterial overgrowth by E.4 ± 0.1 1.C. J.. cultivating and determining haemolytic reactions of fastidious pathogens. J. Bact. J. Bacteriol.G. 46(4) 211-217. 40. E. The inhibition of the spreading growth of Proteus and other bacteria to permit the isolation of associated streptococci. Pathol. C. The medium may be prepared as a pre-reduced anaerobic sterilised medium (PRAS) by the Holdeman and Moore method. pH: 7. Push the swab down one third of the depth of the medium and cut the stick. 1. 78. opaque. R. organisms: Enterococcus faecalis NCIMB 50030 Staphylococcus aureus NCIMB 50080 Escherichia coli NCIMB 50034 (inhibition) Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. M. Sodium azide as an inhibition of Gram-negative bacteria. J. 330-334.H. H.L. Lichstein. New York. 42(5) 653-664.N.C.2 Minimum Q. Diepen.Infect. R. anearobically or under conditions of increased CO2 (5-10%) for 18-24 and 40-48 hours of incubation. Path. 283-288. The diagnosis of Streptococcus mastitis by cultural methods.0 5. Appearance: Light to medium amber. Pathol. American Public Health Association (1978) Standard Methods for the Examination of Dairy Products. Salmonella and Shigella after 49 days and Yersinia pestis up to 75 days storage at 28°C. J.3 grams of powder and disperse in 1 litre of deionised water. 113. Sodium azide has a bacteriostatic effect on the majority of Gram-negative bacteria but permits the growth of Gram-positive bacteria such as streptococci and some strains of staphylococci.A. 67. Matthew.1 New Media Products Azide Blood Agar Base NEW Packer. cool to 47°C and add sterile defibrinated blood to a concentration of 5%.M. 88.5 5.0 0. Syndar. Wood.D.2 ± 0..A. For transport of fastidious anaerobic bacteria prepare the medium as directed and fill into long narrow screw capped tubes. and Stuart. (1965). APHA Inc. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. (1940). (1943). Survival of Shigella and Salmonella in a new transport medium for shipment of clinical samples. Vet. 431-438. cotton-tipped swab on wooden sticks to collect the specimen.09 5. 218. R. which can occur in other transport medium containing sodium glycerophosphate. Dis. Bohm. (1971) Z. K. Cary and Blair reported recovery of cholera vibrios up to 22 days.2 grams of powder and disperse in 1 litre of deionised water.G.0 3.0 Cary-Blair Medium NEW LAB 505 Description Cary-Blair medium is a transport medium for the collection and shipment of clinical specimens based on the formulation of Cary and Blair. (1940) J. Fusillo. C. Bact. D. References Edwards. M. Citrobacter freundii and Klebsiella aerogenes. 43. Distribute into bijou bottles and sterilise by immersing in free-steam for 15 minutes. Anaerobic incubation will enhance haemolysis reactions. Inoculation: Use sterile. and Synder M. Bact. (1957).bl. slightly opalescent gel.1 . E.1.L.S. swirl to mix then bring to the boil.6 Method for reconstitution Weigh 30. J. With 5% blood. (1933). Appl. Crookes. M. Incubation: 35°C ± 2°C aerobically. Some strains of Gram-positive bacteria may be encountered that fail to grow or grow poorly on this medium. pH: 8.C. and Lichstein H. Orig.J. S. Infect. (1959) J. organisms: Shigella sonnei ATCC® 25931 Vibrio furnissi NCTC 11218 Storage of Prepared Medium: Store away from light at 2-8°C or at room temperature (22-25°C) for up to 19 months.J. Inoculation: Streak for isolation of single colonies. H. Ther. Screw the cap firmly on the bottle.. Comp.B.H. Survival of Vibrio parahaemolyticus has been reported after a 35-day period at 70-80°F. Stuart. Formula Tryptose Beef Extract Sodium chloride Sodium Azide Agar g/litre 10. Pour in to sterile Petri dishes.M. Method for reconstitution Weigh 13. and Blair. or to the neck of the Bijou bottle. 96-98. Dis 67(2) 113-115. and Harkins. Allow the medium to cool and tighten the screw caps to prevent water loss.2 12. (1941). Bact.0 0. Res 26. 20(2) 265-272. with mixing.S. LAB 523 Description This is a selective medium for the isolation of Gram-positive organisms from clinical and non-clinical specimens.van and De Bruin A. Formula Disodium hydrogen phosphate Sodium thioglycollate Sodium chloride Calcium chloride Agar g/litre 1. The medium can be supplemented with blood making it suitable for isolating. Clin. S. Am. 14th Edn. References Cary. (1964). 1303-1308. Label the bottle and send to the testing laboratory without delay.. (1959) Public Health Reports 74. 228-231. Allow the mixture to soak for 10 minutes. medium is cherry red. allow the medium to set and dry the surface prior to inoculation. The low oxidation-reduction potential of the medium ensures bacterial survival over long periods. coli. Azide blood agar is recommended by the American Public Health Association for the isolation of streptococci from cheese. 294-296. To prepare azide blood agar. Appearance: Colourless soft gel. Allow the mixture to soak for 10 minutes.L. to dissolve the agar. J.2 Minimum Q. Bakt. J. Cary.1.C. R. The use of sodium azide (NaN3) as an inhibition substance of Gram-negative bacteria. Precautions: Proteus and Escherichia species may not always be inhibited on this formulation.D.A. Dale. Interpretation: Examine plates for growth and haemolytic reactions. S.

0 2. Path. A.. B. (1970). Lecithin is included to neutralise quaternary ammonium compounds and Tween® 80 neutralises phenols. Bromocresol purple allows detection of growth via a colour change from purple to yellow when organisms ferment the glucose contained in the medium. Allow to soak for 10 minutes. References Roberts. Bromocresol purple allows detection of growth via a colour change from purple to yellow when organisms ferment the glucose contained in the medium. D/E Neutralising Broth contains thioglycollate to neutralise mercurial compounds. F. Developed by Dey and Engley.Neumann D. Practical Food Microbiology. and Greenwood. Med.0 7. this agar neutralises a broad spectrum of antimicrobial chemicals.1. Graf. Cool to 47°C and pour into sterile Petri dishes and allow to set. Method for reconstitution Weigh 39. 195-201. Whereas. Complete neutralisation is required to prevent false results arising from disinfectant carryover. ISBN 0 901144 36 3. Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. G. formalin.2 Minimum Q.10. and Dey. 57. i. Interpretation: Count all colonies for total counts. therefore the potency of the disinfectant is not diminished after addition to the medium. section 5. Curry.02 Method for reconstitution Weigh 54. Allow the mixture to soak for 10 minutes.2 . Benenson.5 2. L. 56th Mid Year Meeting. swirl to mix and dispense into final containers.W. and McEwen Jr.6 ± 0. Toiletry and Fragrance Association.N.0 2. Sterilise by autoclaving at 121°C for 15 minutes. It can also be used to test disinfectants by a disc diffusion method. ethanol. D.0 5. J.C (1975) Anaerobe Laboratory Manual. M. Incubation: 37°C aerobically for 24-48 hours.0 5. J. When used with D/E Neutralising Broth Base the action of the antimicrobial agent can be assessed..5 2. Appearance: Purple opaque gel. and Engley Jr. The procedure is based upon D/E Neutralising Broth Base being deficient of all neutralising agents. Sterilise by autoclaving at 121°C for 15 minutes.P. Virginia Polytechnic Institute Anaerobe Laboratory. sodium thiosulphate to neutralise iodine and chlorine and sodium bisulphite to neutralise formaldehyde and gluteraldehyde. Thioglycollate Medium and Neutralising Buffer. pH: 7.0 5. Clin. producing better results than those obtained using alternatives such as Letheen Agar.02 15. D.0 grams of powder and disperse in 1 litre of deionised water.. Wren. when disinfectant is added to the D/E Neutralising Broth..V.0 6. and combined with lecithin. N.) (1993) CFTA Microbiology Guidelines.0 6.A.0 0. Microbiol. Formula Glucose Lecithin Sodium thiosulphate Tween® 80 Tryptone Sodium bisulphite Yeast extract Sodium thioglycollate Bromocresol purple g/litre 10.W.5 1.5 1. 14:21-25. 3rd Ed. D/E Neutralising Agar contains thioglycollate to neutralise mercurial compounds. and Moore.D. A universal neutralising medium for antimicrobial chemicals. (1995).C. Microbiol. Hubster. Butler & Tanner.S.B. M. W. W. Complete neutralisation is required to prevent false results arising from disinfectant carryover. J. B. (1971). F. J.e. hexachlorophene. The Cosmetic.. sodium thiosulphate to neutralise iodine and chlorine and sodium bisulphite to neutralise formaldehyde and gluteraldehyde.C.T.B. Formula Glucose Lecithin Sodium thiosulphate Tween® 80 Tryptone Sodium bisulphite Yeast extract Sodium thioglycollate Bromocresol purple Agar g/litre 10. hexachlorophene. Inoculation: Consult appropriate references as this product is used in several procedures. Comparison of Dey and Engley (D/E) Neutralising medium to Lethhen medium and Standards Methods Medium for recovery of Staphylococcus aureus from sanitised surfaces. E.0 7. ethanol. its activity is neutralised allowing for the detection of any bacteria presence.0 0. Presented at the Chemical Specialities Manufacturing Association (CSMA) Proceedings. Washington. Lecithin is included to neutralise quaternary ammonium compounds and Tween® 80 neutralises phenols. Hooper.P. Engley Jr. (ed. Methods for the examination of food for micro-organisms of public health significance.E.0 D/E Neutralising Broth (Dey & Engley) NEW LAB 187 Description D/E Neutralising Broth is used to neutralise and determine the bacteriocidal activity of antiseptics and disinfectants. formalin. M.. and Tuan.N. Non-acid producing colonies are grey to colourless. D/E Neutralising Agar is used as the plating medium when testing disinfectants using D/E Neutralising Broth and D/E Neutralising Broth Base. Formulation is very hygroscopic. 2nd edition. Holdeman. Ind.. 10.0 5. Developed by Dey and Engley. D/E Neutralising Agar (Dey & Engley) NEW LAB 188 Description D/E Neutralising Agar is used to neutralise and determine the bactericidal activity of antiseptics and disinfectants. 1. and combined with lecithin. Dey.. whether it is bacteriostatic or has bactericidal properties. keep container tightly closed after use.G. count yellow colonies for differential acid producer count. producing better results than those obtained using alternatives such as Letheen Broth. Am.0 grams of powder and disperse in 1 litre of deionised water. (1995). organisms: Bacillus subtilis NCIMB 8054 Escherichia coli NCIMB 12210 Pseudomonas aeruginosa NCIMB 12469 Salmonella typhimurium NCIMB 13284 Staphylococcus aureus NCIMB 12702 Storage of Powder: Store at 2-8°C in the dark. D/E Neutralising Broth neutralises a broad spectrum of antimicrobial chemicals.

Appearance: Purple opaque liquid. pH: 7.6 ± 0.2

autoclaving at 121°C for 15 minutes. Appearance: Purple clear liquid. pH: 7.6 ± 0.2

Minimum Q.C. organisms: Bacillus subtilis NCIMB 8054 Escherichia coli NCIMB 12210 Pseudomonas aeruginosa NCIMB 12469 Salmonella typhimurium NCIMB 13284 Staphylococcus aureus NCIMB 12702
Storage of Powder: Store at 2-8°C in the dark. The formulation is very hygroscopic therefore keep the container tightly closed after use. Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. Inoculation: Consult appropriate references as this product is used in several procedures. Incubation: 37°C aerobically for 24-48 hours. Interpretation: Examine all tubes for increased turbidity, formation of a pellicle or a colour change from purple to yellow, indicating bacterial growth.

Minimum Q.C. organisms: Bacillus subtilis NCIMB 8054 Escherichia coli NCIMB 12210 Pseudomonas aeruginosa NCIMB 12469 Salmonella typhimurium NCIMB 13284 Staphylococcus aureus NCIMB 12702
Storage of Powder: Store at 2-8°C in the dark. Formulation is very hygroscopic, keep container tightly closed after use. Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. Inoculation: Consult appropriate references as this product is used in several procedures. Incubation: 37°C aerobically for 24-48 hours. Interpretation: Examine all tubes for turbidity, indicating growth.

References References
Roberts D., Hooper, W. and Greenwood, M., (1995). Methods for the examination of food for micro-organisms of public health significance, 2nd edition, section 5.10, Practical Food Microbiology. Butler & Tanner. ISBN 0 901144 36 3. Engley Jr., F.B. and Dey, B.P. (1970). A universal neutralising medium for antimicrobial chemicals. Presented at the Chemical Specialities Manufacturing Association (CSMA) Proceedings, 56th Mid Year Meeting. Dey, B.P. and Engley Jr., F.B. (1995). Comparison of Dey and Engley (D/E) Neutralising medium to Lethhen medium and Standards Methods Medium for recovery of Staphylococcus aureus from sanitised surfaces. J. Ind. Microbiol. 14:21-25. Curry, A.S., Graf, J.G. and McEwen Jr., G.N. (ed.) (1993) CFTA Microbiology Guidelines. The Cosmetic, Toiletry and Fragrance Association, Washington, D.C. Roberts, D., Hooper, W. and Greenwood, M., (1995). Methods for the examination of food for micro-organisms of public health significance, 2nd edition, section 5.10, Practical Food Microbiology. Butler & Tanner. ISBN 0 901144 36 3. Engley Jr., F.B. and Dey, B.P. (1970). A universal neutralising medium for antimicrobial chemicals. Presented at the Chemical Specialities Manufacturing Association (CSMA) Proceedings, 56th Mid Year Meeting. Dey, B.P. and Engley Jr., F.B. (1995). Comparison of Dey and Engley (D/E) Neutralising medium to Lethhen medium and Standards Methods Medium for recovery of Staphylococcus aureus from sanitised surfaces. J. Ind. Microbiol. 14:21-25. Curry, A.S., Graf, J.G. and McEwen Jr., G.N. (ed.) (1993) CFTA Microbiology Guidelines. The Cosmetic, Toiletry and Fragrance Association, Washington, D.C..

D/E Neutralising Broth Base
(Dey & Engley)
NEW

EC Medium
(Escherichia coli Medium)
NEW

LAB 171

LAB 186

Description
EC Medium (Escherichia coli Medium) is a selective enrichment broth designed for the isolation of coliforms, including E. coli, from water and food samples. It is the recommended medium of the American Public Health Association (APHA) and the AOAC. EC Medium is made selective for coliforms by the inclusion of Bile Salts No.3 in the dehydrated medium. The selective nature of this medium ensures that the growth of non-coliform bacteria is minimised. The medium is buffered by the addition of potassium phosphates and osmotically balanced by sodium chloride. The medium is used at 37°C for coliform organisms and 45.5°C is recommended for the isolation E. coli. Formula Tryptone Lactose K2HPO4 KH2PO4 Sodium chloride Bile Salts No. 3 g/litre 20.0 5.0 4.0 1.5 5.0 1.5

Description
D/E Neutralising Broth Base is a nutritious medium deficient of all neutralising agents. Therefore when a test disinfectant is added to the broth, the potency is undiminished. Developed for use with Dey and Engley’s Neutralising Broth (LAB 187), incorporating D/E Neutralising Broth Base into the test procedure allows the user to differentiate between bacteriostatic and bactericidal activity, and to detect viable organisms that remain after treatment. Its use is recommended in disinfectant evaluation, environmental sampling and water-miscible cosmetics in accordance with Cosmetic, Toiletry and Fragrance Association (CTFA) guidelines. Formula Glucose Tryptone Yeast extract Bromocresol purple g/litre 10.0 5.0 2.5 0.02

Method for reconstitution
Weigh 17.5 grams of powder and disperse in 1 litre of deionised water. Allow the mixture to soak for 10 minutes, swirl to dissolve, and dispense into final containers. Sterilise the medium by

1.1.3

Method for reconstitution
Weigh 37.0 grams of powder and disperse in 1 litre of deionised water. Allow the mixture to soak for 10 minutes, swirl to mix. Dispense into tubes of appropriate volume and, where applicable, add Durham tubes. Sterilise by autoclaving at 121°C for 15 minutes. Appearance: Clear straw broth. pH: 6.9 ± 0.2

Formula Tryptose Soy Peptone Dextrose L-Cystine Sodium chloride Sodium sulphite Agar

g/litre 15.0 5.0 5.5 0.7 4.0 0.2 15.0

Minimum Q.C. organisms: Escherichia coli NCIMB 50034 Enterococcus faecalis NCIMB 50030 (inhibition) Bacillus subtilis NCIMB 13061 (inhibition)
Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. Inoculation: Coliforms: Follow the methods and procedures as stated in Standard Methods for the Examination of Water and Wastewater and Compendium of Methods for the Microbiological Examination of Foods. Incubation: 45.5°C for 18-24 hours aerobically for E. coli and 37°C for 18-24 hours, aerobically for coliforms. Interpretation: Turbidity of broth and gas collection in the Durham tube indicates the presumptive growth of organisms from the coliaerogenes group. All broths should be sub-cultured onto selective media whether turbid or not.

Method for reconstitution
Weigh 45.4 grams of powder and disperse in 1 litre of deionised water. Allow the mixture to soak for 10 minutes, swirl to mix and then sterilise by autoclaving at 121°C for 15 minutes. Cool to 47°C before the addition of supplements or pouring into sterile Petri dishes. Appearance: Light amber clear gel, may contain a slight precipitate. pH: 7.0 ± 0.2

References
American Public health Association, (1980). Standards Methods for the Examination of Water and Wastewater, 15th Edition, American Public Health Association, Inc., Washington, D.C. American Public health Association, (1976). Compendium of Methods for the Microbiological Examination of Foods, American Public Health Association, Inc., Washington, D.C. Association of Official Analytical chemists. (1995). Bacteriological analytical manual, 8th ed. AOAC International, Gaithersburg, MD. Perry and Hajna, (1943). American Journal of Public Health, 33:550. Perry and Hajna, (1944). American Journal of Public Health, 34:735.

Minimum QC organisms: Aspergillus niger NCIMB 50097 Candida albicans NCIMB 50010 Lactobacillus fermentum ATCC® 9388 Streptococcus pyogenes NCIMB 13285
Storage of Prepared Medium: Plates can be stored up to 7 days at 2-8°C in the dark. Inoculation: For the examination of clinical specimens for bacteria and fungi refer to the appropriate published references. For the examination of food for the examination of bacteria and fungi refer to standard methods. Incubation: 35°C ± 2°C for up 72 ± 4 hours for bacteria. 30°C ± 2°C for up 72 ± 4 hours for fungi. Interpretation: Refer to appropriate references and procedures.

References

Eugon Agar
(Eugonic Agar)
NEW

Vera, H.D. (1947). The ability of peptones to support surface growth of lactobacilli. J. Bacteriol. 54:14. MacFaddin, J.D. (1985). Media for the isolation-cultivationidentification-maintenance of medical bacteria. 301-303. vol. 1. Williams & Wilkens, MD. Niven (1949). J. Bacteriol. 58:633. Harrison, A.P.Jr. and Hansen, P.A. (1950). The bacterial flora of the cecal feces of healthy turkeys. J. Bacteriol. 59. 197. Frank, H.A. (1955). The influence of various media on spore count determinations of a putrefactive anaerobe. J. Bacteriol. 70:269. Vanderzant, C. and Splittstoesser, D.F. (ed.). (1992). Compendium of methods for the microbiological examination of food, 3rd ed. American Public Health Association, Washington, D.C. Isenberg, H.D. (ed.) (1992). Clinical microbiological procedures handbook, American Society for Microbiology, Washington, D.C. Murray, P.R. et al (ed) (1995). Manual of Clinical Microbiology, 6th ed. American Society for Microbiology, Washington, D.C. Association of Official Analytical Chemists. (1995). Bacteriological analytical manual, 8th ed. AOAC International, Gaithersburg, MD.

LAB 525

Description Eugon Agar is used for the cultivation of a wide variety of microorganisms, particularly in mass cultivation procedures. The medium is prepared according to the formulation of Vera and was developed to obtain eugonic (luxuriant) growth of fastidious microorganisms. The medium can be used with additions to enhance its performance with certain microorganisms, e.g. Eugon Agar supplemented with 5% sterile defibrinated blood will enable the growth of pathogenic fungi such as Nocardia, Histoplasma and Blastomyces. Niven reported Eugon Agar for the detection of lactic acid bacteria in cured meats and recommended it for investigating spoilage in meats. Harrison and Hansen employed the medium for plate counts of the intestinal flora of turkeys and Frank showed its use for the germination of anaerobic spores pasteurised at 104°C. Eugon Agar is also specified in the APHA Compendium of Methods for the Microbiological Examination of Food. The high sugar content of this medium dictates that it not suitable as a base for haemolytic reactions.

1.1.4

Eugon Broth
(Eugonic Broth)
NEW

Lauryl Tryptose Broth
(Lauryl Sulphate Broth, LTB, LSB)

LAB 526
Description

NEW

LAB 196

Description This is the broth version of Eugon Agar (LAB525) for the cultivation of a wide variety of microorganisms, particularly in mass cultivation procedures. The medium is prepared according to the formulation of Vera and was developed to obtain eugonic (luxuriant) growth of fastidious microorganisms. The medium can be used with additions to enhance its performance with certain microorganisms e.g. Eugon Broth supplemented with 5% sterile defibrinated blood the medium will support the growth of pathogenic fungi such as Nocardia, Histoplasma and Blastomyces. Formula Tryptose Soy Peptone Dextrose L-Cystine Sodium chloride Sodium sulphite g/litre 15.0 5.0 5.5 0.7 4.0 0.2

Lauryl Tryptose Broth is a selective medium for the detection of coliforms in water, dairy products and other foods. The American Public Health Authority (APHA) recommend Lauryl Tryptose Broth for the Most Probable Number Presumptive Test of coliforms in waters, effluent or sewage and as a confirmation test of lactose fermentation with gas production from milk samples and for the detection of coliforms in foods. Lauryl Tryptose Broth is prepared according to the formulation of Mallmann and Darby. Mallmann and Darby showed that tryptose at a concentration of 2% increased the early logarithmic growth phase when compared to meat peptone. These researchers added phosphate buffers and sodium chloride, which improved gas production by "slow lactose fermenting" organisms. Sodium lauryl sulfate was incorporated as a selective agent for the inhibition of noncoliform organisms. This medium can also be used with the addition of MUG (4methylumbelliferyl-β-D-glucuronide) according to the ISO Standard 11866-2 to give enhanced detection of Escherichia coli. Formula Tryptose Lactose Sodium chloride Dipotassium hydrogen phosphate Potassium dihydrogen phosphate Sodium lauryl sulphate g/litre 20.0 5.0 5.0 2.75 2.75 0.1

Method for reconstitution
Weigh 30.4 grams of powder and disperse in 1 litre of deionised water. Allow the mixture to soak for 10 minutes, swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. Cool before the addition of enrichments and aseptically dispense into appropriate containers. Appearance: Light amber solution, may contain a slight precipitate. pH: 7.0 ± 0.2

Minimum QC organisms: Aspergillus niger NCIMB 50097 Candida albicans NCIMB 50010 Lactobacillus fermentum ATCC® 9388 Streptococcus pyogenes NCIMB 13285
Storage of Prepared Medium: Store the prepared medium at 2-8°C. Inoculation: For the examination of clinical specimens for bacteria and fungi refer to the appropriate published references. Incubation: 35°C ± 2°C for up 72 ± 4 hours for bacteria. 30°C ± 2°C for up 72 ± 4 hours for fungi. Interpretation: Refer to appropriate references and procedures.

Method for reconstitution
Weigh 35.6 grams of powder and disperse in 1 litre of deionised water. Allow the mixture to soak for 10 minutes, swirl to mix and dispense into tubes or bottles containing inverted Durham tubes. Sterilise by autoclaving at 121°C for 15 minutes. Appearance: straw, clear liquid. pH: 6.8 ± 0.2

Minimum Q.C. organisms: Escherichia coli NCIMB 50034 Enterobacter aerogenes NCIMB 50029 Staphylococcus aureus NCIMB 50080 (inhibition)
Storage of Prepared Medium: Store the prepared medium at room temperature (18-22°C), in the dark. Inoculation: Inoculate the medium in accordance with standard methods or laboratory policy. Incubation: 35°C ± 2°C for 24 and 48 hours. Interpretation: After incubation at 35°C for 24 hours examine for turbidity and gas production. If no gas has formed incubate for a further 24 hours and re-examine. Turbidity in the medium accompanied by the formation of gas within 48 hours is a presumptive result for the presence of coliforms. The results should be confirmed by standard testing methods.

References
Vera, H.D. (1947). The ability of peptones to support surface growth of lactobacilli. J. Bacteriol. 54:14. MacFaddin, J.D. (1985). Media for the isolation-cultivationidentification-maintenance of medical bacteria. 301-303. vol. 1. Williams & Wilkens, MD. Isenberg, H.D. (ed.) (1992). Clinical microbiological procedures handbook, American Society for Microbiology, Washington, D.C. Murray, P.R. et al (ed) (1995). Manual of Clinical Microbiology, 6th ed. American Society for Microbiology, Washington, D.C.

