MILK AND MILK PRODUCTS/Microblology of Cream and Butter 1445 ‘Table 7 Suggested microbiological standards for dried mik products Contaminant ‘Skimiwhole mk powder Cassin/caseinates Whey powder ‘Total viable count 3x 10%chug* 3x10'cug* 5x 10'ctug* Colitorms 48 Used in dessarts and whipped in gateaux Cltted cream 255 Used as spread on scones in conjunction with frit reserves High-fat roams (plastic crear) 70-80 For ice cream manufacture ‘Table 2. Food and Agriculture Organization classification of Product type Fat content (6) Cream 18-26 Light croam (or cream with additional terms >10 ‘such as coffee crear) Whipping cream >28 Heavy cream >96 Double cream >4s Homogenization of Cream Homogenization is an extra treatment and hence increases the risk of con- tamination of cream. Homogenization increases the viscosity, which is preferred by consumers, but also increases the potential for light-induced rancidity (manifested as oxidized flavour) owing to the increased surface area resulting from homogenization, It is used only for some types of creams, such as half cream or single cream, to prevent fat separation. Production of Milk on theFarm This should be done in as hygienic a manner as possible, since although vegetative cells may be killed by subsequent heat treat- ment, spores and organisms such as Bacillus cereus ‘can survive and cause subsequent spoilage of the milk. ‘Transport to the Dairy Milk should be transported at 5°C. It is a common practice to hold the milk at 5°C for up to 48h in creameries. Storage of Milk in the Dairy Milk should be stored at less than 5*C in silos or suitable tanks until cream manufacture, Separation and Standardization of Cream Milk is heated at 44-55°C for separation of the cream. Tem- peratures less than 40°C yield a product with high viscosity and the potential for development of off flavours due to lipolytic activity, whereas tem- peratures above 55°C cause the cream to thicken rapidly and excessively during storage. The cream separation is carried out continuously in centrifugal separators which have separate ports for skimmed milk and cream. The mechanics of keeping cream separated from skimmed milk depends on the type of centrifugal separator used. The fat content in the cream is controlled by restricting the cream outlet by ‘means of a throttling valve or ‘cream screw’. Double cream may also be lightly homogenized. ‘Whipping creams in general are not homogenized since inhibition in the formation of a stable foam is encountered. Homogenization is carried out after standardization at 65°C and 17 MPa. Heat Treatment of Cream Cream is a high-moisture product and has a short shelf life, Heat treatment extends the shelf life by inhibiting the growth of pathogenic and spoilage organisms and denaturing the indigenous lipases which may promote rancidity. Heat treatments must conform to one of the following minima: © pasteurization at 63°C for at least 30min or not less than 72°C for 15s; temperatures up to 80°C are also used sterilization at 108°C for 45 min © ultra-high temperature (UHT) treatment at 140°C for 2s. Pasteurization causes reduction in the viscosity of the cream, and also produces some sulphurous notes which disappear on storage. Higher temperatures result in cooked flavours, and may impair cream quality by possibly activating bacterial spores. A major defect of pasteurized cream that has not been homogenized is the formation of ‘cream plugs. This is attributed to the free fat which welds the globules together, and in extreme cases may even solidify the cream into gel. The fat composition and rate of cooling of the cream are other factors that affect plug formation.MILK AND MILK PRODUCTS/Microblology of Cream and Butter 1447 ‘The efficacy of the heat treatment must be checked by testing for Escherichia coli and phosphatase, Application of the phosphatase test to pasteurized creams presents special problems owing to reacti- vation of phosphatase on storage. Packaging Pasteurized cream for domestic con- sumption is packed in plastic pots or cardboard cartons. Polystyrene containers should be avoided because this material can cause taints; polypropylene pots are generally preferred. These plastic packaging materials are generally used for holding about 5-101 of cream. Sterilized cream is mostly produced in cans. Cans are sterilized with superheated steam, while aerosol cans are sterilized by hydrogen peroxide. Bulk ‘quantities of cream (2000-15 0001) are transported in stainless steel tankers, Storage and Distribution of Cartoned Cream A tem- perature of not more than 10°C during storage and distribution is recommended, with 5°C being pre- ferred. Cream should not be stored in places that house odoriferous materials such as disinfectants, paints, varnishes, scents or strong-smelling foods, since the cream may be rendered inedible, Sale - Possibly a Multistage Operation Cream pres- ents more problems than mille owing to distribution methods and the requirements for longer keeping quality. Sales are erratic, depending on the