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Anaerobic Microbiology Lab Techniques
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Anaerobic Microbiology Lab Techniques

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Questions and Answers

What is the primary purpose of inoculating a PEDS bottle in fluid culture?

  • To detect anaerobic bacteria
  • To detect yeast and other microorganisms
  • To incubate the sample for 5 days
  • To load the sample onto the BacT (correct)

    • What type of agar is specifically used for Vibrio species?

  • BA plate
  • XLD agar
  • MacConkey's agar
  • TCBS agar (correct)

    • What is the purpose of making a cytospin gram in fluid culture?

  • To centrifuge the sample (correct)
  • To plant the sample to agar
  • To detect anaerobic bacteria
  • To examine the sample for viscosity

    • What type of media is required for Campylobacter species?

    <p>Skirrow's media</p> Signup and view all the answers

    What is the purpose of using a gram stain in fluid culture?

    <p>To directly plant the sample to agar</p> Signup and view all the answers

    Which of the following is NOT an example of anaerobic bacteria?

    <p>E.coli</p> Signup and view all the answers

    What is the primary purpose of using a BA plate in fluid culture?

    <p>To detect multiple types of microorganisms</p> Signup and view all the answers

    What type of agar is used for stool cultures?

    <p>All of the above</p> Signup and view all the answers

    What is the primary purpose of using differential media in a microbiology lab?

    <p>To differentiate between various types of bacteria</p> Signup and view all the answers

    What is the correct order of inoculation in a microbiology lab?

    <p>Supportive, Enrichment, Differential, Selective</p> Signup and view all the answers

    What is the purpose of streaking plates in a microbiology lab?

    <p>To isolate a specific bacteria from a mixed population</p> Signup and view all the answers

    What is the most common type of media used in a microbiology lab?

    <p>MacConkey's agar</p> Signup and view all the answers

    What is the purpose of the three-phase streaking pattern, known as the T-Streak?

    <p>To isolate a specific bacteria from a mixed population</p> Signup and view all the answers

    What is the correct way to inoculate a culture plate from a swab or other source?

    <p>Roll the swab or other source across the media in a zigzag motion</p> Signup and view all the answers

    What is the purpose of using a sterile loop or swab in inoculating a culture plate?

    <p>To prevent contamination of the culture</p> Signup and view all the answers

    What is the purpose of Step 1 in the streaking process?

    <p>To cover approximately 30% of the plate with the inoculum</p> Signup and view all the answers

    What is the primary purpose of performing quality control tests on prepared media?

    <p>To assess the growth support properties of the product</p> Signup and view all the answers

    What information should be included on the label of a prepared medium before storage?

    <p>Type of media, lot number, and expiry date</p> Signup and view all the answers

    What is the primary function of enrichment media?

    <p>To enhance the growth of pathogens while inhibiting normal flora</p> Signup and view all the answers

    Which type of media is used to differentiate between various microorganisms?

    <p>Differential media</p> Signup and view all the answers

    What is the purpose of performing stability tests on prepared media?

    <p>To evaluate the shelf life of the product</p> Signup and view all the answers

    Which of the following is an example of selective media?

    <p>Salmonella-Shigella agar</p> Signup and view all the answers

    What is the primary purpose of using supportive media?

    <p>To provide minimum requirements for growth</p> Signup and view all the answers

    Which of the following media is used for sputum and stool cultures?

    <p>Salmonella-Shigella agar</p> Signup and view all the answers

    What type of environment is created in a Microaerophilic Jar?

    <p>5% oxygen environment</p> Signup and view all the answers

    What is the primary purpose of the Lag Phase in the growth curve of bacteria?

    <p>Adjustment period to a new medium and environment</p> Signup and view all the answers

    What type of specimen has priority when inoculating plates?

    <p>Surgical specimens</p> Signup and view all the answers

    What is the purpose of using MacConkey's plate in urine cultures?

    <p>To detect Enterobacteriaceae family</p> Signup and view all the answers

    What is the primary purpose of incubating culture plates for 18-24 hours?

    <p>To allow for colonies to appear</p> Signup and view all the answers

    What type of agar is used for throat swabs?

    <p>BA plate</p> Signup and view all the answers

    What is the primary purpose of using a calibrated loop in urine cultures?

    <p>To quantify the colonies</p> Signup and view all the answers

    What is the primary purpose of using a Candle Jar?

    <p>To create a carbon dioxide enriched environment</p> Signup and view all the answers

    What is the primary purpose of using Amies Transport Media with charcoal?