1.1.5

Mallmann. Methods for the examination of food for micro-organisms of public health significance. clear liquid. The American Society for Testing Materials (ASTM) specifies the use of Letheen Broth in the Standard Test Method for Preservatives in Water Containing Cosmetics.6 . Hlth.2 Minimum Q. APHA Inc. Formula Dextrose Tryptone Beef extract Lecithin Tween® 80 Agar g/litre 1. or the cfu of the area swabbed (typically 25cm2). Association of Analytical Chemists. Interpretation: Count all colonies and calculate the number of cfu per ml of sample allowing for dilution factors. M. Pub. Standard Test Method for Preservatives in WaterContaining Cosmetics. Appearance: Straw. Standard Test Method for Preservatives in Water-Containing Cosmetics. Soap and Sanit. (1995). pH: 7. and is used with Letheen Broth to determine the suitability of preservatives for use in cosmetic formulations.0 5. section 5. section 6.0 5. Philadelphia. swirl to dissolve and dispense into final containers. Official methods of analysis. Incubation: 37°C aerobically for 24-48 hours. subculture to Letheen Agar using a pour plate technique. APHA Inc. L. formalin and (with lecithin) ethanol. Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. Allow the mixture to soak for 10 minutes. Formulation is very hygroscopic.0 1.C. ISBN 0 901144 36 3.0 ± 0. D. (1995). Inoculation: Preservative testing . Formula Peptone Beef extract Sodium chloride Lecithin Tween® 80 g/litre 10. when used with a hygiene swabbing protocol and will ensure against disinfectant carry-over from the swabbing diluent/medium.10. The addition of Tween® 80 means Letheen Agar also neutralises phenols. A modification of FDA Broth. Formulation is very hygroscopic. 127-134.From the dilutions of product in Letheen Broth. (1948). Practical Food Microbiology. J.0 ± 0. Hooper.0 7. Roberts.0 Letheen Broth (AOAC) NEW LAB 184 Description Letheen Broth is primarily used for the assessing the bactericidal activity of quaternary ammonium compounds..R..2 Method for Reconstitution Weigh 25. Washington DC.1. 1.0 Method for reconstitution Weigh 32. Washington. Chem. Letheen Agar is a modification of Tryptone Glucose Extract (TGE) Agar.. C. Appearance: Straw. Washington DC. APHA Inc. E640-78. 24: 134-139.C. Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. D. American Public Health Association (1976) Standard Methods for the Examination of Foods. It is also used in hygiene swabbing protocols where it is necessary to neutralise quaternary ammonium compounds. opalescent gel. Washington DC.References American Public Health Association (1980) Standard Methods for the Examination of Water and Wastewater. keep container tightly closed after use. Inoculation: There are a variety of methods which use Letheen Broth and the appropriate references should be consulted. organisms: Escherichia coli NCIMB 9517 Staphylococcus aureus NCIMB 9518 Storage of Powder: Store at 2-8°C in the dark. Relative efficiencies of quaternary inhibitors. Letheen Agar also allows calculation of colony forming units to be assessed. and for determining the phenol co-efficient of cationic surfactants as recommended by the Official Methods of Analysis of the Association of Official Analytical Chemists (AOAC). American Public Health Association (1978) Standard Methods for the Examination of Dairy Products.7 5. Allow the mixture to soak for 10 minutes. plate or surface inoculation to allow calculation of the colony forming units (cfu) for the area swabbed.0 5. 16th edition. Letheen Broth is easily prepared and has a clear appearance aiding in visual inspection for growth.Association of Official Analytical Chemists. 15th Edn. Letheen Agar (AOAC) (Tryptone Glucose Extract Agar with Lecithin and Polysorbate 80) NEW LAB 185 Description Letheen Agar is used for evaluating the bactericidal activity of quaternary ammonium compounds. keep container tightly closed after use.0 grams of powder and disperse in 1 litre of deionised water.0 15. pH: 7.0 0. American Society for Testing Materials. and Greenwood. 14th Edn. hexachlorophene.L.W. W. (1941) Am.7 grams of powder and disperse in 1 litre of deionised water.C. Hygiene swabbing – Subculture from the swab diluent using a pour Minimum Q. PA. ISO Standard 11866-2 Milk and Milk Products –Enumeration of presumptive Escherichia coli – part 2: Most probable number technique using 4-methyl umbelliferyl-β-D-glucuronide. hexachlorophene. Butler & Tanner. (1998). For example: Phenol co-efficient testing – Subculture from disinfectant dilutions into 10ml volumes of Letheen Broth. swirl to mix.A.0 3. W. the importance of which was first described by Weber and Black in 1948. Letheen Broth contains lecithin to neutralise quaternary ammonium compounds and Tween® 80 to neutralise phenols. and Darby. organisms: Escherichia coli NCIMB 9517 Staphylococcus aureus NCIMB 9518 Storage of Dehydrated Medium: Store at 2-8°C in the dark. and Black. G. References Weber. Sterilise by autoclaving at 121°C for 15 minutes. 31. 2nd edition. and then sterilise by autoclaving at 121°C for 15 minutes. Cool to 47°C and pour into sterile Petri dishes and allow the medium to set. formalin and ethanol (in the presence of lecithin). or surface inoculation. as specified by the American Society for Testing and Materials (ASTM). and is formulated to neutralise quaternary ammonium compounds used in testing of germicidal activity. Annual Book of ASTM Standards. 15th Edn.

Hooper. Association of Analytical Chemists.1. wide zone of β haemolysis Cream. C. Mix well. Incubation: Aerobically at 37°C for 24-48 hours. 1. Washington. monocytogenes in ready to eat foods by up to 22%. (1995). Interpretation: Examine all tubes for turbidity or as stipulated in the method. monocytogenes is the only important human pathogen among the species of Listeria currently recognised. seeligeri and L. very β haemolysis No growth No growth weak References Johansson. 7.Hygiene swabbing – Swab measured area or specific equipment and place in 10ml volume of Letheen Broth. Methods for the examination of food for micro-organisms of public health significance. LBMA is a cost effective method by which to specifically isolate and enumerate L. Cool to 47°C. E640-78.V. seeligeri Enterococcus faecalis Escherichia coli Shape and Surface Colour & Haemolysis C. Standard Test Method for Preservatives in Water-Containing Cosmetics. monocytogenes and is an animal rather than human pathogen. However. 2 vials of XO72 supplement and 2 vials of XO72N supplement.0 10. Appearance: Opaque and blood red. C. Listeria innocua NCTC 11288.V. Philadelphia.V. Escherichia coli NCIMB 50034. sore throat. Annual Book of ASTM Standards. innocua L. 215-222. (1996).0 3. ivanovii. seeligeri is rarely isolated from foods and produces very weak haemolysis.2 Listeria Monocytogenes Blood Agar (LMBA) NEW LAB 172 Description Listeria monocytogenes Blood Agar (LMBA) has been developed for the specific detection and enumeration of L. due to immune suppression. 0. ISBN 0 901144 36 3. Studies have shown that the most commonly isolated species of listeria. Inoculation: Surface plating.V. The use of citrated sheep blood prevents this and allows differentiation of L. J. Prevalence of Listeria monocytogenes in foods: incidence in dairy products. monocytogenes from other haemolytic Listeria species e. no haemolysis Cream. J. and Fisher. narrow zone of β haemolysis Cream.1ml of neat or 10-1 dilution of the food sample is spread over the entire surface of the plate.C. in meat and poultry products. streaking out to single colonies. and aseptically add citrated sheep blood to 5%. Formula Tryptone Soy peptone Sodium chloride Lithium chloride Magnesium sulphate (3/4H2O) Agar g/litre 15. The use of sheep blood is standard methodology for Listeria testing. Allow the mixture to soak for 10 minutes.g. International Journal of Food Microbiology. dairy. 40.E. L. section 5. D. Use multiple plates if the volume required is greater than 0. swirl to mix then sterilise at 121°C for 15 minutes. from the enrichment broth. Method for Reconstitution Weigh 53. ivanovii produces wide zones of haemolysis compared to the narrow zone of L. The symptoms of infection with this organism include fever. innocua on LMBA using colonial appearance and haemolysis. It is distinguished from L. Cream. Kozak. other that L.M. and Greenwood.. septicaemia and meningitis may develop. R. monocytogenes in food samples. Area to be swabbed and volume of medium may vary depending upon swabbing protocol used. 2nd edition. due to its distinctive haemolytic pattern. monocytogenes Minimum Q.0 5. 16th edition. Official methods of analysis.0 5. monocytogenes L. ivanovii L. Interpretation: Organism L.E. L. Incubation: 37°C aerobically for 24-48 hours. In severe cases pneumonia. M. L. monocytogenes from food and processing environments is L. Enhanced detection and enumeration of Listeria monocytogenes from foodstuffs and food processing environments.E. Food Control. Storage of Prepared Medium: Plates can be stored up to 7 days at 2-8°C in the dark. ingredients in selective agars can result in partial lyses or darkening of the blood supplement. whilst L. or meningitis of the new born.7 .10. and pour into sterile Petri dishes and allow to set.C.. Balmer. still birth. For enumeration. K. (1995).G. T (1998). PA. (1996).g. Jay.0 References American Society for Testing Materials. D. Practical Food Microbiology. The addition of donated sheep blood (defibrinated with sodium citrate) to LMBA allows differentiation between haemolytic and nonhaemolytic stains of Listeria. 77-85. This medium has been shown to improve the isolation rate of L. monocytogenes can be found in all main categories of products e.1 ml. Butler & Tanner. Pregnant women are particularly susceptible to listeriosis. 209-214. monocytogenes can cross the placenta causing abortion.8 15. LMBA contains lithium chloride in concentrations that inhibit the growth of enterococci yet allow good haemolysis by L. Association of Official Analytical Chemists. Prevalence of Listeria spp.. Enterococcus faecalis NCIMB 50030.8 grams of powder and disperse into 1 litre of deionised water. C. Food Control.G.E. diarrhoea and abdominal pains. W. L. generalised aches and pains. Roberts. (1998).G. Byrne. innocua. meat and poultry.G. 7. L. pH: 7.2 ± 0. LBMA is supplemented by polymyxin plus ceftazidine (X072) and nalidixic acid (X072N) to suppress competing flora such as members of the bacillus group and staphylococci. organisms: Listeria monocytogenes NCIMB 50007. section 6. monocytogenes. T.

swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. swirl to mix and bring to the boil.5 0. Interpretation: Count all colonies and calculate the number of colony forming units.A.02 12.0 10. Dispense into tubes and sterilise by autoclaving at 121°C for 15 minutes. by means of lactose fermentation.3 ± 0.04 0. 3cm in depth. Allow the mixture to soak for 10 minutes. 1 Yeast Extract Glucose L-Lysine Ferric Ammonium Citrate Sodium thiosulphate Bromocresol Purple Agar No.0 1. Lecithin is included to neutralise quaternary ammonium compounds and Tween® 80 is incorporated to neutralise phenols. Allow the mixture to soak for 10 minutes. Enteric organisms that do not decarboxylate lysine yield an alkaline slant over an acid butt (yellow). P.0 5. P. keep container tightly closed after use. Minn. Thus no distinction between Shigella and E. however the use of Bismuth Sulphite Agar with subculture into Lysine Iron Agar allows determination of their presence. reducing "preservative carryover". Appearance: Straw opalescent gel. Formula Balanced Peptone No. formalin and with lecithin. Formulation is very hygroscopic. and Ewing.5 grams of powder and disperse in 1 litre of deionised water. Edwards. phenolic compounds and formalin. equipment and work areas treated with disinfectants or other sanitisers. Appearance: Clear purple gel. Incubation: 37°C aerobically for 24-48 hours. Incubation: 37°C aerobically for 18-24 hours. Tween® 80 Lecithin Agar No. 1. ethanol.C. The addition of Lecithin and Tween® 80 in the formula inactivates some preservatives that may inhibit bacterial growth. Cool to 47°C and pour into sterile Petri dishes and allow the medium to set. with frequent stirring to dissolve completely.Lysine Iron Agar NEW References LAB 54 Edwards.R.7 ± 0. Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. M. coli is possible and Triple Sugar Iron Agar (LAB53) is recommended in parallel.7 grams of powder and disperse in 1 litre of deionised water. (cfu) per ml of sample allowing for dilution factors. together with blackening due to sulphide production. The formulation is recommended for Aerobic Plate Count (Microbial Limit Test) for water miscible cosmetic products containing preservatives. 9:478-480.1. Appl.7 15. W.0 Method for reconstitution Weigh 31. The medium is a modification of Tryptone Soy Agar with added neutralising compounds lecithin and Tween® 80. Storage of Powdered Medium: Store at 2-8°C in the dark. Proteus and Providencia cultures characteristically produce a distinctive red slant over an acid butt since these organisms deaminate lysine but without sulphide production. Inoculation: Subcultures for further identification are picked from the centre of isolated colonies on selective media and streaked across the slant and stabbed into the butt of tubes of Lysine Iron Agar. Inoculation: Consult the appropriate references as this product is used in several procedures. Salmonella strains (including Salmonella arizona) which ferment lactose and produce black colonies on Bismuth Sulphite Agar (LAB13) can be recognised by the alkaline reaction (purple colour) produced throughout the medium. Staphylococcus aureus ATCC 6538P.2 Growth Characteristics Organism Salmonella arizona Salmonella Salmonella paratyphi Enterobacter aerogenes) Klebsiella Hafnia Serratia Citrobacter Escherichia coli Shigella Proteus Providencia ) Alkaline Alkaline ) ) Acid Acid Acid Acid Alkaline Alkaline ‘red’ ‘red’ + Acid (NC) Alkaline Butt Slant Sulphide Production + + Alkaline Alkaline Alkaline Alkaline Acid Acid Minimum Q. Salmonella arizona strains which produce pink to red colonies on bile salt media are often overlooked in outbreaks of food poisoning. Burgess Publishing Co. 2 g/litre 5. pH: 7.8 . and Fife.0 5. (1964). Lysine iron agar in the detection of Arizona cultures.R. pH: 6.0 0.2 Storage of Prepared Medium: Tightly capped containers – up to 3 months at 15-20°C in the dark. It is recommended for determining the hygiene status of containers.H.0 5. (1961). Microbiol. Description This is a differential medium for the detection of salmonellae and other enteric pathogens.2 Method for Reconstitution Weigh 45. hexachlorophene. Identification of Enterobacteriaceae.0 3.0 Microbial Content Test Agar (MCA) (Tryptone Soy Agar with Lecithin and Tween® 80 (TSALT)) (Casein Soy Peptone Agar with Lecithin and Polysorbate 80) NEW LAB 189 Description The use of Microbial Content Test Agar (MCA) is recommended for the detection of microorganisms on surfaces sanitised with quaternary ammonium compounds.0 0. organisms: Escherichia coli ATCC 11229. lysine decarboxylase activity and hydrogen sulphide production. Formula Tryptone Soy Peptone Sodium Chloride g/litre 15. Cool in a slanted position such that slopes are formed over deep butts approx.

D. However. Cool to 47°C. NY.0 2. and is well recognised as a pathogen amongst immuno-compromised patients. ORSIM (Oxacillin Resistant Staphylococci Isolation Medium) NEW LAB 192 PEMBA (Bacillus Cereus Medium) NEW Description The over-prescription of therapeutic antibiotics in recent years is thought to be a contributing factor in the rising numbers of multiresistant bacteria being encountered. Storage of Prepared Medium: up to 7 days at 2-8ºC in the dark.2 ± 0. Marcel Dekker. Mix well. Brummer. and pour into sterile Petri dishes. so all negative plates should be re-incubated for a further 24 hours*.5 0. Once confirmed.References Orth. Cool to 47ºC and add two vials of X193 and 50ml of Egg Yolk Emulsion (X073) mix well and pour the plates.9 . The selectivity is provided by the polymyxin B supplement (X193) and provides excellent results for the majority of sample types. MRSA (multi resistant Staphylococcus aureus). App.2 Method for Reconstitution Weigh 103.1. cereus. Brummer. 1. This combination produces intense blue colonies as presumptive MRSA..0 15. D. Inoculation: Take a swab sample from a suspected infection and apply the swab end directly to the surface of a supplemented plate of ORSIM and streak out for single colonies. In this situation. Minimum Q. Inoculation: Surface spreading or streaking for single colonies. Microscopic examination of presumptive B.5 grams of powder and disperse into 1 litre of deionised water. Microbiol. Influence of possible disinfectant transfer on Staphylococcus aureus plate counts after contact sampling. and the introduction of Lithium chloride at 5g/L.0 55.0 0.C. Environ. To complete the medium. organisms: Staphylococcus aureus (MRSA Strain) NCTC 11940 Staphylococcus aureus (MSSA Strain) NCIMB 50080 (inhibition). This chemical mixture still provides the required inhibition towards competing organisms. swirl to mix then sterilise at 121°C for 15 minutes. pH: 7. To complement this.C. This media is an improved version of the highly regarded Mannitol Salt Agar (LAB007) and incorporates an enhanced indicator system using aniline blue and mannitol fermentation. Inc. early detection is vital to ensure the individual’s survival. organisms: Bacillus cereus NCIMB 50014. (1993) Handbook of Cosmetic Microbiology. This formulation specifically enhances egg yolk precipitation and sporulation of Bacillus cereus. Escherichia coli NCIMB 50034 (inhibition). cereus colony can confirm identity by presence of lipid globules in vegetative cells.0 10. The bromothymol blue pH indicator gives clear visualisation of alkaline mannitol nonfermenting colonies and egg yolk precipitation indicative of B. 32:80-84. which are unmistakable amongst mixed cultures and easily visualised against the media background.S..8 9.2 Minimum Q. Dry the agar surface before inoculation. oxacillin to inhibit multi sensitive Staphylococcus aureus (the cause of false positives).5 LAB 193 Description This medium is based on the highly specific and sensitive PEMBA medium. and aseptically add 2 vials of X192 supplement. (1993) Handbook of Cosmetic Microbiology. and polymyxin B to suppress other halophillic bacteria such as Proteus spp. the selective supplement X192 is included. Microbiol. examine the plate for intense blue colonies and confirm using either coagulase/latex agglutination and Penicillin binding protein 2’ test (PBP2’). Inc. B. even at low numbers. (1976). pH: 7. Environ. allow the mixture to soak for 10 minutes.S. whilst ensuring optimal recovery of MRSA. NY.2 12. One such bacterium. Allow the mixture to soak for 10 minutes. (1976). New York. all positive plates should be discarded safely. is particularly prevalent within the hospital environment. Storage of Prepared Medium: Plates can be stored up to 7 days at 2-8°C in the dark. Incubation: Aerobically at 37°C for 24 and 48 hours.2 ± 0. derived from a reduction in the salt level to 55g/L.0 0. 32:80-84.0 5. B. This contains two antibiotics. *Typical strains of MRSA will be detected within 24 hours on this medium. Appearance: Straw/grey gel.0 Method for Reconstitution Weigh 41g of powder and disperse in 950ml of deionised water. Appearance: Yellow and opaque. Incubation: 30ºC aerobically for 24-48 hours. Marcel Dekker. Formula Peptone Mannitol Sodium chloride Magnesium sulphate Disodium hydrogen phosphate Bromothymol blue Sodium pyruvate Agar g/litre 1. ORSIM possesses a refined selectivity.0 10. Influence of possible disinfectant transfer on Staphylococcus aureus plate counts after contact sampling.12 10.1 2. It is used for the isolation and enumeration of Bacillus cereus. References Orth. Formula Peptone Yeast Extract Mannitol Sodium chloride Lithium chloride Aniline blue Agar g/litre 11. some strains may require longer incubation. Interpretation: After incubation for 24 hours. swirl to mix and sterilize by autoclaving for 15 minutes at 121ºC for 15 minutes. New York. App.

Susceptibility Testing ‘Iso’ Agar is produced having a stable mineral content. CV. 21(1) 100-103. 22. aueus Storage of Prepared Medium: Plates can be stored up to 7 days at 2-8°C in the dark. and Harmon.W.O. 78-82. 3rd ed 623-635.M.0 2. References Holbrook. cereus B. Rhodehamel. perfringens. The AST is performed to detailed standards. Arlington. Andrews.C. 521-526. Interpretation: Count all black colonies with or without a halo as presumptive C..J. Appl. Appl. colour Blue white halo Yellow Yellow Yellow Blue (swarms) Yellow References CV. Metal ions can exert known antagonistic effects on a number of antibiotics. perfringens produces black colonies and the egg yolk emulsion incorporated into the media detects the lecithinase activity of this bacteria.D F. A. Allow the mixture to soak for 10 minutes.CR.Rz. 27. MD. and Hilsheimar R. licheniformis Proteus spp. Appl. (1971). and Furguson. Appl.0 5. A. E. mix well and pour into sterile Petri dishes. 26(7) 753-759.2 Minimum Q.0 1. et al (1977). Microbiol. The antimicrobial susceptibility test is utilised in epidemiological studies and in determining the appropriate usage of antimicrobials in the clinical environment. Method for reconstitution Weigh 46.D F. and Harmon.H. staphylococci. (1971). and the detection of microbial resistance. in which the undefined elements are maintained at minimum levels. Labbe. perfringens. pH: 7.1ml aliquots of an appropriate serial dilution of the homogenised test sample and overlay if required. E. and Peeler.D F.J.D.E. a major factor is the medium on which it is performed. Bacteriological Analytical Manual 8th ed. 15(3) 650-653. S. (1958). faecalis E.CR.0 shape & surface F.CR. Further confirmation should be carried out according to standard method protocols e. Microbiol. 688-692. Microbiol.10 . coli S.0 1. Inoculation: For a spread plate inoculate the agar plate with 0.g. Harmon.A. E. & Anderson. have a detrimental effect on results obtained.6 ± 0. Donovan. S.A. Compendium of methods for the microbiological examination of foods..C. However not all strains produce lecithinase and therefore black lecithinase positive and black lecithinase negative colonies should be considered as presumptive Cl. A. J. certain organisms such as some streptococci. H. (1995) Official methods of analysis AOAC International 16th ed. the presence of minimum antagonistic elements.GROWTH CHARACTERISTICS Organism B.G. The presence of antagonists in the medium e. G. & Jongerius. The addition of thymidine for the growth of dependant strains antagonises the antimicrobial action of Trimethoprim and sulphonamides and results in false resistance results.G. Gaithersberg. Microbiol. AOAC International. M. swirl to mix and sterilise by autoclaving at 121°C for 10 minutes. VA.D F.W. Incubation: 35°C ± 2°C anaerobically for 18-24 hours. thymidine.0-4. lactose fermentation. Enterobacteriaceae and Neisseria may require the addition of intrinsic growth factors e. W. (1992).M.A. This medium will support the growth of the majority of pathogens requiring susceptibility testing.0 Susceptibility Test ‘ISO’ Agar NEW LAB 170 Description Susceptibility Testing ‘Iso’ Agar is a semi-defined medium for antimicrobial susceptibility (sensitivity) testing (AST). (1973). Microbiol.CR.0 0.06 AOAC International.0 14. Appearance: Straw.A. Microbiol. coagulans B. organisms: Clostridium perfringens NCIMB 50027 Escherichia coli NCIMB 50034 (inhibition) 1. W. nitrate reduction. gelatin liquefaction and absence of motility.5 no growth 1. American Public Health Association. Bacteriol. Can. The egg yolk emulsion is omitted for preparation of Egg Yolk Free TSC Agar and Egg Yolk Free TSC Agar should be used for an overlay medium. Sodium metabisulphite and ferric ammonium citrate are used as an indicator of sulphite reduction by Clostridium perfringens. 16.0 2. Hauschild. R.T.. D. K. colony size (mm) 3. 500-506. clear gel or pale yellow opaque gel. Mossel. Washington. Hauschild A. without the addition of supplements.0 5. However. Kauttar.g. thymidine and metal ions. However these supplements can introduce errors as they can affect the activity of certain antibiotics and consequently their affects must assessed before use.0 5. Hauschild.H. Bacteriol. D.E. J. S. (1967). Appl.R. lysed horse blood. D. (1995). Perfringens Agar Base (TSC) (Tryptose Sulphite Cycloserine (TSC) Agar) NEW LAB 194 Description Perfringens Agar Base is a nutrient medium to which egg yolk emulsion (X073) and cycloserine (X194) are added for the preparation of Tryptose Sulphite Cycloserine (TSC) Agar.M. the results of which must be reproducible. S. 884-892..M. Appl. J. and a constant isotonic pH (preventing the blocking or enhancement of antimicrobials). allows for precise and rapid treatment. For full details refer to appropriate references and standard method protocols.0 2. The reduction of sulphite by Cl. R. 27. The response of clinical isolates to antimicrobials.0 grams of powder and disperse in 1 litre of deionised water.1. Allow the medium to cool to 47°C and supplement with 2 vials of X194 (cycloserine) and 50ml of egg yolk emulsion (X073). Koopman. J. R. thiamine and menadione.01-16. (1973). Formula Tryptose Soy Peptone Beef extract Yeast extract Sodium metabisulphite Ferric ammonium citrate Agar g/litre 15. J.0 1.0-3. J. 21.g. subtilis B. Yellow white halo Shahidi. and Hilsheimar. thereby ensuring production of optimum zones of microbial inhibition. (1980). Can. For a pour plate mix 1ml aliquots of an appropriate serial dilution of the homogenised test sample with approximately 20 ml of TSC plus egg yolk emulsion. 23. Therefore the anion and cation content of the medium must be regulated to prevent adverse effects on performance. 1-119.

0 7.03 0. Antimicrobial Chermother. Marshall (ed..M. D.. Compendium of methods for the microbiological examination of foods. K. 2 MUG.. Appearance: Light purple-violet clear gel.A.1 1. G. pH: 7.P. Hawkey. 2: 135-138. Capped containers up to 1 month at 15-20°C in the dark.D. American Public Health Association. Waterworth. Non-lactose fermenters produce pale colonies.M. 22: 534-538.8 0.11 . and polymyxin B. Cool to 47°C.. I. Acta. Storage: Capped containers . Interpretation: Examine plates for growth and fluorescence. Allow the mixture to soak for 10 minutes. Hitchens.B. and Todd. C. P. by bringing to the boil. R.3 0. Effect of divalent cations in bacteriological media on the susceptibility of Xanthomonas maltophila to imipenem with special reference to zinc ions. with frequent mixing. J. coli. Washington.0 5. Kerr.8°C in the dark.1 Method for Reconstitution Weigh 35. 247-269.4 0. Antibiotic Sensitivity Testing.S.D.) Standard methods for the microbiological examination of dairy products.. Cool to 47°C and if required add 5-7% sterile lysed horse blood. (1971).C.. If held molten in a water bath.A. Pour into sterile petri dishes and allow to set.M. Duncan.0 Violet Red Bile Agar with MUG (V.B.0 1.2 Minimum Q. Amato. Vanderzant and D. Hartman. R.. (1969). Formula Yeast extract Balanced peptone No. pH: 7. Washington. Antimicrobial Agents and Chermother. J.. Susceptibility of 1500 isolates of Pseudomonas aeruginosa to gentimicin. A. K. Swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. mix well and dispense into petri dishes. Inoculation: Surface. Calcium and Magnesium in Mueller Hinton Agar and their influence on disc diffusion susceptibility results. which hydrolyses MUG to yield the fluorescent compound methylumbelliferone. Minimum Q.C.0 0.G.5mm in diameter as coliforms and all fluorescent colonies as presumptive E. Thornberry.2 References Ericsson.F. Birkenhead. swirl to mix and then sterilise the medium. Pathol. and Roth. Incubation: As stipulated in the BSAC methodology. Hyde. Appearance: Straw clear gel. J. H.A. 4°C for 10 days for psychrotrophs.. Microbiol.C. Sherris. W. P. Inoculation: Pour plate method (with or without overlay) or surface spread.0 0.5 10. with MUG) NEW LAB 573 Description Violet Red Bile Agar with MUG (Methylumbelliferyl-β-Dglucuronide) is a medium for the simultaneous enumeration of coliform organisms and Escherichia coli in food and dairy products.. Plates .C.C. American Public Health Association.6 grams of powder. Clin. D. carbenecillin. 31: 181183. (1974). Sect B Suppl. Report of an International Collaborative Study.M.1. 16th ed. 32°C for 24-48 hours for mesotrophs. 1. (1979). (1993). inoculum as described by standard methods. References Christen. Lactose fermenters produce red/purple colonies often surrounded by a halo of bile precipitate.002 12. Coliform and other indicator bacteria. coli in food and dairy products by fluorescence. C. (1992) ColiformsEscherichia coli and its toxins. 4 methylumbelliferyl-ß-D-glucuronide g/litre 3. (1993).R. Count all red/purple colonies > 0.4 ± 0. colistin. Pathol. 3 Lactose Neutral red Crystal violet Agar No.). 217: 1-90. J.22 0..T.R.E. Splittstoesser (ed. P. 3rd ed. detectable by long-wave UV light.0 0.up to 7 days at 2 . D. Newton. Davidson.3 ± 0.9 grams of powder and disperse in 1 litre of deionised water. Calculate the number of coliforms and E. organisms: Escherichia coli (as recommended by the NCTC 12241 British Society for Staphylococcus aureus Antimicrobial NCTC 12981 Chemotherapy (BSAC)) Pseudomonas aeruginosa NCTC 12934 Enterococcus faecalis NCTC 12697 Haemophilus influenzae NCTC 12699 Streptococcus pneumoniae ATCC 49619 Neisseria gonorrhoeae NCTC 12700. Cool to 47°C and distribute into bottles or tubes. Scand.078 12. P. 42°C for 18 hours for thermotrophs.0 2.0 1. L.up to 7 days at 2-8°C. Effect of medium composition on the apparent sensitivity of pseudomonas aeruginosa to gentimicin.C. E. coli in the original sample. L.Formula Peptone Mixture Glucose Starch Sodium chloride Na2HPO4 Sodium glycerophosphate Sodium gluconate Sodium acetate Uridine Defined Chemical Mixture Agar g/litre 16. Current Microbiol. McAllistair.up to 3 months at 15-20°C in the dark. Incubation: 37°C for 18-24 hours for ‘coliforms’. use within 3 hours.L. Standard Methods procedures specify VRBA with MUG for detecting E. disperse in 1 litre of deionised water.. Method for reconstitution Weigh 38. organisms: Escherichia coli NCIMB 50034 Enterobacter aerogenes NCIMB 50029 Staphylococcus aureus NCIMB 50080 Storage of Prepared Medium: Plates . F. in the dark.0 2.A. The selectivity of the medium is due to the presence of bile salts and crystal violet. (Jan) 9-15. Escherichia coli produce red/purple fluorescent colonies due to the fermentation of lactose and production of the enzyme glucuronidase.1 Sodium chloride Bile Salts No. Garrod.

