Basic Microbiology.

The central principle of infection control is that of control of the growth of microorganisms which potentially cause us harm. To underpin the concepts of infection control it is useful to understand some basic microbiology regarding the types of organisms involved, how they may grow and multiply, and the kinds of diseases they may be responsible for. Basically there are three groups of micro-organisms: • Bacteria • Fungi • Viruses

There are many different types of bacteria in existence, and by far the greater percentage of them are harmless to us. Bacteria are also essential to our daily lives, ensuring that we remain healthy, helping the development of drugs, and also in the development of new foods. Bacteria can be grouped visually, depending upon their shape and arrangement : • Cocci – all round or spherical • Bacilli – rod-shaped • Spiral-shaped. (coccobacillus falls between coccus and bacillus). Each group may be further sub-divided : Cocci Monococcus –arranged singly Diplococcus –2 together Staphylococcus –haphazard Streptococcus –chains Sarcina – clumps in cubes Gaffkya –groups of four Staphylococcus is found normally in the body in three formss. aureus in the nasal passages s. epidermis on the skin s. albus in the mouth. Neisseria are similar to diplococcus, but are kidney-shaped. These are usually found in pus, i.e. N. gonorrhoea, N. meningitides.

Bacilli Found as single rods or as chains. Single rods are randomly arranged, and may be either long and thin or short and fat. E. coli is an example, normally found in the bowel. Salmonella is another example.

Lysozymes attack the polysacchyaride ‘backbone’ of the muerin. The cell wall is more susceptible to mechanical damage. Teicoic acid (another polysaccharide) is also associated with the cell wall. a layer of phospholipids. which is why these bacteria do not take up the Gramm stain. and is also the site where enzymes occur. a peptidoglycan. This forms a very rigid framework for the cell wall. The cell wall determines the shape of the bacteria. This occurs because all bacterial cell walls contain muerin. Muerin consists of parallel strands of polysaccharide. forming a net around the bacteria. The cell membrane of the bacteria is differentially permeable.Spiral. With these bacteria there are further layers around the muerin consisting of lipid material. completely destroying the cell wall in Gramm positive bacteria. and also dyes. These layers also provide resistance to phagocytosis. The cell wall is a good target for antimicrobial drugs. and we utilise this to make it possible to identify the bacteria by immunological means. The individual layers of muerin are connected by peptide cross bridges. sited on the mesosomes. In Gramm positive bacteria the cell wall has a muerin region thicker than Gramm negative bacteria. which is dependent upon the structure of the cell wall. In Gramm negative bacteria the lipid-constituent layer . such as penicillin. Spirilla –move via flagella Vibrio – Spirochetes – Structure of bacteria. cross-linked by short polypeptide chains to give a rigid structure. and bacteria can also be classified by how they take up the Gramm stain. Gram negative bacteria contain less muerin and have no teicoic acids. and next to the muerin is a layer of phophslipid. These act as a barrier to substances passing from the outside into the cell. therefore the bacteria are resistant to lysozymes. These are layers of polysaccharide.

The capsule surrounds the cell wall. particularly those of the Bacillus and Clostridium groups. forming a circular chromosome. the bacterial cell will remain alive. If the autoclave is working correctly. but a mechanism which aids survival. These are round or oval bodies. but conditions improve the endospore will germinate and produce a new bacterial cell. mitochondria. Within the bacteria there is no nucleus. The spores are put into an autoclave. and they do not have a constant shape. will be killed. Bacterial motility. They are not reproductive structures. and after the cycle is complete they are allowed to germinate. They are almost are exclusive to Gramm negative organisms. and usually is comprised of polysaccarhide. The capsule also protects the bacteria from being phagocytised. Some bacteria can survive without a cell wall. then all spores.g. and appear much shorter than flagellae. but without the capsule the bacteria is less likely to produce the disease. Spores are formed mainly by the bacilli-type organisms.e. They are not found on cocci. then this spore will survive the raised temperature. Endospores. and is loosely attached to the cell wall. formed inside the bacterial cell in adverse conditions to help the bacteria survive. Genetic material can be exchanged between bacteria at the site of the pili.remains unharmed. i. e. Spores can be found in soil dust and air. The usual structures to aid motility are the flagellae. but are on most bacilli and some spirilla. whilst the Clostridium group are responsible for botulism. Diplococcus pneumoniae will produce pneumonia if it has it’s capsule. However. This then indicates that the autoclave is faulty. but the muerin layer will be destroyed. endoplastic reticulum. . The mycoplasma is the smallest known bacteria to survive outside living cells. Bacillus stearothermophilis is a heat resistant spore that we can use in a clinical situation. They are usually formed from a protein material. and some ribosomes are also present. and will germinate successfully. The Bacillus group are responsible for anthrax and tetanus. including this one. lysosomes or membranous structures. They can be arranged in a number of ways : Monotrichous Amphitrichous Lophotrichous Peritrichous A pili is a structure used for adhesion found on the outside of some bacteria. which are sited on the outside of the cell membrane. There is a single strand of DNA. If the autoclave does not reach the correct temperature. There is a direct correlation between the possession of a capsule and virulence of the organisms. Endospores may remain viable for many years through adverse conditions. they are pleomorphic.

