Microbial Growth and The Control of Microbial Growth (Chapter 6 & 7)
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1 Microbial Growth and The Control of Microbial Growth (Chapter 6 & 7) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Eastern Campus Primary Source for figures and content: Tortora, G.J. Microbiology An Introduction 8th, 9th, 10th ed. San Francisco: Pearson Benjamin Cummings, 2004, 2007, Amy Warenda Czura, Ph.D. 1 SCCC BIO244 Chapter 6 & 7 Lecture Slides
2 Chapter 6: Microbial Growth Microbial requirements for growth: 1. Physical A. Temperature Optimal growth temperature Permissible range human pathogens optimal = 37 C B. ph most organisms optimal ph C. Osmotic pressure Most require isotonic solutions human blood = 0.9% NaCl, isotonic human skin = ~3-6% NaCl, hypertonic 2. Chemical A. Carbon B. Nitrogen C. Sulfur D. Phosphorus (all above: to build organic molecules) E. Trace elements: K +, Mg 2+, Ca 2+, Fe cofactors for enzymes Amy Warenda Czura, Ph.D. 2 SCCC BIO244 Chapter 6 & 7 Lecture Slides
3 F. Oxygen aerobe, anaerobe, facultative G. Organic factors: vitamins, amino acids... (essential organic compound an organism is unable to synthesize) Microbial Growth -for unicellular organisms, growth = increase in cell number (population) not individual cell size -generation time = time required for a cell to divide -both growth and death of the population is exponential, graphed on a log scale Amy Warenda Czura, Ph.D. 3 SCCC BIO244 Chapter 6 & 7 Lecture Slides
4 Bacterial Growth Curve both growth and death occur at a constant rate 1. Lag Phase: little to no growth, bacteria acclimate to new environment 2. Log Phase: period of exponential growth with constant generation time 3. Stationary Phase: cell growth is equal to cell death 4. Death Phase: cell death exceeds cell growth Amy Warenda Czura, Ph.D. 4 SCCC BIO244 Chapter 6 & 7 Lecture Slides
5 Chapter 7: Control of Microorganisms Sterilization = destruction or removal of all forms of microbial life, including endospores Commercial sterilization = (canning) enough heat to kill endospores of Clostridium botulinum (thermophilic species survive) Disinfection = destruction of vegetative pathogens on inert surfaces (disinfectant) Antisepsis = disinfection of living tissue (antiseptic) Degermation = mechanical removal of microbes in a limited area Sanitization = lower microbe counts to a safe level to minimize disease transmission suffix cide = kill e.g. biocide, fungicide suffix -stat or -stasis = stop inhibit growth: when agent removed, growth can resume e.g. fungistat Amy Warenda Czura, Ph.D. 5 SCCC BIO244 Chapter 6 & 7 Lecture Slides
6 sepsis = bacterial contamination present asepsis = absence of significant bacterial contamination (not sterile) in clinical terms, absence of pathogens Microbial Death Rates: -microbial populations that are heated or treated with chemicals will die at a constant rate (exponential decline) Amy Warenda Czura, Ph.D. 6 SCCC BIO244 Chapter 6 & 7 Lecture Slides
7 Factors that influence effectiveness of an antimicrobial treatment: 1. Number of microbes -more cells, more time needed to kill all 2. Environmental influences -organics often inhibit chemical agents -temperature -ph 3. Time of exposure -same agent may need longer on resistant organisms or spores -with heat, lower temps require longer to kill 4. Microbial characteristics -resistance genes, protective structures (e.g.capsules) etc. can inhibit action -bioflims prevent penetration Amy Warenda Czura, Ph.D. 7 SCCC BIO244 Chapter 6 & 7 Lecture Slides
8 Actions of Microbial Control Agents: 1. Alteration of membrane permeability: -damage to proteins or lipids of membrane = leak lysis, death 2. Damage to proteins and/or nucleic acids -denature enzymes (no reactions) -prevent replication, transcription, or translation Physical Methods of Microbial Control: -to disinfect objects, food, and solutions -common methods: -temperature: kill or inhibit growth -filtration: physical removal -desiccation: inhibit growth -radiation: kill Amy Warenda Czura, Ph.D. 8 SCCC BIO244 Chapter 6 & 7 Lecture Slides
9 For methods that involve heat: Thermal Death Point (TDP) = lowest temp at which all microbes in liquid suspension will be killed in 10 min Thermal Death Time (TDT)= minimal length of time for all microbes in liquid suspension to be killed at given temp *Both are different for different species due to microbial variation in heat tolerance Concept of equivalent treatments: With any heat treatment, the higher the temperature used the shorter the exposure time needed to achieve the same effect Moist heat will always kill faster than dry heat at the same temperature Examples of physical methods: (on handout) Amy Warenda Czura, Ph.D. 9 SCCC BIO244 Chapter 6 & 7 Lecture Slides
10 autoclave Amy Warenda Czura, Ph.D. 10 SCCC BIO244 Chapter 6 & 7 Lecture Slides
11 low temperature Amy Warenda Czura, Ph.D. 11 SCCC BIO244 Chapter 6 & 7 Lecture Slides
12 filtration osmotic pressure Amy Warenda Czura, Ph.D. 12 SCCC BIO244 Chapter 6 & 7 Lecture Slides
13 Radiation Chemical Methods of Microbial Control -used to disinfect surfaces and people Chemical antimicrobial agents should: 1. be effective against wide range of microbes 2. be effective in presence of organic material 3. be toxic to microbe but non-toxic to people and non-corrosive to surfaces -hard to find all in one agent: usually have to select best agent based on use application -most chemical agents that are safe do not sterilize: they reduce microbe levels, but only if used at the correct concentration Amy Warenda Czura, Ph.D. 13 SCCC BIO244 Chapter 6 & 7 Lecture Slides
14 Use Dilution Test: -determine conc. of chemical agent necessary to kill 95% test organisms in 10 min -used to determine effectiveness of disinfectant and antiseptics against various microbes Disk-Diffusion Method -determine efficacy of a chemical agent -score for zone of no growth Chemical Agents (take notes on handout) Amy Warenda Czura, Ph.D. 14 SCCC BIO244 Chapter 6 & 7 Lecture Slides
15 Microbial characteristics and microbial control -some characteristics provide resistance: *Pseudomonas & Burkholderia: -porins in cell wall prevent chemical entry -can live on any organics, even in detergents & antiseptics *Mycobacterium: waxy mycolic acid prevents chemical entry *Endospores and cysts: thick coats prevent chemical entry *Enveloped viruses more easily damaged than non-enveloped *Prions are resistant to most chemical methods and even autoclaving Typical resistance to biocides Amy Warenda Czura, Ph.D. 15 SCCC BIO244 Chapter 6 & 7 Lecture Slides
16 Amy Warenda Czura, Ph.D. 16 SCCC BIO244 Chapter 6 & 7 Lecture Slides
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