Damare. Hansen. MD. W.Colony count by inoculation in a nutrient agar medium. J. AOAC International. The main value of colony counts lies in the detection of changes in water supply quality from those expected.W.. and Shaffer. Peng.2 Minimum Q. M. Detection of β-D-glucuronidase in lactose fermenting members of the family Enterobacteriaceae and its presence in bacterial urine cultures. A sudden increase in the numbers can be a warning of pollution and can call for immediate remedial action Formula Tryptone Yeast Extract Agar g/litre 6.0 3. Appl. and Bulow. Microbiol. Rippey. Clinical Microbiol. Bacteriological Analytical Manual. Interpretation: Count all colonies and calculate the number of organisms (or 'colony forming units' c.Hitchens.f.D.0 15.12 . Appearance: Pale straw coloured. Membrane filtration differentiation of E. D. (1995). Escherichia coli and the coliform bacteria. Environ...M. Water Plate Count Agar (ISO) NEW LAB 197 Description A nutritious non-selective medium which conforms to ISO 6222:1999(E) Water quality . coli from coliforms in the examination of water.S. B.Enumeration of culturable microorganisms .C. P. Watkins. P. Mates. Brill. Campbell. E. 50:1736-1746. They also give an indication of the cleanliness and integrity of the distribution system. thus avoiding contamination of the product with spoilage organisms.) per ml of sample allowing for dilution factors. based on frequent long term monitoring. Chang.0 Method for reconstitution Weigh 24. and Chandler. Kilian. Cool to 47°C before use. J. Pathol. Colony counts are useful for assessment of ground water integrity and the efficiency of water treatment processes.0 grams of powder and disperse in 1 litre of deionised water. (1984).. and Yourassowsky. L. Allow the mixture to soak for 10 minutes. Microbiol. Appl. A. organisms: Staphylococcus epidermidis NCIMB 50082 Escherichia coli NCIMB 50034 1. Environ. Microbiol. (1985).Colony count by inoculation in a nutrient agar culture medium. M. Incubation: Aerobically at 36°C ± 2°C for 44 ± 4 hours and 22°C ± 2°C for 68 ± 4 hours. Proportion of β-D-glucuronidase negative Escherichia coli in human fecal samples. (1982). clear gel.A. Inoculation: Pour plate technique or surface inoculation.. Gaithersburg. 67:343-346. Simplified direct plating method for enhanced recovery of Escherichia coli in food. swirl to mix then sterilise at 121°C for 15 minutes.W.u.1. 84:245-251. R. R. 55:335-339. S.Enumeration of culturable microorganisms . (1976). Scand.C. Microbiol. Feng. 4. References ISO 6222 (1999) Water quality . Sect. Journal of Food Science.F. J. P. G.29.01-4.2 ± 0. They can be used to assess the suitability of a water supply for the preparation of food and drink. and Hartman.A.R. Storage of Prepared Medium: Plates can be stored up to 7 days at 2-8°C in the dark. Appl. J. A. Acta. Fluorogenic assays for immediate confirmation of Escherichia coli. 20:1177-1179. and Johnston. (1989). W.D. P. (1989). pH: 7. and Lum. 43:1320-1329. 8th ed. Rapid diagnosis of Enterobacteriaceae. The estimation of overall numbers of microorganisms can be used for the assessment and surveillance of water quality.

V.2.0 . Inoculation: Clinical: Streak for single colonies after selective enrichment in Selenite Broth. The medium is based on D.0 .2 ± 0.G C. most media are highly non-specific and consequently place a heavy burden on the laboratory in terms of biochemical and serological confirmation of suspect colonies. a New Chromogenic Agar for Selective Isolation of Salmonella spp. However.CR.. Harlequin™ chromogenic media provide a more specific identification by detection of specific enzymes produced by certain groups of bacteria.D Colourless (Fishy Odour) Colourless (Green) References Perry. aeruginosa. pp .E. producing green colonies that are easily distinguished from the black or colourless colonies of other organisms. The first substrate. ABC Medium.E.up to 7 days at 2 ..G Green Other (Black if β-galactosidase +ve) Shigella spp.2 Minimum QC organisms: Salmonella typhimurium NCIMB 50076 Escherichia coli NCTC 9111 Storage of Prepared Medium: Plates . J.2. 1.V.3 0.0 .0 C. M. aerogenes Proteus spp.V. Chromogenic substrates act as the substrate for specific enzymes and change colour due to the action of the enzyme.0 0.0 C.. coli K. Most Enterobacteriaceae are β-galactosidase positive and these produce black colonies on Salmonella ABC. A.5 5. saving labour and reducing consumable costs. This medium will also detect Salmonella typhi and paratyphi.8°C in the dark.1. Traditional culture media generally rely on the fermentation of sugars or other biochemical reactions for presumptive identification of bacteria. There is also an increase in interest for chromogenic Salmonella media as traditional tests have a very poor specificity resulting in many false positive results. Incubation: 37°C aerobically for 18 .0 12. The second substrate.E. 37: 766-768. disperse in 1 litre of deionised water. Food: Streak for single colonies after selective enrichment. Clin.08 0.24 hours Harlequin™ Salmonella ABC (Freeman Formulation) NEW HAL 1 GROWTH CHARACTERISTICS Organism Salmonella spp. The β-D-glucuronide enzyme is present in approximately 95% of E. which give a blue-green colour on cleavage. pH: 7. is hydrolysed by Salmonella spp. The use of chromogenic media simplifies Salmonella testing and saves much time in unnecessary confirmation tests.G CV. swirl to mix and then sterilise the medium by bringing to the boil.E. Harlequin™ Chromogenic Media The Harlequin™ chromogenic microbiological culture media range has been developed to improve the isolation and identification of a range of microorganisms. Description Salmonella spp. developed for the isolation of Salmonella spp.5 .1.5 . Jones.0 8.5 grams of powder.0 C.. R. J. mix well and dispense into Petri dishes.G C. With improved specificity.V.03 Method for reconstitution Weigh 36. (1999). Allow the mixture to soak for 10 minutes. by culture remains the most reliable method of detection. Gould. 2.D.A Hynes and hence utilises sodium desoxycholate and sodium citrate as inhibitors. F. Chromogenic media are most rapidly gaining acceptance as an indicator for Escherichia coli. Appearance: Translucent straw gel. The advantage of this type of media is that it can eliminate or reduce the need for subculture and the performance of confirmatory biochemical tests to determine the identity of some microorganisms. is enzymatically cleaved by βgalactosidase producing organisms giving black colonies in the presence of iron. X-α-Gal. J.E. This medium.G Colourless (Black if β-galactosidase +ve) Black Black (No Growth) (Mucoid) E.2. utilises a dual chromogen system to visualise these enzyme activities. Freeman.V.2. can be differentiated from other members of the family Enterobacteriaceae by their ability to produce α-galactosidase in the absence of β-galactosidase. from food and clinical samples. the ABC medium dramatically reduces the need for ‘false positive’ screening. Formula Beef Extract Peptone Sodium citrate Sodium desoxycholate Agar X-α-Gal CHE-β-Gal Ferric ammonium citrate IPTG g/litre 5. This type of test is now used widely for water and food microbiology..2. P. Micro.0 5. Cool to 47°C. Some of the CHE derivatives are used in Harlequin™ media along with indolyl derivatives (5-bromo-4-choro-3-indolyl). LAB M have their own patented chromogenic compounds called the novel CHE (cyclohexenoesculetin) substrates which give the bacterial colony a black non-diffusing colouration when hydrolysed by the enzyme involved in the presence of iron salts.5 0. CHE-β-Gal.5 0.5 1.C. coli and is uncommon in the other Enterobacteriaceae.K. Isolation of Salmonella spp. Colony Shape & Colour Size (mm) Surface 1. Taylor..0 0. Ford. DO NOT REMELT OR AUTOCLAVE THIS MEDIUM.1 .

and is detected by the MUG reagent in other formulations. coli. a red halo will appear in the medium around the colony. The 9th Meeting of the Microbiological Methods Innovations Forum.2 Minimum QC organisms: L. A simple indole test can be performed by placing one drop of Kovac’s reagent onto a colony and if positive. Appearance: Yellow. (1984). Letters in Applied Microbiology. and Bottolfsen. Evaluation of a Fluorogenic Assay for Detection of E. Description This is a selective identification medium for the isolation of Listeria spp. Inoculation: Surface plating. Scand. coli without the need for membranes. App & Env. clear gel. CHE-glucoside. Further dilution may be necessary if large numbers of E. or when large numbers are present on the plate. and it is concentrated within the colony.0 1. Acta Path. If negative. It is based upon the formulation of Tryptone Bile Agar.0 5. B 92 261-264. Cool to 47°C and pour in to Petri dishes.5 grams of powder. Interpretation: Count all blue/green colonies as presumptive E.E. Holroyd. using naturally contaminated food samples. K. pH: 7.075 15.2 ± 0. In a comparative study.Microbiol. and ferrous gluconate. to ensure colonies can be easily counted. Bacillus spp.20 (6) 1177-1179. Appearance: Straw. Mellors.3 X-glucuronide Agar g/litre 20. Robinson. then the halo will be white.2 Method for reconstitution Weigh 36.0 Method for reconstitution Weigh 73. Colony Shape & Size (mm) Surface 0. C.5 0. from food and clinical materials based on the Oxford formulation. coli (inhibition) NCIMB 50034 Storage of Prepared Medium: Plates .1.5 .A.up to 7 days at 2-8 °C in the dark.L. J.0 Harlequin™ TBGA (TBX) (Tryptone Bile Glucuronide Agar) NEW HAL 3 Description A medium developed for the simple enumeration of E.0 0. W. A notable exception is E.1. LAB 072. Lithium chloride is incorporated as an inhibitor of enterococci and further selectivity is achieved by adding the antibiotic supplement X122 or X123 after sterilisation of the medium.2 .1. or pre-incubation on Minerals Modified Glutamate Medium.0 5.E. facilitating easier enumeration in the presence of other organisms. add 1 vial of selective supplement X122 or X123.Harlequin™ Listeria Medium NEW References HAL 2 Smith.5 0. Microbiol. GROWTH CHARACTERISTICS Organism L. A new chromogenic medium for the isolation of Listeria spp. streaking out to single colonies. coli are present. Dry the surface prior to inoculation.G 2. A. Sect. coli. slightly opaque gel.0 ± 0. the Listeria Novel Substrate Medium gave a 34% increase in positive isolations over the traditional Listeria Isolation Medium (Oxford Formulation). B.5 ml of a 1:10 dilution of the sample and spread over the entire surface of the plate. (2001). W.0 CV. Incubation: 30°C aerobically for a minimum of 22 hours (samples should not be considered negative until plates have been incubated for a minimum of 44 hours). Detection of ß-glucuronidase in Lactose Fermenting Members of the Family Enterobacteriaceae and its Presence in Bacterial Urine Cultures. coli strains positive. monocytogenes Minimum QC organisms: Escherichia coli NCIMB 50034 (blue/green) Enterobacter aerogenes NCIMB 50029 (cream) Inoculation: Inoculate 0.0 1. calculate the cfu/g in the original material. Formula Tryptone Bile Salts No. Hansen. coli*.5 grams of powder.. Formula Peptone mix Meat extracts Starch Sodium chloride Lithium chloride Ferrous gluconate Sodium pyruvate CHE-glucoside Agar g/litre 30. disperse in 1 litre of deionised water. *96-97% of E.CR.J. (1984). swirl to mix and then sterilise by autoclaving at 121°C for 15 minutes.. Poster presentation.0 CV.0 .p .0 15. coli 0157:H7. 1. Immunol. mix well and pour into Petri dishes.. the medium has been modified by the addition of a chromogenic substrate to detect the ß-glucuronidase enzyme.CR. Swirl to mix and sterilise at 121°C for 15 minutes. P.7 1.D Grey centre with buff periphery Buff Pale Grey Usually no growth Gram +ve cocci enterococci p. D. disperse in 1 litre of deionised water and allow the mixture to soak for 10 minutes. The aesculin hydrolysis reaction is replaced by a combination of the novel chromogen. Incubation: 30°C for 4 hours. The retention of the pigment against the clear agar makes screening for suspect colonies very easy. Evaluation of Rosco Diagnostic ß-glucuronidase Tablets in the Identification of Urinary Isolates of Escherichia coli. and Yourassowsky.L. Allow the medium to cool to 47°C. Allow the mixture to soak for 10 minutes. followed by 18 hours at 44°C. The advantage of the chromogenic substrate is that it requires no UV lamp to visualise the reaction. Micro.D colonies References Dibb.G Colour Black Other Draughtsman or CV. which is highly specific for E.5 CV. This reaction results in the formation of an intense black pigment that is retained within the Listeria colonies. Clin. Campden and Chorleywood Food Research Association. pH: 7.25 15. monocytogenes NCIMB 50007 E. (1984). 78-82.48 (2) 285-288. E. Gray. 32.

In Harlequin™ C. Berrocal.5 5. Consequently this medium can distinguish between non-O157 sorbitol negative E.D.2 LB Agar NEW HAL 4 Refer to the Biomolecular Section Minimum QC organisms: Escherichia coli O157:H7 NCTC 12900 (non-toxigenic) Escherichia coli NCIMB 50034 Enterococcus faecalis NCIMB50030 (inhibition) Inoculation: From O157 Broth LAB 165.'s include Staphylococcus aureus. E. Klebsiella pneumoniae. is a chromogenic medium for the improved isolation and differentiation of urinary tract pathogens.Perez.g. coli O157. often encountered. 28 (4) 803-805. Temperature Range for Growth of Escherichia coli Serotype 0157:H7 and Selected Coliforms in E.I. Pseudomonas aeruginosa.E. and Lattuada.. Traditional C.G. To our knowledge these isolates are limited to a small geographical area in Germany. Appearance: Pale red. Formula Peptone Sorbitol Bile Salts No.G. The majority of other E. pH: 7. Rose.E.D..5-4. This medium is a modification of Sorbitol MacConkey Agar (SMAC). This medium can also be useful for the detection of other VTEC producing E. surface streak for single colonies. O145).0 1. The superior growth qualities of Harlequin™ C. J.L. Examine plates for sorbitol negative. Evaluation of a Commercial ß-glucuronidase Test for the Rapid and Economical Identification of Escherichia coli.R. Method for reconstitution Weigh 48.T.E. and Citrobacter spp. and Matches. with other Gram-negative organisms such as Proteus mirabilis.App. J. Pathogenicity of the organism is linked to the production of verocytotoxins (VT1 and VT2). if they possess the ß- 2. A.J.'s. coli colonies (96-97% of strains are positive for ß-glucuronidase. Staphylococcus saprophyticus. coli O157:H7 have been isolated and appear as sorbitol positive and β-glucuronide positive on this medium. Dry the surface prior to inoculation. β-glucuronide negative colonies.. coli O157:H7 sorbitol -ve β-glucuronide -ve E. and Berrocal. Most workers recommend the use of CT-supplemented medium alongside unsupplemented medium to ensure maximum isolation of E. Morganella spp.T.E.L. and that strains from other serovars can be toxin producers (e. C.3 . swirl to mix and sterilise by autoclaving at 121°C for 15 minutes.J.Clin. Allow to soak for 10 minutes. Refer to the Biomolecular Section Organism GROWTH CHARACTERISTICS colony size (mm) 2.E. coli are β-glucuronidase positive and sorbitol negative and thus appear as blue/green colonies on this medium. the enzyme required to split this chromogenic compound). coli O157:H7 is typically sorbitol negative and β-glucuronidase negative producing pale translucent colonies on this medium. (1990) Use of 5-Bromo-4-Chloro-3-Indoxyl-β-D-Glucuronide in MacConkey Sorbitol Agar to Aid in the Isolation of Escherichia coli O157:H7 from Ground Beef..0 Harlequin™ SMAC-BCIG (Sorbitol MacConkey Agar with BCIG) NEW shape & surface CV. J. Grampositive organisms involved in U.I. B. The addition of the chromogenic substrate BCIG (5-bromo4-chloro-3-indoxyl-β-D-glucuronide) improves the specificty of the medium. Raghubeer. 61 541-545.6 grams of powder and add to 1 litre of de-ionised water.0 10. coli strains are β-glucuronidase positive and sorbitol positive (pink/red colonies). enterococci. E. which allows clear differentiation of many of the key pathogens involved.E. However this does not allow discrimination between E.Bacteriol. the primary serovar associated with haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS). Confirm as O157:H7 by serology. C. This reduces the number of unnecessary confirmation tests that are performed.1 ± 0.D. Micro.5-5. The medium can be made more selective by the addition of Cefixime Tellurite supplement X161 to prepare CT-SMAC.0 0. add 2 vials of X161 CT supplement and pour plates. Escherichia coli is responsible for the majority of urinary tract infections (U.. coli sorbitol -ve β-glucuronide +ve 2.L. References 1) Okrend. (1986). coli Medium. LB Top Agar NEW HAL 5 Incubation: 37°C aerobically for 18-24 hr.T. O103.E. O111. coli O157 has been associated epidemiologically with food poisoning outbreaks involving beef burgers and cold cooked meats. (Cystine Lactose Electrolyte Deficient) formulations rely upon Lactose fermentation to differentiate organisms.G.G. Cool to 47°C. E. Description This is a specific substrate medium for the isolation of Escherichia coli O157:H7. (1990). light violet tinge. coli and the other coliforms that cause U. F. J. O26.P.I.L.D.E. J.001 12.I.03 0. L.Food Protection 53 (11) 941-943 Harlequin™ CLED NEW HAL 7 Description Harlequin™ C.'s). coli sorbitol +ve β-glucuronide +ve E. Enterobacter spp. mean that it can also be used to improve differentiation of mixed growth in specimens where more fastidious organisms may be expected. but it should be noted that not all strains of O157 produce verocytotoxins.0 Pink/red or purple centre Green or translucent with green centre 2. coli in conjuncion with specifically targetted IMS particles (Captivate™). Bolton. and Lancefield Group B streptococci.0 CV.5-4. (commercial kits or antiserum available). Note: Sorbitol positive toxigenic E. A small percentage of E. (1995) Personal Communication. coli and the genuine toxigenic E. coli O157:H7.1 0. colour Translucent HAL 6 E. Other coliforms will produce colourless colonies or. X-glucuronide is incorporated to produce characteristic green E. O113. 3 Sodium Chloride BCIG Neutral Red Crystal Violet Agar g/litre 20. lactose has been replaced by a double chromogen system.0 CV.L..

5 0. LAB 072.G.C. J. CV. formulation and an alternative chromogenic formulation(1). GROWTH CHARACTERISTICS Organism Escherichia coli Method for reconstitution Weigh 36.0-2.E.2.G. pH: 7.-1.E. Typical coliforms.5-3.CR. coli strains possess both enzymes and produce blue-purple colonies (a combination of the blue and magenta pigments produced by the cleavage of the chromogenic compounds).0 Inoculation: Surface inoculation. Appearance: Tan. producing brown colonies (may also have a brown halo surrounding). GROWTH CHARACTERISTICS Organism Escherichia coli Salmonella spp Staphylococci Pseudomonas Aeruginosa Enterococci Group B streptococci 1.5 0. however.075 0.1 15.D.E.0 4.G. and related organisms. pH: 7. Glucosidase positive organisms include enterococci and Klebsiella spp. Typical E.5 CV. or using a 1µl loop for semi-quantitative enumeration.0 1.0-2. coli and coliforms on a single medium. Proteus spp.E.5-3.E. Allow the medium to set.L. In parallel trials Harlequin™ C. Formula Tryptone Bile Salts No.G.2 ± 0.2 0. isolated more clinically significant isolates than traditional C. CV.G Colour Blue-Purple Rose-Pink Other Enterobacter aerogenes Pseudomonas aeruginosa Candida spp. coli and coliforms a quick and simple procedure.G. This colour is due to the action of this enzyme on the second chromogenic substrate CHE-glucoside.D. Citrobacter spp p.3 ± 0.0 7. Formula Balanced Peptone No.-2.4 .5 0. The colony types are easily distinguishable. black colonies will be produced.5 p. possess only the β-galactosidase enzyme and produce red-magenta colonies. one to detect the β-glucuronidase enzyme (X-glucuronide) and another to detect the β-galactosidase enzyme (magenta-β-gal).5 . Colourless Cream/white Colourless Black Cream Green (if ßglucuronidase +ve) Colony Shape & Size (mm) Surface 1.0 0.D CV.0 1. the medium has been modified by the addition of two chromogenic substrates.0 1.3 X-glucuronide Magenta-β-galactoside Agar g/litre 20.G.E. disperse in 1 litre of deionised water and allow the medium to soak for 10 minutes. colourless surround Buff No growth No growth Enterococcus faecalis Staphylococcus aureus 2. Swirl to mix and sterilise at 121°C for 15 minutes. 1.0-2.-2. or when large numbers are observed.D.0 grams of powder.up to 7 days at 2 . Enterobacter spp. CV. The different colony types are simple to distinguish allowing rapid counting of both E.D. disperse in 1 litre of deionised water and allow the mixture to soak for 10 minutes.2 Colony Shape & Size (mm) Surface 1.5-1.0 p.11 0.E.E.CR.E.0 CV. CV.0 1.E.p.075 0.0 Method for reconstitution Weigh 43.24 hours at 37° C References 1) Perry.5 .2.1. Dry the surface prior to inoculation.E. Cool to 47°C and pour in to sterile Petri dishes. Incubation: 18 . .5 CV. Based upon the formulation of Tryptone Bile Agar. Cool to 47°C and pour into sterile Petri dishes. F. coli/Coliform Medium NEW HAL 8 Description This dual chromogenic substrate medium has been developed for the simultaneous enumeration of Escherichia coli and coliforms in food and environmental samples. making simultaneous enumeration of E.E.L.0 .1 Beef Extract Meat Peptone Yeast Extract L-Cystine CHE-glucoside X-glucuronide Ferrous Gluconate Phenylalanine Agar g/litre 6. organisms: Escherichia coli NCIMB 50034 Staphylococcus aureus NCIMB 50080 Storage of Powdered Medium: Plates .2 Minimum Q.6 grams of powder.G.G.D/ CV.up to 7 days at 2 .5 0. Appearance: Straw.p. Black Brown (halo) Minimum Q.E.5 CV. Further discrimination is achieved by the addition of phenylalanine which is deaminated by Proteus spp.0 1.95 0.glucosidase enzyme.8°C in the dark.E.5 4. White Black Black Black centre. organisms: Escherichia coli NCIMB 50034 Enterobacter aerogenes NCIMB 50029 Staphylococcus aureus NCIMB 50080 (Inhibition) Storage of Powdered Medium: Plates . CV.2 20.C.5-1.Personal Communication (data on file) Harlequin™ E.G F. Colour Green Other Colourless (if ß-glucuronidase -ve) Mucoid Colourless if weak phenylalanine reaction Klebsiella spp.E. even in the presence of other organisms.0 CV.8°C in the dark. clear gel. Dry the surface prior to inoculation. Swirl to mix and sterilise at 121°C for 15 minutes. clear gel. either spreading for single colonies.p. Allow the medium to set.G.0-2.G CV.

coli) from water potentially contaminated with sewage.G CV. Swirl to mix and sterilise at 121°C for 15 minutes. The medium has been modified from the mLSB formulation by the incorporation of X-glucuronide. M.5 . With waters expected to contain low numbers of coliforms. Cool to 47°C and pour into sterile Petri dishes. Incubation: 4 hours at 30 °C followed by 14 hours at 37 °C Interpretation: Count all green-blue colonies as presumptive E.G Yellow Red Description Traditionally.5 . The volume and dilution of test sample should be chosen so as the number of colonies on the membrane lies between 20 and 80.4 ± 0.E.5 CV.up to 7 days at 2-8°C in the dark. coli.5 Non-lactose fermenters Staphylococcus aureus 0. Letters in Applied Microbiology 15.5 . Storage of Powdered Medium: Plates .E.5 CV. No. pH: 7. X-glucuronide is incorporated to allow for the presumptive isolation of E.L. and calculate cfu/g.2 Minimum Q. D. coli and coliform counts can be performed on the same sample of water. G. disperse in 1 litre of deionised water and allow the mixture to soak for 10 minutes. Further dilution may be necessary if large numbers of E. & Howard. membrane Lauryl Sulphate Broth (mLSB) has been used as the standard media for isolating coliforms (including E. (1999). sodium pyruvate and agar. coli. Inoculation: E. coli by a single membrane filtration technique in The Microbiology of Drinking Water 2002 (previously Report 71).. organisms: Escherichia coli NCIMB 50034 Enterobacter aerogenes NCIMB 50029 Staphylococcus aureus NCIMB 50080 (inhibition) 2. Count all blue-purple and magenta colonies as presumptive coliforms. Applied and Environmental Microbiology 56. Comparison of a range of Chromogenic media for enumeration of total Coliforms and Escherichia coli in foods. 273-276. coli and/or coliforms are present.24 hours at 37° C Interpretation: Count all blue-purple colonies as presumptive E.1. Harlequin™ mLGA (Membrane Lactose Glucuronide Agar) NEW Organism Escherichia coli* colony shape & size (mm) surface 0. clear gel. and all green-blue and yellow colonies as presumptive coliforms. coli by a single membrane filtration technique. The enumeration of coliform bacteria and E. Incubation: 18 .0 No growth (suppressed) *96-97% of E. This medium is recommended for the enumeration of coliform bacteria and E. coli 0157:H7 References Sartory. A notable exception is E. GROWTH CHARACTERISTICS References 1) Baylis. (1992). Method for reconstitution Weigh 87. C. coli strains positive.P.0 0. Improved membrane filtration method incorporating catalase and sodium pyruvate for detection of chlorine stressed coliform bacteria.2 1.0 grams of powder. Leatherhead International Technical Notes.Environment Agency.C. Calabrese.The enumeration of coliform bacteria and E.2 10.E. & Bisssonette. coli.0 0. (1990).0 30. J.1. sodium pyruvate aids the recovery of chlorine stressed organisms and agar is incorporated to remove the need for absorbent pads.Inoculation: Inoculate 0. Harlequin™ membrane Lactose Glucuronide Agar (mLGA) is a modification of mLSB aimed at reducing costs by reducing the number of filters used per test sample and aiding in the recovery and identification of coliforms and E. a sample of 100ml should be filtered.0 6. Patrick. Appearance: Red.K. and calculate the cfu/g.5 0. to ensure colonies can be easily counted.P.5 . coli. Microbiology of Drinking Water 2002 section 4 B . A medium detecting B-glucuronidase for the simultaneous membrane filtration enumeration of Escherichia coli and coliforms from drinking water. 3558-3564.1.G colour Green other Yellow if glucuronidase -ve HAL 9 Lactose fermenters 0.135: 99. Formula Peptone Yeast Extract Lactose Phenol Red Sodium Lauryl Sulphate Sodium Pyruvate X-Glucuronide Agar g/litre 39. L.5 ml of a 1:10 dilution of the sample and spread over the entire surface of the plate. For full methodology refer to The Microbiology of Drinking Water 2002 section 4 B . coli by a single membrane filtration technique.

