CONTROLLING MICROBIAL GROWTH IN THE ENVIRONMENT

BASIC PRINCIPLES OF MICROBIAL CONTROL

TERMINOLOGY

2 major types: sterilization (eliminate all) & disinfection/antisepsis (destroy vegetative (nonspore) & many viruses) Sterilization: destroys all viable microbes, including bacterial endospores & viruses, although not prions, in/on an object; guarantees all pathogens are eliminated Aseptic: environment/procedure free of contamination by pathogens Disinfection (inanimate objects only, private): use of physical/chemical agents (disinfectants) to inhibit/destroy microorganisms (especially pathogens) on inanimate objects; disinfectants include UV light, heat, alcohol, bleach, ozone, chlorination, etc; doesnt guarantee all pathogens eliminated, as disinfectants alone cant inhibit endospores or some viruses; more concentrated than antiseptic & can be left on surface for longer Antisepsis: inhibition/killing of microorganisms on skin/tissue using chemical antiseptic; often same components as disinfectant Modifications of disinfection Degerming: removal of microbes from surface by scrubbing; soap/alcohol used, but the actual scrubbing is more important Sanitization (public): disinfecting surfaces/utensils used by public to reduce pathogens to meet public health standards Pasteurization: use heat to kill pathogens & reduce number of spoilage microorganisms in food/beverages Suffixes -stasis/-static: chemical/physical agent inhibits microbial metabolism/growth, but doesnt necessarily kill microbes (refrigeration is bacteriostatic since it inhibits growth, but they can resume metabolism when optimum temperature is restored) -cide/-cidal: agent destroys/permanently inactivates a particular microbe (virucides inactivate viruses, bactericides kill bacteria, fungicides kill fungal hyphae/spores/yeast, germicides destroy pathogenic microorganisms in general)
MICROBIAL DEATH RATES

Microbial death: permanent loss of reproductive ability under ideal environmental conditions Microbial death rate: measurement of efficacy of antimicrobial agent Microbicidal agents dont simultaneously kill all cells, rather they kill a constant percentage of cells over time (indicated by straight line)
ACTION OF MICROBIAL AGENTS

Alternation of cell walls & membranes If cell wall is disrupted by physical/chemical agents, it cant prevent cell from bursting as waster moves in by osmosis Damange to membranes proteins/phospholipids by agent allows cellular content to leak out Enveloped viruses: envelope is membrane of proteins/phospholipids thats responsible for attaching the virus to target cell, thus damaging envelope fatally interrupts viral replication; lack of envelope in nonenveloped viruses account for greater tolerance to antimicrobials Damage to proteins & nucleic acid Certain chemicals/extreme heat denature proteins, bringing about cell dfeath Chemicals, radiation, heat also alter (and even destroy) nucleic acids; since genes of cell/virus are composed of nucleic acids, disruption can produce fatal mutations Portion of ribosomes that catalyze protein synthesis is a ribozyme (enzymatic RNA molecule), so agents that interfere w/ nucleic acids also stop protein synthesis
SELECTION OF MICROBIAL CONTROL METHODS
FACTORS AFFECTING EFFICACY OF ANTIMICROBIAL METHODS

Site to be treated Harsh chemicals/intense heat cant be used on humans, animals, fragile objects (artificial heart valves, plastic utensils) Level of control also based on site of procedure (needles/scalpels have greater infection risk so theyre sterilized, surface contact just has to be disinfected) Relative susceptibility of the microbes Scientists/medical usually select method to kill hardiest microbes present (assuming more fragile will be killed as well) Most resistant microbes: bacterial endospores (especially bacillus & clostridium); species of mycobacterium (have lots of waxy lipids); protozoa cysts; prions more resistant than any living thing Germicide effectiveness classified as high/intermediate/low depending on proficiency inactivating/destroying microbes on medical instruments that cant be sterilized w/ heat - High: kill all pathogens, including endospores; sterilize invasive instruments (catheters/implants/heart-lung machines) - Intermediate: kill funal spores, protozoan cysts, viruses, pathogenic bacteria, but not endospores; disinfect noninvasive instruments that come in contact w/ mucous membranes (respiratory equipment , endoscopes) - Low: eliminate vegetative bacteria, fungi, protozoa, some viruses; disinfect items that only contact skin (furniture, electrodes) Environmental conditions Temperature & pH affect microbial death rates/efficacy of methods Warm disinfectants work better than cool (chemicals react faster at higher temperatures)

 Antimicrobial effect of heat enhanced by acidic conditions; some chemical disinfectants (household bleach) more effective at low pH Organic materials (fat, feces, vomit, blood, biofilm secretions) interfere w/ penetration of heat, chemicals, some radiation (in some cases they inactivate chemical disinfectants); therefore its important to clean objects before sterilization/disinfection so antimicrobial agents can thoroughly contatct all surfaces
METHODS FOR EVALUATING DISINFECTANTS & ANTISEPTICS

Longer exposure time, higher concentrations & fresher solutions more effective than more dilute/older ones Phenol coefiicient: evaluate effectiveness by comparing agents efficacy to that of phenol; coefficient greater than 1.0 indicates agent is more effective than phenol Use-dilution (US standard): evaluate effectiveness against specific microbes in which most effective agent is the one that entirely prevents microbial growth at highest dilution (dip metal cylinders in bacteria broth, dry, immerse each in different dilution of evaluating disinfectants, 10 minutes later remove/incubate) Kelsey-sykes capacity (europe standard): determines capacity of chemical to inhibit bacterial growth (chemicals added to bacteria broth cultures, turbidity measured after 48 hours an indication of growth inhibition) In-use (realistic conditions): more accurate determination of proper strength/application procedure of agent for each specific situation; swab objects (operation room equipment) before/after application of agent, inoculate (appropriate growth media), incubate

TERM Sterilization Disinfection - Degerming - Sanitization - Pasteurization Antiseptis Aseptic -cide/-cidal -stasis/-static

DEFINITION Destroys all microbes/viruses in/on object Destroys most microbes/viruses on nonliving tissue Remove microbes by mechanical means Remove pathogens from objects to meet public health standards Heat destroys pathogens & reduces number of spoilage microbes in food/beverages Reduce number of microbes/viruses (particularly pathogens) on living tissue Environment or procedure free of pathogenic contaminants Suffixes indicating destruction of type of microbe Suffixes indicating inhibition, not complete destruction of type of microbe

EXAMPLE Microbiological culture media & canned food Phenolics, alcohols, aldehydes, soaps Handwashing, alcohol swabbing at injection site Washing tableware in scalding water in restaurants Pasteurized milk, fruit juices Iodine, alcohol Preparation of surgical field, handwashing, flame sterilization of equiptment Bactericide, fungicide, germicide, virucide Bacteriostatic, fungistatic, virustatic

COMMENTS Achieved by steam under pressure, incineration, ethylene oxide gas Term used in relation to pathogens Chemicals arent as important as scrubbing (mechanical removal) Standards vary among jurisdictions Heat treatement brief (to maintain taste/nutrients), microbes still there Usually disinfectants w/ reduced strength to make them safe Aseptic techniques Germicides include ethylene oxide, propulene oxide, aldehydes Germistatic agents include some chemicals, refrigerate, freezing