weather, hholiday seasons, local activities and other such factors. Cream should be dispatched from the manu- facturing dairies in chilled distribution vehicles to larger retailers who may then supply the product under identical conditions to smaller retailers or their shops via wholesalers. A typical flow sheet for the manufacture of ster- ilized cream is shown in Figure 1. Apart from clotted ‘cream, most creams are produced by means of mech- anical separators. Clotted cream has a very high vis- cosity, a golden creamy colour and granular texture. Figure 2is a flow sheet for the manufacture of cloteed cream. In whipping creams, air is incorporated at the air— ‘water interface and there is a disruption of the milk fat globule membrane. Some important factors for whipping cream are: © the amount of beating required to form a stable aerated structure © overrun, expressed as the percentage volume increase of the cream due to air incorporation © the stiffness of the whipped © serum leakage from whipped cream which is due to overwhipping and leads to sogginess if used in cakes. ae ‘Cream separators (Cream with approx. 30% tt Y Pestourized at 62°C for 308 or 72°C for 158, (Coole and standardize to requited fat content Adktion of stailaing ‘sats such as Na: HPO, + NaHCO. andseaium cate at 005% to prevant sane dae: Prehested 19140" tor 28 (to destroy spre-orming bacteria) Homogenization a 45 ~60°C “wo stages wih pressures OITT=195MPa nthe fest Single stage at ‘stage and3.5MPaintne —19.5MPS sccond tage “ , Cream ling in cans ‘Steilzation at 115-120°C for 18-S5min in-cans in bach or continuous retorts (Sepending onthe typeof retort and can size) Figure 1. Flow sheet of typical manufacturing process for ster- zed cream. Factors that affect whipping properties of cream are the fat content, temperature (should be below 10°C) and the distribution of the size of fat globules and ‘membrane structure. Whipping creams can also be foamed by aerosols. In this process, crcam is filed into hermetically sealed cans that are prefilled with an inert gas such as nitrogen. Although the foam stability in aerosol-foamed creams is low, it can be compensated for with stabilizers; this also prevents microbial spoilage. Sour cream is made by inoculating cream with cultures of lactic acid-producing bacteria such as Lactococcus lactis subspp. lactis and cremoris, and flavourproducing bacteria such as Leuconostoc mesenteroides subspp. cremoris and dextranicum, Souring takes place at 20°C and avoids spoilage by thermophilic organisms. The creams are processed in different ways and sold accordingly. For example, sterilized cream has a distinct caramelized flavour due to the in-can ster- ilization process, is usually sold in cans and has a shelf life of about 2 years. The temperatures employed are 110-120°C for 10-20 min This severe heating brings1448 MILK AND MILK PRODUCTS/Microblology of Cream and Buttor w TTadional athe Mik {Stained ino shatow pans [lows to stand for 814 foreream separation Heated over a waterbath fo 82-01°C for 40-50min Cooling of ereamtor 2h 1 CCarlu! separation of crust having 65-70% fat ® © Float method [Scala method Double cream fostad Cream having ‘on mikikim ili approximatoly large jacketed ways ‘sees, Y Heat to fo-asec for 40-600in by staam or hot water intrays , cooing flowed by refrigeration 12 Cooled below 76 emer fort2-tan 1 1 ‘Secoping ol the Cutting off tho cotod lotied eeam ‘ream tom the ays , , Packaging Packaging in polystyrene or potprepylene pots Figure 2 Flow sheet of typical manufacturing processes for clotted cream. (A) Traditional process. (B, C) Commercial pro- about protein denaturation, Maillard browning and fat agglomeration which collectively modify the texture and flavour of the cream. A process for rapid sterilization of cream, known as the ATAD friction process, consists of preheating the cream to about 70°C and then heating to 140°C for 0.548. This process can be applied successfully to creams ranging in fat content from 12% to 33%. Double, whipping, single and half cream may be UHT treated, or frozen after adequate pasteurization. Ultra-high temperature sterilization at 135-150°C for 3-Ss followed by aseptic packaging does not induce chemical changes in the same way as in-container sterilization, but creaming and fat agglomeration do take place on storage. In this process the shelf-life is limited by biochemical rather than microbiological con- siderations. Since all forms of microorganisms are destroyed, the cream can be stored indefinitely without refrigeration. Calcium-casein interactions destabilize the emulsion, and any proteases surviving the heat treatment may bring about gelation. The development of a stale or ‘cardboardy’ flavour gen- erally limits the shelf life to 3-6 months. Problems arise in controlling the UHT method for high-fat creams. Bulk storage of surplus cream may be done by freezing after pasteurization. The temperatures employed are -18°C to ~26°C. A shelf life of 2-18 months (average 6 months) is achieved. The cream is frozen in rotary