Microbial Sterilization Methods Quiz
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Questions and Answers

What is the main mechanism by which dry heat sterilization kills microorganisms?

  • Oxidation (correct)
  • Desiccation
  • Denaturation of proteins
  • Freezing
  • Which sterilization method uses Cobalt-60 radioisotope?

  • Ionizing radiation (correct)
  • Filtration
  • Nonionizing radiation
  • Hot-air sterilization
  • What is the pore size range typically used for membrane filters to remove bacteria?

  • 1.0 – 5.0 µm
  • 0.01 – 0.1 µm
  • 0.2 – 0.4 µm (correct)
  • 0.5 – 1.0 µm
  • Which method is used as 'cold pasteurization' for food?

    <p>Ionizing radiation (A)</p> Signup and view all the answers

    What is one effect of nonionizing radiation, such as UV light, on actively dividing cells?

    <p>Formation of thymine dimers (B)</p> Signup and view all the answers

    What is the main disadvantage of using microwaves for sterilization?

    <p>Heats solid food unevenly (B)</p> Signup and view all the answers

    Which sterilization technique uses temperature of 170˚C for effective treatment?

    <p>Hot-air sterilization (C)</p> Signup and view all the answers

    What effect does osmotic pressure have on microorganisms?

    <p>Induces plasmolysis (A)</p> Signup and view all the answers

    What is the primary objective of sterilization?

    <p>To kill all microbial life (C)</p> Signup and view all the answers

    Which of the following methods is commonly used for commercial sterilization?

    <p>High heat treatment (A)</p> Signup and view all the answers

    What best describes pasteurization?

    <p>Reduction of pathogenic microorganisms (C)</p> Signup and view all the answers

    What is the primary difference between a disinfectant and an antiseptic?

    <p>Disinfectants are used on surfaces, while antiseptics are used on living tissues. (A)</p> Signup and view all the answers

    The microbial death curve illustrates which of the following?

    <p>The constant rate of microbial death over time (D)</p> Signup and view all the answers

    Which sterilization method is specifically effective for plastic labware?

    <p>Radiation sterilization (C)</p> Signup and view all the answers

    What is a key disadvantage of using dry heat sterilization compared to moist heat methods?

    <p>All of the above. (D)</p> Signup and view all the answers

    What is the primary action of a biocide?

    <p>To kill all bacterial cells (D)</p> Signup and view all the answers

    What is the thermal death point (TDP)?

    <p>The lowest temperature at which all cells in a culture are killed in 10 minutes. (C)</p> Signup and view all the answers

    Which of the following statements about autoclaving is correct?

    <p>Steam must contact the material directly to be effective. (C)</p> Signup and view all the answers

    What is the primary purpose of pasteurization?

    <p>To significantly reduce spoilage and pathogenic organisms. (D)</p> Signup and view all the answers

    Which method effectively eliminates prions during sterilization?

    <p>132°C for 4.5 hours. (C)</p> Signup and view all the answers

    What does the Decimal Reduction Time (DRT) indicate?

    <p>Time required to kill 90% of a population at a specific temperature. (C)</p> Signup and view all the answers

    Which pasteurization method achieves sterilization?

    <p>Ultra High Temperature (UHT) at 140°C for less than 1 second. (B)</p> Signup and view all the answers

    Which of the following is NOT an action of microbial control agents?

    <p>Alteration of lipid mobility. (B)</p> Signup and view all the answers

    How does the presence of organic matter affect heat treatments?

    <p>It may interfere with heat treatments and chemical controls. (B)</p> Signup and view all the answers

    What is the lowest temperature at which all cells in a culture are killed in 10 minutes?

    <p>Thermal death point (B)</p> Signup and view all the answers

    Pasteurization is a process that sterilizes a substance.

    <p>False (B)</p> Signup and view all the answers

    What is the name of the process used to kill bacteria with high pressure in liquids?

    <p>High pressure processing or pascalization</p> Signup and view all the answers

    Give two examples of bisphenols used as disinfectants.

    <p>Hexachlorophene and triclosan</p> Signup and view all the answers

    Alcohols are effective against all types of microorganisms, including spores.

