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Questions and Answers
What is the maximum temperature that psychrophiles can grow at?
What is the maximum temperature that psychrophiles can grow at?
At what temperature do mesophiles thrive best?
At what temperature do mesophiles thrive best?
Which of the following examples is a psychrophile?
Which of the following examples is a psychrophile?
What type of organism grows best at pH levels between 5.5 and 8.0?
What type of organism grows best at pH levels between 5.5 and 8.0?
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Which method is used for obtaining a pure culture?
Which method is used for obtaining a pure culture?
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What is the thermal death time?
What is the thermal death time?
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Which organism is classified as an alkalophile?
Which organism is classified as an alkalophile?
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Which of the following is NOT a characteristic of liquid culture media?
Which of the following is NOT a characteristic of liquid culture media?
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Which type of media would be used for the growth of fastidious organisms?
Which type of media would be used for the growth of fastidious organisms?
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What does Blood Agar Plate (BAP) help to distinguish?
What does Blood Agar Plate (BAP) help to distinguish?
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Which medium differentiates E.coli from other lactose fermenting bacteria?
Which medium differentiates E.coli from other lactose fermenting bacteria?
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What type of media incorporates antibiotics to inhibit the growth of unwanted organisms?
What type of media incorporates antibiotics to inhibit the growth of unwanted organisms?
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Which medium is specifically selective for Vibrio?
Which medium is specifically selective for Vibrio?
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What does HEA identify?
What does HEA identify?
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Which component can be added to a selective medium to inhibit swarming bacteria?
Which component can be added to a selective medium to inhibit swarming bacteria?
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What is the function of the Antimicrobial Removal Device in blood culture systems?
What is the function of the Antimicrobial Removal Device in blood culture systems?
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What does the automated BACTEC system detect to indicate microbial growth?
What does the automated BACTEC system detect to indicate microbial growth?
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What are the possible appearances of colonies on agar plates?
What are the possible appearances of colonies on agar plates?
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How do some bacteria exhibit chromogenesis?
How do some bacteria exhibit chromogenesis?
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What characteristic is NOT observed in colony margins?
What characteristic is NOT observed in colony margins?
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What does turbid growth in a nutrient broth indicate?
What does turbid growth in a nutrient broth indicate?
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Which feature is observed in the optical characteristics of colonies?
Which feature is observed in the optical characteristics of colonies?
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In gelatin slants, how does bacterial growth usually appear?
In gelatin slants, how does bacterial growth usually appear?
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What kind of odors may bacteria produce during growth?
What kind of odors may bacteria produce during growth?
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What characterizes obligate aerobes?
What characterizes obligate aerobes?
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Which organism is an example of an obligate anaerobe?
Which organism is an example of an obligate anaerobe?
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Facultative anaerobes are significant in which way?
Facultative anaerobes are significant in which way?
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Microaerophiles require which concentration of oxygen for optimal growth?
Microaerophiles require which concentration of oxygen for optimal growth?
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Capnophiles prefer an environment with increased levels of which gas?
Capnophiles prefer an environment with increased levels of which gas?
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Which statement is true about phototrophs?
Which statement is true about phototrophs?
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What is the optimal temperature range for most clinically significant bacteria?
What is the optimal temperature range for most clinically significant bacteria?
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Heterotrophs differ from autotrophs in that they:
Heterotrophs differ from autotrophs in that they:
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What is the primary use of semi-solid medium?
What is the primary use of semi-solid medium?
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Which of the following is NOT a characteristic of solid medium?
Which of the following is NOT a characteristic of solid medium?
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Which type of medium has known components and is designed for research purposes?
Which type of medium has known components and is designed for research purposes?
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What is the function of enrichment media?
What is the function of enrichment media?
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What type of medium is Hela 229 cells grown in?
What type of medium is Hela 229 cells grown in?
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Which example is associated with solid medium?
Which example is associated with solid medium?
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What characterizes synthetic or defined medium?
What characterizes synthetic or defined medium?
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Alkaline Peptone Water is best known for its use with which species?
Alkaline Peptone Water is best known for its use with which species?
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Study Notes
Oxygen Requirement
- Obligate aerobes require oxygen for growth and thrive in room air (15-20% oxygen, 1% carbon dioxide).
- Examples: Bordetella, Brucella, Mycobacterium, Neisseria, and Pseudomonas
- Obligate anaerobes cannot survive in the presence of oxygen and die after prolonged exposure.