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(1972). Cold Spring Harbour New.3 0. The CHE-galactoside replaces the traditional X-gal substrate. Cold Spring Harbour New . and Maniatis. Cold Spring Harbour Laboratory.03 2. Incubation: 37°C aerobically. Biomolecular Products LB Agar LB Broth Harlequin™ LB agar* Harlequin™ LB Top* LB Agar (Lennox) LB Broth (Lennox) Superbroth Terrific Broth YPD Agar YPD Broth Superbroth with Agar 2 x YT Agar 2 x YT Broth NZY Broth NZCYM Broth Luria Bertani Agar (Hi-Salt) water.0 1. J. Simply add water to the powder and autoclave. Pour into sterile Petri dishes. simplifying the technique as there is no preparation of stock solutions in dimethyl formamide or dimethyl sulphoxide and surface application of the chromogen to the medium. More importantly. Cold Spring Harbour New.0 5. Interpretation: Examine for the presence of cream colonies. Formula Tryptone Yeast Extract Sodium chloride CHE-galactoside IPTG Ferric ammonium citrate Agar g/litre 10. Sambrook. Fritsch. fast and easy way to differentiate between lac+ and lac.ß-gal reaction Escherichia coli DH5a (ATCC® 53868) Lac Z+ve (black) Escherichia coli DH5a. References Miller.0 5. safe and unambiguous detection of phage transformed bacteria.0 0. These products are formulated to promote the growth of the recipient and donor cells used in DNA insertion technology. J. Fritsch. E. J. LAB M offer a range of media types.colonies.5 12. The intense black colour of the colonies gives a sharper contrast between lac. References Miller. Lac Z -ve (remains cream) Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. LAB M have formulated unique versions of LB Agar and LB Top Agar which incorporate the patented water soluble chromogen CHE-β-gal into the complete medium.0 ± 0. *Available from LAB M in the UK only. Sambrook. 2nd ed..2 ) Minimum QC organisms . T. This improves colour definition of α-complemented colonies compared to the standard X-β-gal plate and removes the need for hazardous chemicals in the preparation of the medium. Appearance: Straw. to indicate inactivation of α-complementation. Cold Spring Harbour Laboratory. 2nd ed. Molecular Cloning: A Laboratory Manual.03 0..1 . safe and unambiguous detection of plasmid transformed bacteria.F. and Maniatis. (1972). As different applications have varied requirements of the culture medium used. Cold Spring Harbour Laboratory.. This variety allows the researcher to choose the appropriate medium for the application being used. Cool to 47°C and add appropriate filter sterilised antibiotic if required. Cold Spring Harbour Laboratory. The colour of the colonies will substantially increase with prolonged incubation (up to 24 hours).0 0. York.and lac+ colonies.2 Storage of Prepared Medium: Storage of capped medium in bottles for up to 1 month at room temperature in the dark.0 10. giving improved colony detection compared to blue X-gal stained colonies.0 Harlequin™ LB Agar NEW Tryptone Yeast Extract HAL 4 Sodium chloride CHE-galactoside IPTG Ferric ammonium citrate Agar Description A nutritious molecular biology medium containing the novel chromogen CHE-galactoside to enable rapid. for 16-18 hours. Therefore we have produced a safe. such as X-gal and our patented CHE-gal. they are formulated to provide optimum conditions for plasmid retention or bacteriophage reproduction and survival. Molecular Cloning: A Laboratory Manual. Cold Spring Harbour New. clear gel. which indicates a successful insertion of the target DNA. Method for reconstitution Weigh 37.F. Cool to 47°C before seeding the medium with required strains. Harlequin™ LB Top Agar NEW HAL 5 Description A nutritious molecular biology medium containing the novel chromogen CHE-galactoside to enable rapid. (1989).York. Experiments in Molecular Genetics. clear gel. pH: 7.0 10.0 ± 0. Inoculation: Typically surface spread over plate to detect cream colonies indicating disruption of β-complementation.8 grams of powder and disperse in 1 litre of deionised 3. E.0 4.3. Formula g/litre 10. J.H. Experiments in Molecular Genetics. Appearance: Straw.H. (1989). pH: 7. Swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. LAB M’s Biomolecular products form the basis of gene reporter assays that employ enzyme substrates. Swirl to mix and sterilise by autoclaving at 121°C for 15 minutes.0 Method for reconstitution Weigh 32 grams of powder and disperse in 1 litre of deionised water. allow the medium to set and dry the surface prior to inoculation. All products are available directly from LAB M in the UK. T. spread for single colonies if required.. Some are of standard formulation whilst others are modified to enhance the performance of specific applications.York. Alternatively. Outside the UK Harlequin LB agar is available from Sigma-Aldrich as S-Gal LB agar (C4478). York.

. Minimum QC organisms . 1. F. Fritsch. Vol.D.. T. Alternatively. Lac Z -ve (remains cream even in the presence of CHE-gal) Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. If a chromogenic substrate is used. Experiments in Molecular Genetics. (1972).0 NEW LAB 174 Description A nutritious medium designed for rapid bacterial growth.0 5. E.M. the colour of the colonies will substantially increase with prolonged incubation (up to 24 hours). Cold Spring Harbour Laboratory.0 15. spread 40µl as a surface layer over the top of the agar and allow to dry. 190. Allow the mixture to soak for 10 minutes. pH: 7.G. Inoculation: Typically surface spread over plate to detect cream colonies indicating disruption of ß-complementation. if a chromogen is included.LB Agar NEW LB Agar (Lennox) LAB 168 Description This is a nutritionally rich medium containing half the sodium chloride level of LB agar (LAB168).York. 2nd ed. typically used in molecular biology procedures e. 2nd ed. Cool to 47°C and add filter sterilised antibiotic as required.. 3. Incubation: 37°C aerobically. Interpretation: Using the base medium alone.2 Addition of Substrate Prepare the X-Gal solution by dissolving in DMF. York. Lac Z -ve (remains cream even in the present of CHE-gal) Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark.. Cold Spring Harbour New. pH: 7. E. J. examine for the presence of cream colonies. Transduction of linked genetic characters of the host by bacteriophage P1. (1955). J. Smith. This medium can also be used for plasmid replication experiments. References Miller. Cold Spring Harbour New. Dry the surface prior to inoculation. which indicates a successful insertion of the target DNA.0 5.F. J.E.. which already contains the enzyme substrate and inducer.F. R.0 grams of powder and disperse in 1 litre of deionised water. (1994). Current protocols. Appearance: Straw. Brent. J. clear gel. and Struhl. New York. which is deficient in B vitamin production. Cold Spring Harbour New .0 15.. Sambrook. Cold Spring Harbour Laboratory. Formula Tryptone Yeast Extract Sodium chloride Agar g/litre 10. for 16-18 hours. Addition of Substrate Prepare the X-Gal solution by dissolving in DMF. J. D. Kingston. Fritsch.0 Method for reconstitution Weigh 40. in the detection of phage or plasmid transformed bacteria and the maintenance of recombinant strains.. This eliminates the potentially hazardous use of DMF and prevents variation in the colour of ß-complemented colonies due to differences in substrate concentration. Interpretation: Using the base medium alone.ß-gal reaction: Escherichia coli DH5a (ATCC® 53868) Lac Z+ve (black if CHE-gal is present in the medium) Escherichia coli DH5a. which indicates a successful insertion of the target DNA. E. and Maniatis.Y. (1989). Appearance: Straw. if a chromogen is included. Virology 1. Once dissolved. to give a concentration of 20mg/ml Once dissolved. Cool to 47°C and add filter sterilised antibiotic if required. Current protocols in molecular biology. References Lennox... Alternatively.0 ± 0. examine for the presence of cream colonies. York.2 . all colonies will appear cream. clear gel. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. either spread over entire surface for colony count or streaking for single colonies. Ausubel. Alternatively.0 ± 0.S. Molecular Cloning: A Laboratory Manual. Also spread 4µl of a solution of IPTG (200mg/ml). Formula Tryptone Yeast Extract Sodium chloride Agar g/litre 10. Seidman. Moore.ß-gal reaction: Escherichia coli DH5a (ATCC® 53868) Lac Z+ve (black if CHE-gal is present in the medium) Escherichia coli DH5a. This agar contains the required concentration of sodium chloride to promote replication of plasmids. Sambrook.H.0 10. Swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. spread 40µl as a surface layer over the top of the agar and allow to dry. Pour into sterile Petri dishes and allow the medium to set.g. R. Incubation: 37°C aerobically for 16-18 hours. Molecular Cloning: A Laboratory Manual. spread for single colonies if required. use Harlequin™ LB agar complete (HAL004). Nutritionally rich media are required for molecular biology applications as the strains used are often derived from Escherichia coli K12. Inoculation: Surface. T. N.0 grams of powder and disperse in 1 litre of deionised water. Also spread 4µl of a solution of IPTG (200mg/ml). (1989).2 Minimum QC organisms . Pour into sterile Petri dishes and allow the medium to set. Cold Spring Harbour Laboratory. Alternatively.A. all colonies will appear cream. and Maniatis.0 5. This allows the researcher to select the optimum salt concentration for his experiment. Method for reconstitution Weigh 35. to give a concentration of 20mg/ml. Dry the surface prior to inoculation.

0 5.0 5. Incubation: 37°C aerobically for 16-18 hours. Incubation: 37°C aerobically. Method for reconstitution Weigh 25. Brent.0 ± 0.0 grams of powder and disperse in 1 litre of deionised water. pH: 7.M. Formula Tryptone Yeast Extract Sodium chloride g/litre 10. J. Current protocols. Virology 1. Inoculation: As per normal techniques. T. F.F. J. (1989). and Maniatis. York. This broth is formulated to LB Broth (LAB169). (1972). Vol. This broth contains a high level of sodium chloride to aid the maintenance of plasmids.A. N.. Sterilise by autoclaving at 121°C for 15 minutes. indicating growth. Current protocols in molecular biology. If working with temperate bacteriophages.0 10. Sambrook.0 5. Cold Spring Harbour New.0 grams of powder and disperse in 1 litre of deionised water. Incubation: 37°C aerobically for 16-18 hours.LB Broth NEW LAB 169 Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark.0 10. Formula Tryptone Yeast Extract Sodium chloride g/litre 10. R.F. Kingston.D. Fritsch. Chloramphenicol can be added to achieve high plasmid copy number by inhibiting chromosomal replication. pH: 7. York. clear gel.G. Molecular Cloning: A Laboratory Manual. Sambrook. J. (1955). E.. E. New York.0 References Miller.. Appearance: Straw.S.E. Interpretation: Examine all tubes for turbidity. D.g. E. phage P1. Appearance: Straw.. clear liquid. Moore. swirl to mix and dispense into final containers. and Maniatis. Molecular Cloning: A Laboratory Manual.Y. Description A nutrient broth primarily used for the growth and maintenance of Escherichia coli.0 References Lennox.H. Cold Spring Harbour Laboratory.. Allow the mixture to soak for 10 minutes. Smith. using a pure culture of donor/recipient cells. J. LAB 191 Description A nutritious medium designed for rapid bacterial growth. Interpretation: Examine all tubes for turbidity.. This medium can also be used for plasmid replication experiments. this allows for the addition of calcium chloride.5 ± 0. when further work is to be performed on LB Agar.0 grams of powder and disperse in 1 litre of deionised water.0 Method for reconstitution Weigh 20.7H2O) at 2 grams per litre is recommended to promote phage absorption. using a pure culture of donor/recipient cells.. such as lambda. typically used in the detection of phage or plasmid transformed bacteria. Cold Spring Harbour Laboratory.. pH: 7. R. Cold Spring Harbour New .. the addition of magnesium sulphate (MgSO4. Cold Spring Harbour Laboratory. Method for reconstitution Weigh 25. 2nd ed. T. Used as the primary propagation step for donor or recipient cells. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes.York. Description This is a nutrient broth containing half the sodium chloride level of LB Broth (LAB169). Inoculation: As per normal techniques. and Struhl. Cold Spring Harbour New. Experiments in Molecular Genetics. 190.2 Luria Bertani (Hi-Salt) Broth NEW Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. (1994). Formula Tryptone Yeast Extract Sodium chloride g/litre 10. Ausubel. 1. Transduction of linked genetic characters of the host by bacteriophage P1.3 . indicating growth.2 LB Broth (Lennox) NEW LAB 173 Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Plates up to 7 days at 2-8°C in the dark.0 5. Allow the mixture to soak for 10 minutes. required in some applications for efficient phage adsorption to the cell e. Allow the mixture to soak for 10 minutes. clear liquid. Fritsch. Inoculation: Dependent upon application. 2nd ed.2 3. Appearance: Straw. J.5 ± 0. (1989). swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. but has a higher pH for different applications. Seidman.

Inoculation: Mix a fresh overnight culture of host cells with bacteriophage and use to inoculate NZCYM Broth. as binding of phage particles to membrane fragments will occur because of increased LamB density. et al. pH: 7. Mol.2% maltose (prepare a 20% solution and add 1ml per 100ml of medium). Sterilise by autoclaving at 121°C for 15 minutes. clear liquid. Appearance: Straw. swirl to mix and dispense into final containers. Inoculation: Mix a fresh overnight culture of host cells with bacteriophage and use to inoculate NZY Broth.0 20.. which is designed for increased bacteriophage lambda replication and yield..0 2. it is recommended that 0.. F. Superbroth is predominantly used for producing liberated phage stocks. Superbroth NEW LAB 177 Description A broth medium incorporating high levels of nutritious peptone and yeast extract.0 References Blattner. pH: 7. used for the producing high yields of bacteriophage Lambda (λ). Formula Tryptone Yeast extract Sodium chloride g/litre 35. Biol..NZCYM Broth NEW LAB 182 Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. 161.0 ± 0.0 5. et al. which promotes expression of LamB (lambda receptor).0 grams of powder and disperse in 1 litre of deionised water.2% maltose be added (prepare a 20% solution and add 1ml per 100ml of medium). Inoculation: Mix fresh overnight culture of host cells with bacteriophage and use to inoculate the Superbroth.. Formula Enzymatic casein digest Yeast extract Magnesium Sulphate Sodium chloride g/litre 10.0 ± 0. D. swirl to mix and dispense into final containers.5 ± 0. To encourage multi phage insertion into the host cell. Appearance: Straw. Incubation: 37°C aerobically. NZY Broth (NZYM) NEW Incubation: 37°C aerobically until lysis occurs.2 Sodium chloride Agar 3.0 20. Formula Enzymatic casein digest Acid hydrolysed casein Yeast extract Magnesium Sulphate Sodium chloride g/litre 10.4 . (1977) Science 196. clear liquid. Description This is an improved medium for increased yields of the phage Lambda. et al. If maltose is added.0 1.0 5. Appearance: Straw. Formula Tryptone Yeast extract g/litre 35.0 Method for reconstitution Weigh 23.2 Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. References Bolstein. To encourage multi phage insertion into the host cell. F.0 Method for reconstitution Weigh 22. do not use this medium to create phage stocks.0 15. the medium should not be used to create phage stocks. LAB 181 Interpretation: Examine tubes for lysis (clearing of broth) and concentrate liberated phage by normal procedures. Allow the mixture to soak for 10 minutes.0 Superbroth with Agar NEW LAB 178 Description An agar version of the extremely rich Superbroth medium. 439. Allow the mixture to soak for 10 minutes. Weigh 60. (1975) J.0 5. 91. If maltose is added. Description Designed for increased replication of phage Lambda.2 Method for reconstitution Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. clear liquid. pH: 7. (1977) Science 196. 161. which promotes expression of LamB (lambda receptor). Allow the mixture to soak for 10 minutes. References Blattner.0 5. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. as binding of phage particles to membrane fragments will occur because of increased LamB density.0 5. Sterilise by autoclaving at 121°C for 15 minutes.0 2. Incubation: 37°C aerobically for 16-18 hours.0 grams of powder and disperse in 1 litre of deionised water. This formulation includes a higher concentration of essential elements for increased bacterial growth.0 grams of powder and disperse in 1 litre of deionised water.0 5. it is recommended to add 0.

using a pure culture of strain to be cultivated. clear liquid. D. Allow the mixture to soak for 10 minutes. Formula g/litre 20. Pour into sterile Petri dishes and allow to set. Glucose is included to promote rapid growth. Cool to 47°C and add filter sterilised antibiotic as required. J. swirl to mix and dispense into final containers. Focus. where a solid base is required. clear gel. (1989).0 ml of glycerol.4 2..5 ± 0. Method for reconstitution Weigh 50. Glucose is included to promote rapid growth. Improved media for growing plasmid and cosmid clones. 3.0/8. Formula Tryptone Yeast extract Di Potassium phosphate Potassium di phosphate Glycerol (added after autoclaving). Allow the mixture to soak for 10 minutes. Cold Spring Harbour New. Mol. g/litre 12. either spread over entire surface for colony count or streaking for single colonies.0 grams of powder and disperse in 1 litre of deionised water. Incubation: 37°C aerobically for 16-18 hours. pH: 7. Interpretation: Yeasts will grow as cream colonies. Appearance: Dark straw. 439.0 Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. T. Biol.2 4.0 24. Dry the surface prior to inoculation. Fritsch.0 Method for reconstitution Weigh 47. Cool to 47°C and add filter sterilised antibiotic as required. Pour into sterile Petri dishes and allow to set. Incubation: 37°C aerobically.0 10.2 References Tartoff.2 Minimum QC organisms: Escherichia coli DH5 (ATCC® 53868) Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark.0 9. Allow the mixture to soak for 10 minutes. Sterilise by autoclaving at 121°C for 15 minutes. C.F. pH: 7. (1987). YPD Broth NEW LAB 175 Description A nutritious broth base recommended for the maintenance and propagation of yeasts widely used in gene insertion techniques. Allow the mixture to soak for 10 minutes. et al.2 NEW Terrific Broth LAB 183 Description A nutritious medium that will support high bacterial cell densities. Incubation: 37°C aerobically for 16-18 hours. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. swirl to dissolve and sterilise by autoclaving at 121°C for 15 minutes. Bethesda Res. York. Appearance: Straw. clear liquid.D. Method for reconstitution Weigh 67. Cold Spring Harbour Laboratory. The formulation requires the addition of glycerol to complete the formulation. indicating growth. Formula Tryptone Yeast extract Glucose g/litre 20.6 grams of powder and disperse in 1 litre of deionised water. Inoculation: Dependent upon application. pH: 6. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. Appearance: Straw.2 YPD Agar NEW LAB 176 Description A nutritious medium used as an alternative to YPD Broth..0 grams of powder and disperse in 1 litre of deionised water.0 ± 0. pH: 6. Molecular Cloning: A Laboratory Manual.0 ml Minimum QC organisms: Saccharomyces cerevisiae Inoculation: Surface. C.0 or 8. swirl to mix and dispense into final containers.Method for reconstitution Weigh 75.A. 91.0 10. size dependent upon inoculum density. Lab. Dry the surface prior to inoculation.5 ± 0.5 . Add 4. E.0 20.0 20. Inoculation: As per normal techniques. Interpretation: Examine all tubes for turbidity.5 ± 0. (1975) J. Sambrook. 9:205. Incubation: 37°C aerobically. 2nd ed. and Maniatis. clear liquid.. Tryptone Yeast extract Glucose Agar References Bolstein. Minimum QC organisms: Saccharomyces cerevisiae Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. and Hobbs. Appearance: Dark straw.0 grams of powder and disperse in 1 litre of deionised water.0 17. Inoculation: Inoculate with a pure culture of the host strain containing the required recombinant plasmid. usually resulting in increased yields of DNA and recombinant proteins.

0 15.0 10.0 Method for reconstitution Weigh 46. Storage of Prepared Medium: Plates – up to 7 days at 2-8°C in the dark. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes. Formula Tryptone Yeast extract Sodium chloride g/litre 16.2xYT Agar NEW LAB 180 Description An agar version of 2xYT broth.0 5. 2xYT Broth NEW LAB 179 Description A nutritious liquid medium formulated to promote the growth of host cells. pH: 7. The resulting bacteriophage stock can be stored at +4°C or -20°C. Storage of Prepared Medium: Capped containers – up to 3 months at 15-20°C in the dark. Dry the surface prior to inoculation. swirl to mix and sterilise by autoclaving at 121°C for 15 minutes.0 10. organisms: Escherichia coli DH5 (ATCC® 53868).2 Minimum Q. thereby encouraging increased replication and yield from filamentous single stranded bacteriophages (such as the M13 phage).0 5.0 ± 0. Inoculation: dependent upon application. Formula Tryptone Yeast extract Sodium chloride Agar g/litre 16. Allow the mixture to soak for 10 minutes.6 . Appearance: Straw. Cool to 47°C.0 grams of powder and disperse in 1 litre of deionised water. clear gel. 3. clear liquid. Incubation: 37°C aerobically. the M13 phage. Inoculation: Mix fresh overnight culture of host cells with bacteriophage and use to inoculate 2xYTBroth. being careful not to disturb the pellet formed. organisms: Escherichia coli DH5 (ATCC® 53868).2 Minimum Q.0 ± 0.0 Method for reconstitution Weigh 31.0 grams of powder and disperse in 1 litre of deionised water. Incubation: 37˚C aerobically.C. Interpretation: Concentrate bacterial cells by centrifugation and transfer supernatant containing bacteriophage to a fresh tube. typically for 4-5 hours to reduce risk of selecting deletion mutants. Allow the mixture to soak for 10 minutes. Appearance: Straw.g. pour into sterile Petri dishes and allow the medium to set.C. pH: 7. for the growth of host cells of filamentous single stranded bacteriophages e.

5µm average size. The beads are manufactured by a high speed blending process and typically cover a size diameter range of 1-4 µm. Buffered Peptone Water (LAB 46) plus VCC (X546)5-8. results can be achieved 24 hours earlier than standard protocols. affinity purified and absorbed polyclonal antibodies to cell surface antigens.g.2. It is also recommended that a further IMS and inoculation of SMAC plates is performed after incubation of the sample for 24 hours. coli O157:H7.g.09% azide as preservative. reducing the effect of the matrix and allowing more efficient bead recovery. Thorough mixing of the particles and sample allied with efficient recovery of the beads from the sample matrix is paramount to the success of this technique. To counter-act this interference.0 0. The bead/microorganism complexes are then removed from the sample by placing the sample in a magnetic concentrator device. With careful antibody selection. Store in the dark and use within one month.e. beverages. samples can also be diluted in PBS-Tween® e. 5) Carefully aspirate the supernatant from the tube and cap without removing particles. E. animal feeds. Reacts with target organism.5 µm (typical range 1-8µm) Particles are suspended in PBS plus1% BSA pH 7.5 . The particles help to concentrate O157:H7 cells in mixed culture reducing the probability of missing low numbers or overgrowth of O157:H7 colonies by competing flora. Purified antibodies to surface components of the target microorganism are covalently coupled to the bead. 4) Insert tube into magnetic separator rack for 3 minutes to concentrate the beads to a pellet.5 and 0. The recommended protocol for the isolation of E. remove 800µl) and continuation of the wash protocol as described can minimise bead losses. A sample is taken from a filter stomacher bag and incubated with the Captivate™ beads for 30 minutes. coli O157:H7 employs a 6 hour enrichment step at 42°C in modified Tryptone Soy Broth (mTSB. particulate and viscous samples) can interfere with bead recovery.5ml volume). Care must be taken not to aspirate the sample vigorously as this can result in the loss of captured target organisms.5 Captivate™ Product Specification Working concentration: Fe3O4 content: Antibody: Typically 5mg/ml 29-33% w/w Particles coated with high avidity. coli O157:H7 is the primary serovar associated with food borne gastrointestinal infection. The organism itself is associated with raw meats and unpasteurised milk1. streaking for single colonies. 3) Cap tube tightly and rotamix the suspension for 30 minutes at room temperature. a highly specific separation system for microorganisms is produced.15 0. coli O157:H7 from food. This separates them from the background organisms and interfering materials. 6) Remove magnet from rack or tubes from the rack and add 1ml of wash. Cap and resuspend particles by inverting several times. In fact. coli O157:H7.2 Dissolve the components in deionised water and check the pH. that can lead to serious disease conditions such as haemorrhagic colitis. after the initial magnetic separation the incomplete removal of the sample (i. 8) Remove 50µl of the complexed. 8°C (may be shipped at ambient) 2 years. 1:2-1:4. haemolytic uraemic syndrome (HUS) and thrombotic thrombocytopaenic purpura (TTP). Specificity: Average size: Formulation: Storage: Shelf life: pH: 7. resuspended particles to the plating media. These products can also serve as a capture system for rapid detection systems. Incubate plates at 37°C for 18-24 hours and examine for typical colonies. Alternatively with problem samples. Gently invert the rack several times to aid pelleting of the beads. The beads have a magnetite core and a "ceramic" zirconium oxide coating. Sterilise the solution by autoclaving at 121°C for 15 min. resulting in self-limiting diarrhoea. This patented technology consists of microscopic paramagnetic particles. Large outbreaks have been recorded in the United States from consumption of unpasteurised apple juice (apple cider) possibly as a result of using apples which have fallen to the ground where the potential for contamination with the organism exists3. Generic Captivate™ IMS Procedure 1) Add 20µl of well mixed Captivate™ particles to a suitable micro-tube (1.3-7. There are important factors that affect the performance of IMS techniques.0 ± 0. with a 2. The complexes are then washed using a PBS/Tween® 20 wash buffer to remove non-specifically bound material. Alternative enrichment protocols using different media have been described e.4. The IMS technique will increase the sensitivity of the methodology and. The pre-coated beads are designed for the IMS of target bacteria from enrichment cultures. in most circumstances. Allow the solution to cool and check the pH.1 . very fatty. Enrichment Protocol for E.020 1. g/litre 8. 2) To this add 1ml of the enrichment culture taking care to avoid transfer of sample debris.g. Finally resuspend particles in 100µl of wash. Certain sample types (e. The beads can then be plated out onto the appropriate selective agar media and incubated as described. Formula Sodium chloride Potassium chloride Disodium hydrogen phosphate Potassium dihydrogen phosphate Tween® 20. 4. Phosphate Buffered Saline plus Tween®. taking care to avoid splashing. 7) Repeat separation and wash steps 4-6 twice more. Special Notes on IMS Techniques. LAB 165) plus novobiocin (X150) followed by IMS (see below) and plating onto Sorbitol MacConkey Agar (LAB 161 or HAL006) supplemented with or without the addition of cefixime and potassium tellurite (X161)5-8. 2. coli O157:H7 from food and environmental samples. probably due to the implication of farm animals and particularly cattle as carriers of E. Captivate™ O157 NEW CAP 1 Captivate™ Description Captivate™ O157 are magnetisable particles coated with specific antibody intended for the isolation of E.4. Captivate™ is a range of antibody coated paramagnetic particles for the specific ImmunoMagneticSeparation (IMS) of microorganisms. immunomagnetic separation is now regarded as the gold standard method for isolation of E.2 0. pharmaceutical or environmental samples.