    <p>False (B)</p> Signup and view all the answers

    What is the term for the action of heavy metals against microorganisms?

    <p>Oligodynamic action</p> Signup and view all the answers

    The use-dilution test is used to determine the effectiveness of antiseptics.

    <p>False (B)</p> Signup and view all the answers

    Plasma sterilization is most suitable for sterilizing porous materials.

    <p>False (B)</p> Signup and view all the answers

    What are the main types of microbial control agents?

    <p>Physical and chemical</p> Signup and view all the answers

    Identify two factors that are essential for effective disinfection.

    <p>Concentration and time</p> Signup and view all the answers

    Match the antimicrobial agent to its associated effect.

    <p>Aldehydes = Disrupt cell membranes Halogens = Oxidation Heavy metals = Oligodynamic action Alcohols = Denature proteins Surface-active agents = Denaturation and disruption of cell membranes Plasma = Free radicals Hydrogen peroxide = Inactivate proteins and DNA UV light = Thymine dimers</p> Signup and view all the answers

    Which group of microbes thrives at temperatures above 80°C?

    <p>Thermophiles (C)</p> Signup and view all the answers

    What impact does a hypertonic environment have on microbial cells?

    <p>Causes plasmolysis (C)</p> Signup and view all the answers

    What are halophiles specifically adapted to thrive in?

    <p>High salt concentrations (B)</p> Signup and view all the answers

    Which of the following elements is most abundant in the dry weight of microbial cells?

    <p>Carbon (A)</p> Signup and view all the answers

    What term is used to describe the time taken for the population of bacteria to double?

    <p>Generation time (C)</p> Signup and view all the answers

    Which media type provides all the chemical requirements for microbial growth in known quantities?

    <p>Chemically defined media (D)</p> Signup and view all the answers

    Which method is commonly used to isolate pure cultures of bacteria?

    <p>Streak plating (A)</p> Signup and view all the answers

    How do aerobes typically protect themselves from toxic forms of oxygen?

    <p>Generating superoxide dismutase (A)</p> Signup and view all the answers

    What phase of bacterial growth is characterized by the maximum rate of cell division?

    <p>Exponential phase (D)</p> Signup and view all the answers

    Which method is commonly used for counting viable bacterial cells?

    <p>Plate count (C)</p> Signup and view all the answers

    What does turbidity indicate when measuring microbial growth?

    <p>Cell number (C)</p> Signup and view all the answers

    What is a disadvantage of using direct microscopic counts for microbial growth measurement?

    <p>It cannot distinguish between living and dead cells. (B)</p> Signup and view all the answers

    Which of the following is NOT a direct method of measuring microbial growth?

    <p>Turbidity measurements (A)</p> Signup and view all the answers

    What does the lag phase in bacterial growth represent?

    <p>Adaptation to the new environment (B)</p> Signup and view all the answers

    Which measurement method allows for estimating microbial populations based on metabolic activity?

    <p>Indirect measurement (B)</p> Signup and view all the answers

    What is the primary purpose of differential media in microbiology?

    <p>To change in a recognizable manner for the identification of specific microbes (A)</p> Signup and view all the answers

    Which of the following techniques is essential for obtaining pure cultures?

    <p>Streak-plate method (B)</p> Signup and view all the answers

    Which phase of bacterial growth is characterized by the balance between cell division and cell death?

    <p>Stationary phase (D)</p> Signup and view all the answers

    Which method allows for the selective growth of phenol-degrading bacteria from a mixed culture?

    <p>Enrichment culture (A)</p> Signup and view all the answers

    What is the generation time of E. coli under optimal growth conditions?

    <p>20 minutes (C)</p> Signup and view all the answers

    How can bacterial cultures be preserved for several years?

    <p>By lyophilization or deep-freezing (D)</p> Signup and view all the answers

    What does a colony represent in microbiological terms?

    <p>A population arising from a single cell or spore (C)</p> Signup and view all the answers

    What temperature range is typical for deep-freezing bacterial cultures?

    <p>-50° to -95°C (C)</p> Signup and view all the answers

    Which of the following statements about binary fission is correct?