drum freezers: a rotating drum containing recirculating refrigerant is immersed in a vat of cream to form a frozen film. The process can be made continuous and ensures rapid freezing with less damage to the cream than plate freezing. In plate freezing, plates containing the refrigerant are arranged vertically in parallel. The cream is poured into gaps between the plates, and this causes instantaneous surface freezing which progresses towards the centre. Cryogenic freezing using liquid nitrogen coming in contact with cream in a countercurrent manner in an insulated tunnel has also been shown to be a useful technique. However, for a good freeze-thaw stability of frozen creams, care must be taken to preserve the natural milk fat globule membrane. Itis for this reason that homogenization should be avoided for frozen creams. Additives for stabilization and improvement of whipping properties of cream are permitted in many countries. Gelatin and carboxymethylcellulose serve mainly to increase the viscosity, while the alginates and carrageenan interact with calcium-casein-phos- phate complex to enhance whipping properties. Emul- sifiers and stabilizers improve the freeze-thaw stability of cream. Low-molecular-weight sugars such as glucose and sucrose also impart freeze-thaw stability. ‘Nutritive sweeteners and characteristic flavouring and colouring ingredients are also sometimes used. Cream powders, and imitation creams produced by emul- sifying edible oils and fats in water, are other products available for industrial use. ‘The keeping quality of creams can be enhanced by following Good Manufacturing Practices which take into account the hygienic conditions of the plant and quality control of the products. Some steps that can ensure this quality assurance to the manufacturer and the consumer are as follows: © sanitizing all items that come into contact with cream at any stage by removing any dirt and killing residual organisms by heat or chemical dis- infectants such as chlorine compounds © ina manually operated plant, ensuring good super- vision by managerial staff © controlling the contamination of the air around the fillers - this is often neglectedMILK AND MILK PRODUCTS/Microbiology of Cream and Butter 1449 ‘© packaging creams in rooms that are separate from processing activities © using water of acceptable bacteriological quality (5p.p.m. available chlorine is generally used) © in-line testing of cream equipment. Mlcrofiora of Retail Cream ‘The main source of microorganisms in butter made under excellent sanitary conditions is cream. Fat glob- ules in the raw milk carry microbial pathogens as well as spoilage organisms which originate from the udder of the cow or from the hides and milking lines used in the processing. Bacteria found in milk may be placed in three groups from the point of view of the kceping quality of milk: © thermolabile organisms such as Pseudomonas spp. and coliforms which ate destroyed during pas- tenrization @ thermoduric non-spore-formers such as Emtero- coccus faecalis, which survive pasteurization © aerobic spore-formers such as Bacillus cereus. Generally, Gram-negative organisms, yeasts and moulds are destroyed during the pasteurization con- ditions used in processing cream. Psychrotcophic Bacillus and Clostridium spp. survive cream pas- teurization and so do heat-resistant microbes such as some strains of Lactobacillus, Enterococcus and spores of Bacillus and Clostridium. Bacillus spp., par- ticularly B. cereus, have been shown to be more important contributors to product failure in late summer and early autumn than at other times of the yeat. Bacillus cereus is also of particular importance in creams, since it can reduce methylene blue and hence lead to failure in the official Public Health Laboratory Services (PHLS) test. Bacillus lichen- iformis and B. subtilis are other species that produce spores and cause thinning and bitterness in cream. Yeasts are occasionally involved in the spoilage of pasteurized whipping cream containing added sucrose: Candida lipolyticum and Geotrichum candidum are of greatest importance. Controversy exists about the survival of Mycobacterium para- tuberculosis during cream pasteurization. In unpas- teurized cream, Streptococcus agalactiae, S. pyogenes, Staphylococcus aureus and Brucella abortus survive for varying periods of time and find their way into butter, Pasteurization at 62.8°C for 30 min of butter ‘made from contaminated eream can eliminate these ‘organisms. Spoilage of UHT cream is normally due to failure of packaging systems and entry of post- processing contaminants. Endospores of Bacillus species may survive both UHT and in-container ster- ilization. In pasteurized clotted cream, the microflora depends on the nature of the process, the degree of control and the standard of hygiene. In most cases, Bacillus spp. are dominant, although non-endospore- forming thermoduric species such as Enterococcus are present where lower cooking temperatures are used. Development of flavour defects in cream is