Chapters 6 and 7: Microbial Growth and Control - Vocabulary Flashcards

Chapter 6: Microbial Growth

  • Key terms

    • Bacteriostatic: curbs the replication rate of bacteria without killing them. Example: refrigeration slows enzyme activity needed for replication; bacteria remain viable but replicate slowly.

    • Bactericidal: kills bacteria. Example: boiling water to make it safe to drink.

  • Physical requirements for growth

    • Temperature

    • Temperature optimizes enzyme activity; different bacteria have preferred ranges.

    • Classifications among Eubacteria and Archaea:

      • Psychrophiles: cold-loving

      • Mesophiles: moderate temperatures (overlap with psychrophiles to some extent)

      • Thermophiles: heat-loving (including moderate and extreme thermophiles in the Eubacteria)

      • Beyond the extreme thermophiles, Archaea are extremophiles

    • pH

    • Most bacteria prefer near-neutral pH: roughly 6.5 \leq pH \leq 7.5

    • Acidophiles: acidic environments

    • Neutrophiles: near-neutral environments (most bacteria fall here)

    • Alkalophiles: basic environments

    • Examples of ranges:

      • Escherichia coli (E. coli): optimum around pH = 6-7 (neutrophile); tolerates pH = 4.4\text{ to } 9.0

      • Staphylococcus aureus: optimum pH \approx 7-7.5; tolerates pH = 4.2\text{ to } 9.3

    • The ranges illustrate the wide tolerance of some bacteria to pH conditions

    • Osmotic pressure (water activity, tonicity)

    • Isotonic environments preferred for most bacteria

    • Halophiles require salt:

      • Mild halophiles: 16\%\,salt\ (w/v)

      • Moderate halophiles: 6-15\%\,salt

      • Extreme halophiles (often Archaea): 15-30\%\,salt

    • Note: some bacteria require salt for growth; most eubacteria fall between mild and moderate halophiles

  • Chemical requirements for growth

    • Carbon and nutritional types (energy source vs carbon source)

    • Phototrophs: energy from light; carbon source not specified here

    • Photoautotrophs: energy from light; carbon source is CO₂

    • Photoheterotrophs: energy from light; carbon source is organic compounds

    • Chemoautotrophs: energy from inorganic compounds; carbon from CO₂

    • Chemoheterotrophs: energy from organic compounds; carbon from organic compounds

    • Concept: break down names to identify energy source (first part) and carbon source (second part):

      • Photo-: energy source is light

      • Auto-: carbon source is CO₂

      • Hetero-: carbon source is organic compounds

    • Essential elements and trace elements

    • Primary elements: nitrogen, sulfur, phosphorus

      • Nitrogen: incorporated into amino acids, nucleotides, coenzymes, etc.

      • Sulfur, phosphorus: various cellular roles (not enumerated in detail here)

    • Trace elements: iron, chlorine, and others; present in small amounts; commonly serve as enzyme cofactors

    • Oxygen requirements and terminology

    • Aerobic vs anaerobic growth

    • Facultative: can grow with or without oxygen

    • Obligate: requires a specific condition (either aerobic or anaerobic depending on organism)

    • Aero-tolerant anaerobes: fermentatively produce ATP regardless of oxygen presence; always rely on fermentation; least efficient mode of ATP production

  • Culture media for growing bacteria

    • Generalized media

    • Example: Nutrient Agar

    • Purpose: support growth of a wide variety of bacteria when the species is unknown (e.g., swabbing a tabletop)

    • Specialized media

    • Selective: favors growth of a particular group and inhibits others (e.g., salt concentrations favor Gram-positive bacteria)

    • Differential: allows differentiation between colonies (often by color changes)

    • Some media are both selective and differential

    • Example: Eosin Methylene Blue (EMB) agar

      • Contains dyes that indicate lactose fermentation by Gram-negative bacteria

      • Greater lactose fermentation yields darker dye coloration; darker color => higher fermentation activity

    • Enrichment cultures

    • Liquid media designed to promote growth of a particular organism from a mixed culture, often by providing a specific composition