Different atmospheres will also benefit different organisms : • Obligate aerobes – these carry out aerobic respiration and therefore require oxygen from the atmosphere. Microbial growth is controlled by physical and chemical means. suitable pH 5. devitalised tissues) these organisms can flourish. Antisepsis refers to chemical disinfection of the skin. suitable temperature 4. ‘disinfection’. Sterilisation is the process of destroying all forms of microbial life. Heat and autoclaves represent the physical method. growth will resume. Disinfection is the process of destroying vegetative pathogens. suitable nutrients 2. and ‘germicide’ all refer to the destruction of micro-organisms. and magnesium. • Strict anaerobes – unable to grow and reproduce in the presence of oxygen. Disinfectants tend to reduce or inhibit microbial growth. in particular oxygen. whilst disinfectants represent the chemical method. Essential minerals indlcude calcium.Bacterial growth. suitable atmosphere 3. nitrogen. The terms ‘bacteriostasis’ and ‘asepsis’ refer to the suppression of microbial growth. In areas where there is no oxygen present (i. and usually die if oxygen is present. but not necessarily endospores or viruses. together with zinc. It is absolute. copper.e. If oxygen levels are too high. Bacteria require a readily available supply of organic nutrients. Bacteriostasis is a condition in which bacterial growth and multiplication are inhibited. but not necessarily their spores. but not killed. The terms ‘sterilisation’. but normally do not sterilise. Controlling the growth of micro-organisms. and sulphur. including spores. iron. • Microaerophilic – obligate aerobes which only require a tiny amount of oxygen. cobalt. or other living tissues. They cannot survive the absence of oxygen. ‘antisepsis’. hydrogen. carbon. The maximum size of a bacteria is the size at which maximum efficiency occurs (it grows in volume but the increase in surface area is at a much slower rate. If removed. suitable osmotic potential 6. therefore the cell eventually becomes inefficient). A germicide is a chemical agent that kills microbes rapidly. commonly producing gangrene in areas of damaged tissue. The conditions required for bacterial growth are: 1. • Facultative anaerobes – can grow with or without the presence of oxygen. mucous membranes. mosture. phosphorous. potassium and magnesium. they will be destroyed. and those that respire anaerobically in the absence of oxygen and aerobically if oxygen is present. Individual bacteria at maximum size will split by binary fission into two bacteria. Asepsis is the absence of . These are sub-divided into two groups – those that respire anaerobically whether oxygen is present or not. and there are no degrees of sterilisation.

and is often used to needles or forceps. In it’s dry form is kills by the oxidation effect. and all types of organism can be destroyed. Heat is more effective under acidic conditions. Flaming is 100% effective. Moist heat methods include boiling and autoclaving. Endospores are much more resistant than vegetative cells. Gramm positive organisms are more susceptible to disinfectants and antiseptics than Gramm negative organisms. denaturing the cell’s proteins.g. Other agents work by damaging the proteins or nucleic acids within the cell. Physical methods of destruction. Anti-microbial agents work by altering the permeability of the cell membrane. whilst physically hot water has a greater heat content than air at the same temperature.pathogens from an object or area. but with an increase in age comes an increase in resistance. It can be used in either a dry or moist form. incineration. Botulinum will survive after 5½ hours of boiling. Dry heat methods include flaming. Pseudomonads are resistant to chemical agents. denaturing the protein by a different method. killing of micro-organisms is not instantaneous. agent used must be able to affect the micro-organisms directly. Heat. and will actively grow in some disinfectants and antiseptics. They can also survive in simple saline solutions. the temperature. but it has limited use . If these proteins are destroyed then the cell is unable to synthesise proteins. as are some chemical agents.e. spores of C. These apply to all methods of control of microbial growth : 1. the item must be cleaned so as to remove extraneous soil. Micro-organisms are also more susceptible to chemical agents during their growth. and hot air sterilisation. the nature of the organism b. 4. Aseptic technique is designed to prevent the entry of pathogens into the body. leading to cell death. Basic principles of controlling microbial growth. i. and the time required depends upon : a. when the normal flora has been suppressed during abti-biotic therapy. e. the numbers of organisms present d. 2. 3. With moist heat spores and bacteria are killed more rapidly at a given temperature for two reasons – chemically the proteins are denatured due to the breaking of the hydrogen bonds. This is rapid and penetrates objects as well as clumps of micro-organisms. moisture is essential for the action of chemical agents. In it’s moist form because the presence of the water allows the hydrogen bonds in the proteins to be broken down. They are a major cause of problems in hospitals as they are opportunistic pathogens in the absence of the normal flora of the body. causing leakage of the contents and affecting growth. the nature of the agent used c. and are resistant to many antibiotics.