ass0- ciated with high numbers (>10 per milllitee) of psychrotrophs due to post-processing contamination of milk or cream, These produce lipolytic enzymes which cause hydrolytic rancidity. The rancid flavour is caused mainly by fatty acids of C, to Cz while long- chain acids of Ci, to Cis make little contribution. due to the lower threshold of short-chain fatty aci in fat than in water compared with long-chain fatty acids. Processing steps such as agitation of milk at S— 10°C or 37°C results in a severalfold increase in lipase activity associated with the cream. The transferred enzyme is bound to the milk fat globule membrane wherein it has enhanced heat stability. This redis- tribution is of relevance in butter manufacture. Hom- ogenization of the cream at high pressure, slow cooling and subsequent storage at higher tem- peratures, slow freezing and repeated freeze-thawing also promote lipolysis. Difficulties have been experi- enced in churning cream made from rancid milk. The cream foams excessively and may take up to five times longer than normal cream to churn. Lipolysed milk and products prepared therefrom may slow down the manufacture of fermented products (in this case, sour cream) owing to the inhibitory effects of free Fatty acids developed during the hydrolytic rancidity. Pro- teolytic enzymes can originate from germination of heat-resistant spores of Bacillus subtilis in sterilized cream. This may produce bitterness and also cause coagulation, The lipolytic enzymes in these products cause thinning of the cream. These enzymes produce a ‘bitter’ taint, which reflects the use of poor-quality raw milk rich in spores or the use of dirty equipment. Presence of non-spore-forming organisms in sterilized cream indicates contamination after sterilization, and in canned cream indicates a defective or leaking can, Proteus may cause bitterness and thinning due to attack on proteins to produce peptones and poly- peptides. Associated growth of organisms may also be responsible for bitterness. This is exemplified by the production of bitterness by Rhodotorula muc- ilaginosa after souring by lactic organisms such as Lactococcus lactis. Coliforms may produce gas, and cocci may give rise to acid curdling. All these are common water-borne organisms. Yeasts such as Torula cremoris, Candida pseudotropicalis and Toru- lopsis sphaerica may produce fruity flavours in ‘whipped cream containing added sugar. Herbage-derived substances that are fat-soluble may also produce flavour taints in cream. This can be readily removed by the steam distillation process1450 MILK AND MILK PRODUCTS/Microblology of Cream and Butter developed in New Zealand, which pasteurizes the cream and also removes the undesirable volatiles. Other types of chemical taints have been reported to be caused by cows eating certain plants such as garlic and decaying fruits. Butter Manufacture and Typical Microfiora of Fresh Butter Butter is a water-in-oil emulsion with fat as the con- tinuous phase, obtained by the phase reversal of cream during the churning process in its manufacture, The ‘manufacture of butter is shown in Figure 3 and the steps involved along with their process conditions and significance are outlined in Table 3. Cream for butter manufacture should have at least undergone a heat treatment of 74-76°C for 15s, should have a fat content of about 40% so as to be amenable for con- tinuous butter-making, and should be cooled to 4- 5°C for at least 4h (preferably overnight) to enable sufficient crystallization of the liquid fat. This is fol- owed first by ageing to bring about an equilibration of the a and B forms of fat crystals, and then by agitation in tanks, washing, salting, packaging, storage and then repackaging. Butter may be either sweet cream butter which may or may not be salted, or ripened cream butter in which lactic acid bacteria ferment the citrate in the cream to flavour-imparting compounds such as acetoin and diacetyl. Sweet cream butter is bland in taste but has Faw mile ‘Warmed, separated standardleed to dested tat content Cream Pastoured ‘Sweet ream = Ripened utter cream buttor cool to 18-2116 i 4% inoculum, ‘davon facie | 19-20, hols ‘sed bactana | unt eeches t ‘about pH Ceoled ane 2-8, 4h patil fe erystalized s-re.an — [Chumning Separation at terme t Potable water {Washing _] > Waste water ¥ Tadiion feat and Salting water to ve 2% at in fal product E i Maing (knescing) ¥ Buter z >[ Rata pactaging (Gal storage ater sting Distetion Figure 3 Flow sheet for manufacture of butter.MILK AND MILK PRODUCTS/Microblology of Cream and Butter 1451 ‘Table 3 Steps in butter manufacture Stop, Process ‘Sionticance 1. Concentration of fat Using cream separators, For separation and standardization of resultant ‘phase in the mike Cream to the desired fat content 2. Crystalization of the fat phase 3, Phase separation and formation of water in it ‘emulsion (a) For sweet crear - cooling at 5°C for at least 4h after pasteurization of cream at 86°C for 80 min (b) for ripened cream - addition of lactic acid bacteria