- Examples: Clostridium and Bacteriodes
- Facultative anaerobes grow in both the presence and absence of oxygen, but thrive better with oxygen.
- Examples: Enterobacteriaceae, Staphylococcus, Streptococcus
- Aerotolerant anaerobes can survive in the presence of oxygen but cannot use it for metabolism.
- Examples: Lactobacillus, Clostridium perfringes, and Cutibacterium
- Microaerophiles require low oxygen concentrations (2-10%) and can be grown in special jars or bags.
- Examples: Campylobacter, Helicobacter, and Treponema
- Capnophiles require increased carbon dioxide concentration (5-10%) and 15% oxygen.
- Examples: Neisseria, HACEK
Carbon Dioxide Requirement
- Capnophiles require increased carbon dioxide (5-10%).
- Most aerobic and facultative aerobic bacteria need 0.03% carbon dioxide.
- Example: Haemophilus influenza
Carbon Source
- Autotrophs utilize carbon as their sole carbon source.
- Heterotrophs obtain reduced organic material from other organisms.
Energy Source
- Phototrophs use light as their energy source.
- Chemotrophs obtain energy from the oxidation of organic or inorganic compounds.
Electron Source
- Lithotrophs obtain electrons from inorganic molecules.
- Organotrophs require organic substances like carbohydrates and lipids for growth and multiplication.
Temperature Requirement
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Optimum temperature for most clinically significant bacteria: 35°C to 37°C.
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Psychrophiles/Cryophiles: Grow well at 0°C to 20°C.
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Examples: Listeria monocytogenes, Yersinia enterocolitica
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Mesophiles: Grow between 20°C to 45°C, with an optimum of 37°C.
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Most commonly encountered pathogenic bacteria in clinical labs.
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Thermophiles/Hyperthermophiles: Grow between 50°C to 60°C.
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Examples: Geobacillus stearothermophilus, Thermus aquaticus, Sulfolobus, Pyrococcus, Pyrodictum
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Extremophiles: Prokaryotes that thrive in unusual conditions, like low oxygen, high temperatures, or underground environments.
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Example: Bacillus infernus
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Thermal death time: The minimum time required to kill organisms at a constant temperature.
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Thermal death point: The lowest temperature required to kill organisms in a constant time.
pH Tolerance
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pH scale: Measures hydrogen ion concentration.
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Acidophiles: Grow between pH 0 to 5.5.
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Example: Lactobacillus acidophilus
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Neutrophiles: Grow between pH 5.5 to 8.0.
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Most clinically significant bacteria.
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Alkalophiles: Grow between pH 8.5 to 11.5.
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Example: Vibrio
Culture and Culture Media
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Pure culture: Contains a single species.
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Used for identification and antimicrobial susceptibility testing.
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Methods: Streak/spread plate, pour plate, selective medium, animal inoculation.
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Mixed culture: Contains more than one species.
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Stock culture: Contains several species in separate culture media (one species per medium).
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Cultivated in large volumes of broth, divided into small vials for long shelf life (up to 1 year).
Classification of Culture Media by Consistency
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Liquid media: No agar; for rapid culture; supports growth of aerobes, anaerobes, and facultative anaerobes.
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Examples: Brain heart infusion, trypticase soy broth, thioglycolate broth
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Semi-solid media: Contains 0.5% to 1% agar; used to observe bacterial motility and detect indole and sulfide production.
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Example: Sulfide Indole Motility (SIM) medium
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Solid media:
- Liquefiable: Contains 2% to 3% agar, liquefies when heated.
- Non-liquefiable: Doesn't contain agar.
- Biphasic: Contains both liquid and solid components.
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Examples: Triple Sugar Iron (TSI) agar, MacConkey agar, Blood agar plate (BAP), chocolate agar plate (CAP).
Classification of Culture Media by Composition
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Synthetic/Defined media: Components are known; used for research purposes; can be liquid or solid.
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Non-synthetic media: Some components are unknown (peptones, meat, yeast extracts).
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Tissue culture media:
- HELA 229 cells: Human cervical tissue cells.
- McCoy cells (W138 cells): Fibroblasts.
- Used for isolating Chlamydia.
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Embryonated cells: Used for propagating Rickettsia.
Classification of Culture Media by Dispensing/Distribution
- Tube media: Prepared as liquid, slant, butt and slant, or butt.