    • Purpose: allow low-abundance organisms to outgrow others (e.g., enrich for E. coli from soil)

    • Viruses and culture media

    • Viruses cannot be grown on standard culture media; require living cells for propagation

    • Discussion on viral culture is reserved for Chapter 13

  • Preservation of bacterial samples

    • Lyophilization (freeze-drying)

    • Bacteria are frozen, then dried under heat and vacuum to remove water from cytoplasm

    • Used for long-term storage; cultures become dormant but viable

    • Revival: add nutrient source to resume metabolism and replication

    • Cryopreservation

    • Bacteria frozen in liquid nitrogen (~-196°C)

    • Long-term storage; kept until thawed for revival

  • Bacterial growth dynamics

    • Reproduction: asexual binary fission

    • Generation time (doubling time): time required for population to double

    • If needed: N(t) = N0 \cdot 2^{\frac{t}{td}} where t_d is the doubling time

    • Growth curve phases (in a closed system)

    • Lag phase: cells adjust to new environment; little to no observable growth

    • Log (exponential) phase: rapid cell division; growth rate exceeds death rate (some cell death occurs)

    • Stationary phase: growth rate equals death rate; nutrients depleted and waste products accumulate, often causing pH to drop

    • Death (death) phase: death rate exceeds growth rate; nutrients exhausted and pH becomes highly acidic

    • Closed system: no addition of nutrients and no removal of waste

  • Measuring bacterial growth

    • Direct counts: membrane filtration (colony counting)

    • Process: filter sample through a membrane; place membrane on nutrient agar; incubate and count colonies

    • Statistical significance: select plates with 30–300 colonies for reliable counting

    • From colony counts, calculate the original cell concentration and plot growth curves

    • Indirect counts: spectrophotometry

    • Principle: measure light transmission through a sample; more cells absorb more light, so less light passes through

    • Result: a range (e.g., 100–300 cells) rather than an exact count

    • Pros: rapid, provides immediate results

    • Cons: cannot distinguish viable from nonviable cells; organic debris can affect readings

    • Viability testing recommended: plate a sample after spectrophotometry and compare; growth after incubation confirms viability

  • Chapter 7: The control of microbial growth (condensed overview)

  • Why control microbial growth?

    • Prevent food spoilage and ingestion of contaminated food

    • Sterilize medical equipment to ensure patient safety

  • Key control terms

    • Sterilization: kills all microbes, including vegetative cells and endospores

    • Vegetative cell: actively replicating form of the cell

    • Disinfection: kills vegetative cells only (does not kill endospores)

    • Antisepsis: kills vegetative cells on living tissue (human host)

    • Sanitation: reduces microbial counts to public health standards (e.g., glasses, utensils) without achieving sterility

  • Variables affecting microbial death rate

    • Number of microbes present: more organisms take longer to kill

    • Presence of endospores: highly resistant to many methods

    • Presence of organic material: organic debris can shield microbes from treatment (e.g., UV light may be absorbed by debris, protecting organisms underneath)

  • Methods of control

    • Physical methods

    • Temperature: hot and cold can kill microbes; hot denatures proteins; freezing and thawing can cause ice crystal damage

    • Filtration: physically removes cells

    • Desiccation: dehydration inhibits microbial metabolism

    • Ultraviolet (UV) light: damages DNA, creating mutations; severe damage prevents repair and replication

    • Chemical methods

    • Alcohols: dissolve lipids (disrupt membranes and proteins)

    • Soaps: act as surfactants and aid in mechanical removal

    • Preservatives (nitrates and nitrites): inhibit growth or kill microbes

  • Exceptions and notable organisms

    • Pseudomonas species: can survive and grow in disinfectants; a problem in medical facilities due to contamination of cleaning agents and equipment

    • Endospores: extremely resistant to many physical and chemical control methods; remain a major challenge in sterilization

  • Summary

    • Chapter 6: covers microbial growth physics, biochemistry, growth measurement, and preservation techniques

    • Chapter 7: covers rationale and methods for controlling microbial growth, including definitions of sterilization, disinfection, antisepsis, sanitation, and considerations of when these controls fail due to resistant organisms or protective organic matter

Chapter 6: Microbial Growth
  • Key terms

    • Bacteriostatic: curbs the replication rate of bacteria without killing them.