    <p>It results in exponential growth of bacteria (C)</p> Signup and view all the answers

    What role do trace elements play in microbial growth?

    <p>They serve as cofactors for enzymatic reactions. (C)</p> Signup and view all the answers

    Which type of medium is specifically known for having all its chemical components defined?

    <p>Chemically defined media (D)</p> Signup and view all the answers

    What is a characteristic feature of biofilms?

    <p>They are sheltered from harmful factors. (B)</p> Signup and view all the answers

    What chemical is used in anaerobic culture methods to remove oxygen?

    <p>Thioglycollate (B)</p> Signup and view all the answers

    What defines a capnophile?

    <p>A microorganism that requires high levels of carbon dioxide. (D)</p> Signup and view all the answers

    What temperature range allows agar to solidify?

    <p>40-60°C (A)</p> Signup and view all the answers

    Which of the following best describes how biofilms form?

    <p>By attachment of planktonic bacteria to surfaces. (C)</p> Signup and view all the answers

    What form of oxygen can harm microorganisms by causing oxidative damage?

    <p>Superoxide free radicals (A)</p> Signup and view all the answers

    What type of culture medium selectively suppresses the growth of undesired microorganisms?

    <p>Selective medium (C)</p> Signup and view all the answers

    What is the initial step in the formation of biofilms?

    <p>Environmental attachment (A)</p> Signup and view all the answers

    Flashcards

    Antimicrobial treatment effectiveness

    Depends on the time it takes to eliminate a population of microbes, which is affected by the number of microbes, microbial species' susceptibility (e.g., endospores), organic matter interfering with treatments, and exposure time.

    Thermal death point (TDP)

    The lowest temperature that kills all cells in a culture within 10 minutes.

    Thermal death time (TDT)

    The time needed to kill all cells in a culture at a specific temperature.

    Decimal Reduction Time (DRT)

    Time required to kill 90% of a microbial population at a set temperature.

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    Moist heat sterilization

    A highly dependable sterilization method using steam under pressure that denatures proteins.

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    Autoclave

    A device that uses steam under pressure for sterilization.

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    Pasteurization

    A heat treatment process that significantly reduces the number of spoilage and pathogenic microorganisms, but does not sterilize.

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    Prion destruction

    Prion destruction requires higher temperatures (132°C) and longer durations (4.5 hours) than typical autoclave conditions to fully eliminate them.

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    Sterilization

    Complete elimination of all microbial life, including spores, from an object or surface.

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    Disinfection

    Process that eliminates most pathogenic microbes, but not necessarily all microbes or spores.

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    Antisepsis

    Application of antimicrobial agents to living tissue to prevent infection.

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    Sanitization

    Reduction of microbial populations to safe levels, often used for food preparation surfaces.

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    Microbial Death Curve

    Graphical representation of the logarithmic decline in microbial population over time when exposed to a lethal agent.

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    Filtration

    Physical removal of microbes by passing a liquid or gas through a filter with pores too small for microbes to pass.

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    Dry heat sterilization

    A sterilization method that kills microorganisms through oxidation.

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    Hot-air sterilization

    A sterilization method that uses dry heat at high temperatures (170˚C) for a period of time.

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    Autoclave sterilization

    A sterilization method that uses moist heat (121˚C) under pressure for a short time (15 minutes).

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    Filtration sterilization

    A sterilization method using filters to remove microbes (e.g. HEPA filters).

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    Low temperature sterilization

    Slows down microbial growth by inhibiting enzymatic reactions, damaging microbes with ice crystals.

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    High pressure sterilization

    A process that denatures bacterial proteins in liquids, preserving food flavor.

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    Desiccation

    A process that prevents microbial metabolism by drying out the microorganism.

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    Ionizing radiation sterilization

    A sterilization method that uses high-energy radiation to damage microbial DNA.

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    Microbial Growth

    An increase in the number of microbial cells, not their size.

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    Optimum Growth Temperature

    The temperature at which a microbe grows best.

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    Psychrophiles

    Microbes that thrive in cold temperatures, with an optimum growth temperature below 15°C.