J. P. et al (1986) Isolation of Escherichia coli from cattle associated with two cases of hemolytic syndrome. 12 67-71. Pathogenesis and Methods for Detection in Food. Chapman. The optimisation of media used in the immunomagnetic separation methods for the detection of Escherichia coli O157 in foods.A. M. but not provided. (1996) E. Crozier. 5)Wright. (1995) New technical approaches to Escherichia coli O157. M. 555-565. & MacRae. coli O157:H7 outbreak in USA traced to apple juice. product code CAP001-250. R.Mic. Dig. 373-379. D. 3) Besser.A. (2001). Detection of Escherichia coli O157 and other VTEC in food.. JAMA 259 2217-2220 4) McCarthy. Immunomagnetic separation as a sensitive method for the isolation of Escherichia coli O157 from food samples.D. L.V.Prot. M.. include phosphate buffered saline-Tween® 20. PHLS Microbiol. Lancet 348 1299. (1997). N. Hepburn. et al (1993) An outbreak of diarrhoea and hemolytic uremic syndrome from Escherichia coli O157:H7 in fresh pressed apple cider. Epidemiology and Infection 113. product code CAP001-050 and 250 test.Interpretation: Examine the SMAC and CTSMAC plates for typical E. C. J. 1-3 mm in diameter that are colourless to pale orange.. 537-551.P. C. (1992). M. coli O157 non-sorbitol fermenting colonies that are smooth and circular.. Journal of Applied Microbiology.K. and Doyle.. Williamson. 2) Martin.J. stomacher machine and bags..Food. pipettes and tips.L. Lancet ii 1043.E. 31-39.F. 91. magnetic separator rack and culture media. 4. Product Presentation Captivate™ O157 is available in packs of 50 test. Appl. and Siddons. Escherichia coli O157:H7: Epidemiology. Confirm the colony identity with commercially available latex agglutination kits or antisera.. Magnetic separating racks (CAP-100-12P) and rotating mixers (CAP101-58) are also available from LAB M. 8) Ogden. N.J. Reference 1) Padhye. Materials required. 6) Bolton. 55. (1994). 7) Vernozy-Rozand. F. J. I.2 . 82.

Prada.. This gives an excellent high avidity broad spectrum IMS reagent for the capture of salmonellas. Beutin. Zimmerman. Montenegro. Vol 16. 4. in combination with the Captivate™ O157 reagent. Close association of verocytotoxin (shigalike toxin) production and enterohaemolysin production in strains of Esherichia coli. Therefore. Unlike E. coli (VTEC). the "top five" VTEC as identified by the World Health Organisation can be targeted with these reagents. 711-718. Therefore using a screening isolation medium such as Sorbitol MacConkey Agar does not help in the isolation of these organisms. The IMS step should greatly increase the chance of isolating these organisms. World Journal of Microbiology and Biotechnology. Journal of Clinical Microbiology 27. coli plating medium such as Tryptone Bile Glucuronide Agar (HAL 3) or TBA (LAB 72) plus MUG (MC406).. The particles are coated with affinity purified polyclonal antibody directed towards common somatic and flagellar antigens. L. Current microbiological methods do not permit the sensitive isolation of nonO157-VTEC. Bearing this in mind. (2000). O111 and O145 to aid the isolation of the other common serotypes of verotoxigenic E.A. which is sorbitol negative. Due to the large variation in Salmonella serotypes and antigen expression there is naturally strain dependent variation in the capture efficiency. coli react with.3 . & Karch. we suggest users try the same enrichment protocol that we recommend for E. which the majority of E. Orskov. M.Captivate™ O26 NEW Captivate™ Salmonella CAP 3 Description NEW CAP 2 Captivate™ O111 NEW CAP 4 Captivate™ O103 NEW This product is designed to capture and concentrate the common serotypes of Salmonella involved in human and animal disease from enriched samples. and Stephan. Verocytotoxin production has been shown to be closely linked with the enterohaemolytic phenotype. R.. To address this basic problem LAB M has developed a quartet of individual immunomagnetic separation reagents.. coli O157 at 37°C and 42°C with a generic E. I.com There is now growing concern that VTEC’s could be playing a more significant role in human disease than currently estimated. 2559-2564. CAP 5 Custom Coating Service A coating service is available for coating our IMS reagent with alternative antibodies. References Bielaszewska. (1989). O26. D. O103. Enrichment methods are currently being developed by researchers and consequently there are no standard protocols to recommend. Some workers have recommended the use of "Enterohaemolysin agar" sheep blood agar for detection of VTEC. 8-9. Captivate™ Description O145 NEW CAP 6 For more information please contact: Dr. this group does not appear to have any common distinct biochemical properties. J. M. H. S. Prices will be calculated on an individual basis.S. Non-O157:H7 Shiga toxin (verocytotoxin)-producing Escherichia coli strains: epidemiological significance and microbiological diagnosis. SMAC-BCIG (HAL 6) has also been used for this application as it contains the glucuronide chromogen. Illingworth. coli O157:H7. Tel: +44 (0)161 797 5729 Fax: +44 (0)161 762 9322 Email: sillingworth@idgplc.

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5 Stannard. Suitable for use with LAB 90 Fastidious Anaerobe Agar. For this reason any unused. Add aseptically to sterilised medium cooled to 47˚C together with other additives. Add aseptically to sterilised medium cooled to 47˚C.T. molten medium. Once rehydrated the stability of antibiotics varies greatly and will determine the shelf life of the prepared agars and broths. A.E. Clin. from clinical material.W. and other anaerobes. When used with other blood agar bases. rehydrated.5. The shelf life of supplemented media is governed by the stability of the added components. Larger and smaller volumes are indicated for relevant products. London. The shelf life of freeze-dried supplements is 2-3 years provided they are stored in a refrigerator at 2-8˚C. Clin. To ensure the correct level of selective supplements the entire vial contents must be added to the stated volume of cooled. 5. J. followed by immediate disposal of the syringe into an approved container. mg/litre 10 30 Addition Most antibiotics are heat labile. withdrawn and added to the medium in a single process.W. Add aseptically to sterilised medium cooled to 47˚C together with other additives. and so to prevent a reduction of potency the medium should be cooled to 47˚C by holding in a water bath set at this temperature. as even deep-freezing may not prevent the rapid degradation of the antibiotics. most Bacteriodes fragilis strains and some anaerobic cocci. Add aseptically to sterilised medium cooled to 47˚C together with other additives. VANCOMYCIN for the isolation of Gram negative anaerobes from clinical material. mix gently and pour. The Tween® 80 may be added before sterilisation at a concentration of 0. When added to blood agar the resulting medium will allow the growth of clostridia. Path. X290 NALIDIXIC ACID. menadione and sodium pyruvate is beneficial.D.A. Multiple Selective Media for the isolation of anaerobic bacteria. Personal Communication. Reference: Anaerobes X090. Suitable for use with LAB 90 Fastidious Anaerobe Agar. Path. rehydrate the supplement using a sterile pipette. to the medium is required for N. For information on the shelf life of prepared media consult the individual product listings in the previous section of the manual. supplement should be discarded. mg/litre 10 2. Reference: Wren. and for the majority of the supplements each vial is sufficient for 500ml of medium. medium. Selective Supplements Presentation and Shelf Life LAB M lyophilised supplements are presented in packs of 10 vials. Final Concentration Nalidixic acid Add 1 vial X091 to 500ml medium Add 1 vial X291 to 1 litre medium mg/litre 10 Rehydration Vials should be rehydrated aseptically using a sterile needle and syringe charged with 5ml of the specified diluent for the particular supplement being added. Final Concentration Nalidixic acid Vancomycin Add 1 vial X090 to 500ml medium Add 1 vial X290 to 1 litre medium Rehydrate contents of vial with 5ml sterile deionised water. Final Concentration Metronidazole Nalidixic acid Add 1 vial X092 to 500ml medium. the rubber stopper may be completely removed and. When used with other blood agar bases. e. X092 METRONIDAZOLE. 33: 61-65. Failure to do this will result in a range of concentrations in the plates/bottles and consequent inconsistency in results. LAB 1 Columbia Agar. mix gently and pour.D. J.. X215 NEOMYCIN 75 for the isolation of Clostridium spp. mix gently and pour. Under no circumstances attempt to re-sheath an exposed needle. If sterile needles and syringes are not readily available. enhances the growth of anaerobic cocci. e...g. Rehydrate contents of each vial with 5ml sterile deionised water. M. mix gently and pour. further enrichment of the medium with haemin and menadione is beneficial. which. The metronidazole will suppress the growth of most other anaerobes. 1980. The supplement should be rehydrated. The addition of Tween® 80. X291 NALIDIXIC ACID for the isolation of non-sporing anaerobes from clinical material. X015. further enrichment of the medium with haemin. Once the supplement has been added the medium must be gently but thoroughly mixed to ensure that the selective agents are evenly distributed. using careful aseptic technique. X091. LAB 1 Columbia Agar.g. 33: 61-65. NALIDIXIC ACID for the isolation of Actinomyces spp. Final Concentration Neomycin Add 1 vial X015 to 500ml medium Add 1 vial X215 to 1 litre medium Reconstitute each vial by the addition of 5ml of sterile deionised water. M. Multiple Selective Media for the isolation of anaerobic bacteria. mg/litre 75 Reference: Wren.1%. and is generally shorter than unsupplemented agars and broths. 1980.1 . Suitable for use with LAB 90 Fastidious Anaerobe Agar. Rehydrate contents of each vial with 5ml sterile deionised water. National Hospital for Nervous Diseases.

(1971). Reassessment of Selective Agars and Filtration Techniques for Isolation of Campylobacter Species from Faeces.. References: Lowbury. A selective plate medium for Cl. Final Concentration Polymyxin B Add 1 vial X193 to 500ml medium Rehydrate contents of vial by the addition of 5ml of sterile deionised water. Eur. welchi. J. Suitable for the preparation of LAB 73 Bacillus cereus Medium (P.X016 NEOMYCIN 100 for the selective isolation of Clostridium spp. A. mix well and pour. supplemented with lysed horse blood.J.M. Egg Yolk Emulsion.2 . Final Concentration Kanamycin Add 1 vial X018 to 500ml medium Reconstitution as X015. C. & Bact.F. Microbiol.L. D. Final Concentration Neomycin Add 1 vial X016 to 500ml medium Reconstitution as X015. Spore Research ed. PEMBA Bacillus cereus Medium. X212 CEFOPERAZONE. J.000i. For addition to LAB 193. along with other additives. Ed.. AMPHOTERICIN for the isolation of Campylobacter spp. (1985). Collee. Belmont.. Bacillus cereus X074 POLYMYXIN for the isolation of B. The addition of X073 sterile egg yolk emulsion is also required. Wadsworth Anaerobic Bacteriology Manual 4 ed. Add aseptically to sterilised medium cooled to 47˚C together with egg yolk emulsion. 7: 155-160.S. Citron.R. Add aseptically to sterilised medium cooled to 47°C. and other anaerobes. Finegold. Watt.E. published by Univ. Path. Wren. cereus from foods.N. D. F. Clin. Reference: Micro-organisms in Food. 5.L.. Cefoperazone Amphotericin Add 1 vial X112 to 500ml medium Add 1 vial X212 to 1 litre medium Rehydrate contents of vial with 5ml of sterile deionised water. Reference: Bolton. Path. Clarke. M. Barkeer. Add aseptically to sterilised medium cooled to 47˚C. Infect.A. mix gently and pour.. mg/litre 10 2500 iu/litre 5 Add 1 vial X074 to 500ml medium Rehydrate contents of vial with 5ml of sterile deionised water.. S.W. F. Sutter. TRIMETHOPRIM.. 33: 61-65.. (1980).J. Thatcher. H. D. 70: 105. of Toronto Press.u/litre X214 VANCOMYCIN. Multiple selective media for the isolation of anaerobic bacteria from clinical specimens. environmental and food samples. (1977) British Medical Journal 2 11-9. Final Concentration Vancomycin Polymyxin Trimethoprim Add 1 vial of X214 to 1 litre of medium Rehydrate contents of vial with 5ml sterile deionised water. M. from clinical. Reference: Skirrow.B. Suitable for use with LAB 112 Campylobacter Selective Medium (blood free) or with blood agar media. N. mix gently and pour. V.P. (1988). (1955).G. The addition of X073. J. Clin. J. mg/litre 100 NEW X193 POLYMYXIN B for the isolation of Bacillus cereus from foods. When added to egg yolk medium this supplement will allow the growth of clostridia whilst inhibiting other lecithinase producing organisms. to make Skirrow’s medium for the isolation of Campylobacter spp. Final Concentration Polymyxin B 8mg/litre = 64. Kanamycin is more inhibitory to anaerobic cocci. Dis. B. mix gently and pour.D.).000 IU X018 KANAMYCIN 75 for the selective isolation of Clostridium spp.C. POLYMYXIN. is also required.M. California. X016. Add aseptically to sterilised medium cooled to 47˚C. Parker. G. M. Changing approaches to the sporing anaerobes in medical microbiology. mg/litre 100. Edelstein. Final Concentration mg/litre 32 10 mg/litre 75 Campylobacter species X112. Incubation at 37˚C gives better results than at 42˚C and is generally more convenient. An alternative to X015. Hutchinson. Lilly.A. Suitable for use with LAB001 Columbia Agar or other blood agar bases. Star publishers.

Add aseptically to sterilised medium cooled to 47°C.M. mix gently and dispense. Suitable for use with LAB 90 Fastidious Anaerobe Agar. Final Concentration D-Cycloserine Cefoxitin Add 1 vial X093 to 500ml medium Rehydrate contents of vial with 5ml of water.5 10. V. Campylobacter Enrichment Broth... Preston.C. Add aseptically to sterilised medium cooled to 47˚C. Rehydrate contents of vial by the addition of 5mls of sterile deionised water. Developed for use with LAB 135 Campylobacter Enrichment Broth. Final Concentration Cefoperazone Vancomycin Trimethoprim Natamycin Add 1 vial X132 to 500ml medium Rehydrate contents of vial by the addition of 5ml of sterile deionised water.P.H. For use with LAB 109 Perfringens agar to prepare O. Rehydrate contents of vial with 5ml sterile deionised water. environmental and clinical samples.V. VANCOMYCIN.000 i. CEFOXITIN for the isolation of Clostridium difficile from clinical materials. Selective and differential medium for isolation of Clostridium difficile. Gives higher isolation rates than Preston broth and does not require modified atmosphere incubation.. NATAMYCIN. mix gently and pour. Add aseptically to sterilised medium cooled to 47˚C together with other additives. For the isolation of Campylobacter spp./litre Add 1 vial X109 and 1 vial X110 to 500ml medium Rehydrate contents of vials with 5ml of sterile deionised water. 559-570.J. POLYMYXIN (X110). Reference: Handford.. W. OLEANDOMYCIN PHOSPHATE. D.M. CYCLOHEXIMIDE. For use with LAB 194.V. For the addition to LAB165 O157 Broth MTSB Final concentration mg/litre 20 Clostridium difficile X093 CYCLOSERINE.3 .T. CEFOPERAZONE.N. For addition to LAB 135. Bact. Sutter. TRIMETHOPRIM. mix gently and dispense into sterile containers. VANCOMYCIN. 37.T. mg/litre 250 8 Novobiocin Add 1 vial of X150 to 500ml of O157 Broth MTSB. P.L. X110 SULPHADIAZINE (X109). mg/litre 20 20 20 25 Escherichia coli X150 NOVOBIOCIN for the enrichment of E. Add aseptically to sterilised medium cooled to 47°C.. coli O157:H7 from food.P.u. Finegold. Reference: Bolton. S. Reference: George. 5. J.S.S. Perfringens Agar Base (TSC). mg/litre 400 C. mg/litre 20 20 20 50 Clostridium perfringens X109. (1976). mix well and pour. for the selective isolation of Clostridium perfringens from foodstuffs. Personal communication. P.L. from food and environmental samples by the enrichment broth technique. (1974). Add aseptically to sterilised medium cooled to 47˚C. CEFOPERAZONE. mix gently and pour. An alternative natamycin based supplement for the selective enrichment broth culture of Campylobacter spp. F. Final Concentration Cefoperazone Vancomycin Trimethoprim Cycloheximide Add 1 vial X131 to 500ml medium Rehydrate contents of vial with 5ml of sterile 50% alcohol. NEW X194 NEW X132 D-CYCLOSERINE supplement for the isolation of Clostridium perfringens from foods. Final Concentration D-Cycloserine Add 1 vial of X194 to 500mls medium.L. Add aseptically to sterilised medium cooled to 47˚C. Appl.X131 C. mix gently and pour. Final Concentration Sulphadiazine Oleandomycin Polymyxin mg/litre 100 0. TRIMETHOPRIM. Citron. (1989).

Clin. Add aseptically to sterilised medium cooled to 47˚C. CEFSULODIN.N. Path.L. Haemophilus influenzae X260 BACITRACIN for the isolation of Haemophilus influenzae. Drakeford. NALIDIXIC ACID for the isolation of G. NALIDIXIC ACID for the preparation of Columbia C. “A new culture medium for medical bacteriology. coli 0157:H7 from food and other samples. together with any other additives. Microbiol.4 . mix gently and pour. 5. Suitable for addition to LAB 1 Columbia Agar or LAB 15 Blood Agar Base No. mix well and pour. (1966). vaginalis from clinical material. C. Add aseptically to sterilised medium with heated blood cooled to 47˚C.0 0.D. Final Concentration Bacitracin mg/litre 75 Gardnerella vaginalis X011 COLISTIN. mix well and pour. Final Concentration Colistin Nalidixic acid Add 1 vial X011 to 500ml medium Rehydrate contents of vial with 5ml sterile deionised water. J. Vasi. Add aseptically to sterilised medium cooled to 47˚C. medium. mg/litre 8. for the isolation of Helicobacter pylori.5 Gram Positive Cocci X012 COLISTIN. F. E. J. together with any other additives. X546 V.I. environmental and clinical samples. Nalidixic Acid Agar. For addition to Helicobacter pylori medium LAB 140 Final Concentration Cefsulodin Vancomycin mg/litre 10 10 20 Reference: Goldberg. Washington. mix gently and pour.C.0 Reference: Ellner.” Amer.25 litres of LAB 46 Rehydrate the contents of one vial with 20ml of sterile deionised water. mg/litre 10 15 Add 1 vial of X260 to 1 litre of medium. along with other additives. Add aseptically to sterilised medium cooled to 47˚C. Amphotericin Add 1 vial to 500ml medium Rehydrate contents of vial with 5ml sterile deionised water. Final Concentration Colistin Nalidixic acid Add 1 vial X012 to 500ml medium mg/litre 10 10 Add 1 vial of X161 to 500 ml of Sorbitol MacConkey Agar (SMAC) or HAL 6 (BCIG-SMAC) Rehydrate contents of vial with 5ml sterile deionised water. Supplement for the selective enrichment of E. For the addition to LAB 161 Sorbitol MacConkey Agar (SMAC) or HAL 6 (BCIG-SMAC). Add aseptically to sterilised medium cooled to 47˚C. Add aseptically to sterilised medium cooled to 47˚C. 4(3): 245.A. Clin. Rehydrate contents of vial with 5ml sterile deionised water. Stossel. AMPHOTERICIN. Mix well and dispense into 225ml aliquots.. Suitable for use with Columbia blood agar base and other blood agars supplemented with heated (“chocolated”) blood. “Comparison of Isolation of Haemophilus vaginalis (Corynebacterium vaginalae) from Peptone-Starch-Dextrose Agar and Columbia.C.05 10. P.05 2.A. 45: 502.. Rehydrate contents of vial with 5ml sterile deionised water. Colistin. 2 to produce a selective isolation medium.X161 CEFIXIME TELLURITE supplement for the isolation of E. A medium selective for Gram positive cocci is obtained when this antibiotic mixture is added to LAB 1 Columbia Agar. II (1976). J. Final concentration Cefixime Potassium tellurite mg/litre 0. coli O157:H7 from food. R... Helicobacter pylori X040 VANCOMYCIN. For use with Buffered Peptone Water LAB 46 Final Concentration Vancomycin Cefixime Cefsulodin Add 1 vial of X546 to 2. mix well and pour.

(1989). clinical and food samples. Add aseptically to sterilised medium cooled to 47˚C.0 X138 N. (1985). et al. Food Protect. mix gently then pour. For addition to LAB 122 Listeria Isolation Medium or HAL 2 Harlequin™ Listeria Medium. 63: 299-304. supplement for the enrichment and isolation of Listeria spp from food and environmental samples. Add aseptically to sterilised medium cooled to 47˚C. Appl. Listeria monocytogenes in raw milk: detection incidence and pathogenicity. Rapid and automated detection of Salmonella by electrical measurements. X164. (1987). Final Concentration Nalidixic acid Cycloheximide Acriflavine Add 1 vial of X138 to 500ml medium Reconstitute contents of vial by the addition of sterile 50% ethanol in water. Lett.M.M. Final Concentration Cefotetan Cycloheximide Colistin Fosfomycin Acriflavine Add 1 vial of X122 to 500ml of LAB 122.A. For the addition to LAB 144 Palcam Broth and Lab 148 Palcam Agar Listeria X122 C. Some modifications to the media for rapid automated detection of salmonellas by conductance. J. to T. Add aseptically to sterilised medium cooled to 47˚C. X144 P. Final Concentration T. I.C. Odgen.C. mix gently then pour. A selective differential medium for the isolation of Listeria monocytogenes.A. (1987). Swirl to mix then dispense into sterile containers.A.O.A.5 10 Reference: Curtis. for Listeria isolation from food and environmental samples and LAB 139 Buffered Listeria Enrichment Broth. NALIDIXIC ACID.C. (Trimethylamine-N-oxide) Sodium biselenite Add 1 vial X137 to 100ml medium Reconstitute contents with 5ml of sterile deionised water.F. Gibson. 94: 245-262.A. D.. for the isolation of Listeria monocytogenes from environmental.M. The growth of Salmonella in the medium reduces T.O. FOSFOMYCIN. D.M.A. mg/litre 40 50 15 References: Easter. For addition to LAB 164 Fraser Broth Base Final Concentration Ferric ammonium citrate Acriflavine Nalidixic acid mg/litre 500 12. and in so doing.A. CYCLOHEXIMIDE for the selective enrichment broth culture of Listeria monocytogenes. 63: 359-464. along with other additives. For addition to LAB 138 Listeria Enrichment Broth recommended by the F. Bacteriol.M. A modified conductance medium for the detection of Salmonella spp.O. X564 1/2 FRASER supplement for the primary enrichment of Listeria spp from food and environmental samples.C. J.. mix well and pour. D. Selenite for inclusion in Easter and Gibson Salmonella Detection Medium LAB 137. Add 1 vial of X164 to 450ml of Fraser Broth Base Add 1 vial of X564 to 2. CEFOTETAN. The incorporation of sodium biselenite makes the medium selective for salmonellae. Reconstitute contents of vial by the addition of sterile 50% ethanol in water. Add aseptically to sterilised medium cooled to 47˚C. (1987). M. ACRIFLAVINE. Bacteriol. 50: 188-192.D. 8: 95-98.C. g/litre 5. Reference: Lovett et al.0 4. 5. significantly increases the conductivity of the medium.25 litres of Fraser Broth Base Rehydrate contents of vial with 2ml 50% methanol (5ml for X564).M. in Appl. Hyg.5 . H. Add aseptically to sterilised medium cooled to 47˚C.C.D. Microbiol. Add 1 vial of X122 to 1 litre of HAL 2. Cann. CYCLOHEXIMIDE. Appl. mg/litre 2 400 20 10 5 Final concentration Polymyxin Acriflavine Ceftazidime mg/litre 10 5 20 Add 1 vial of X144 to 500ml of Palcam Broth or Agar Rehydrate contents of vial with 5ml sterile deionised water. J. COLISTIN. ACRIFLAVINE.Impedance Microbiology X137 T.A. mix well and pour. Gibson.

Add 1 vial X072N and 1 vial of X072 to 500ml medium. ACRIFLAVINE. Final Concentration Nalidixic acid Acriflavine Natamycin Add 1 vial of X139 to 500ml medium. Add aseptically to sterilised medium cooled to 47°C. mix gently and pour. For addition to LAB 122 Listeria Isolation Medium. X155. mix well and pour NEW X072N NEW X123 COLISTIN. Add aseptically to sterilised medium cooled to 47˚C. Add 1 vial of X539 to 2.X539 N. For addition to LAB 172. Final Concentration Cefotetan Natamycin Colistin Fosfomycin Acriflavine Add 1 vial X123 to 500ml medium Final Concentration Ferric ammonium citrate Acriflavine Nalidixic acid mg/litre 500 25 20 NEW mg/litre 2 25 20 10 5 X165 FRASER supplement for the secondary enrichment of Listeria spp from food and environmental samples.N.25 L medium. For addition to LAB 172. Rehydrate contents of vial by the addition of 5 ml of sterile deionised water.NEW X072 X156 UVMII. Add 1 vial of X156 to 500ml of UVM Broth Base Rehydrate contents of vial with 5ml sterile deionised water. mix well and pour. Rehydrate contents of vial by the addition of 5ml of sterile deionised water (10ml for X539). Add 1 vial of X165 to 500ml of Fraser Broth Base Rehydrate contents of vial with 2ml 50% methanol.N. CEFOTETAN. Add aseptically to sterilised medium cooled to 47°C. For addition to LAB 164 Fraser Broth Base Rehydrate the contents of vial by the addition of 5ml of sterile deionised water. NALIDIXIC ACID supplement for the isolation of Listeria monocytogenes.A. FOSFOMYCIN. Add aseptically to sterilised medium cooled to 47°C. Rehydrate contents of vial by the addition of 5ml of sterile deionised water. mix gently and pour. mix well and pour. Add aseptically to sterilised medium cooled to 47˚C.F. Add aseptically to sterilised medium cooled to 47˚C.6 . NATAMYCIN.C. Supplement for the primary enrichment of Listeria spp from food and environmental samples. NATAMYCIN. Add aseptically to sterilised medium cooled to 47°C. mg/litre 40 15 25 Add 1 vial of X155 to 500ml of UVM Broth Base Add 1 vial of X555 to 2. CEFTAZIDIME supplement for the isolation of Listeria monocytogenes. An alternative natamycin supplement for the isolation of Listeria spp. For the addition to LAB 155 UVM Broth Base POLYMYXIN B. Supplement for the secondary enrichment of Listeria spp from food and environmental samples.25 litres of UVM Broth Base Rehydrate contents of vial with 5ml sterile deionised water (10ml for X555). clinical and food samples. from environmental. Buffered Listeria Enrichment Broth. X139.A. Listeria Enrichment Broth and LAB139. An alternative natamycin based supplement for the selective enrichment broth culture of Listeria spp. 5. mix gently and pour. mix gently and dispense. For addition to LAB 155 UVM Broth Base Final Concentration Nalidixic acid Acriflavine mg/litre 20 12 For addition to LAB 138. LMBA Final Concentration Nalidixic acid mg/litre 40 C. NALIDIXIC ACID. ACRIFLAVINE. X555 UVMI. LMBA Final Concentration Polymyxin B Ceftazidime mg/litre 10 20 Final Concentration Nalidixic acid Acriflavine mg/litre 20 25 Add 1 vial X072 and 1 vial of X072N to 500ml medium.