Viruses and larger proteins can also be removed. . Filtration. i. antibiotics.g. Iodine – inhibits protein These agents may act alone or as a function and is a strong component of inorganic/organic oxidiser compounds. The following table illustrates some of the commonly used chemical agents. but this works very slowly. organic material. This method is used especially for scalpel blade sterilisation. Disrupts the plasma No longer widely used due to its membrane and denatures irritating properties. Incineration rapidly kills micro-organisms and is reliable. but not cloth or rubber. thymol. This is the passing of a gas or liquid through a filter with pores small enough to prevent bacteria passing through. and also for air-filtration. and also items made from glass or metal. teratogenic. destroying pathogens. It’s effects are also cumulative. They are both effective Chlorine – forms antiseptics. 120oC of dry heat will take 8 hours to achieve sterilisation. It is the standard for measuring the effectiveness of other disinfectants (phenol co-efficient). and may also be proteins. Phenolics Disrupts the plasma These are derivatives of phenol and membrane and denatures are reactive even in the presence of proteins. Ultra-violet light may also be used. especially for vaccine sterilisation. acting on the cell ineffective against endospores and membrane. as higher temperatures are required for longer periods of time to ensure cell death. Examples include cresol. Used for skin swabbing and also wiping instruments. This method is used for liquids which would be destroyed by heat. a strong oxidiser which alters cellular components. Used to disinfect surfaces and instruments as well as skin surfaces and mucous membranes. Gamma rays will penetrate cells. Radiation. killing them by destroying their DNA. Chemical agents should be selected so as to kill the organisms as quickly as possible. Halogens. Sterilisation by this method is also effective for enclosed containers. Lysol. xylenol. but solvent. Alcohols. but they do reduce the microbial population to safe levels. Agent Action Uses. The use of hot ovens is not as efficient as moist heat. e. Phenol. This is reduced to 20 minutes at 180oC.e. non-enveloped viruses. Chemical methods of destruction. Very few chemical agents actually achieve sterility. hypochlorous acid.otherwise. Dehydrates and is a lipid Bacvtericidal and fungicidal.

Bactericidal. preventing their use by pathogens. and as food preservatives to inhibit mould growths. bacteriostatic. found commonly in the nose and anus. mouth. The major constituent of the normal flora is S. The normal flora of the body. Commensals benefit from living on the body. i. (QAC) Germicidal and antiseptic in action. and virucidal against enveloped viruses.e. Hydrogen peroxide is used as an antiseptic in deep wounds. Very effective agents. They also maintain certain physiological conditions within the body. Many organisms inhabit the body without causing us any harm. denature ammonium proteins and disrupt plasma compounds membranes. odourless. Surface-active agents. making up around 90% of the population. but we gain no benefit directly from their presence. Acids and alkalis. Oxidising agents. The normal flora will utilise any nutrients that are available. Ozone can be used to replace chlorine for disinfecting water. Those that have no exterior connection have no flora. Quarternary Inhibit enzymes. making it impossible for pathogens to flourish. Cause hydrolysis and denaturing of proteins. Oxidise cellular components of cells. Interfere with cellular oxidation. Inactivate proteins. Mutuals are bacteria which actually aid some process in the body. Aldehydes. Another common organism is S. Flora on the epithelial layers occupy specific attachment sites.Heavy Denature proteins metals and their compounds. They are colourless. aureus. fungicidal. and is some cases are positively beneficial and essential for health. tasteless. These mostly live on the skin. Will also inactivate micro-organisms and vaccines. such as vitamin K-producing bacteria in the gut. albus. Used to preserve and embalm specimens. The areas on the body which mainly have normal flora are those areas which have an exterior connection. stable and easily diluted. and once these attachment sites are occuplied there is no-where for pathogens to attach and establish themselves. such as pH. Used for skin antisepsis and washing/disinfecting instruments. They can also produce secretions which inhibit invading micro-organisms. Antiseptic and antifungal. Diphtheroids occur cutaneously. and commonly inhabit oily areas such as the . Dyes. and rubber goods. non-toxic. Used on the skin to control dermatophytes. utensils. Common example id formalin.