to pasteurized cream after cooling to 16-21°C untl a pH of .0is reached and then followed by cooling to $-5°C (Churning and working a proper blend of solid and Tiquid fat, usually at 57°C. Develops an extensive network of stable fat crystals {a) Disrupts membranes on mk fat globules, ‘followed by effective clumping that further ‘causes butterfat to harden {b) Enhances diacety| production in ripened ‘cream butter Removes excess buttermilk Inhibits microbial growth Protection from air, workers, plant environment ‘and temperatures that may promote spolage 4. Washing Rinsing with water 5. Salting Using finely ground sat or brine containing 26% ‘w/w salt, oF suis of salt in saturated brine Containing 70% sodium chloride 6. Packaging Cardboard boxes ined with vegetable parchment, ‘lurrinium fol or plastic fms for bulk packaging 7. Storage 15°C to -20°C 18. Repackaging or retail outlets nutty or boiled milk favour, this is preferred in America, Australia and New Zealand. In Europe, Latin America and Asia the preference is for intense flavour which can be developed by use of milk cul- tures. In ripened cream butter, diacetyl formation can be enhanced by incorporation of air by intensive stir- ring of the cream, using ripening temperatures below 15°C, maintaining an optimal pH below 5.2 and adding 0.15% citric acid to cream. The level of diacetyl in ripened cream butter is 0.5-2.0mg kg" butter. Diacety! also inhibits Gram-negative bacteria and fungi. Another type of butter is whey cream butter, which is processed from whey cream and is indistinguishable from sweet cream butter. Whey cream in turn is obtained from milk fat recovered from the whey produced during cheese-making. In the manufacture of ripened cream butter, pasteurized cream is cooled to 6-8°C for 2h or more to initiate fat crystallization followed by warming to 19-21°C and then inoculation with pure or mixed strains of Lactococcus lactis subspp. lactis, cremoris and diacetylactis, and Leuconostoc mesenteroides. In some areas of Europe Candida krussi has been tried in mixed cultures. Ripening occurs for 4-6h until a PH of 4.6-4.7 is achieved, and the product is then cooled. Spoilage microorganisms are primarily con- trolled through the bacteriostatic effect of lactic acid produced in the fermentation. During the churning and working of the cream, most of the starter culture is retained in the but™ termilk; however, about 0.5-2.0% of the cultures remain in the butter. The NIZO method for manufacture of cultured butter is used in many factories in western Europe. In this method, starter culture is not added to the cream, but instead a mixture of diacetyl-rich permeate and starter cultures is worked into butter. The permeate is itself produced by the fermentation of delactolized whey or other suitable medium. The method has advantages of greater control over the manufacturing process, lower risks of oxidative defects, lower chances of hydrolytic rancidity, less need of starter cultures, better quality of butter even after 3 years of cold storage, and elimination of pumping problems often encountered with viscous ripened cream. The PH of the butter made with this process is also easier to adjust to the desired range of 4.8-5.3. Salt is added to butter after removal of the excess buttermilk, It should distribute evenly in the moisture phase of the product and can inhibit microbial growth, Salt can be used in finely ground form or as. a slurry in saturated brine solutions containing up to 70% sodium chloride. The microbial quality of water used for washing or for preparation of brine is of great importance: Listeria is known to survive in a saturated brine solution at 4°C for 132 days, hence ‘water used to prepare brine must be free of Listeria. Paychrotrophic organisms multiply in salted butter stored at temperatures as low as ~6°C, owing to the fact that salt lowers the freezing point of water and allows the growth of these organisms. Addition of herbs for herb butters is also practised in certain countries to increase the variety of foods.1452 MILK AND MILK PRODUGTS/Microbiology of Cream and Butter ‘Table 4 Most common spollage and defects encountered in butter ‘Nature of spoilage or defect Causative factors ‘Spoilage Bacterial spoilage Contaminated water supplios Improper distribution of salt ‘Ternperature abuse in sweet cream buttor Putriity or ‘surface taint” and hydrolytic rancidity Mould growth producing musty favour Pseudomonas spp. such as P fragi, Shewanella putrefaciens and P.fuorescans hich grow on butter surfaces at 4-7°C and produce proteases and lipases Growth of Rhizopus, Geotrichum, Penicillium and Cladosporium which may cause hyoroiytiorancidity Humilty above 70% Improper personal hygiene of workers in the manufacturing plant ‘Matty flavour, skunk-lke odour and black ecoloration Colour changes ‘Acid production Detects Matalic taste and smelt Sospy taste and smell Short, brite structure ‘Salve-tko, greasy structure Growth of Lactococcus lactis var. matigenes, Pseudomonas mephitica and ‘Alteromones nignfaciens ‘Surface growth of various fungi which produce coloured spores. Growth of yeasts euch as Saccharomyces, Candida mycoderma, Tonulopsis holt (Over-acification of cream, high level of metalic ions in wash water, defects of ‘tinned utensis ‘Contamination with cleansing agent residues Butterfat too hard, improper cooling during ripening “Too much liquid fat in fat lobules, defects in ripening, too high buttering and ‘kneading temperature ‘Streaky, marbled appearance Flat or insipid flavour in freshly made butter Uneven salt distribution, blending of butter Excessive washing of butter grains during manufacture, ution of cream with ‘water, initial stages of bacterial deterioration Medicinal favour in butter Soft butters that are spreadable at 5°C can be pro- duced by the following ways: 1. Using cream from the summer period, which is the softest. 2. Subjecting the butter to extra working. 3. Incorporating about 10% soft vegetable far with milk fat to give a normal butter composition (80% fat minimum, 16% moisture). 4, Reducing the percentage fat below the traditional 80% to as little as 50% fat. This low-fat dairy spread contains about 11-15% milk solids and emulsifers in order to maintain a stable water-in- fat emulsion. However, traditionally and legally, butter must contain not less than 80% of only milk fat. The products described in (3) and (4) above cannot be called butter, but are dairy-type spreads. ‘At least six different continuous processes of butter manufacture are available, each of which utilizes spe- cialized processing equipment and technology. Con- tinuous processes for butter manufacture are generally economically advantageous and are finding increasing, use. Possible Problems During Storage All commercial butter is produced from pasteurized cream. If properly performed, pasteurization destroys all pathogens and more than 99.9% of organisms present in milk or Use of medicaments for treating cows, presence of chlorine compounds in milk cream, The only avenue for infection and spoilage during storage is post-pasteurization carelessness. Microbiologically induced flavours developed prior to pasteurization may be carried over into butter. The introduction of stainless steel equipment has elim- inated many flavour problems, particularly those due to yeasts and moulds. Spollage of Butter ‘The low temperatures (below ~10°C) employed for bulk storage of butter are inhibitory to the growth ‘of most microorganisms, and a general decline in numbers is expected. The lethal effect of temperature is, however, selective. Survival of Micrococcus spp. and yeasts is generally greater than that of the Enterobacteriaceae. Microorganisms that enter during reworking and packaging and those that survive low temperatures are capable of growing during retail and domestic storage at temperatures greater than 0°C. ‘Various types of spoilages and defects have been encountered in butter (Table 4). All of the Pseudo- ‘monas groups found in butter are psychrophiles, and have been traced to wash water. They grow well at refrigerator temperatures and produce putrid or lipolytic flavours in 5-10 days. These psychrophiles also produce extracellular phospholipases, which are important, since they have the ability to degrade the phospholipids of the milk fat globule membrane,MILK AND MILK PRODUCTS/Microblology of Cream and Butter 1453 Table 5 Suggested microbiological standards for cream and butter Product Test (Count or results ‘Raw cream for direct consumption “Total bacterial count ‘<30000 (10.000) por mit ‘Total coliform count <30 (10) per mi E coll faecal typo) 1 (10) per mi ‘Methylane blue reduction time (at 36°C) Not less than 7h ‘3h resazurin test fat 36°C, Lovibond disc No. 4/2) Not loss than 4 ‘Staphylococeus aureus (coagulsse-postve) ‘<10(1) por mi ‘Somatic cell count -<5000000 (250000) per mi Pastourzed cream “Total bacterial count -<90000 (6000) per mi ‘Total colforms <1 0.1) per mi E coll aecal type) ‘Absent in 4 mi Butter ‘Contaminating organisms (non-lactio-acid bacteria) -< 10000 (6000) per 9 “Total bacterial count (non-cultured butter only) +<50000 per g Total coliforms <10(1) perg. E.coli faecal type) Absent int 9 ‘Staphylococcus aureus (coaguiase-positive) ‘Absent int g Yeasts and moulds <10perg Proteolytic organisms <100 per g Upotytc organisms <50perg “Figures in parentheses are target values. thereby increasing the susceptibility of milk fat to lipolytic attack. Most lactic starters used in the manu- facture of fermented milk products have a weak lipo- lytic activity. While itis the natural milk lipase that accounts for hydrolytic rancidity in milk and cream, microbial lipases assume greater significance in stored products. The off flavours developed as a result of hydrolytic rancidity are described variously as ‘bitter’, ‘unclean’, wintry’, ‘butyric’ or ‘rancid’. These defects are sometimes evident during manufacture, but may also develop during storage. Butters made from creams that have undergone hydrolytic rancidity may not show this defect, since the rancid