Presentation on theme: "Chapter 5 Microbial growth control"— Presentation transcript:
1Chapter 5 Microbial growth control Physical methodsChemical agentsFood Preservation
2Control of microorganisms by physical and chemical agents frequently used terms Sterilization: the process by which all living cells, viable spores, virus, and viroids are either destroyed or removed from an object or habitat.Disinfection: the killing, inhibition, or removal of microorganisms that may cause disease.Sanitization: the microbial population is reduced to levels that are considered safe by public health standards.Antisepsis: the prevention of infection or sepsis-cide: kill; -static: stopgermicide: kill pathogens but not necessarily endospores. Bactericide; fungicide; algicide; viricide; bacteriostatic; fungistatic
3Conditions influencing the effectiveness of antimicrobial agent activity 1. Population size.2. Population composition. Spore, young/mature3. Concentration or intensity of an antimicrobial agent.4. Duration of exposure.5. Temperature. higher6. local environment.eg. 1. Heat kill more readily at an acid pH.2. Organic matter protect microorganisms.3. Biofilm protection.
4Microbial Growth Control Physical waysHeat sterilization (including autoclave and pasteurization)Radiation (microwaves, UV, X-rays, g-rays and electrons)Filtration (depth filter, membrane filter and nucleation track (nucleopore) filter)
5Heat(terms)TDP: the lowest temperature at which a microbial suspension is killed in 10 minutes.TDT: the shortest time needed to kill all organisms in a microbial suspension at a specific temperature and under defined conditions.Decimal reduction time (D) or D value: the time required to kill 90% of the microorganisms or spores in a sample at a specified temperature. D121
6Moist heat sterilization Autoclave: a device somewhat like a fancy pressure cooker.The air initially present is forced out the chamber is filled with saturated stream121oC or 15 pounds min1. All air must be flushed out of the chamber, or it will not reach 121oC even though it may reach pressure of 15 pounds.2. The chamber should not be packed too tightly.
7PasteurizationMilk, beer, and many other beverages are treated with controlled heating at temperatures well below boiling. Not sterilize, just pasteurized.Past : 63oC, 30minHTST: 72oC, secUHT: oC, 1-3sec
8Dry heat sterilization oC hSuitable for glass petri dishes and pipettesNot suitable for heat-sensitive materials like many plastic and rubber items.
9Filtration Heat-sensitive solution Depth filters: diatomaceous; unglazed porcelain.Membrane filters: membranes with pores about 0.2m in diameter are used to remove most vegetative cells, but not viruses.Air: 1. Surgical masks and cotton plugs on culture vessels.2. Biological safety cabinets.
10Radiation1. Ultraviolet radiation: 260nm is quite lethal but does not penetrate glass, dirt films, water and other substances.uv lamps: the ceilings of roomsbiological safety cabinetsattention! uv lamp must be off2 . Ionizing radiation: not always as effective against viruses.Co60: cold sterilization for antibiotics, hormones, sutures.
11Microbial Growth Control Chemical ways (germicides)Agents that kill organisms are often called cidal agents (bactericidal, fungicidal and viricidal agents)Agents that do not kill but only inhibit growth are called static agents (bacteriostatic, fungistatic, and viristatic agents)Disinfectants are chemicals that kill microorganisms and are used on inanimate objectsAntiseptics are chemical agents that kill or inhibit growth of microorganisms and that are sufficiently nontoxic to be applied to living tissues.
12Ideal disinfectant1. Must be effective against a wide variety of infectious agents at high dilutions and in the presence of organic matter.2. Toxic for infectious agentsno toxic to people or corrosive for common materials.3. Odorless or with a pleasant odor, soluble in water and low surface tension.4. Relatively inexpensive
13PhenolicsDenaturing proteins and disrupting cell membranes. Phenol, orthocresol(邻甲酚)Advantages:1. Tuberculocidal2. Effective in the presence of organic material.3. Remain active on surfaces long after application.Disadvantages:1. Disagreeable odor2. Cause skin irritation
14alcohols Bactericidal and fungicidal, not sporicidal 70-80% ethanol and isopropanolDenature proteins and dissolve membrane lipids.10-15min soaking
15Halogens Iodine and chlorine Iodine: oxidizing cell constituents iodinating cell proteins.2% or more iodine in a water-ethanol solution of potassium iodide.Iodophor(碘载体): iodine is complexed with an organic carries to form. water soluble, nonstaining, release iodine slowly to minimize skin burns and irritation.Chlorine: municipal water suppliesswimming pools Not sporesCl2, HClO, Ca(OCl)2, oxidation
16Heavy metalsMercury (Hg), silver (Ag), arsenic(As), zinc (Zn), copper (Cu)1% AgNO3: the eyes of infants to prevent ophthalmic gonorrheaSilver sulfadizine is used on burns.CuSO4: algicide in lakes and swimming pools.
17AldehydesCombine with nucleic acids and proteins and inactivate them, probably by cross-linking and alkylating molecules.SporicidalFormaldehyde; 2% glutaraldehyde(戊二醛)
18Sterilizing gases Ethylene oxide gas 10-20% mixed with CO2 or dichlorodifluoromethane(二氯二荧光甲烷)Relative humidity: 40-50%38oC 5-8h or oC 3-4hEtO con: 700mg/literToxic! Removed before use.
19Evaluation of antimicrobial agent effectiveness The phenol coefficient test:The test bacteria: Salmonella typhiStaphylococcus aureusa series of dilutions of phenol and the experimental disinfectant.bacteria + agent; 5min, 10min, 15min; subculture to fresh medium, respectively; incubate 2-3 days.The highest dilutions that kill the bacteria after a 10mins exposure, but not after 5mins, are used to calculate the phenol coefficient.phenol coefficient =disinfect max dilution/phenol max dilutionIf >1: more effective than phenol
20Microbial Growth Control: Chemotherapeutic agents Chemotherapeutic agents: to be used internally for control of infectious disease, they must have selective toxicity, these include:Growth factor analog:Sulfa drugs (磺胺药)Amino acid analogsVitamin analogsDNA base, RNA base analogs
21Chemotherapeutic agents: Antibiotics Antibiotics are chemical substances produced by certain microorganisms that inhibit and kill other microorganisms, they are natural products rather than synthetic chemicalsGram-positive bacteria are usually more sensitive to Gram-negative bacteriaIn bacteria, the important targets of antibiotic action are the cell wall, the cytoplasmic membrane and the biosynthetic processes of protein and nucleic acid synthesisb-lactam (内酰胺) group, which includes the penicillins and related compounds, has major clinical significance
25Food Preservation Sterilization Low temperature (best -20oC or 80oC) pH or acidityLow water availabilityCanningChemical food preservation
26Questions • several concepts Conditions influencing the effectiveness of antimicrobial agent activity?What are the physical- and chemical ways to halt microbial growth?• On what sites do antibiotics act on cells?• How to preserve food?