aspergillus. e. Oral and upper respiratory tract – pharynx and trachea contain many potentially pathogenic organisms. c. Urogenital tract – depends upon age. • Mechanical damage to the skin or tissues. and this is often the site of the initial colonisation of pathogens. will multiply. whereas only 1 rickittsia will cause Scrub Typhus. but α-haemolytic streptococci exist in the mouth and can spread to the skin. d. but in addition particles of dust and other materials can be trapped which contain spores and bacteria. • Debilitated patients. thrush. especially those recovering from major surgery. portal on entry – the organisms enters the wrong portal it cannot cause disease. . must attach to epithelial surfaces. penicillium. numbers of micro-organisms – salmonella poisoning requires over 1 million organisms. and hormone levels. f.sebaceous glands. Nail flora – this is generally similar to that of skin. They are associated with Gramm negative bacteria and tend to be non-specific. cancer. Disease production depends upon a number of factors : 1. ability to multiply. thereby killing all bacteria. pH. Streptococci are seldom found in normal flora. b. • Large amounts of alcohol. Microbial invasiveness and disease production. Intestinal tract – the concentration of organisms increase towards the end of the small intestine. e.g. They result in headache. a high percentage of which are anaerobic (more than 90%). such as fungi and yeasts. There is a high concentration in the colon. There are certain conditions in which normal flora may cause infection : • when taking antibiotics that are not specific to one type of bacteria. needs correct metabolic and nutritional situation. and tend to occur in moist areas only as they cannot survive in dry areas. This destroys the normal flora and the other organisms. production of a capsule for protection b. causing infection. actual species – only certain micro-organisms cause disease. toxin production – endotoxins are part of the bacterial cell wall which is released on lysis or destruction. diabetes or chest conditions. 2. Gram negative bacilli form a very small proportion of the flora. host/microbial factors – a. and possible drugs of abuse. • Transmission of normal flora to another region. • Extreme and prolonged fatigue . • The use of drugs such as steroids. eg. specific microbial factors – a.

dystentery Secondary Mixed Acute Chronic Localised Generalised Bacterial pneumonia. clostridium tetani produces a toxin which results in Lockjaw. leprosy Staphylococcal boil Gramm negative . haemolysins – released by bacteria to break down red blood cells leukocidins – released by bacteria to destroy leukocytes coagulase – produced by some staphylococci to promote blood clotting fibrinolysis – produced by streptococci and breaks down a blood clot by destroying the fibrin hyaluronidase – breaks down hyaluronic acid. h. have the correct nutrients. e. g. f. States of bacterial infection. and produce substances such as capsules. d. They must have the ability to attach to epithelial surfaces. toxins and enzymes necessary for their survival and growth. enabling bacteria to get in between the host’s cells collagenase – breaks down collagen in connective tissues penicillinase – breaks down penicillin and other similar substances. i.g.c. and it is the exotoxin which causes the disease. temperature and environmental conditions for growth. and nausea. brief duration Prolonged duration Confined to a small area or an organ Disseminated to many Examples Asymptomatic gonorrhea in women and men Typhoid carrier Anthrax Candida infection Primary Shigella. They are mainly associated with Gramm positive bacteria. Summary of factors necessary for disease to occur : The correct species must enter the right portal in sufficient numbers for that species to cause disease. Exotoxins are secreted into the surrounding environment and are very powerful. e. viral lung infections Anaerobic abscess Diphtheria Tuberculosis. State of infection Subclinical (apparent) Dormant (latent) Accidental Opportunistic Description No detectable clinical symptoms of infection Carrier state Zoonosis and environmental exposure Infection caused by normal flora or transient bacteria when normal host defences compromised Clinically apparent invasion and multiplication of microbes in body tissues causing local tissue injury Microbial invasion subsequent to primary infection Two or more microbes infecting the same tissue Rapid onset. fever.

Pyogenic Retrograde Fulminant body regions Pus forming Microbes ascending a duct or tube against the flow of secretions or excretions Infections that occur suddenly and intensely bacteraemia Streptococcal infection E.coli urinary tract infection Pneumonic plague .

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