flavours arising due to short-chain fatty acids (C, and C,) are water- soluble and are readily lost in buttermilk. However, if lipolysis occurs after the manufacture of cream, off flavours associated with butters with a low degree of hydrolytic rancidity may also appear to be intense. This discussion is valid only for sweet cream butter. Ripened cream butter is believed to be less prone to hydrolytic rancidity. However, yeasts such as Candida lipolyticum, Torulopsis, Cryptococcus and Rhodo- torula are capable of growth and lipolysis at low temperatures. These are particularly favoured at the low pH of some cultured cream butter. Microbiological Standards for Cream and Butter Microbiological standards for cream are not favoured by many because of the complexity of the factors involved. However, based on total colony count, methylene blue reduction test and coliforms, a dis- tinction can be made between satisfactory, doubtful and unsatisfactory types. Suggested standards for cream satisfactory for buttermaking are counts of less than 1cfum!" for yeasts, moulds and coliforms, and a total colony count of less than 1000 cfu”. Table 5 gives the limits that have been proposed as micro- biological standards for cream and butter. Complying with these standards may be 2 useful tool to give good-quality products. Bacteriological standards for cream in some countries have been laid down as follows: © Northern Ireland: untreated — bacterial count < 50000 per gram pasteurized — no coliforms in 1g © Canada: ‘count less than 50000 per gram ‘no coliforms in 1g phosphatase negative © Sweden: count <10000 per gram coliforms <10 per gram aerobic spores < 100 per gram. Public Health Concerns The incidence of documented food poisoning asso- ciated with butter consumption was low even before the widespread pasteurization of cream for manu- facture of butter. Early outbreaks of diphtheria (caused by Corynebacterium diphtheriae) and tuber- culosis (caused by Mycobacterium tuberculosis or M. bovis in naturally contaminated cream) in the USA and Europe, and of typhoid fever (caused by Sal- monella typhi) in the USA from 1915 to 1927, have been reported to be caused by butter, Butter con- taminated by a convalescent carrier of S. typhi was1454 MILK AND MILK PRODUCTS/Microbiology of Cream and Butter responsible for 35-40 cases (including 6 deaths) of typhoid fever in Minnesota in 1913. Salmonella typhi can survive for 2-4 weeks in butter prepared from contaminated cream and hence acts as a vehicle of infection in outbreaks. Some major public health con- cerns with respect to cream and butter are outlined below. Aftatoxins Aflatoxins are secondary metabolites produced by ceftain moulds, namely Aspergillus flavus, A. para- siticus and A. nomius, and are recognized as extremely potent liver carcinogens for both animals and humans. Four types of aflatoxin designated AFB1, AFB2, AFGI and AFG2 are currently recognized, of which AFBI is the most potent and comes from contaminated feeds. Ingestion of aflatoxin-con- taminated animal feed leads to the excretion of the less toxigenic AFM1 in milk within 12-24h. While many countries have legislation regarding aflatoxin limits in animal feed, the USA and many European countries also have legislations for maximum levels of AFM1 in milk and other dairy products. Present evidence indicates the level of AFM1 in milk and dairy products to be relatively unaffected by pasteurization, sterilization, fermentation, cold storage, freezing, con- centration or drying. However, treatment with hydro- gen peroxide, benzoyl peroxide, ultraviolet light, bisulphites, riboflavin or lactoperoxidase have been shown to be effective in reducing the levels of AFM1 in experimental trials. Only about 0.4-2.2% of the ingested AFB1 appears in milk as AFM1. Further- more, since AFMI is water-soluble it partitions nat- ually during manufacture of cream and butter ‘Typically, about 10% of the AFM1 in milk appears in cream and about 2% appears in butter. Rigid moni- toring of animal feed for AFB1 can control the AFM1 levels in dairy products including cream and butter. Brucellosis Brucellosis remains one of the most widespread and costly diseases afflicting humans and animals and is, acquired by direct or indirect contact with infected animals harbouring any of three of the six bacterial species belonging to the genus Brucella. Human bru- cellosis ranges from a mild, flu-like illness to a severe disease. The severity depends on the species involved, with B. melitensis being the most pathogenic for humans, followed by B. suis and B, abortus. Osteo- :myelitis is the most common complication of B. miel- itensis infection, followed by skeletal, genitourinary, cardiovascular and neurological complaints. Cream and butter are unusual sources for Brucella spp. with only $ out of 916 cream samples being positive in one outbreak-related survey. Both these products can extend survival of B. melitensis and B. abortus for 4 6 weeks stored at 4°C. These microorganisms can survive even longer in refrigerated