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Questions and Answers
What does the term 'generation time' refer to in microbial growth?
What does the term 'generation time' refer to in microbial growth?
Which phase of microbial growth is characterized by the cells being in the healthiest state?
Which phase of microbial growth is characterized by the cells being in the healthiest state?
What is the main process by which bacterial cells replicate?
What is the main process by which bacterial cells replicate?
Which of the following is NOT one of the four phases of microbial growth in a closed system?
Which of the following is NOT one of the four phases of microbial growth in a closed system?
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How does generation time vary among different microbes?
How does generation time vary among different microbes?
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During which phase does the growth rate of a microbial population become zero?
During which phase does the growth rate of a microbial population become zero?
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What is the principal characteristic of the lag phase in microbial growth?
What is the principal characteristic of the lag phase in microbial growth?
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In a traditional sigmoid growth curve, what is indicated on the y-axis?
In a traditional sigmoid growth curve, what is indicated on the y-axis?
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What is one advantage of directly counting cells using automated counters?
What is one advantage of directly counting cells using automated counters?
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What is a significant disadvantage of using direct microscopic examination for total cell count?
What is a significant disadvantage of using direct microscopic examination for total cell count?
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Which method is primarily used to measure the viable count of bacteria?
Which method is primarily used to measure the viable count of bacteria?
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What is a key requirement when using the viable count method?
What is a key requirement when using the viable count method?
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What type of measurement is represented by most probable number (MPN)?
What type of measurement is represented by most probable number (MPN)?
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What is a significant disadvantage of the pour-plate method in viable counting?
What is a significant disadvantage of the pour-plate method in viable counting?
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In the context of indirect measurement, what does dry weight measurement involve?
In the context of indirect measurement, what does dry weight measurement involve?
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Which of the following is NOT a method for direct measurement of microbial growth?
Which of the following is NOT a method for direct measurement of microbial growth?
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Which type of bacteria can tolerate oxygen but cannot use it for growth?
Which type of bacteria can tolerate oxygen but cannot use it for growth?
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What enzyme catalyzes the inactivation of hydrogen peroxide in bacteria?
What enzyme catalyzes the inactivation of hydrogen peroxide in bacteria?
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Which organisms can only utilize oxygen at reduced levels compared to air?
Which organisms can only utilize oxygen at reduced levels compared to air?
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What is the primary nitrogen source for most bacteria?
What is the primary nitrogen source for most bacteria?
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Which component is essential for forming DNA, RNA, and ATP in bacteria?
Which component is essential for forming DNA, RNA, and ATP in bacteria?
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What is the role of superoxide dismutase in bacteria?
What is the role of superoxide dismutase in bacteria?
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Which type of bacteria captures energy from inorganic compounds?
Which type of bacteria captures energy from inorganic compounds?
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What type of salts dissociate to provide sulphate ions in bacteria?
What type of salts dissociate to provide sulphate ions in bacteria?
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What primarily causes the death phase in microbial growth?
What primarily causes the death phase in microbial growth?
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What does the equation $N_{final}=(N_{initial}) \times 2^n$ represent in microbial growth?
What does the equation $N_{final}=(N_{initial}) \times 2^n$ represent in microbial growth?
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Why is exponential growth in batch conditions limited to a few generations?
Why is exponential growth in batch conditions limited to a few generations?
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How does a chemostat maintain constant bacterial growth?
How does a chemostat maintain constant bacterial growth?
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What purpose does a turbidostat serve in continuous culture?
What purpose does a turbidostat serve in continuous culture?
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What is the importance of achieving a steady state condition before sampling?
What is the importance of achieving a steady state condition before sampling?
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What does the term 'washout' refer to in continuous culture systems?
What does the term 'washout' refer to in continuous culture systems?
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In continuous culture, what does the dilution rate (D) depend on?
In continuous culture, what does the dilution rate (D) depend on?
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What is a significant disadvantage of measuring turbidity for microbial growth?
What is a significant disadvantage of measuring turbidity for microbial growth?