    <p>To neutralize materials toxic to sensitive pathogens</p> Signup and view all the answers

    What type of specimen requires a sterile container for collection?

    <p>Urine for C&amp;S</p> Signup and view all the answers

    What is the purpose of using Cary Blair Transport Medium?

    <p>To transport stool and rectal swabs</p> Signup and view all the answers

    Which of the following specimens typically has normal flora?

    <p>Throat</p> Signup and view all the answers

    What should be written on the requisition during specimen collection?

    <p>All of the above</p> Signup and view all the answers

    What should be avoided when collecting a specimen for C&S?

    <p>Using laxative, enemas, or antibiotics for one week prior</p> Signup and view all the answers

    What is the purpose of using Amies Transport Media?

    <p>To maintain microorganisms without overgrowth</p> Signup and view all the answers

    Which of the following specimens is considered sterile?

    <p>Blood</p> Signup and view all the answers

    Facultative anaerobes can only grow in the presence of oxygen.

    <p>False</p> Signup and view all the answers

    Bacteria require 50% humidity to grow.

    <p>False</p> Signup and view all the answers

    The anaerobic jar is used to create an environment with 5% oxygen.

    <p>False</p> Signup and view all the answers

    The pH range for bacterial growth is between 6.0 and 8.0.

    <p>False</p> Signup and view all the answers

    Bacteria require 90% water to grow.

    <p>False</p> Signup and view all the answers

    The incubation temperature for bacterial growth is between 30-40°C.

    <p>False</p> Signup and view all the answers

    Capnophilic bacteria grow in the presence of 10-15% CO2.

    <p>False</p> Signup and view all the answers

    Methylene Blue is used as an indicator to detect aerobic environments.

    <p>False</p> Signup and view all the answers

    The primary purpose of using enrichment media is to selectively isolate microorganisms that require specific nutrients.

    <p>True</p> Signup and view all the answers

    Chocolate agar is a type of selective media used for the growth of fastidious microorganisms.

    <p>False</p> Signup and view all the answers

    The lag phase of the bacterial growth curve is the period of rapid cell division.

    <p>False</p> Signup and view all the answers

    A microaerophilic environment is one that is completely devoid of oxygen.

    <p>False</p> Signup and view all the answers

    The correct order of inoculation is supportive, enrichment, differential, and then selective media.

    <p>False</p> Signup and view all the answers

    Incubation of culture plates at 37°C is necessary for the growth of all microorganisms.

    <p>False</p> Signup and view all the answers

    The streaking process is used to isolate microorganisms from a mixed culture.

    <p>True</p> Signup and view all the answers

    MacConkey's agar is a type of supportive media used for the growth of all microorganisms.

    <p>False</p> Signup and view all the answers

    A Microaerophilic Jar creates a 10% oxygen environment.

    <p>False</p> Signup and view all the answers

    The Log phase of a growth curve is a period of slow growth for bacteria.

    <p>False</p> Signup and view all the answers

    Blood cultures must be incubated at 37 degrees C.

    <p>False</p> Signup and view all the answers

    Blood cultures are collected using a sterile loop or swab.

    <p>False</p> Signup and view all the answers

    Candle Jar is used to create a carbon dioxide enriched environment.

    <p>True</p> Signup and view all the answers

    Patients weighing more than 27 kg must have 10 mL of blood drawn from two different venipunctures or sites within one hour.

    <p>False</p> Signup and view all the answers

    The Stationary phase of a growth curve is a period of rapid growth for bacteria.

    <p>False</p> Signup and view all the answers

    Each blood culture bottle can hold up to 20 mL of blood.

    <p>False</p> Signup and view all the answers

    Culture plates are typically incubated for 48 hours or more before colonies appear.

    <p>False</p> Signup and view all the answers

    Blood cultures are collected in a microaerophilic jar with 20% O2.

    <p>False</p> Signup and view all the answers

    Throat swabs are inoculated onto MacConkey's plate.

    <p>False</p> Signup and view all the answers

    Aerobic bottles are filled before anaerobic bottles when collecting blood cultures.

    <p>True</p> Signup and view all the answers

    Urine cultures are inoculated using a calibrated loop of 1 ml.

    <p>False</p> Signup and view all the answers

    Blood cultures can be collected from a single venipuncture site.

    <p>False</p> Signup and view all the answers

    Surgical specimens have a low priority when inoculating plates.