butter, persisting for 6 months and 13 months in salted and unsalted buteer respectively. Dairy-related brucellosis out- breaks have been virtually eliminated as a result of immunization of livestock, slaughtering of infected animals and mandatory pasteurization of milk. Listeriosis Listeria monocytogenes, the causative agent of this disease, has recently emerged as a serious food-borne pathogen which can cause abortion in pregnant ‘women, and meningitis, encephalitis and septicaemia in newborn infants and immunocompromised adults. ‘The organism is a Gram-positive, non-spore-forming, facultatively anaerobic, short, diphtheroid-like rod- shaped bacterium that occurs singly or in short chains. Listeria infections are devastating, with a mortality rate of 20-30%. Dairy-related outbreaks of listeriosis, two in Switzerland and one in the USA in the 1980s have been linked to consumption of various products including pasteurized milk. Cream has been impli- cated in a major outbreak of listeriosis in Fialle, East Germany, during the period 1949-1957. Listeria monocytogenes can attain populations of 10° cfu ml"! in whipping cream after 8 days of storage at 8°C. It has occasionally been recovered from commercially produced butter, with survival up to 70 days also being reported in butter prepared from inoculated Salmonellos! ‘The causative agent is Salmonella typhi. It produces infections ranging from a mild, self-limiting form of gastroenteritis to septicaemia and life-threatening typhoid fever. Salmonellae are short, Gram-negative, facultatively anaerobic, rod-shaped bacteria and can grow at 545°C. Inadequate pasteurization and post- processing contamination have occasionally resulted in milk and cream that test positive for Salmonella, evidenced by various outbreaks of salmonellosis. The numbers of salmonellae decrease in fluid milk prod- uucts and butter prepared from inoculated cream during extended storage at 7°C or below. Staphylococcal Poisoning Staphylococcal poisoning results from ingesting a pre- formed, heat-stable toxin (enterotoxin) which is pro- duced by the bacterium Staphylococcus aureus. The bacteria are facultatively anaerobic, non-motile, small Gram-positive cocci growing at temperatures of 10— 45°C and a pH of 4.2-9.3. Ten serologically distinct enterotoxigenic proteins known as enterotoxin types A, B, C1, Co, Cs, D, E, BG and H are recognized inMILK AND MILK PRODUCTS/Microbiology of Cream and Butter 1455 S. aureus. The severe intoxication is of short duration and develops 1-6 h after ingestion of the enterotoxin. ‘The common symptoms are nausea, vomiting, diar- thoea, abdominal cramps and mild leg cramps. Between 1951 and 1970 cream has been implicated in six outbreaks of staphylococcal poisoning in the USA involving 131 cases. Large numbers of S. aureus are seldom found in butter since the product com- position and storage conditions severely limit its growth. However, when cream was inoculated with S. aureus, incubated for 24h at 37°C and then churned to butter, the finished product contained at least 1 ug of enterotoxin per 100g, or approximately 10% of the enterotoxin present originally in the cream. Since 0.1 jg of enterotoxin can induce symp- toms of staphylococcal poisoning, ingesting such a dose poses a potential health hazard. This has been demonstrated by butter-related outbreaks. See also: Aspergillus: Introduction; Aspergilus flavus Bacillus: Bacilus cereus. Food Potsoning Outbreaks. Heat Treatment of Foods: Ultra-high Temperature (UHT) Treatments; Principles of Pasteurization. Lac- tococeus: Lactococcus lactis Sub-species lactis and cremoris. Leuconostoc. Listeria: Listeria mono- cytogenes. Milk and Milk Products: Microbiology of Liquid Mik; Microbiology of Dried Mik Products. Pack= aging of Foods. Pseudomonas: Introduction. Sal- ‘monelia: Salmonella typhi. Spoilage Problems: Problems Caused by Bacteria. Staphylococcus: intro- duction; Detection of Staphylococcal Enterotoxins. Ultra sonic Imaging. Further Reading Early R (ed,) (1992) The Technology of Dairy Products. Glasgow: Blackie. Fox PF (ed.} (1983) Developments in Dairy Chemistry 2: Lipids. London: Applied Science. Lapides DN (ed.) (1977) McGraw-Hill Encyclopedia of Food, Agriculture and Nutrition. New York: McGraw- Hill. Marth EH and Steele JL (eds) (1998) Applied Dairy Micro- biology. New York: Marcel Dekker. Robinson RK (ed.) (1990) Dairy Microbiology. Vol. 1: The Microbiology of Milk, 2nd edn. London: Elsevier. Robinson RK (ed.) (1990) Dairy Microbiology. Vol 2: The Microbiology of Milk Products, 2nd edn. London: Elsevier, Robinson RK (ed.) (1994) Modern Dairy Technology. Vol 1: Advances in Milk Processing, 2nd eda, London Chapman & Hall. Spreer E (1998) Milk. and Dairy Product Technology. New York: Marcel Dekker. ‘Varnam AH and Sutherland JP (1994) Milk and Milk Prod- ucts Technology, Chemisiry and Microbiology. London: ‘Chapman & Hall. [_ Millet see Fermented Foods: Beverages from Sorghum and Milt ] [_Mineral Metabolism soe Metabolic Pathways: Metabolism of Minerals and Vitamins, |