T.000 I. For addition to LAB 123 Kirchner’s T.C. Add aseptically to sterilised medium cooled to 47°C.N. P. J. TRIMETHOPRIM for Thayer Martin Medium. H. VANCOMYCIN.T.T.C.0 0. with the amphotericin omitted to permit the growth of yeasts.T.C.A. mix gently and pour. J. (1978). AMPHOTERICIN. For addition to GC agar base LAB 67. agar. Rehydrate contents of vial with 5ml sterile deionised water.C. Add aseptically to sterilised medium cooled to 47°C together with other additives. X069.C.N.Y.7 . 100 10 10 The addition of trimethoprim in V. X271 GROWTH SUPPLEMENT.C. gonorrhoeae because of the emergence of vancomycin sensitive strains.5 Thayer.C.N.A. which occasionally make interpretation difficult.T. Final Concentration L-cystine L-cysteine Thiamine HCl Ferric nitrate Co-Carboxylase NAD Guanine HCl Adenine L-glutamine PABA Vitamin B12 Add 1 vial to 1 litre of medium mg/litre 11 259 0.2 1 1.T. 54: 36-40. POLYMYXIN B. agar but can readily be substituted for V. Dis. Add aseptically to sterilised medium cooled to 47˚C together with other additives. mg/litre 1 6 1 6.A. NYSTATIN. LINCOMYCIN.N. L.T.A.B. Medium.N. Final Concentration Vancomycin Colistin Nystatin Trimethoprim Add 1 vial X068 to 500ml medium Add 1 vial X268 to 1 litre medium Rehydrate contents of vial with 5ml sterile deionised water.T. is often preferred to X068 V.5 12. Concentrations and rehydration as L. Add 1 vial X069 to 500ml medium Add 1 vial X269 to 1 litre medium 5. or V. meningitidis.C. LINCOMYCIN.Mycobacterium tuberculosis NEW X124 X068. mix well and pour. Ven. from clinical material. mg/litre 200. and Martin. (1966).3 10 100 0. Rehydrate contents of vial by the addition of 5 ml of sterile deionised water. COLISTIN.T. for the isolation of N.1 Reference: Young. mg/litre 3 7.A.C. TRIMETHOPRIM for the isolation of Neisseria spp.E. Brit. Cultural Diagnosis of Gonorrhoea with modified N. L. along with other additives.D.T. TRIMETHOPRIM. A variant of L. Improved medium selective for the cultivation of N. Medium. X268 V. gonorrhoeae and N. TICARCILLIN. Public Health rep. COLISTIN. Final Concentration Polymyxin B Ticarcillin Trimethoprim Amphotericin Add 1 vial of X124 to 500ml medium.C.U. X269 L. inhibits the swarming of Proteus spp.T. Journ. 81: 559-562.13 0.C. from selective media. X270 L. is quoted as the selective agent for New York City G.5 5 Reference: Neisseria gonorrhoeae X070. Add aseptically to sterilised medium cooled to 47˚C. to improve the isolation of Neisseria spp.C.A. AMPHOTERICIN supplement for the isolation of Mycobacterium tuberculosis from clinical samples. The antifungal agent amphotericin is more readily soluble and therefore a more active antifungal than nystatin.03 0. mix gently and pour.C. COLISTIN. Final Concentration Lincomycin Colistin Amphotericin Trimethoprim Add 1 vial X070 to 500ml medium Add 1 vial X270 to 1 litre medium Rehydrate contents of vial with 5ml sterile 25% alcohol in water. in Thayer Martin G.T. TRIMETHOPRIM. mix gently and dispense.C.

Pseudomonas species
X108
MODIFIED C.F.C. – CEPHALOTHIN, FUCIDIN, CETRIMIDE for the selective isolation of Pseudomonas spp. When added to LAB 108 Pseudomonas Agar, to prepare C.F.C. medium this supplement can be used to select pseudomonads from food and environmental samples. Final Concentration Cephalothin Fucidin Cetrimide Add 1 vial X108 to 500ml medium Rehydrate contents of vial with 5ml of sterile 50% alcohol. Add aseptically to sterilised medium cooled to 47˚C, mix gently and pour. mg/litre 50 10 10

Pre-Incubation Test (P-INC)
X019, X219
PENICILLIN, NISIN, CRYSTAL VIOLET, for accelerated shelf life determination of dairy products. The Pre-incubation test uses a selective mixture to inhibit Gram positive organisms whilst allowing the growth of Gram negative bacteria, the main cause of post-pasteurisation contamination and a major factor in determining the shelf life of the product. The technique is also useful for monitoring plant hygiene. Final Concentration Penicillin Nisin Crystal violet mg/litre 20,000iu/litre 40,000iu/litre 2.0

Add 1 vial of X019 to 200ml of Milk Agar LAB019 Add 1 vial of X219 to 1 litre of Milk Agar LAB019 Rehydrate contents of 1 vial with 5ml sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C, mix thoroughly and pour plates.

Reference:
Mead, G.C. and Adams, B.W. (1977). Br. Poult. Sci. 18: 661-667

X107
C.N. CETRIMIDE, NALIDIXIC ACID for the isolation of Pseudomonas aeruginosa. Suitable for use with LAB 108 Pseudomonas Agar to make the medium selective for Ps. aeruginosa. Final Concentration Cetrimide Nalidixic acid Add 1 vial X107 to 500ml medium Rehydrate contents of vial with 5ml of sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C, mix gently and pour. mg/litre 200 15

Method A
Pre-incubate test material at 21˚C for 24hr. Prepare suitable dilution series, and inoculate Milk Agar plates containing P-INC supplement. Incubate at 21˚C for 24hr, and count all colonies (some may be small, use of a hand lens is recommended). Calculate the CFU/ml and using the tables of Griffith’s et al the shelf life can be determined.

Method B
Rehydrate X219 with 1ml of deionised water only, add 0.1ml to the test material and incubate at 20˚C for 24hr. Prepare suitable dilution series, and inoculate Milk Agar plates. Proceed as for Method A above.

References:
Griffiths, M.W., and Phillips, J.D. (1985) J.Appl.Bact. 57, 107. Griffiths, M.W., and Phillips, J.D., and Muir, D.D. (1980) J. Soc. Dairy Technol. 33, 8. Griffiths, M.W., and Phillips, J.D., and Muir, D.D. (1981) J. Soc. Dairy Technol. 34, 142. Griffiths, M.W., and Phillips, J.D., and Muir, D.D. (1984) J. Soc. Dairy Technol. 37, 22. Griffiths, M.W., and Phillips, J.D., and Muir, D.D. (1984) Rapid detection of post-pasteurised contamination. Hannah Research Inst. Bulletin No.10. Griffiths, M.W., and Phillips ,J.D., and Muir, D.D. (1984) Dairy Ind. Int. 50 (3) 25 Griffiths, M.W., and Phillips, J.D., and Muir, D.D. (1984) Postpasteurisation contamination - the major cause of failure of fresh dairy products. Hannah Research Inst. Griffiths, M.W., and Phillips, J.D., and Muir, D.D. (1986) Aust. J. Dairy Technol. 41, 77-79.

Reference:
Goto, S., Enomoto, S. 1970. Jap. J. Microbiol. 14: 65-72.

X140
TICARCILLIN, POLYMYXIN, for the isolation of Burkholderia (Pseudomonas) cepacia Suitable for use with LAB 108 pseudomonas selective agar, or specific selective bases such as that described by Gilligan et al. Final Concentration Ticarcillin Polymyxin Add 1 vial to 500ml of medium Rehydrate contents of vial with 5ml sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C, mix well and pour. mg/litre 100 300,000 iu/litre

Reference:
Gilligan, P.H., Gage, P.A., Bradshaw, L.M., Schidlow, D.V., DeCicco, B.T. (1985) Isolation medium for the recovery of Pseudomonas cepacia from respiratory secretions of patients with cystic fibrosis. J.Clin.Microbiol. 22 (1) 5-8.

5.8

Salmonella
X150
NOVOBIOCIN, for the isolation of Salmonella using semi-solid technology. For addition to LAB 150 MSRV and LAB 537 Diassalm Final Concentration Novobiocin Novobiocin Add 1 vial to 500ml (MSRV) Add 1 vial to 1 litre (Diassalm) Rehydrate contents of vial with 5ml sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C, mix well and pour.
NEW

X207
METHICILLIN, for the isolation of Methicillin Resistant S.aureus (MRSA) Suitable for use with LAB 7 Mannitol salt agar. Final Concentration Methicillin Add 1 vial of X207 to 1 litre of medium Rehydrate contents of vial with 5ml sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C, mix well and pour. mg/litre 4

mg/litre 20 (MSRV) 10 (Diassalm)

X192

References:
De Smedt, J.M., and Bolderdijk, R.F., (1986) Dynamics of salmonella isolation with modified semi-solid Rappaport Vassiliadis medium. J.Food Protection 50 658-661 Van Netten, P., Van Der Zee H., and Van Der Moosdijk A., (1991) The use of diagnostic selective semi-solid medium for the isolation of Salsmonella enteritidis from poultry. Proceedings of the 10th symposium on the quality of poultry meat. Spelderholt Beckbergen 56-67.

OXACILLIN, POLYMYXIN B supplement for the isolation of Methicillin Resistant Staphylococcus aureus (MRSA). For addition to LAB 192, ORSIM (Oxcacillin Resistant Staphylococcus Isolation Medium). Final Concentration Oxacillin Polymyxin B Add 1 vial of X192 to 500ml medium. Rehydrate contents of vial by the addition of 5ml of sterile deionised water. Add aseptically to sterilised medium cooled to 47°C, mix gently and dispense. mg/litre 2 25,000 I.U

Staphylococci
X085
EGG YOLK TELLURITE A sterile emulsion of egg yolk and potassium tellurite for use as a selective and differential agent in Baird-Parker Medium Base LAB 85. The complete medium is selective for S.aureus, and the addition of egg yolk tellurite aids differentiation of this organism from others capable of growing on the agar. Presented in 100ml bottles with a tellurite concentration of 0.2% to give a final concentration in the complete medium of 0.01% (w/v). Add 50ml to 1 litre of Baird-Parker Medium Base.

Streptococci
X013
COLISTIN, OXOLINIC ACID for the selective isolation of streptococci from clinical material. When added to LAB 1 Columbia agar or LAB 15 Blood Agar Base No. 2, X013 renders the medium selective for streptococci. Alteration in haemolysis patterns may occur when azide or crystal violet are employed as selective agents but this does not occur with X013. Final Concentration Colistin Oxolinic acid Add 1 vial X013 to 500ml medium mg/litre 10 5

NEW

X086

RPF: BOVINE FIBRINOGEN, RABBIT PLASMA, TRYPSIN INHIBITOR, POTASSIUM TELLURITE supplement for the isolation of Staphylococcus aureus. For addition to LAB 85 Baird-Parker Medium. Final Concentration Bovine Fibrinogen Rabbit Plasma Trypsin Inhibitor Potassium Tellurite Add 1 vial of X086 to 90ml medium. Rehydrate contents of vial by the addition of 10ml of sterile deionised water. Add aseptically to sterilised medium cooled to 47°C, mix gently and pour. mg/litre 0.375 2.5ml 2.5 2.5

Rehydrate contents of vial with 5ml of sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C together with other additives, mix gently and pour.

Reference:
Petts, D. (1984). Colistin - Oxolinic Acid - Blood Agar: a new selective medium for streptococci. J. Clin. Microbiol. 19: 4-7.

5.9

Yeasts and Moulds
X009, X209
CHLORAMPHENICOL for the selective isolation of yeasts and moulds from food, environmental and clinical specimens. Chloramphenicol’s broad antibiotic spectrum suppresses most contaminating bacteria allowing the yeasts and moulds to grow. It can be added to such media as LAB 9 Sabouraud Dextrose Agar, LAB 36 Rose Bengal Chloramphenicol Agar, LAB 37 Malt Extract Agar and LAB 117 Dermatophyte Test Medium to increase their selectivity whilst not lowering the pH. Reduction of pH will increase the selectivity of a yeast and mould medium but will also inhibit some yeasts as well as having a deleterious effect on the agar gel. Final Concentration Chloramphenicol Add 1 vial X009 to 500ml medium Add 1 vial X209 to 1 litre medium Rehydrate contents of vial with 5ml of Ethyl or Methyl alcohol. Add aseptically to sterilised medium cooled to 47˚C, mix gently and pour. mg/litre 100

Yersinia
X120
C.I.N. - CEFSULODIN, IRGASAN, NOVOBIOCIN for the isolation of Yersinia spp. from clinical and environmental material. For addition to LAB 120 Yersinia C.I.N. Agar Base used in the selective isolation of Y. enterocolitica. Final Concentration Cefsulodin Irgasan Novobiocin Add 1 vial X120 to 500ml medium Rehydrate contents of vial with 5ml of 30% sterile alcohol. Add aseptically to sterilised medium cooled to 47˚C, mix gently and pour. mg/litre 15 4 2.5

References:
Schiemann, D.A. (1979). Synthesis of a selective medium of Yersinia enterocolitica. Can. J. Microbiol. 25 (2): 1298. Schiemann, D.A. (1980). Isolation of toxigenic Yersinia enterocolitica from retail pork products. J. Food Prot. 43: 360. Schiemann, D.A. (1982). Development of a two-step enrichment procedure for recovery of Yersinia enterocolitica from food. Appl. Microbiol. 43 (1): 14.

References:
Jervis, B. (1973). Rose Bengal Chlortetracycline agar with other media for the selective isolation and enumeration of moulds and yeasts in foods. J. Appl. Bact. 36 Pages 723-727.

X089
OXYTETRACYCLINE for O.G.Y.E. medium. For use with LAB 89 Oxytetracycline Glucose Yeast Extract Agar for the enumeration of yeasts and moulds from foodstuffs. Highly proteinaceous foods and incubation above 30˚C will inactivate oxytetracycline. Final Concentration Oxytetracycline Add 1 vial X089 to 500ml medium Rehydrate contents of vial with 5ml sterile deionised water. Add aseptically to sterilised medium cooled to 47˚C, mix gently and pour. mg/litre 100

References:
Mossel, D.A.A., et al. (1970). O.G.Y.E. for the selective enumeration of moulds and yeasts in food and clinical material. J. Appl. Bact. 35: 454-457.

5.10

5% Absent < 10 3/g < 2/g Agars A range of agars are offered to suit all microbiological applications. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Total Amino Nitrogen cream/white powder total clear and colourless 6.3 at 340nm >0. Peptones and Extracts Like agars.01% < 0.5-7. Typical Analysis Gel strength (Nikan) Colourimetry (1.5 6. A firm gel is obtained at working concentrations of 1. Growth parameters are obtained by classical microbiological techniques and by automated growth rate analysis.0 ± 0. Koch originally used gelatin to solidify culture media.5-7.28 at 340nm > 0.5% < 0. They provide the amino acids and peptides required by micro-organisms for growth as well as other vital growth factors such as minerals. LAB M can select specific peptones for special purposes such as vaccine production and fermentation processes.5 8. This agar is suitable for all bacteriological purposes including sensitivity testing and pour plate techniques. Agar No.5% soln at 65˚C) 650-1000g/m2 > 0. Acid Hydrolysed Casein MC 7 A soluble protein hyrolysate obtained by digesting casein with hot acid.0 to 1.5% soln at 65˚C) 650-1000g/m2 > 0.1% < 3.5 Typical Analysis Gel strength (Nikan) Colourimetry (1.02% < 1. Chemical and physical properties are also closely monitored. No significant precipitation is observed on reheating or prolonged holding at 65˚C.1% ± 0. Extracts & Other Media Constituents Sourcing The Lab M range of media constituents are selected on the basis of quality and performance from the world’s leading suppliers.5%.01% < 0. It is a deliberate policy not to invest in our own peptone manufacturing facility in order to allow our microbiologist freedom to choose the best ingredients available on the international market. and these qualities make it suitable for use as a protein source in media for antibiotic and vitamin assays.02 at 525nm Melting point Setting point pH Moisture Total ash Calcium Magnesium Sodium chloride Iron Insoluble ash Sulphate Salmonella TVC Spores > 85˚C 32-35˚C 6. 2 MC 6 A bacteriological agar which gives a firm gel at working concentrations of 1. This agar is recommended for all culture media except sensitivity testing media and those where absolute clarity is advantageous. Agars.04 at 525nm Appearance Melting point Setting point pH Moisture Total ash Calcium Magnesium Sodium chloride cream/white powder >85˚C 32-35˚C 6.5% which is reasonably clear.5% < 0.4 < 10% < 3% < 0.3% ± 0. 1 MC 2 A high clarity agar with good gelling properties and a low concentration of metal ions.0% < 0. If more information is required on special services please contact LAB M or your local agent. peptones and extracts are biologically variable products requiring careful selection.6. Peptones.02% < 0.1% 1.4 <12% <3. Iron Insoluble ash Sulphate Salmonella TVC Spores 6.1% <1.0% Absent < 10 3/g < 2/g Agar No. vitamins and antagonists. It is almost free from growth factors. vitamins and nucleic acid fractions. Careful selection of agars is vital as they can interact with nutrient components in a beneficial or deleterious manner.0 to 1. but the superior properties of agar resulted in its universal adoption as the gelling agent of choice.1 .0% < 0. To ensure we use only the best available peptones and extracts these materials are exhaustively tested.

4 has been selected specifically for use as a gelling agent in plant tissue culture techniques.5% 88-91˚C 32-33˚C 80 7. a parameter of particular importance for this application.2 12. light brown colour 7. Beef extract can be used as a direct replacement for meat peptones and.0% ± 0.5 pH of 2% solution Total Nitrogen Amino Nitrogen Bile Salts No.1.6% ± 0. and then tested to ensure it meets the parameters set by a major plant producer. allowing maximum selection of organisms of enteric origin at relatively low concentrations (0. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear. pale straw colour 7. MacConkey Agar No. This fraction of bile is highly active.2 ± 0.5 LAB M Agar No.0 ± 0. as it contains no carbohydrates. Typical analysis Ash Acid Insoluble Ash Calcium Magnesium Iron Total Nitrogen Recommended Concentration Melting Point Setting Point Mesh pH (1.018% 0.5% at 20˚C) Gel Strength (1.5 Bacteriological Peptone MC 24 An economical source of nutrients provided by a balanced mixture of meat peptones and tryptone. Typical Analysis Appearance Solubility in water at 5% Clarity light brown powder total clear. 1 MC 4 MC 29 A rich mixture of tryptone and meat peptones which fulfills the nutritional demands of a wide variety of micro-organisms.30% 0.5 5% ± 0.5 6. comprising mainly sodium cholate and sodium desoxycholate.5 1. can be used as a component of media for fermentation studies. It is used as a selective agent in culture media such as Violet Red Bile Agar for the enumeration of coliforms.2 ± 0.1% ± 0.Agar No.4 Plant Tissue Culture Grade Balanced Peptone No.0 ± 0.2 >700g/cm2 Total Nitrogen Amino Nitrogen beige powder total clear.2 12.16% 0. 3 for the isolation and differentiation of enteric organisms. peptides and proteoses in this mixture. pale straw colour 7.8% ± 0. The growth requirements of most non fastidious organisms will be fulfilled by the range of amino acids. The product is selected primarily on gel strength. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution 2.5 5.2 12% ± 0.31% 0. This peptone is used in many LAB M culture media formulations. 3 MC 25 A refined bile salt.5% W/V) Beef Extract MC 19 This complements the nutritive properties of peptones in culture media and is often used as an added enrichment. as well as customer’s own formulations. The agar contains no nutrients for plant growth and is designed to be incorporated into classical formulations such as Murashige and Skoog.12% 0. Typical Analysis Appearance Solubility in water at 2% Clarity pH of a 2% solution white powder total clear 8.15% 0.15%).2 .75 . and SS Agar for the isolation of enteric pathogens.0 ± 0.

as well as yeasts and filamentous moulds.0 ± 0. Lactalbumin hydrolysate is a pancreatic digest of these proteins. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen light brown powder total clear.0 .0 9.5.3 11. pale straw colour 7.13.14. Description A collagenous protein used for the solidification of culture media and for the detection and differentiation of certain proteolytic bacteria.5 5.5 . Due to its capacity to stimulate sugar metabolism in saccharolytic organisms it is perfectly suited for the growth of a broad range of organisms such as Clostridium. 6. Lactalbumin Hydrolysate NEW MC40 FMV (Foot and Mouth Vaccine) Peptones NEW Description Lactalbumin is a protein removed from the whey.2 10.5 .2% MC402 Description IPTG is used as an inducer of the lac Z operon.0 . Other uses include growth of lactobacilli.0 ± 0. straw colour 7.7.5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen Lactose NEW Gelatine NEW MC20 Description MC15 Lactose for use in microbiological culture media. This combination of compounds is especially useful in transformation experiments involving Escherichia coli where disruption of the lac Z operon is used as a marker for DNA insertion. Typical Analysis: Appearance Solubility in water at 2% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear. meat and milk protein hydrolysates developed to provide the nutritional requirements for a wide range of organisms.5% 2.5% 4.0 .5 ± 0.Columbia Peptone NEW MC12 IPTG (Isopropyl thiogalactopyranoside) dioxane free NEW Description A blend of plant. homo and hetero-fermentative Lactic acid bacteria. Typical analysis: Appearance Solubility in water at 5% cream powder total clear.2 – 6. enhances the colour development of organisms capable of fermenting lactose. Typical Analysis: Appearance pH buff crystalline powder 5. clostridial spores and certain fermentation procedures. Typical analysis: Appearance Solubility in water at 2% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear. or equivalent chromogenic compound. Incorporation of this compound into media containing X-β-galactoside (MC405).2 12. Bacillus. left after removal of casein from milk. It can be used for tissue culture media and for production of vaccines of viral origin.5 MC33 Description A special blend of peptones developed for use in the production of Foot and Mouth vaccine.7 .7. brown colour 6.6.11.0% ± 0.0% Glucose (Dextrose) NEW MC13 Description Glucose for use in microbiological culture media. containing high levels of essential amino acids. pale straw colour 6.3 .7 Liver Digest MC 34 Liver digest is prepared by the controlled hydrolysis of liver. It is rich in vitamins and essential amino acids and has excellent nutritional properties and especially favours the growth of strict anaerobes and other fastidious microorganisms.2 .2% 4. Leuconostoc.

coli. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Maltose Other Carbohydrates Protein yellow/brown powder total clear.Malt Extract MC 23 A water soluble extract of malted barley suitable for use in the cultivation of yeasts and moulds. Mannitol NEW Amino Nitrogen MC14 Description D-Mannitol for use in microbiological culture media. pale straw colour 5.5 1.3 11. salmonellae. light straw colour 7. such as Brilliant Green Bile 2% Broth (LAB 51).4 .1 13% ± 0. Ox Bile NEW MC10 Meat Peptone NEW Description MC18 Ox bile is a dehydrated. light brown colour 5. Bacterial growth is inhibited by the low pH of this peptone.0% ± 0. MUG can be incorporated into a range of culture media to enhance detection of E.0% Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear.0 . Haemophilus and Pasteurella species.2 ± 0.2 12.5 .7 .5 Proteose Peptone A MC 11 An enzymatic digest of meat adapted to encourage the production of toxins by Corynebacterium diphtheriae. This product is available in 1g amounts.5 5.13. staphylococci.4 ± 0. and clostridia.5. Malt extract has a very high carbohydrate content and consequently is very sensitive to over heating which will cause a darkening of the medium. Description A highly nutritious enzymatic digest of meat for use in microbiological culture media Typical Analysis: Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear.5 Description Maltose for use in microbiological culture media. 6. purified fresh bile used as a selective agent in bile media. Maltose Monohydrate NEW Typical Analysis MC22 Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen beige powder total clear. pale straw colour 6. MUG reagent is cleaved by the enzyme glucuronidase to release fluorescent 4-methylumbelliferone that can be detected under long wave UV light (366nm) as a blue/green fluorescence. Mycological Peptone MC 9 A mixture of peptones with a high carbohydrate content suitable for the rapid growth and colonial development of yeasts and moulds.7.2 55% 40% 5% MUG (4-methylumbelliferyl-β-Dglucuronide) NEW MC406 Description MUG is a fluorogenic compound used for the specific detection of E. coli in bacteriological culture media. This peptone is highly nutritious and suitable for use in culture media for fastidious organisms such as Neisseria.4% ± 0.8% ± 0.0% 2.0 ± 0.

5 total <5% <20 ppm <2% MC 5 An enzymatic hydrolysate of casein.7 .3 ± 0.13. Typical Analysis Appearance pH of 2% solution Solubility in water at 2% Moisture Heavy metals Sodium cholate white powder 8.0 12. pale straw colour 6. and that this could be enhanced by the removal of magnesium ions by chelating with sodium citrate. These components comprise the selective agents in DCA. DCA (Hynes) and DCLS. this peptone is a rich source of nutrients with a high carbohydrate content.5 .6% ± 0.2 9.2 ± 0. high quality peptone providing superior growth characteristics.5 1. Irritating to eyes. Typical Analysis Appearance Clarity Total Nitrogen Lactose white powder opaque white suspension 5.0% ± 0.5 4.0% ± 0.5 48. respiratory system and skin.9% ± 0. Special tryptone is a high quality source of peptides produced by enzymatic digest of casein. derived from deconjugated bile salts. It is a uniform.3% ± 0. Wear suitable protective clothing and eye/face protection. with very high solubility and clarity in solution. This peptone can be utilised by most bacteria as a growth substrate. Typical Analysis: Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear.5 Soy Peptone MC 3 Prepared using the enzyme papain to digest soyabean meal. drench with water.5 13. rich in peptones and amino acids (including tryptophane). Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen cream powder total clear.5 .5 . Obtain immediate medical attention.5 6.P.S. Recommended working concentration 10%. IRRITANT Keep away from oxidising agents.0% ± 0. Used in Milk Plate Count Agar LAB 115 and in media for diagnostic tests involving the digestion or coagulation of casein and the fermentation of lactose.7. For eyes give prolonged irrigation with water.5 Sodium Chloride NEW Amino Nitrogen MC17 Description Sodium chloride for use in microbiological culture media. Special tryptone is a refined hydrolysate. Special Tryptone NEW MC35 Description Sodium Thioglycollate NEW MC16 Description Sodium thioglycollate for use in bacteriological culture media.5% 5. This peptone conforms to the U. requirements for a pancreatic digest of casein. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen cream powder total clear. straw colour 7. It is recommended for laboratory media and fermentation. It is used to lower the oxidation-reduction potential of the medium and for the neutralisation of mercurial compound preservatives. Leifson showed that desoxycholic acid had the most inhibitory effect on bacterial growth. Most organisms will grow rapidly in this peptone but some bacteria will produce high levels of acid leading to auto-sterilisation unless an adequate buffering system is incorporated.Skim Milk Powder MC 27 A bacteriological grade of thermophile free spray dried skim milk. pale straw colour 7.1 ± 0.7.0% Tryptone Sodium Desoxycholate MC26 Sodium desoxycholate is a specific bile acid. especially where high clarity of solution is required. In case of contact.