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    Mesophiles

    Microbes that grow best at moderate temperatures, with an optimum growth temperature between 20°C and 45°C.

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    Thermophiles

    Microbes that thrive in hot temperatures, with an optimum growth temperature between 45°C and 70°C.

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    pH and Microbial Growth

    Most bacteria grow best at a neutral pH (6.5-7.5), while some thrive in acidic or alkaline environments.

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    Osmotic Pressure

    The pressure exerted by the concentration of solutes in a solution, impacting water movement into or out of microbial cells.

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    Hypertonic Environment

    An environment with a higher solute concentration than the inside of a cell, causing water to move out, potentially leading to plasmolysis.

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    Obligate Halophiles

    Microbes that require high salt concentrations to survive.

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    Facultative Halophiles

    Microbes that can tolerate high salt concentrations but don't require them.

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    Carbon's Role in Microbial Growth

    Carbon is essential for microbial growth, making up about half of a microbe's dry weight.

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    Chemoheterotrophs

    Microbes that obtain their energy from organic compounds like sugars and proteins.

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    Nitrogen's Role in Microbial Growth

    Nitrogen is crucial for building amino acids and proteins, which are essential for microbial structure and function.

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    Sulfur's Role in Microbial Growth

    Sulfur is essential for building certain amino acids and vitamins necessary for microbial growth.

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    Phosphorus's Role in Microbial Growth

    Phosphorus is crucial for building nucleic acids (DNA and RNA) as well as ATP (the energy currency of cells).

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    Oxygen and Microbial Growth

    Microbes have different oxygen requirements, some needing it, some tolerating it, and some avoiding it.

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    Aerobes

    Microbes that require oxygen to survive.

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    Anaerobes

    Microbes that don't require oxygen and may even be harmed by it.

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    Facultative Anaerobes

    Microbes that can grow with or without oxygen, but prefer oxygen.

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    Toxic Forms of Oxygen

    Certain oxygen-containing molecules can be harmful to cells, such as singlet oxygen, superoxide radicals, and hydroxyl radicals.

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    Biofilms

    Complex microbial communities encased in a sticky matrix, often attached to surfaces.

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    Quorum Sensing

    A communication system used by bacteria to coordinate their behavior and create biofilms.

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    Culture Media

    Nutrients prepared for microbial growth, typically sterilized to avoid contamination.

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    Chemically Defined Media

    Culture media with a precisely known chemical composition.

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    Complex Media

    Culture media containing extracts or digests of yeast, meat, or plants, with an unknown exact composition.

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    Agar

    A complex polysaccharide used as a solidifying agent in culture media, allowing microbes to grow on a solid surface.

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    Anaerobic Culture Methods

    Techniques used to grow microbes that cannot tolerate oxygen, typically using reducing media or anaerobic jars.

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    Capnophiles

    Bacteria that require high CO2 levels, often resembling conditions found in body tissues.

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    Selective Media

    Culture media containing additives that suppress the growth of unwanted microbes and encourage the desired ones.

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    Differential Media

    Culture media that allow for the differentiation of different types of microbes based on visible changes in colony appearance.

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    Enrichment Media

    Culture media that encourages the growth of a desired microbe while suppressing others, often used for isolating specific microbes from a mixed population.

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    Pure Culture

    A culture containing only one species or strain of microbe, separated from other organisms.

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    Streak Plate Method

    A technique used to isolate individual colonies of bacteria from a mixed culture by streaking the sample across an agar plate.

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    Colony Formation

    A visible cluster of microbes on a solid medium arising from a single cell or group of attached cells.

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    Preserving Bacterial Cultures

    Methods used to store bacterial cultures for extended periods, such as deep-freezing or freeze-drying.

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    Binary Fission

    The most common method of bacterial cell division, where a single cell divides into two identical daughter cells.

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    Generation Time

    The time it takes for a microbial population to double in size.

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    Bacterial Growth Curve

    A graphical representation of the stages of microbial growth in a closed culture system.

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    Direct Microscopic Count

    A method to directly count microbes using a microscope and a counting chamber.

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    Serial Dilution

    A technique used to dilute a sample of microbes repeatedly, allowing for accurate counting of colonies on agar plates.