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Which temperature classification includes organisms that can grow best at temperatures above 45ºC?
Which temperature classification includes organisms that can grow best at temperatures above 45ºC?
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Which type of microorganism is best suited for growth in highly acidic environments?
Which type of microorganism is best suited for growth in highly acidic environments?
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What is the primary effect of placing cells in a hypotonic solution?
What is the primary effect of placing cells in a hypotonic solution?
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What role does hydrostatic pressure play in microbial growth?
What role does hydrostatic pressure play in microbial growth?
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Which of the following statements is true regarding facultative organisms?
Which of the following statements is true regarding facultative organisms?
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What cardinal temperature refers to the maximum temperature at which a microorganism can grow?
What cardinal temperature refers to the maximum temperature at which a microorganism can grow?
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What is typically the optimal pH range for most bacteria?
What is typically the optimal pH range for most bacteria?
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What is the defining characteristic of halophilic microorganisms?
What is the defining characteristic of halophilic microorganisms?
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Which of the following is true about extremophiles?
Which of the following is true about extremophiles?
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Study Notes
Microbial Growth
- Microbial growth refers to an increase in population size rather than the size of a single microbe.
- Microbial cells replicate by various methods, including binary fission (bacteria) and budding (yeast).
Generation Time
- Generation time is the time it takes for a microbial population to double in size.
- Different microbes have different generation times, ranging from 20-30 minutes for some bacterium to 10-30 days for Mycobacterium leprae.
Microbial Growth Cycle in Batch Culture
- A batch culture is a closed system with a fixed volume where nutrients are not renewed.
- Four phases characterize the growth curve of microbial populations in batch culture.
- Lag Phase: The time between inoculation and the start of growth where microbial cells are adjusting to their environment.
- Log or Exponential Phase: The period of most rapid growth where microbial cells are in their healthiest state.
- Stationary Phase: Growth rate equals death rate, resulting in a stable population due to nutrient depletion and waste accumulation.
- Death Phase: The number of cell deaths exceeds the number of new divisions, due to a lack of nutrients and increasing waste products.
Continuous Culture
- Maintaining constant environmental conditions over extended periods requires continuous culture.
- Two primary continuous culture methods include chemostats and turbidostats.
Chemostat
- A chemostat is a continuous culture device that maintains a constant volume by:
- Continually adding fresh medium from a sterile reservoir.
- Removing spent medium through an overflow drain.
- Matching the rate of growth to the removal rate of bacterial cells and waste products.
- The rate of growth in a chemostat is directly proportional to the rate of addition of fresh medium.
Turbidostat
- A turbidostat continuously monitors and adjusts the growth environment:
- Maintaining constant turbidity through manipulation of medium feed rates.
- Increasing feed rate when turbidity rises to dilute the culture.
- Decreasing feed rate when turbidity falls to encourage growth and restore turbidity.
Important Terminology in Continuous Culture
- Dilution rate (D): the ratio of flow rate (F) to volume (V): D= F/V.
- Growth rate (µ): Constant growth rate in continuous culture.
- Maximum growth rate (µmax): The highest growth rate achievable under given conditions.
- Steady state: a stable condition where the rate of growth is balanced by the removal rate, keeping population size constant.
- Washout: where the dilution rate exceeds the growth rate, leading to the complete removal of cells from the culture.
Measuring Microbial Growth
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Direct Measurement:
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Total Cell Count: Directly counting cells using methods like:
- Microscopic examination with special slides.
- Automated counters (flow cytometry)
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Viable Count: Determining the number of living, reproducing cells using methods like:
- Plating Techniques: Spread-plate method and Pour-plate method. - Serial Dilutions: Diluting the sample to achieve countable colonies, typically 25-250 colonies. - Filtration: Filtering large volumes to concentrate microbial cells for counting.