Xylose NEW MC32 Description Xylose for use in microbiological culture media.2 ± 0. peptides.Tryptose MC 8 A blend of peptones suitable for the cultivation of most fastidious organisms including Neisseria gonorrhoeae. In combination with IPTG (MC402).6 . vitamins and carbohydrates making it suitable for many applications. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen yellow powder total clear.0 ± 0.2 12.5 4. X-gal can be used in transformation experiments involving Escherichia coli where disruption of the lac Z operon is used as a marker for DNA insertion.5 X-β-Galactoside (X-gal) NEW MC405 Description X-β-galactoside (X-gal or 5-Bromo-4-chloro-3-indolyl-β-Dgalactoside) is used as a chromogenic substrate for the detection of organisms capable of fermenting lactose (lac Z positive organisms). especially where rapid or profuse growth is required such as in blood culture media and blood agars.2 10. This extract provides a mixture of amino acids.5 Yeast Extract Powder MC 1 Prepared by the autolysis of Saccharomyces cerevisae under thermostatically controlled conditions to protect the B vitamins and other heat labile constituents.3% ± 0. 6.5% ± 0.9% ± 0. light straw colour 7.5 5. Streptococcus milleri and Brucella spp.5% ± 0. Typical Analysis Appearance Solubility in water at 5% Clarity pH of 2% solution Total Nitrogen Amino Nitrogen beige powder total clear. pale yellow 7.

8% 37.3 138.1 44.7 .0 58.5 29.0 82.6 52.1 51.7 22.6 77.0 8.3% 50.2 55.0 9.6 50.3 9.9 2.0 75.0 34.8 41.3 22.7 3.7 49.0 14.4% 11.2 10.1 15.0 65.7 27.4 35.38 52.3 8.0 46.1 19.1% 73.1 40.8 36.0 _ 62.0 21.5% 5.9 27.2 15. 1 12.1% 40.0 52.3 55.2 26.7 31.2 74.0 26.0 37. Total Amino Acid Assay (mg/g) Lysine Histidine Arginine Aspartic Acid Threonine Serine Glutamic Acid Proline Glycine Alanine Cystine Valine Methionine Isoleucine Leucine Tyrosine Phenylalanine Tryptophane 53.3 19.4% 1.3 27.0 3.8 22.6-1.0 1.6 26.1% MC 11 Proteose Peptone A 12.5 18.3 23.0 73.9 7.0 110.7 22.0 32.5 62.5 13.9 49.5 30.7 17.4 6.6 63.0% MC 3 Soy Peptone MC 5 Tryptone MC 8 Tryptose MC 1 Yeast Extract 10.0% 5.0% 41.8% 17.1 1.9 17.3 23.0 9.5 105.8 31.0 40.7% MC 4 Balanced Peptone No.0 34.8 38.2 59.7 55.7 32.1 74.0 6.0 25.0 12.6 23.9 61.9 12.8% 4.0% 5.0 27.0 73.0 29.2 4.9 12.0 33.5 38.7 79.5 83.6 30.2 12.1 5.5 1.2 1.2 11.0 45.1 16.0 12.0 69.8 4.6 34.3% 6.9% 38.5 14.7 6.8 20.6 43.5% 5.0 58.8 65.3 26.7% 4.9 30.4 21.5 71.0-9.0 28.7 42.2 39.0 130.6% 1.0 32.9 2.4 27.1 45.1 23.0 160.0 45.5 36.0 69.7% 6.2 37.6 83.3 161.2 37.0 40.4% 9.4 31.4 12.9 48.0 1.0 73.2 25.2 60.3 184.0 25.1 5.1 35.2 100.0 51.0 39.0 39.9 38.5% 12.1 43.LAB M Peptones TYPICAL ANALYSIS MC 7 MC24 Acid Hydrolysed Bacteriological Casein Peptone Total Nitrogen Total AminoNitrogen Amino N/Total N.9 50.0 33.2 103.8% MC 9 Mycological Peptone 12.0 44.0 52.0 8.7 26.0 12.0 18.6 16.8% 48.7% 12.5 74.0 22.4% 12.

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Description:(100x5ml) A modified version of Stuarts’s transport medium. All sterile reagents should be stored at 2-8ºC away from light. Cooked Meat with Nutrient Broth P002 Description:(100x10ml) A medium for the growth of a wide range of organisms especially anaerobic bacteria. Resuscitation Broth P081 Description:(100x10ml) A general medium for the resuscitation of microorganisms. aseptic preparation. e.1 . food and sewage. which will not affect the performance of the product. Trichomonas Medium P018 Description:(100x8ml) A medium for the cultivation of Trichomonas vaginalis and Candida spp. Transport Medium (Amies Charcoal) P013 Nutrient Agar Slopes P007 Description:(100x3ml) A medium for the short term maintenance and transport of a wide range of organisms. 7. Cooked Meat with FAB P069 Description:(100x10ml) A culture medium useful for the recovery of fastidious anaerobic bacteria.7. Each product has been prepared with an appropriate sterilisation procedure. Selenite Broth P011E Description:(34x10ml) A medium for the selective enrichment of salmonellae from faeces. filtration or irradiation. The ready prepared media offer a cost effective alternative to preparation of large quantities of small volume dispensed media. Serum Broth P017 Description:(100x4ml) Can be used as a basal medium for sugar fermentation tests for Neisseria. Sterile Additives and Ready Prepared Media Availability Sterile additive products are offered in ready-to-use format. Peptone Water P009 Description:(100x4ml) A general purpose growth medium.g. Nutrient Broth P008 Description:(100x4ml) A medium for the growth of a wide range of organisms. This medium is also useful for prolonged storage of cultures and for the growth of a wide range of organisms. Urea Agar Slope P015 Description:(100x3ml) A medium for the detection of urease activity by Proteus spp. Selenite Cystine Broth P012E Description:(34x10ml) A modification of the original selenite medium for the isolation of salmonellae.

Egg Yolk Emulsion
X073
Description A sterile emulsion of egg yolks for use in bacteriological culture media. It may be added directly to nutrient media for the identification of Clostridium, Bacillus and Staphylococcus species by their lipase and/or lecithinase activity. Presented in 100ml bottles, add 100ml to 900ml of Bacillus cereus medium, or 50ml to Blood Agar Base LAB 28 containing Fildes extract and serum. Technique For detection of lecithinase activity (especially in the investigation of `bitty cream' conditions) add 0.5 or 1.0ml of the emulsion to 10ml of sterile Blood Agar Base (LAB 28) or Nutrient Broth No.2 (LAB 14). In order to clear the medium, raise the final salt concentration by the addition of 1% of sodium chloride. After incubation for up to 5 days at 35ºC, lecithinase-producers render the broth opalescent, whilst, on the solid medium, the colonies are surrounded by opaque zones.

Potassium Tellurite Solution 3.5%
X027
Description A sterile solution of 3.5% potassium tellurite. A selective agent for addition to Hoyles’s Medium (LAB 27), for the selective isolation and differentiation of Corynebacterium diphtheriae. Directions Add 1 vial (2ml) to 200ml of Hoyle’s Medium (LAB 27).

Urea Solution 40%
X130/X135
Description A sterile solution of 40% urea, for addition to Urea Broth Base (LAB 131) and Urea Agar Base (LAB 130) for the detection of urease production by Proteus spp. Directions Add 1 vial X130 (5ml) to 95ml of Urea Broth Base (LAB 131) and Urea Agar Base (LAB 130). X135 contains 100ml of solution for production of larger volumes of Urea Broth or Agar.

Egg Yolk Tellurite Emulsion
X085
Description A sterile egg yolk emulsion containing potassium tellurite for use in Baird-Parker Medium (LAB 85). Baird-Parker Medium is widely used in the food industry for the detection of pathogenic staphylococci. Baird-Parker plates containing Egg Yolk Tellurite Emulsion should be protected from dehydration by storing in vapour proof packaging. Directions Add 50ml to 1 litre of Baird-Parker Medium LAB 85. (50ml Egg Yolk Tellurite Emulsion contains the equivalent of 3ml of 3.5% potassium tellurite. This is the amount recommended for 1 litre of Baird-Parker Medium, i.e. concentration in X085 is 0.21% w/v. Final concentration in Baird-Parker Medium is 0.01% w/v.)

Lactic Acid 10%
X037
Description A sterile solution of 10% lactic acid added to culture media to reduce the pH, in order to suppress bacterial growth. Directions Add 1 vial (5ml) to 1 litre of Malt Extract Agar LAB 37. Add 2 vials (10ml) to 1 litre of Potato Dextrose Agar LAB 98. Addition of X037 should be carried out after sterilisation and cooling the medium to 47ºC.

7.2

NOTES

NOTES

3.3 2. 1. 7.8 1.6 xiv. 1. 3.39. 5.35.1.53 1.3.19.10.13. 1.3 2. 2. 3. 1.8.1. 7.29. 1.19 1. 1.21. 1.48 xxiii.21. 1.3 4.5.5 xxiv.32.14 1.2 1. 4.57 xxiv. 1.36 1. 5.43 xv. 2 Fluorescence Agar Yeast Extract Agar Milk Agar Dextrose Tryptone Agar Reinforced Clostridial Medium (Broth) Reinforced Clostridial Agar Cooked Meat Granules Fluid Thioglycollate Medium USP Hoyle’s Medium Blood Agar Base Desoxycholate Citrate Agar (DCA) MacConkey Agar (with salt) Violet Red Bile Agar (VRBA) XLD Agar Sabouraud Liquid Medium USP Brilliant Green Agar (Modified) TYC Medium Rose Bengal Chloramphenicol Agar LAB 80A Minerals Modified Glutamate Medium LAB 80B Sodium Glutamate LAB 82 LAB 84 LAB 85 LAB 86 LAB 87 LAB 88 Membrane Lauryl Sulphate Broth Single Step Staph Selective Agar (4S) Baird-Parker Medium Rappaport Vassiliadis Medium (RVS) Sugar Free Agar Violet Red Bile Glucose Agar (VRBGA) Oxytetracycline Glucose Yeast Extract Agar Fastidious Anaerobe Agar (FAA) E.61. coli/Coliform Medium Harlequin™ mLGA Columbia Agar Base MacConkey Agar (without salt) DCLS Agar Tryptone Soy Broth USP MacConkey Broth Purple CLED Medium (Bevis-double indicator) Mannitol Salt Agar Nutrient Agar Sabouraud Dextrose Agar Plate Count Agar APHA Tryptone Soy Agar USP Sensitivity Test Agar (STA) 4. xxvii.36 LAB 44A Selenite Broth Base LAB 44B Sodium Biselenite LAB 45 LAB 46 MacConkey Agar No.52 xxvi. 7.50 1.9 1.1.49 xxvii. 2.55 xxiv.37 1. 6.19.40.2. 5.49.10.13.3.0) CAP 1 CAP 2 CAP 3 CAP 4 CAP 5 CAP 6 HAL 1 HAL 2 HAL 3 HAL 4 HAL 5 HAL 6 HAL 7 HAL 8 HAL 9 LAB 1 LAB 2 LAB 3 LAB 4 LAB 5 LAB 6 LAB 7 LAB 8 LAB 9 LAB 10 LAB 11 LAB 12 Captivate™ O157 Captivate™ Salmonella Captivate™ O26 Captivate™ O111 Captivate™ O103 Captivate™ O145 Harlequin™ Salmonella ABC Harlequin™ Listeria Medium Harlequin™ TBGA (TBX) formerly LAB 162 Harlequin™ LB Agar Harlequin™ LB Top Agar Harlequin™ SMAC-BCIG Harlequin™ CLED Harlequin™ E.2 1.1.4. 1.3 4.59 xxv.1. 1. 1.2 xxvii.59 1.57 1.22 xxiii. 1.58 1. 1.41. 1.38 1.31. 3 Buffered Peptone Water 2.10 LAB 90 LAB 91 LAB 92 LAB 93 LAB 94 LAB 89 LAB 52 LAB 53 LAB 54 LAB 59 LAB 60 LAB 61 LAB 62 LAB 63 LAB 64 LAB 65 LAB 66 LAB 67 LAB 68 LAB 69 LAB 71 LAB 72 LAB 73 LAB 74 LAB 75 LAB 78 LAB 79 SS Agar (Modified) Triple Sugar Iron Agar Lysine Iron Agar Kligler Iron Agar Endo Agar Eosin Methylene Blue Agar (Levine) Tryptose Phosphate Broth Tryptone Glucose Extract Agar Thioglycollate Medium (Brewer) Desoxycholate Citrate Agar (Hynes) Anaerobe Identification Medium Base GC Agar Base Nutrient Broth ‘E’ Simmons Citrate Agar Fastidious Anaerobe Broth (FAB) Tryptone Bile Agar Bacillus Cereus Medium (PREP) Nusens Agar Todd Hewitt Broth CEMO Agar WL Nutrient Agar LAB 37 LAB 38 LAB 39 LAB 41 LAB 42 Malt Extract Agar Wort Agar Mueller Hinton Agar II CLED Medium (single indicator) Mueller Kauffman Tetrathionate Broth xxv.6.26 1.3. 1.19 1. 5.12 xxvii.51 xv.53 xxiii.23. 5.2.57.13.5. 1. 1.4 2.8.39. 1. 1.4 LAB 48 LAB 49 LAB 51 Brain Heart Infusion Agar Brain Heart Infusion Broth Brilliant Green Bile 2% Broth 1. 1. 5.15 xv.7 1.29 1.57 1.14 1. xxvii. 1.1 xxiv.3 2.45 1. 5. 5. 1.13.51 xxiv. 5. 1. 2 BP Blood Agar Base No. 7.34. 3.2 1.9.2 1.39 1. 1. 1.14 xv.1 . Broth M17 Agar MRS Agar MRS Broth LAB 24Z Cooked Meat Medium Tablets 8.1. 1. xxvii. 1. 1. 5.62 xxvi.2 1.16 1.51 xxv.2 1.4 2.33. 1.23 1. 1.9 1.1 2. 5. 5.1.E.7 1.50 1. 5. 1. 1. 5. 1. 1.8 1. 4.29 xxiii.63 1. 5.46 1.49.31 1. 5. 1. 1.62 xxv.1.46 1.55 1.3 4.49. 5. xix.3 xxv.8 1.34.33 1.8 1. 1.9 1.9.47.56. 1. 1. 7.12.1.41. INDEXES By product code (V1. 1. 1. 7.64 xxiii.30.4.55 1.10 1.14.15 1.8 xxvii. 1. 1.3.5 1.4 ix.48 xxvi.62 xxiv. 1.20 1. 1. 1.58 xxiv. 1.3 1.30 1.1 4. 1.10 1.38 1.4.55 1.3.20.2 LAB 55A Selenite Cystine Broth Base LAB 13A Bismuth Sulphite Agar Base A LAB 13B Bismuth Sulphite Agar Base B LAB 14 LAB 15 LAB 16 LAB 18 LAB 19 LAB 20 LAB 22 LAB 23 LAB 24 LAB 25 LAB 27 LAB 28 LAB 29 LAB 30 LAB 31 LAB 32 LAB 33 LAB 34 LAB 35 LAB 36 Nutrient Broth No. 1. 1. 4.34.57 1. 1.64 1.3 4. 2.24. 4.2. 1.2. 4. 1.50 1. 1.

6 1. 5.7 xxiv.1.4.3 1.5 3.B.3 1.6 1. 1.7 6. 5. 2 Bacteriological .47 ix. 5. 1.54 xxvii.1.1. 1.25 LAB 159 Malt Extract Broth LAB 160 Brazier’s CCEY Agar LAB 161 Sorbitol MacConkey Agar LAB 162 Now HAL 3 (TBGA) LAB 163 R2A Medium LAB 164 Fraser Broth LAB 165 O157 Broth MTSB LAB 166 Slanetz & Bartley Medium LAB 167 Aeromonas Agar LAB 168 LB Agar LAB 169 LB Broth LAB 170 Susceptibility Test ‘ISO’ Agar LAB 171 EC Medium LAB 172 Listeria Monocytogenes Blood Agar (LMBA) LAB 173 LB Broth (Lennox) LAB 174 LB Agar (Lennox) LAB 175 YPD Broth LAB 176 YPD Agar LAB 177 Superbroth LAB 178 Superbroth with Agar LAB 179 2 x YT Broth LAB 180 2 x YT Agar LAB 181 NZY Broth (NZYM) LAB 182 NZCYM Broth LAB 183 Terrific Broth LAB 184 Letheen Broth (AOAC) LAB 185 Letheen Agar (AOAC) LAB 186 D/E Neutralising Broth Base LAB 187 D/E Neutralising Broth LAB 188 D/E Neutralising Agar LAB 189 Microbial Content Test Agar (MCA) LAB 191 Luria Bertani (Hi Salt) Broth LAB 192 ORSIM (Oxacillin Resistant Staphylococci Isolation Medium) LAB 193 PEMBA (Bacillus Cereus Medium) LAB 194 Perfringens Agar Base (TSC) LAB 196 Lauryl Tryptose Broth LAB 197 Water Plate Count Agar (ISO) LAB 505 Cary Blair Medium LAB 523 Azide Blood Agar Base LAB 525 Eugon Agar LAB 526 Eugon Broth LAB 537 Diassalm LAB 573 Violet Red Bile Agar with MUG MC 1 MC 2 MC 3 MC 4 MC 5 MC 6 MC 7 MC 8 MC 9 MC 10 Yeast Extract Agar No.52 1.5.6 1.1.1.2 1.42. 6.5 1. 1.4 1.1. 1.17 1.9 xxvi. 5.16. 1.52. 5.35 1.2 1.1 1. 1. 1.7 6.54 1. 5. 1.1. 1. 5.9.9 xxv.10. 1. 4. 5.4 1. 6. 1. 7.2 1.5. 1.2 .18 1. 1.27 8.6 xxvi. 5. 1.1 1.1 1. 1.1.33 1.59 1. 6.33.3 1.11 1. 5.4 2. 1.3 1. Purpose Acid Hydrolysed Casein Tryptose Mycological Peptone Ox Bile 1.3.6 xxiv. 1.4 3.22.1.43 xxvi.34.1.47 ix. 1.6 3.44.3.5. 2.40 xxvi.1. 5.6 3.4 LAB 100 Ringer’s Solution 1/4 Strength LAB 100Z Ringer’s Solution 1/4 Strength Tablets LAB 101 Ringer’s Solution (Calgon) LAB 102 Ringer’s Solution (Thiosulphate) LAB 103 Maximum Recovery Diluent LAB 104 Peptone Water LAB 105 China Blue Lactose Agar LAB 106 Kanamycin Aesculin Azide Agar (complete) LAB 107 Kanamycin Aesculin Azide Broth (complete) LAB 108 Pseudomonas Agar Base LAB 109 Perfringens Agar OPSP LAB 110 Hektoen Enteric Medium LAB 111 Sabouraud Maltose Agar LAB 112 Campylobacter Blood Free (modified CCDA) LAB 113Z Salt Meat Broth Tablets LAB 114 Mueller Hinton Broth II LAB 115 Milk Plate Count Agar LAB 116 MLCB Agar LAB 117 DTM Dermatophyte Test Medium LAB 119 Yeast Extract Dextrose Chloramphenicol Agar LAB 120 Yersinia CIN Agar Base LAB 121 Bromocresol Purple Lactose Agar LAB 122 Listeria Isolation Medium (Oxford) LAB 123 Kirchner’s T.27.3 1.LAB 95 LAB 96 LAB 97 LAB 98 LAB 99 DN’ase Agar TCBS Cholera Medium Tetrathionate Broth (APHA) Potato Dextrose Agar Wort Broth 1.25. 6. Enrichment Medium LAB 124 Amies Transport Medium With Charcoal LAB 125 Amies Transport Medium Without Charcoal LAB 126 Lactose Broth LAB 127 Cooked Meat Medium LAB 129 Tryptone Water LAB 130 Urea Agar Base LAB 131 Urea Broth Base LAB 133 Cetrimide Agar LAB 135 Campylobacter Enrichment Broth (Bolton) LAB 136 Easter .1 Tryptone Agar No. 6. 7.23 1. xv.5 xxvi.17. 1.9 1.12 1.47 ix. 1.1.2.45 xxvi.2 xxvi.61.2 1.1.32.5 xxvi. 5.3 1.5 xxiii. 5.34. 1.2 1. 5.33.1. 2.High Clarity Soy Peptone Balanced Peptone No. xxiii-xxvi.9.Gen.1.3 3.32 1.52 1. xxvii.49 1.9. 7.22.1.4 3.7 6.6 xxiv.10 1. 1.7 6.1. 5.24.1.5.7. 5.5.2 3.1.32 1.1.36.63 ix. 1.10. 6.65. 1.1 6.1 1.14 xxiv.3 1.1 3.2 1.11 6.44.48 xiv. 1. 1.7 6.42.47 ix. 5.10 1. 1 Bacteriological . 5. 1.9.7 1. 5. 1. 5.5. 5. 1. 5.40.59 1. 5.10.26. 1.3 1.6 3.37 xxiii.Gibson Pre-enrichment Medium LAB 137 Easter .8 3.10 1.6.60.1.28.5 3.4 3.8 xxvi.6.18.28.41.7 6. 1. 7.1.42.2.40. 5.42.11 xxvii. 6. 5. 1. 1. 5. 6. 1. 5.60.Gibson Salmonella Medium LAB 138 Listeria Enrichment Broth (FDA) LAB 139 Buffered Listeria Enrichment Broth LAB 140 Helicobacter Pylori Agar Base LAB 144 Palcam Broth LAB 147 Orange Serum Agar LAB 148 Palcam Agar Base LAB 149 Plate Count Agar LAB 150 MSRV Medium LAB 155 UVM Base LAB 157 Aseptic Commissioning Broth LAB 158 Liquid Baird-Parker Medium xxv.5 1.8 xxvi.2 1.4 3.5 1.1. 4. 4.8.22.1 6.61. 1. 5.3.2.3 1. 1.2 3.6 1. 1.6 1.5 1. 1.65 1. 6. 1.

Polymyxin Cefoperazone.7.6 1.9.22. 5. 5.1 1.41.8 1. 1.2. 5.1. 7.32. 1.41. 1.22. 2.42. 1.1 7. 5.1. 5.52.8 1.4 7.6. Ceftazimide Nalidixic Acid Egg Yolk Emulsion Polymyxin B Egg Yolk Tellurite 6.3.9. 1.36.6 1. Oxolinic Acid Neomycin 75 Neomycin 100 Kanamycin 75 P-INC Supplement Potassium Tellurite Solution 3.4.48.3 1.1 1. 1.5 6. 5.1. supplement) Urea solution 40% 5ml CVTC CVTN Urea solution 40% 100ml TMAO Selenite NAC (for FDA broth) NAN Burkholderia (Pseudomonas) cepacia Selective Supplement PAC (for Palcam Media) Novobiocin (for MSRV/ Diassalm/O157 Broth) UVM I Supplement UVM II Supplement xxv. 4.33. 5.42. Amphotericin CIN Selective Supplements CCCAF (for Oxford medium) CNCAF (for Oxford medium) PTTA (T. 5. 5. Nalidixic Acid Colistin. 1. 1. 7. 5. 5.6 6. 5.4.2 5.13.2 1. Nalidixic Acid Cycloserine. 5. 1.7 1.2.3 1. 5.61.1.1. 5. 5.1 1.4 Fraser Supplement (1/2 strength) Fraser Supplement (full strength) Oxacillin.1 1.9. 1. 5.22. 1. 5. 1.7 6.6 1.1.4 1. 5.9. 5. 5.5 1.2 1. 5. 5.28. 5.23.7 1.20.7 1.7.10 1. 6.10. 7.61.13. Nalidixic Acid Colistin. 1.7.51.8 5. 5.18. 5. 5.4 6.1. 1.6 6. 1.9 1. 5.20.7 1. 5. 6. 5. 4.31.23. 7.2.5 1.4 6. 1. 5.3 6.51. 5.7 6. 5.3 6.13.44.4.60. 5.1.65.9.1.3 1.17.10. aeruginosa) CFC (for Pseudomonas spp) Sulphadiazine Oleandomycin.9.20.2 1.23.9. 5.1 7.1.5.3 6. 5.2 1.1 7. 5.Plant Tissue Culture Grade Xylose FMV Peptone Liver Digest Special Tryptone Lactalbumin Hydrolysate IPTG X-gal MUG Cooked meat with nutient broth Nutrient Agar Slopes Nutrient Broth Peptone Water Selenite Broth Selenite Cystine Broth Transport Medium (Amies Charcoal) Urea Agar Slope Serum Broth Trichomonas Medium Cooked meat with FAB Resuscitation broth Chloramphenicol Colistin.3. 5.3.2 6.2 1.1 7.5.13.20. 1.20.5 8.13.1 7.23. 5. 7.1 1. 1.1 7.5 6.16.4 6.28. 5.4 1.8 1.1 7.1 7.2 6.6.13.6 Cefixime Tellurite (for SMAC/SMAC-BCIG) xxiv. 5.2 xxv.28.26.40.22.42. 5. 5.5 1. 5.3.3.3 . 1.4 1.1 1.44.1 1. 5. 1.3 6.5 6.44.3 1.10. 1.28.40.4 1.2 1. Cefoxitin CN (for Pseudomonas. Vancomycin Nalidixic Acid Metronidazole. 1. 1.3 1. 5. 1.5.3 6.5% Lactic Acid 10% VCA VCNT (for Thayer Martin) LCT (for New York medium) LCAT (for New York medium) Polymyxin B. Amphotericin Skirrows Neomycin 75 P-INC Supplement Bacitracin VCNT (for Thayer Martin) LCT (for New York medium) LCAT (for New York medium) GC Growth Supplement Nalidixic Acid. 1. 5. 6. 1.44.3.1.2 1.49.6 1.3.9.23. 6.5.10 1.5 6. 5. 5.16.5. 5.3. 1. 4. 5.9 1.8 xxv.9.60.6 4.2.23. 2. 5.5.2 1.2 6.5 6. 5.1 7. 5.47.6 1.B.1 7. 1. 5.2 1. 5.1.65.10. 5. 5. 3 Sodium Desoxycholate Skim Milk Powder Agar No.47.9. 7.5 1. Polymyxin B Polymyxin B Cycloserine Methicillin (for MRSA) Chloramphenicol Cefoperazone.61.1. 1.6 1.9 1. 2. 1.23.5. 5.6 1. 6.2 1. Vancomycin Nalidixic Acid NAN VCC Supplement UVM I Supplement Fraser Supplement (1/2 strength) 1.13.7 1.2 X086 X089 X090 X091 X092 X093 X107 X108 X109 X110 X112 X120 X122 X123 X124 X130 X131 X132 X135 X137 X138 X139 X140 X144 X150 X155 X156 X161 X164 X165 X192 X193 X194 X207 X209 X212 X214 X215 X219 X260 X268 X269 X270 X271 X290 X291 X539 X546 X555 X564 RPF Oxytetracycline (for OGYE) Nalidixic Acid. 1.4 1.7 1.33. 1. 1.MC 11 MC 12 MC 13 MC 14 MC 15 MC 16 MC 17 MC 18 MC 19 MC 20 MC 22 MC 23 MC 24 MC 25 MC 26 MC 27 MC 29 MC 32 MC 33 MC 34 MC 35 MC 40 MC 402 MC 405 MC 406 P002 P007 P008 P009 P011E P012E P013 P015 P017 P018 P069 P081 X009 X011 X012 X013 X015 X016 X018 X019 X027 X037 X040 X068 X069 X070 X072 X072N X073 X074 X085 Proteose Peptone A Columbia Peptone Glucose (Dextrose) Mannitol Gelatine Sodium Thioglycollate Sodium Chloride Meat Peptone Beef Extract Lactose Maltose Monohydrate Malt Extract Bacteriological Peptone Bile Salts No.24.10 1.5 1. 5.3 6.20. 1.9 1.3 6. 4 .7 1. 5. 5.1 7.1.9 1.48.10 1. 5.6 1.3 1. 5.7.4 6.2 1.24.3. 1. 5. 1.65.7 1.28.