    • Example: refrigeration slows enzyme activity needed for replication; bacteria remain viable but replicate slowly.

    • Bactericidal: kills bacteria.

    • Example: boiling water to make it safe to drink.

  • Physical requirements for growth

    • Temperature

    • Temperature optimizes enzyme activity; different bacteria have preferred ranges.

    • Classifications among Eubacteria and Archaea:

      • Psychrophiles: cold-loving

      • Mesophiles: moderate temperatures (overlap with psychrophiles to some extent)

      • Thermophiles: heat-loving (including moderate and extreme thermophiles in the Eubacteria)

      • Beyond the extreme thermophiles, Archaea are extremophiles

    • pH

    • Most bacteria prefer near-neutral pH: roughly 6.5 \leq pH \leq 7.5

    • Acidophiles: acidic environments

    • Neutrophiles: near-neutral environments (most bacteria fall here)

    • Alkalophiles: basic environments

    • Examples of ranges:

      • Escherichia coli (E. coli): optimum around pH = 6-7 (neutrophile); tolerates pH = 4.4\text{ to } 9.0

      • Staphylococcus aureus: optimum pH \approx 7-7.5; tolerates pH = 4.2\text{ to } 9.3

    • The ranges illustrate the wide tolerance of some bacteria to pH conditions

    • Osmotic pressure (water activity, tonicity)

    • Isotonic environments preferred for most bacteria

    • Halophiles require salt:

      • Mild halophiles: 1-6\%\,salt\ (w/v)

      • Moderate halophiles: 6-15\%\,salt

      • Extreme halophiles (often Archaea): 15-30\%\,salt

    • Note: some bacteria require salt for growth; most eubacteria fall between mild and moderate halophiles

  • Chemical requirements for growth

    • Carbon and nutritional types (energy source vs carbon source)

    • Phototrophs: energy from light; carbon source not specified here

    • Photoautotrophs: energy from light; carbon source is CO₂

    • Photoheterotrophs: energy from light; carbon source is organic compounds

    • Chemoautotrophs: energy from inorganic compounds; carbon from CO₂

    • Chemoheterotrophs: energy from organic compounds; carbon from organic compounds

    • Concept: break down names to identify energy source (first part) and carbon source (second part):

      • Photo-: energy source is light

      • Auto-: carbon source is CO₂

      • Hetero-: carbon source is organic compounds

    • Essential elements and trace elements

    • Primary elements: nitrogen, sulfur, phosphorus

      • Nitrogen: incorporated into amino acids, nucleotides, coenzymes, etc.

      • Sulfur, phosphorus: various cellular roles (not enumerated in detail here)

    • Trace elements: iron, chlorine, and others; present in small amounts; commonly serve as enzyme cofactors

  • Oxygen requirements and terminology

    • Aerobic vs anaerobic growth

    • Facultative: can grow with or without oxygen

    • Obligate: requires a specific condition (either aerobic or anaerobic depending on organism)

    • Aero-tolerant anaerobes: fermentatively produce ATP regardless of oxygen presence; always rely on fermentation; least efficient mode of ATP production

  • Culture media for growing bacteria

    • Generalized media

    • Example: Nutrient Agar

    • Purpose: support growth of a wide variety of bacteria when the species is unknown (e.g., swabbing a tabletop)

    • Specialized media

    • Selective: favors growth of a particular group and inhibits others (e.g., salt concentrations favor Gram-positive bacteria)

    • Differential: allows differentiation between colonies (often by color changes)

    • Some media are both selective and differential

    • Example: Eosin Methylene Blue (EMB) agar

      • Contains dyes that indicate lactose fermentation by Gram-negative bacteria

      • Greater lactose fermentation yields darker dye coloration; darker color => higher fermentation activity

    • Enrichment cultures

    • Liquid media designed to promote growth of a particular organism from a mixed culture, often by providing a specific composition