- Most Probable Number (MPN): A statistical method used to estimate microbial populations, often used with water quality testing
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Total Cell Count: Directly counting cells using methods like:
- Indirect Measurement: - Metabolic Activity: Measuring the byproduct of metabolism, such as: - Oxygen consumption. - Carbon dioxide production. - Acid production. - Dry Weight: Weighing the dried cell pellet after centrifuging the liquid culture. Does not distinguish between live and dead cells. - Turbidity: Utilizing a spectrophotometer to measure the cloudiness (turbidity) of the culture, which is related to the number of cells.
Factors Affecting Microbial Growth
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Physical Requirements for Growth:
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Temperature:
- Psychrophiles: Thrive in cold environments (optimum temperature below 15°C)
- Psychrotrophs: Grow at low temperatures (0°C–30°C), causing spoilage in refrigerated foods
- Mesophiles: Optimal growth at moderate temperatures (20°C–45°C), including the majority of human pathogens
- Thermophiles: Grow at higher temperatures (45°C-80°C)
- Extreme Thermophiles (Hyperthermophiles): Live in extremely hot environments, often above 80°C, and some even withstand temperatures above 100°C
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pH:
- Neutrophiles: Favor near-neutral pH (6.5-7.5), the majority of bacteria and protozoa
- Acidophiles: Grow in acidic environments (pH < 4), including some bacteria and fungi
- Alkalinophiles: Flourish in alkaline environments (pH > 9), such as those found in soda lakes
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Osmotic Pressure:
- Plasmolysis: Cell shrinkage due to water loss in a hypertonic environment (high solute concentration).
- Plasmoptisis: Cell swelling and potential bursting due to water entry in a hypotonic environment (low solute concentration).
- Halophiles: Adapted to high-salt environments, typically 30%, including obligate halophiles (require high salt) and facultative halophiles (tolerate high salt).
- Saccharophilic: Microorganisms that thrive in high sugar concentrations.
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Hydrostatic Pressure:
- Barophiles: Live in environments with extreme pressure (high hydrostatic pressure), their enzymes and membranes depend on pressure to maintain their proper shape.
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Oxygen:
- Aerobes: Require oxygen for growth, including Pseudomonas species.
- Anaerobes: Cannot tolerate and may be killed by oxygen, including Clostridium species.
- Facultative Organisms: Can grow with or without oxygen, including E. coli and Staphylococcus species.
- Aerotolerant Anaerobes: Can tolerate oxygen but do not use for metabolism, including Lactobacillus species.
- Microaerophiles: Require low-oxygen levels for growth, including Campylobacter species.
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Temperature:
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Chemical Requirements for Growth:
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Carbon and Energy Sources:
- Chemoautotrophs: Obtain energy from oxidizing inorganic substances and use carbon dioxide as their carbon source.
- Chemoheterotrophs: Obtain both energy and carbon from organic compounds, such as glucose and amino acids.
- Photoautotrophs: Use light as an energy source and CO2 as their carbon source, including plants and algae.
- Photoheterotrophs: Use light as an energy source but obtain carbon from organic compounds, including some bacteria.
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Nitrogen:
- Essential for building amino acids, DNA, and RNA.
- Sources of nitrogen include:
- Protein: common in most bacteria
- Ammonium: found in organic matter
- Nitrogen gas (N2): Fixed by nitrogen-fixing bacteria, both free-living and associated with legumes.
- Nitrates: Inorganic salts that dissociate into NO3-
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Sulfur:
- Essential for forming sulfur-containing amino acids and some vitamins (thiamin and biotin).
- Sources of sulfur include:
- Protein: Major source for bacteria
- Hydrogen sulfide
- Sulfates: salts that dissociate into SO4-
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Phosphorus:
- Essential for building DNA, RNA, ATP, and phospholipids.
- Sources of phosphorus include:
- Inorganic phosphate salts and buffers
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Trace Elements:
- Small amounts of elements are necessary for microbial growth, including iron, copper, zinc, and magnesium.
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Carbon and Energy Sources:
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Description
Explore the fundamentals of microbial growth, including the various methods of replication and the concept of generation time. This quiz delves into the growth cycle phases in batch culture and highlights the characteristics of each phase. Test your knowledge on the dynamics of microbial populations!