1.3 4.3 4.1. Nalidixic Acid Colistin. 1.9 1.3 1.19 1.13. 7.2.1.1 1.1 4.3 Agar No. 1.18.Gibson Salmonella Medium EC Medium Egg Yolk Emulsion LAB 95 LAB 117 LAB 91 LAB 136 LAB 137 LAB 171 Columbia Peptone Cooked Meat Granules Cooked Meat Medium Cooked Meat Medium Tablets Cooked meat with FAB Cooked meat with nutrient broth CVTC CVTN Cycloserine Cycloserine.4 1.2 1. 5. Cefoxitin D/E Neutralising Agar D/E Neutralising Broth D/E Neutralising Broth Base DCLS Agar Desoxycholate Citrate Agar (DCA) Desoxycholate Citrate Agar (Hynes) Dextrose Tryptone Agar Diassalm MC 12 LAB 24 LAB 127 LAB 24Z P069 P002 X131 X132 X194 LAB 188 LAB 187 LAB 186 LAB 3 LAB 29 LAB 65 LAB 20 LAB 537 CIN Selective Supplements CLED Medium (Bevis-double indicator) CLED Medium (single indicator) CN (for Pseudomonas aeruginosa) CNCAF (for Oxford Medium) Colistin. 2. 7.6 xxvi.1.2 1.9.6.1. 5.52.7 6. 5.3 1.2 1. 1.4 X140 LAB 112 LAB 135 CAP 1 CAP 5 CAP 4 CAP 6 CAP 2 CAP 3 LAB 505 X122 5.1. 5.1.10. Nalidixic Acid Colistin. 1.3 1. 5. 1. 1.2. 1.10.10.3.51 1.13.6. 1.51. 1.10. xxvii.9 6.5 Egg Yolk Tellurite Endo Agar Eosin Methylene Blue Agar (Levine) Eugon Agar Eugon Broth Fastidious Anaerobe Agar (FAA) Fastidious Anaerobe Broth (FAB) DTM Dermatophyte Test Medium E.8 xxvi.4. 1. 1.9.15 1. 5. 2 Bacteriological .Gen.8 DN’ase Agar xiv. 5.1.2 MC 7 LAB 167 MC 2 MC 29 LAB 124 LAB 66 LAB 157 LAB 523 LAB 73 LAB 193 X260 MC24 LAB 85 MC4 MC19 MC 25 LAB 13A LAB 13B LAB 28 LAB 15 LAB 49 LAB 160 LAB 34 LAB 51 LAB 121 LAB 139 LAB 46 6. 4 .Gibson Pre-enrichment Medium Easter .1.2 xxv.2 1.23 1.19 1. 6. 2.1 1. 1. 5.18 1.8 1. 1.3. 6.4 1.65.10 X209 1.2 6. 1.2.20.3.16.1 1.10 1.5. 5.3 1.7.1. 7.28.3 1.9. 5. 1. 5. 1.2 1. 5. xxvii.15 1. 7.8 1. 5.10 1. 1. Broth Easter .1 xxiv.2 1.4.10. 1.17 1.16.7 X093 1. 5. 1.19.7 xxiv. 1 Beef Extract Bile Salts No. 1.Plant Tissue Culture Grade Amies Transport Medium With Charcoal Anaerobe Identification Medium Base Aseptic Commissioning Broth Azide Blood Agar Base Bacillus Cereus Medium (PREP) Bacillus Cereus Medium (PEMBA) Bacitracin Bacteriological Peptone Baird-Parker Medium Balanced Peptone No. 5. 5. 5.55 8.13. 1. 4.40.48. 1. 5.20 1.13. 5. 1. 5.8.1.4 1.7.13.21.4. 5.1. 5.12.1. 1. 1. 5.65.E.6 xxvi.1.48.1 6. 1. 1.44. xix. Amphotericin CEMO Agar Cetrimide Agar CFC (for Pseudomonas spp) X212 LAB 78 LAB 133 X108 1.5.9.14 1.5. 5.1 6. 1. 5.1.16.3 1. xxvii. 1.5. 5.10 1.4 6.4. 1.3 4. 1.9.2 1.9 1. 1.5.10. 3 Bismuth Sulphite Agar Base A Bismuth Sulphite Agar Base B Blood Agar Base Blood Agar Base No. Oxolinic Acid Columbia Agar Base Chloramphenicol China Blue Lactose Agar Chloramphenicol LAB 105 X009 Cefoperazone.9.2 X085 LAB 60 LAB 61 LAB 525 LAB 526 LAB 90 LAB 71 xxv. 1. 5.High Clarity Agar No. 2 BPLS Brain Heart Infusion Broth Brazier’s CCEY Agar Brilliant Green Agar (Modified) Brilliant Green Bile 2% Broth Bromocresol Purple Lactose Agar Buffered Listeria Enrichment Broth Buffered Peptone Water Burkholderia (Pseudomonas) cepacia Selective Supplement Campylobacter Blood Free (modified CCDA) Campylobacter Enrichment Broth Captivate™ O157 Captivate™ O103 Captivate™ O111 Captivate™ O145 Captivate™ Salmonella Captivate™ O26 Cary Blair Medium CCCAF (for Oxford medium) Cefixime Tellurite (for SMAC/SMAC-BCIG) Cefoperazone. 5. 1.44.9.4 .1.1.65.5.28.6 1.2. 5. 1.34.2 1.1 1.16 xxvi. Amphotericin X161 xxiv.5 1.11 1. 5.By product name Acid Hydrolysed Casein Aeromonas Agar Agar No.1 xxv.47.11 xxv.2.6.1.7 xxv.4 1. 5.10.1 7.14 7.1.3 1.14 1. 5.9 xiv. 7.3 X073 1.2 5.2. 6.12 1. Purpose MC 6 Amies Transport Medium Without Charcoal LAB 125 LAB 34 xxvi. 5. 7.2. 1.8 1.4 X112 1.1.9.17. 1. 1 Bacteriological .47.7 1.1. 1.2 6.3 4.2 xxvii. 2.22. 4. 5. 1. 1. 1. 1. 1. 6. 1.10 X120 LAB 6 LAB 41 X107 X123 X011 X012 X013 LAB 1 1.3 4. xxvii.8 1.9.20.2. 1.3.14 1. 6.3 xv.3.13. 5. 5.5 1. 1.

22.52 1.5.6 5. 1.4 8.3. 5. 1. 5.4 2. 1.3 3. 5.24.3 1.2 3.29 1.40.3 xxvi. 1. 7.13.32 MC 18 LAB 82 X207 X092 LAB 189 LAB 115 LAB 80A LAB 116 LAB 165 LAB 93 LAB 94 LAB 39 LAB 114 LAB 42 MC 9 X138 X072N X091 X291 X090 X290 X139 X539 X016 X015 X215 X150 LAB 74 LAB 8 P007 P008 LAB 68 LAB 14 6. 5.6 1.6 1.7 1.27. 5.31.5.32 6.Fluid Thioglycollate Medium USP Fluorescence Agar FMV Peptone Fraser Broth Fraser Supplement (1/2 strength) Fraser Supplement (1/2 strength) Fraser Supplement (full strength) GC Agar Base GC Growth Supplement Gelatine Glucose (Dextrose) Harlequin™ CLED Harlequin™ E. xv.8 LAB 150xxvi.61. Nalidixic Acid Microbial Content Test Agar (MCA) Milk Agar Milk Plate Count Agar Minerals Modified Glutamate Medium MLCB Agar Modified Tryptone Soy Broth MRS Agar MRS Broth MSRV Medium Mueller Hinton Agar II Mueller Hinton Broth II Mueller Kauffman Tetrathionate Broth MUG Mycological Peptone NAC (for FDA broth) Nalidixic Acid Nalidixic Acid Nalidixic Acid Nalidixic Acid.33.2 HAL 6 xxiv. 1.1 7.3 1.3. 5.28.13.5. 1.37 1. 1.1. 2.7 1.1.1.55 LAB 16 MC 33 LAB 164 X164 X564 X165 LAB 67 X271 MC 15 MC 13 HAL 7 HAL 8 HAL 4 HAL 5 HAL 2 HAL 9 HAL 1 1. 5. 5. 5. 1.4 1.14.40. 1. 7.38 6.21.28.1. 5. 1.3 1.23.25 1. 1.4.4 6.7 1.33.7 1.1 4.31.3 1. coli/Coliform Medium Harlequin™ LB Agar Harlequin™ LB Top Agar Harlequin™ Listeria Medium Harlequin™ mLGA Harlequin™ Salmonella ABC Harlequin™ SMAC-BCIG Harlequin™ TBGA (TBX) Hektoen Enteric Medium Helicobacter Pylori Agar Base Hoyle’s Medium IPTG Kanamycin 75 Kanamycin Aesculin Azide Agar (complete) Kanamycin Aesculin Azide Broth (complete) Kirchner’s T. 3.1 1. 2 BP LAB 92 LAB 2 LAB 30 LAB 45 LAB 5 MC 23 LAB 37 LAB 159 MC 22 MC 14 LAB 7 1. Enrichment Medium Kligler Iron Agar Lactalbumin Hydrolysate Lactic Acid 10% Lactose Lactose Broth Lauryl Tryptose Broth LB Agar LB Agar (Lennox) LB Broth LB Broth (Lennox) LCAT (for New York medium) LCAT (for New York medium) LCT (for New York medium) LCT (for New York medium) Letheen Agar (AOAC) Letheen Broth (AOAC) Liquid Baird-Parker Medium Listeria Enrichment Broth (FDA) Listeria Isolation Medium (Oxford) Listeria Monocytogenes Blood Agar (LMBA)LAB 172 Liver Digest Luria Bertani (Hi Salt) Broth Lysine Iron Agar LAB 25 xv.13.3.34. 5. 5. 7.7. 5. 5.7 6.2.5 . 5. 1.9.2 6.1.20.3.9 LAB 103 ix. 5. 5.23.2 1. 1.9.5.39. 1.36. 3.1.4 HAL 3 LAB 110 LAB 140 LAB 27 MC 402 X018 LAB 106 LAB 107 LAB 123 LAB 59 MC 40 X037 MC 20 LAB 126 LAB 196 LAB 168 LAB 174 LAB 169 LAB 173 X070 X270 X069 X269 LAB 185 LAB 184 LAB 158 LAB 138 LAB 122 xxiv.22.25. 5.28.5.23.32. 1. 1. 5.5 1.3.1 1. 1. 5.9 1.30 1. 1.35.30.3 1. 5. Vancomycin Nalidixic Acid.23.6 xxvi.6 1.29 1.2 6.27 xxvi.1.23. 1.6 1. 5.10.17. 5. 5.2 2. 5.39.52 xxiv.4 xxv.34. xxiiixxvi.4 1.38 1.5 1.2 1. 1. 5.1.57 7. 1.20.28.22. 5.8 M17 Agar MacConkey Agar (without salt) MacConkey Agar (with salt) MacConkey Agar No.1.9 MC 406 1.33 1.24. 5.1.6 1.3 6. Vancomycin NAN NAN Neomycin 100 Neomycin 75 Neomycin 75 Novobiocin (for MSRV/Diassalm /O157 Broth) Nusens Agar Nutrient Agar Nutrient Agar Slopes Nutrient Broth Nutrient Broth ‘E’ Nutrient Broth No. 5.32.7. 1. 7.6 1.33 1. 6. 1.6 1. 5.7 1.1 1. 5.1.1 1. 5.2 1.3 1.7 1.9. 1. 4. 1. 6.1. 1.1.5 xxiv.6 MC 34 LAB 191 LAB 54 6.2.8 xxiii. 1. 1.44. 1.4 1. 5. 1.3.9 1. 6. 5.3 3. 1. 5.5. 5.23.3 2.3.20. 1. 3.22.2 1. 4. 5.1 1. 2.22. 1.28. 5.1 2. 4. LAB 19 xxiii. 1.26 6.13.5 2. 3 MacConkey Broth Purple Malt Extract Malt Extract Agar Malt Extract Broth Maltose monohydrate Mannitol Mannitol Salt Agar Maximum Recovery Diluent Meat Peptone Membrane Lauryl Sulphate Broth Methicillin (for MRSA) Metronidazole.B. 2.36 1. 6.2 3.1 1. 1.22 6.3 2.5 1. 5.36 1.7 1. 4.6 1.39 1. 1.57 1.20. 4.49.3.20. 1.31 6.36. 5.37 xxvii.29.23 1.5 3.1 1.26. 5.35 xxiv.5.5 2.40.

10 X144 LAB 148 LAB 144 LAB 193 LAB 104 P009 LAB 194 1. 2.4.41. 5.40. 1. 5.57. 6. 5.1 xxvii.5 1. 5. 1.47 xxv. 5. 1. 7.42.3. 1. 1. Polymyxin Orange Serum Agar ORSIM Oxacillin.51 1. 1.9 LAB 9 LAB 33 LAB 111 LAB 113Z P011E LAB 44A P012E LAB 55A LAB 12 1.8 1.61. 4.3 1.1 ix.5 xxiv. 5.5 ix.5 1.5% Potato Dextrose Agar PREP Proteose Peptone A Pseudomonas Agar Base PTTA (T. 5. 7.51.50 6. 1. 5. 5.47 8.42. 5. 6.5 xxvii. 7.53 1.3 LAB 109xxvi.9.1.10 LAB 89 xxv.59 1. Ceftazimide Polymyxin B Polymyxin B Potassium Tellurite Solution 3.2 7.59 1.60.1 xxvi.45 xxvii. 5. 1.51 6.7 1.44.3.50 1. 5. 1.5 1. 1. 1.10 1.2 xxvi. 1. 5.47 ix.2 1. 1. 1. 1.4 4.6 1.42.4.1.6.9 1. 6. 5.60.5 xxv.2.46 1.2 1.60. 7. 5. 1. Supplement) R2A Medium Rappaport Vassiliadis Medium (RVS) Reinforced Clostridial Agar Reinforced Clostridial Medium (Broth) Resuscitation broth Ringer’s Solution (Calgon) Ringer’s Solution (Thiosulphate) Ringer’s Solution 1/4 Strength Ringer’s Solution 1/4 Strength Tablets Rose Bengal Chloramphenicol Agar RPF Sabouraud Dextrose Agar Sabouraud Liquid Medium USP Sabouraud Maltose Agar Salt Meat Broth Tablets Selenite Broth Selenite Broth Base Selenite Cystine Broth Selenite Cystine Broth Base Sensitivity Test Agar (STA) LAB 182 LAB 181 LAB 165 X110 LAB 147 LAB 192 X192 MC 10 X089 3.60.6 1.55 1. 7.57 xv.49. 1.8 1.56. 5.50 UVM I Supplement UVM I Supplement UVM II Supplement VCA VCC Supplement VCNT (for Thayer Martin) VCNT (for Thayer Martin) Violet Red Bile Agar (VRBA) Tryptose Tryptose Phosphate Broth TYC Medium Urea Agar Base Urea Agar Slope Urea Broth Base Urea solution 40% 5ml Urea solution 40% 100ml UVM Base Tryptone Glucose Extract Agar Tryptone Soy Agar USP Tryptone Soy Broth USP Tryptone Water Soy Peptone Special Tryptone SS Agar (Modified) Sugar Free Agar Sulphadiazine Superbroth Superbroth with Agar Susceptibility Test ‘ISO’ Agar TCBS Cholera Medium Terrific Broth Tetrathionate Broth (APHA) Thioglycollate Medium (Brewer) TMAO Selenite Todd Hewitt Broth Transport Medium (Amies Charcoal) Trichomonas Medium Triple Sugar Iron Agar Tryptone Tryptone Bile Agar Sodium Thioglycollate Sorbitol MacConkey Agar 1. 2.3.23.48 1.3 3. 1.24.7 xxv. 5.5 xxiv.1.1. 5.2. 1.47 ix. 1.52. 5.B. 5. 5.1 1.1.22. 5. 1.34. 2. 5.3. 7. 7.5 xxvi.49.4 3.4 3.40 1.2 1.49.58 xxiv.4 xxiv.41.5.8 6. 1. 6.2 1. 1. 5.6 1.23.61.52 xxvii.33.43 1.2 1.6 1.54 1. 1. 1.1 1.9.1. 5. 1. 1. 1.24.2 1.6 1.5.44. 1.NZCYM Broth NZY Broth (NZYM) O157 Broth MTSB Oleandomycin.8 xxiii. 1.10 X086 xxv.1 1. 1.1.7 xxiv.4 1. 1.18.2 6.59 1.42.49 7. 1.7 xxiii. 5.61.7.4 LAB 100 ix. 6. 1. 1.9 6. 5.1.41. 5.9.7 6.2 xxv. 7. 5.1.4 1.2.33.41. 5. 1.7 1. 4.7 1.10. Polymyxin B Ox Bile Oxytetracycline (for OGYE) Oxytetracycline Glucose Yeast Extract Agar PAC (for Palcam media) Palcam Agar Base Palcam Broth PEMBA Peptone Water Peptone Water Perfringens Agar Base (TSC) Perfringens Agar OPSP P-INC Supplement P-INC Supplement Plate Count Agar Plate Count Agar APHA Polymixin C.42.42. 1.34.40.45 1.46 7.54 3.32. 1.57 6.58 xv. 4.59 7. 1. 4.42.5 6.1 1.49 7. 1.1.6 .4 Serum Broth Simmons Citrate Agar Single Step Staph Selective Agar (4S) Skim Milk Powder Skirrows Slanetz & Bartley Medium Sodium Biselenite Sodium Chloride Sodium Desoxycholate Sodium Glutamate P017 LAB 69 LAB 84 MC 27 X214 LAB 166 LAB 44B MC 17 MC 26 LAB 80B MC 16 LAB 161 MC 3 MC 35 LAB 52 LAB 87 X109 LAB 177 LAB 178 LAB 170 LAB 96 LAB 183 LAB 97 LAB 64 X137 LAB 75 P013 P018 LAB 53 MC 5 LAB 72 LAB 63 LAB 11 LAB 4 LAB 129 MC 8 LAB 62 LAB 35 LAB 130 P015 LAB 131 X130 X135 LAB 155 X155 X555 X156 X040 X546 X068 X268 LAB 31 7.1.48 1.47.55 7.3 1.26. 1. 1.53 1.9. 5. 5.61.3 X019 X219 LAB 149 LAB 10 X072 X074 X193 X027 LAB 98 LAB 73 MC 11 LAB 108 X124 LAB 163 LAB 86 LAB 23 LAB 22 P081 LAB 101 LAB 102 LAB 100Z LAB 36 1. 5.3.43 xxiii.33.8.2 1.3. 1.3 1.41.48 1.1 7. 5. 1.13. 6.

12 1.1.6.6 6.3. 5.7 .62 1. 1. 1.6 3. 1.1.5 3.62 1.10 3.63 1.64 8.61. 1.63 6.19.7 xxiii.65.1. xxvii.11 xxiii.65 1.62 Violet Red Bile Agar with MUG Violet Red Bile Glucose Agar (VRBGA) Water Plate Count Agar (ISO) WL Nutrient Agar Wort Agar Wort Broth X-gal XLD Agar Xylose Yeast Extract Yeast Extract Agar Yeast Extract Dextrose Chloramphenicol Agar Yersinia CIN Agar Base YPD Broth YPD Agar 2 x YT Broth 2 x YT Agar LAB 119 LAB 120 LAB 175 LAB 176 LAB 179 LAB 180 1. 6.64 6.5 3.6 xxvi. 6.6 LAB 573 LAB 88 LAB 197 LAB 79 LAB 38 LAB 99 MC 405 LAB 32 MC 32 MC 1 LAB 18 1. 1.

1.7. 1. 3. 5. 1.52 xvii. 1.3. 1.33.2. 1. xxiv. 1.1. 1.11.20.46.35. 7.11. 1.4. 1.64 xvii.By organism Actinomyces Aeromonas Aspergillus niger Bacillus cereus Bacillus stearothermophilis Bacteroides fragilis Blastomyces Campylobacter Candida Citrobacter Clostridium perfringens Clostridium difficile Coccidiodes Coliforms 1. 5. xxvii. 3.4 1. 2. 1.3.1. 1. 1.13. 1.2. xxiv. 1.49. 1. 1.20 1.32. 5.58. 1.40. 1.4 xvii. 1.37. 1. 1. 2. 7.2 1. 1. 1. 1.64. 1. 1.20 1. 5.5.52.42.4.9. xxiii. 1.1 xvii. 1.24.6. 1.5. 1.1.65.4.1.1 xvii. 1.1. 1.1.11.31. 1.64.13. 1.17. 1.11. 1. 1.59. xxii. 5. 1. 1. 1.16.30.10.4.51. xvii.27. 5. 5. 1.5.1 xxi. 3. 1.13.1.16 xxi.1.41.12.10 7.2.54.28.1 1.19.20. 1.18. 5.61.2. 1. 1.61.1.3 1.5. xvii. 1. 1.1.1.62 xvii.31.4 1. 1.48. 1.40. 1.57. 5.13.44. 5. 1.19. xxii.51. 1. 1.6.7. 1.5 xii.1.54 xviii.11. 1.12 xvii.16 xxi. 1. 1. 1.1.15.25. 5.4 1.16 xvii. 1. 1.3 Corynebacterium Dermatophytes Enterobacteriacieae Enterococcus Escherichia coli xvii.2. 1. 1. 2.20.20.8.10.6 xvii.59.53. 1. 1. 1. 1. 1. 1. 1. 1.2.35.54 xvii xii.32.2.20.3. 2. 1.20.5.3.23. 5. 1.1. 1. 1. 1. 1. 1.65.40 1. 1. 1.8 . 1. 1. 1.1 xviii. 1.55. 7.6 vii. 2. 1. 1.6 xvii. 1.36. 7. 1.8 xvii. 1. xvii. 1.44.2 1. xxv. 1.1.1. 7.23.30.2 xvii.13. 1.42. 1.29.34.45.31.12. 1.1.5. 1. 5.4. 5.5 xvii Yersinia Taylorella Trichomonas TVC Vibrio Yeasts and Moulds Staphylococcus epidermidis Streptococcus Salmonella typhi Serratia Shigella Staphylococcus aureus Micrococcus Mycobacterium Neisseria Peptostreptococcus Porphyromonas Propionibacterium Proteus Pseudomonas Saccharomyces Salmonella Lactobacillus Listeria xvii. 4. 1. 1. 1.5. xii. 5. 1.9. 1.34.2.4.3 Escherichia coli O157:H7 Eubacterium Fusobacterium Gardnerella Haemophilus Helicobacter Histoplasma Klebsiella xvii. 1. 1.6.10 8. 1. 5.5. 4. 1. 1. 1.59. 1. 1.4. 1.2 x. 1.3. 1. 1. 4. 5. xxiii. 1.14.9. 1. 7.50.53.9. xxiii.63.17.4.8.47.19. 1.1.13. 3.20 1.3.23.31.1 xvii. 1. 1.36.3. 1. 1.10. 1. 7. 5.1.9.2 xvii. 1. 5.1 1.57.3.3. 1. 1.60. 2.3 xviii xvii xvii.20 1.33.27.1. 1.8. 5.45. 1.24.1. 5.62 xvii.1.6. 1.22. 1.5. 5. 1. 1. xvii.1. 1.26. 5.49.22.9.1.5. 1. 7.32 xvii. 1.14. 1. 1. 3.42.9 1.16.43. 3. xxv. 1. 1. 5. 5.56. 5. 2. 1. 5.1.41. 1. 1.56. 1.15. 1. 1. 5.4. 2. 1. xxiv. 5.57.38.9. 5.2.20 xvii.12.4 xi.5.49 1. 1.

NOTES .

NOTES .

V1. New manual Name: Job Title: ❑ Register for manual updates ❑ Current manual version: Company Name: Company Address: Postcode: Tel. please complete and return the form below to LAB M. Topley House 52 Wash Lane BURY Lancashire BL9 6AU To receive updates to the manual as they are issued. ticking the update box. Forms can be faxed back to +44 (0)161 762 9322 or posted to the address below: Customer Services Department IDG (UK) Limited.Request a copy of the ‘Microbiology Manual’ To request a copy of ‘The Microbiology Manual’. Number: Fax Number: Email address: No. please complete the form below.g. of tests per week? Is Pathogen testing carried out on-site? Yes ❑ No ❑ Who is your current preferred supplier for culture media? .0) at the back of the manual. Please make a note of the version of the manual which you are currently using by entering on the form below the version number which can be found at the top of the Product code index (e.

.

Please make a note of the version of the manual which you are currently using by entering on the form below the version number which can be found at the top of the Product code index (e. V1. New manual Name: Job Title: ❑ Register for manual updates ❑ Current manual version: Company Name: Company Address: Postcode: Tel. Forms can be faxed back to +44 (0)161 762 9322 or posted to the address below: Customer Services Department IDG (UK) Limited. ticking the update box.Request a copy of the ‘Microbiology Manual’ To request a copy of ‘The Microbiology Manual’. Topley House 52 Wash Lane BURY Lancashire BL9 6AU To receive updates to the manual as they are issued. please complete the form below.g. please complete and return the form below to LAB M. of tests per week? Is Pathogen testing carried out on-site? Yes ❑ No ❑ Who is your current preferred supplier for culture media? . Number: Fax Number: Email address: No.0) at the back of the manual.

.

Please make a note of the version of the manual which you are currently using by entering on the form below the version number which can be found at the top of the Product code index (e. ticking the update box.0) at the back of the manual. Number: Fax Number: Email address: No. Forms can be faxed back to +44 (0)161 762 9322 or posted to the address below: Customer Services Department IDG (UK) Limited.g. of tests per week? Is Pathogen testing carried out on-site? Yes ❑ No ❑ Who is your current preferred supplier for culture media? . Topley House 52 Wash Lane BURY Lancashire BL9 6AU To receive updates to the manual as they are issued. New manual Name: Job Title: ❑ Register for manual updates ❑ Current manual version: Company Name: Company Address: Postcode: Tel.Request a copy of the ‘Microbiology Manual’ To request a copy of ‘The Microbiology Manual’. V1. please complete the form below. please complete and return the form below to LAB M.