    • Purpose: allow low-abundance organisms to outgrow others (e.g., enrich for E. coli from soil)

    • Viruses and culture media

    • Viruses cannot be grown on standard culture media; require living cells for propagation

    • Discussion on viral culture is reserved for Chapter 13

  • Preservation of bacterial samples

    • Lyophilization (freeze-drying)

    • Bacteria are frozen, then dried under heat and vacuum to remove water from cytoplasm

    • Used for long-term storage; cultures become dormant but viable

    • Revival: add nutrient source to resume metabolism and replication

    • Cryopreservation

    • Bacteria frozen in liquid nitrogen \approx -196\text{°C}

    • Long-term storage; kept until thawed for revival

  • Bacterial growth dynamics

    • Reproduction: asexual binary fission

    • Generation time (doubling time): time required for population to double

    • If needed: N(t) = N0 \cdot 2^{\frac{t}{td}} where t_d is the doubling time

  • Growth curve phases (in a closed system)

    • Lag phase: cells adjust to new environment; little to no observable growth

    • Log (exponential) phase: rapid cell division; growth rate exceeds death rate (some cell death occurs)

    • Stationary phase: growth rate equals death rate; nutrients depleted and waste products accumulate, often causing pH to drop

    • Death (death) phase: death rate exceeds growth rate; nutrients exhausted and pH becomes highly acidic

    • Closed system: no addition of nutrients and no removal of waste

  • Measuring bacterial growth

    • Direct counts: membrane filtration (colony counting)

    • Process: filter sample through a membrane; place membrane on nutrient agar; incubate and count colonies

    • Statistical significance: select plates with 30–300 colonies for reliable counting

    • From colony counts, calculate the original cell concentration and plot growth curves

    • Indirect counts: spectrophotometry

    • Principle: measure light transmission through a sample; more cells absorb more light, so less light passes through

    • Result: a range (e.g., 100–300 cells) rather than an exact count

    • Pros: rapid, provides immediate results

    • Cons: cannot distinguish viable from nonviable cells; organic debris can affect readings

    • Viability testing recommended: plate a sample after spectrophotometry and compare; growth after incubation confirms viability

Chapter 7: The control of microbial growth (condensed overview)
  • Why control microbial growth?

    • Prevent food spoilage and ingestion of contaminated food

    • Sterilize medical equipment to ensure patient safety

  • Key control terms

    • Sterilization: kills all microbes, including vegetative cells and endospores

    • Vegetative cell: actively replicating form of the cell

    • Disinfection: kills vegetative cells only (does not kill endospores)

    • Antisepsis: kills vegetative cells on living tissue (human host)

    • Sanitation: reduces microbial counts to public health standards (e.g., glasses, utensils) without achieving sterility

  • Variables affecting microbial death rate

    • Number of microbes present: more organisms take longer to kill

    • Presence of endospores: highly resistant to many methods

    • Presence of organic material: organic debris can shield microbes from treatment (e.g., UV light may be absorbed by debris, protecting organisms underneath)

  • Methods of control

    • Physical methods

    • Temperature: hot and cold can kill microbes; hot denatures proteins; freezing and thawing can cause ice crystal damage

    • Filtration: physically removes cells

    • Desiccation: dehydration inhibits microbial metabolism

    • Ultraviolet (UV) light: damages DNA, creating mutations; severe damage prevents repair and replication

    • Chemical methods

    • Alcohols: dissolve lipids (disrupt membranes and proteins)

    • Soaps: act as surfactants and aid in mechanical removal

    • Preservatives (nitrates and nitrites): inhibit growth or kill microbes

  • Exceptions and notable organisms

    • Pseudomonas species: can survive and grow in disinfectants; a problem in medical facilities due to contamination of cleaning agents and equipment

    • Endospores: extremely resistant to many physical and chemical control methods; remain a major challenge in sterilization

  • Summary

    • Chapter 6: covers microbial growth physics, biochemistry, growth measurement, and preservation techniques

    • Chapter 7: covers rationale and