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    Spectrophotometry

    A method to indirectly measure microbial growth by measuring the turbidity (cloudiness) of a culture.

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    Lag Phase

    The initial phase of bacterial growth where cells are adjusting to their new environment and preparing for growth, resulting in little to no increase in population size.

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    Logarithmic (Log) Phase

    The phase of rapid, exponential growth where bacteria divide at a constant rate, doubling in number with each generation.

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    Stationary Phase

    The phase of growth where the rate of cell division equals the death rate, leading to a stable population size.

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    Death Phase

    The phase where bacteria start dying off faster than they're reproducing, leading to a decrease in the population size.

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    Plate Counts

    A method for counting viable bacteria by diluting a sample and plating it on agar, where each colony grows from a single bacterium.

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    What are essential nutrients for bacterial growth?

    Bacteria require essential nutrients like carbon, nitrogen, phosphorus, sulfur, water, and specific inorganic ions like potassium, magnesium, and calcium, for growth. They also need trace elements and organic growth factors like vitamins.

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    What is a biofilm?

    Biofilms are communities of bacteria that stick together and form a thick, slimy layer on a surface. They often form on surfaces like teeth, medical implants, and food processing equipment.

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    What are essential nutrients?

    Essential nutrients are compounds that a microbe cannot synthesize on its own and must obtain from its environment. They include carbon, nitrogen, sulfur, phosphorus, minerals, and sometimes growth factors like vitamins.

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    What is a culture medium?

    A culture medium is a carefully prepared mixture of nutrients designed to support the growth of microbes in a laboratory setting. It can be solid or liquid and contains all the components needed for bacterial growth.

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    What are toxic forms of oxygen?

    Toxic forms of oxygen are highly reactive molecules that can be harmful to cells. These include singlet oxygen, superoxide free radicals, peroxide anions, and hydroxyl radicals.

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    What is an inoculum?

    An inoculum is a small sample of microbes introduced into a culture medium to start a new culture.

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    What is a selective medium?

    A selective medium is a type of culture medium that only allows certain types of microbes to grow, while inhibiting the growth of others.

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    What is a differential medium?

    A differential medium is a type of culture medium that allows different types of microbes to be distinguished based on how they grow on the medium.

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    What are the oxygen requirements for bacterial growth?

    Bacteria have different oxygen requirements. Some need oxygen (aerobes), some don't need it and may be harmed by it (anaerobes), and some can grow with or without it (facultative anaerobes).

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    What does 'sterile' mean in the context of culture media?

    Sterile in microbiology means free from living microbes. Culture media must be sterile to prevent unwanted microbes from contaminating the desired culture.

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    Study Notes

    Chapter 7: Control of Microbial Growth

    • Definitions:

      • Sterilization: Removal of all microbial life.
      • Disinfection: Reduction in the number of pathogenic microorganisms; destroys harmful microorganisms but not all.
      • Antisepsis: Removal of pathogens from living tissue.
      • Sanitization: Reduction in microbial numbers to a safe level (e.g., eating utensils).
      • Biocide/Germicide: Agents that kill microorganisms.
      • Bacteriostasis: Inhibition of microbial growth.
      • Asepsis: Absence of significant contamination.
      • Sepsis: Microbial contamination.
      • Nosocomial: Infections acquired in hospitals.
      • Commercial sterilization: Method of sterilization used for food products. The goal is killing C. botulinum endospores.
      • Bacteriostatic: Inhibits bacterial reproduction.
      • Bactericidal: Kills bacteria.
      • Fungicide/Sporicide/Germicide/Biocide: Agents that kill fungi/bacterial spores/microbes/living things.
    • Microbial Death Curve: The graph shows the constant rate of microbial death when exposed to heat or chemicals. Plotting this logarithmically shows microbial death constant as a straight line.