- Plated media: Distributed in dishes or plates.
Classification of Culture Media by Use
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Simple/General purpose/Supportive media: Support growth of most non-fastidious bacteria; routine in laboratory.
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Examples: Blood agar, nutrient broth, trypticase soy agar
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Enrichment media (Liquid types): Enhance growth of specific bacteria while inhibiting others.
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Alkaline Peptone Water: Used for Vibrio species.
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Thioglycolate broth: General purpose enrichment medium, supports growth of many non-fastidious organisms.
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Gram-negative broth: Enhances growth of Salmonella and Shigella.
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Enriched media and non-selective media (Solid types): Provide additional supplements for fastidious organisms.
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Examples: BAP, CAP (Blood agar plate and chocolate agar plate).
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Blood agar plate (BAP): 5% defibrinated sheep's blood (horse and rabbit blood are second and third choices, respectively).
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Human BAP: Used for Gardnerella.
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Buffered Charcoal Yeast Extract (BCYE): Another enriched medium.
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Differential media: Allow visualization of metabolic differences between bacterial groups.
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Examples:
- MacConkey agar (MAC): Separates lactose fermenters from non-lactose fermenters.
- BAP: Differentiates hemolytic patterns.
- Eosin Methylene Blue (EMB) agar: Differentiates E. coli from other lactose fermenting bacteria.
- Hektoen Enteric (HEA) agar: Identifies Salmonella from Shigella.
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Selective media: Inhibit growth of unwanted bacteria while promoting growth of desired bacteria.
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Examples:
- Gentamicin blood agar: For Streptococcus.
- Bacitracin chocolate agar: For Haemophilus.
- Blood agar plate with ampicillin: For Aeromonas.
- Phenylethyl alcohol agar: For Gram-positive bacteria.
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Inhibitory substances for selective media:
- To inhibit Gram-positive bacteria: Crystal violet/Gentian violet/carbon fuchsin, bile salt
- To inhibit Gram-negative bacteria: Potassium tellurite, sodium azide
- To inhibit swarming of bacteria: Alcohol, chloral hydrate
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Special media: Used to isolate bacteria with specific growth requirements.
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Examples:
- Lowenstein-Jensen (LJ) medium: Protein-rich, composed of whole eggs and malachite green.
- Thiosulfate-citrate-bile-salts-sucrose (TCBS) agar: Selective for Vibrio; sterilized by boiling, not autoclaving.
Blood Culture System
- Blood culture bottles: Used to isolate aerobic and anaerobic bacteria.
- Antimicrobial Removal Devices (ARDs) or resins: Added to the medium to absorb antibiotics present in the blood sample.
- Manually: Biphasic medium containing MacConkey agar (MAC) and Blood agar plate (BAP).
- Automated system (BACTEC): Detects carbon dioxide production by bacteria as an indicator of growth.
Quality Control of Culture Media
- After preparing media, an uninoculated sample should be incubated overnight at 35°C to check for bacterial contamination.
Cultural Characteristics
- Appearance of bacteria on various media: Color of colonies and media, abundance of growth, odor.
Reporting Cultural Characteristics
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Agar plate colonies:
- Size: Pinpoint to large, mucoid, slimy.
- Margin and elevation: Smooth, entire, filiform, lobate, undulate.
- Chromogenesis (Pigmentation): Colored or unpigmented.
- Optical features: Opaque, translucent, shiny, opalescent.
- Odor: Putrid, fruity, ammoniacal, aromatic.
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Growth on slant:
- Amount: Scanty, moderate, abundant.
- Margin or edge: Same as agar plate descriptions.
- Consistency: Butyrous, viscous, dry, brittle.
- Chromogenesis: Pigmented.
- Odor.
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Growth in nutrient broth:
- Turbid: Indicates bacterial growth.
- Viscous: Thick, sticky appearance.
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Growth in gelatin slant:
- Thread-like or beaded-like string pearl pattern: Growth confined to the inoculation zone.
- Liquefaction: Starts evenly from the top of the agar slant, creating a funnel-like appearance.
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Description
This quiz explores the different types of microbial oxygen requirements, including obligate aerobes, obligate anaerobes, facultative anaerobes, aerotolerant anaerobes, microaerophiles, and capnophiles. Learn about the examples of bacteria that fall into each category and their respective oxygen needs for growth. Test your understanding of these essential microbiological concepts.