      • One log decrease = 90% population killed.
    • Factors Affecting Microbial Control Effectiveness:

      • Microbial population size.
      • Microbial characteristics (species, life cycle: endospores are more resistant).
      • Environmental conditions (temperature, organic matter, degree of contact).
    • Actions of Microbial Control Agents:

      • Alteration of membrane permeability.
      • Damage to proteins.
      • Damage to nucleic acids.
    • Physical Methods of Microbial Control:

      • Heat:
        • Moist Heat Sterilization (autoclaving):
          • Steam under pressure.
          • Most reliable sterilization method.
          • Steam must directly contact materials.
          • Normal autoclave conditions: 121.5°C for 15 minutes.
          • Prion destruction: 132°C for 4.5 hours.
          • Disadvantages: Limitations of the autoclave (e.g., can't use heat-sensitive items, care required).
        • Pasteurization:
          • Classic holding method: 63°C for 30 minutes.
          • Flash pasteurization (HTST): 72°C for 15 seconds.
          • Ultra High Temperature (UHT): 140°C for <1 second.
      • Dry Heat Sterilization: Incineration of carcasses, flaming of loops, hot air sterilization.
      • Filtration: HEPA filters for air. Membrane filters for fluids.
      • Low Temperature: Slows enzymatic reactions. Freezing forms ice crystals damaging microbial cells. Refrigeration and lyophilization.
      • High Pressure: Denatures bacterial proteins and preserves flavor.
      • Desiccation: Prevents metabolism.
      • Osmotic Pressure: Causes plasmolysis.
      • Ionizing Radiation:
        • X-rays, gamma rays, electron beams.
        • Dislodge electrons from atoms, producing free radicals.
        • Frequently used Cobalt-60 radioisotope and used to sterilize heat-sensitive materials (drugs, vitamins, herbs, suture materials).
      • Nonionizing Radiation (UV light):
        • Most effective wavelength ~260 nm.
        • Causes thymine dimers.
        • Actively dividing organisms are more sensitive.
        • Useful to limit contamination at a close range (e.g., germicidal lamps).
      • Nonionizing Radiation (Microwave):
        • Wavelength of 1 mm-1m.
        • Water quickly absorbs energy --> heats unevenly in solids.
    • Chemical Methods of Microbial Control:

      • Use-dilution test:
        • Metal rings dipped in bacteria are dried
        • Dried cultures are placed in disinfectant for 10 mins at 20°C.
        • Rings transferred to culture media to see if surviving bacteria.
      • Disk-diffusion method:
        • Disk soaked in chemical placed on inoculated agar plate.
        • In an inoculated agar plate, a zone of inhibition indicates the effectiveness of the chemical's action.
      • Phenol/Carbolic acid: Historic importance.
      • Phenolics(Cresols, Lysol): Disinfectants.
      • Bisphenols (Hexachlorophene, Triclosan): Disrupt plasma membranes.
      • Halogens (Chlorine, Iodine): Disinfectants/Antiseptics. Chlorine is a strong oxidizing agent used in bleaching and disinfecting (broad spectrum), while iodine is a more reactive agent used in wound antiseptics.
      • Alcohols (Ethyl, Isopropyl): Denature proteins, dissolve lipids; ineffective against spores/viruses/fungi. Easy deactivated by organic debris.
      • Heavy Metals (Mercury, Copper, Silver, Zinc): Oligodynamic action. Toxic to microorganisms; bind to sulfhydryl groups.
      • Surface-active agents (Soaps, Detergents): Mechanical removal and wetting. Cationic detergents (quats) are highly bactericidal.
      • Aldehydes (Formaldehyde, Glutaraldehyde): Alkylating agents. Used as sterilants for delicate materials.
      • Hydrogen Peroxide: An oxidizing agent, good for inanimate objects. Inactivated by catalase and not good for open wounds.
      • Chemical Food Preservatives (Sulfur Dioxide, Organic Acids): Inhibit metabolism of bacteria.
    • Plasma: Luminous gas with free radicals destroying microbes, used on instruments/hands.

    • Microbial Characteristics and Resistance:

      • Prions are the most resistant.
      • Endospores of bacteria are highly resistant.
      • Mycobacteria, cysts of protozoa are quite resistant.
      • Gram-negative bacteria are fairly resistant.
      • Fungi (spores) and viruses (without envelopes) are often resistant. Gram-positive bacteria. Viruses with lipid envelopes are the least resistant.

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