    <p>False</p> Signup and view all the answers

    The total blood collection for a blood culture should be at least 20 mL.

    <p>False</p> Signup and view all the answers

    MacConkey Media is used to grow anaerobic bacteria.

    <p>False</p> Signup and view all the answers

    A medium with a pH value outside the specified range should be discarded.

    <p>True</p> Signup and view all the answers

    All media must be stored at room temperature.

    <p>False</p> Signup and view all the answers

    A representative sample of each lot/batch of medium should be incubated for 7-10 days during sterility testing.

    <p>False</p> Signup and view all the answers

    The thermometer used in preparing media should have a range of -20°C to 150°C.

    <p>False</p> Signup and view all the answers

    The label on a prepared medium should include only the type of media and lot number.

    <p>False</p> Signup and view all the answers

    Agar is not a necessary component of MacConkey Media.

    <p>False</p> Signup and view all the answers

    The purpose of pre-incubation is to check for contamination and quality control.

    <p>True</p> Signup and view all the answers

    Match the following types of bacteria with their oxygen requirements:

    <p>Aerobic = Requires 21% oxygen Anaerobic = Grows in the absence of oxygen Facultative anaerobes = Can grow in the presence or absence of oxygen Microaerophilic = Must have 5% oxygen</p> Signup and view all the answers

    Match the following growth conditions with their required humidity levels:

    <p>General bacterial growth = 70% humidity Incubation environment = 70% humidity Campylobacter growth = 5% oxygen Bacterial growth in general = 50% humidity</p> Signup and view all the answers

    Match the following types of jars with their purposes:

    <p>Anaerobic jar = Depletes O2 from the jar Microaerophilic jar = Creates an environment with 5% oxygen Candle jar = Creates an environment with 10% CO2 Autoplak = Not related to bacterial growth</p> Signup and view all the answers

    Match the following bacterial growth requirements with their corresponding values:

    <p>Isotonic environment = 0.9% saline pH range = 7.2-7.6 Temperature = 35-37°C Water requirement = 90%</p> Signup and view all the answers

    Match the following indicators with their purposes:

    <p>Methylene Blue = Detects anaerobic environments Candle jar = Creates an environment with 10% CO2 Autoplak = Not related to bacterial growth</p> Signup and view all the answers

    Match the following types of bacteria with their corresponding CO2 requirements:

    <p>Capnophilic bacteria = 5-10% CO2 Aerobic bacteria = 21% oxygen Facultative anaerobes = Can grow in the presence or absence of oxygen Microaerophilic = Must have 5% oxygen</p> Signup and view all the answers

    Match the following bacterial growth conditions with their corresponding requirements:

    <p>Bacterial growth = No humidity is required</p> Signup and view all the answers

    Match the following specialized growth environments with their purposes:

    <p>Anaerobic environment = Grows anaerobic bacteria Microaerophilic environment = Grows microaerophilic bacteria Candle jar = Creates an environment with 10% CO2 Autoplak = Not related to bacterial growth</p> Signup and view all the answers

    Match the following conditions with the type of environment created:

    <p>42 degrees C = Incubation temperature for bacterial growth 5% O2 = Microaerophilic environment 10-15% CO2 = Capnophilic environment 90% water = Humidity requirement for bacterial growth</p> Signup and view all the answers

    Match the following specimen collection requirements with the corresponding volume:

    <p>Patients weighing more than 27 kg = 40 mL of blood from two different venipunctures Pediatric patients = Collection volumes vary depending on age and weight Blood cultures = 20 mL of blood equally divided into one aerobic and one anaerobic bottle Blood cultures from a single site = Do not place more than 12 mL of blood in a single blood culture bottle</p> Signup and view all the answers

    Match the following growth conditions with the relevant information:

    <p>Aerobic bottle = Filled first to prevent air from the tubing of the butterfly to go into the anaerobic bottle Anaerobic bottle = Filled second to prevent contamination with oxygen Blood culture adapter = Used to collect blood for a blood culture Butterfly method = Used to collect blood from two different venipunctures or sites within one hour</p> Signup and view all the answers

    Match the following growth environments with their respective characteristics:

    <p>Microaerophilic environment = 5% O2 and 95% N2 Anaerobic environment = No oxygen present Aerobic environment = Presence of oxygen Capnophilic environment = 10-15% CO2</p> Signup and view all the answers

    Match the following growth conditions with the relevant information:

    <p>Aerobic environment = Required for growth of bacteria that need oxygen Anaerobic environment = Required for growth of bacteria that do not need oxygen Incubation at 42 degrees C = Optimal temperature for bacterial growth Humidity of 50% = Optimal humidity for bacterial growth</p> Signup and view all the answers

    Match the following incubation conditions with the relevant information:

    <p>Incubation at 42 degrees C = Optimal temperature for bacterial growth Incubation for 18-24 hours = Time required for visible growth of bacteria pH range of 6.0-8.0 = Optimal pH range for bacterial growth Humidity of 50% = Optimal humidity for bacterial growth</p> Signup and view all the answers

    Match the following growth environments with their respective characteristics:

    <p>Aerobic environment = Supports growth of bacteria that require oxygen Anaerobic environment = Supports growth of bacteria that do not require oxygen Microaerophilic environment = Supports growth of bacteria that require low oxygen levels Capnophilic environment = Supports growth of bacteria that require high CO2 levels</p> Signup and view all the answers

    Match the following growth conditions with the relevant information:

    <p>Aerobic environment = Required for growth of bacteria that need oxygen Anaerobic environment = Required for growth of bacteria that do not need oxygen Incubation at 42 degrees C = Optimal temperature for bacterial growth pH range of 6.0-8.0 = Optimal pH range for bacterial growth</p> Signup and view all the answers

    Match the following media preparation steps with their corresponding descriptions:

    <p>Weigh out the dehydrated media = Sterilizing it (usually in an autoclave) Dissolve in distilled water = Weigh out the dehydrated media Sterilizing it (usually in an autoclave) = Pouring the plates Pouring the plates = Dissolve in distilled water</p> Signup and view all the answers

    Match the following quality control tests with their corresponding descriptions:

    <p>pH value = Incubate for 2-5 days at 35-37°C and at room temperature Sterility = Incubate for 2-5 days at 35-37°C and at room temperature</p> Signup and view all the answers

    Match the following media components with their corresponding quantities:

    <p>Peptone = 17.0 g Lactose = 10.0 g Agar = 13.5 g Crystal violet = 0.001 g</p> Signup and view all the answers

    Match the following equipment with their corresponding uses:

    <p>Thermometer = Measuring temperature Balance = Weighing out media components Autoclave = Sterilizing media Funnel = Pouring plates</p> Signup and view all the answers

    Match the following media preparation steps with their corresponding temperatures:

    <p>Incubation = 35-37°C Pre incubation = 25°C Storage = 2-8°C Sterilization = 110°C</p> Signup and view all the answers

    Match the following media components with their corresponding functions:

    <p>Bile salts = Differentiating between microorganisms Neutral red = Selective growth Lactose = Providing energy for microorganisms Agar = Solidifying media</p> Signup and view all the answers

    Match the following media preparation steps with their corresponding durations:

    <p>Incubation = 2-5 days Pre incubation = 18-24 hours Sterilization = 15 minutes Storage = Until expiry date</p> Signup and view all the answers

    Match the following media labels with their corresponding information:

    <p>Lot number = Identification of the media batch Expiry date = Shelf life of the media Type of media = Description of the media composition Storage conditions = Temperature and humidity requirements</p> Signup and view all the answers

    Match the growth phase of bacteria with its description:

    <p>Lag Phase = Rapid growth period for bacteria. Log Phase = Adjustment period to a new medium and environment. Stationary Phase = Balance between cell growth and cell death. Death Phase = Bacteria stop multiplying.</p> Signup and view all the answers

    Match the type of environment with its characteristics:

    <p>Microaerophilic Jar = Creates a 5% oxygen environment. Candle Jar = Creates a 10-15% CO2 environment. Carbon Dioxide enriched environment = Creates a 5% oxygen environment. Anaerobic Jar = Creates an environment with no oxygen.</p> Signup and view all the answers

    Match the type of specimen with the priority order for inoculation:

    <p>Surgical specimens = First priority. Sterile fluids = Second priority. Throat swabs = Third priority. Urine samples = Last priority.</p> Signup and view all the answers

    Match the type of agar with its usage:

    <p>BA plate = Used for throat swabs and looking for Group A Strep. MacConkey's plate = Used for urine cultures and quantitative urine cultures. Chocolate agar = Used for fastidious microorganisms. Candle agar = Used for anaerobic bacteria.</p> Signup and view all the answers

    Match the incubation condition with its description:

    <p>Temperature 30-40°C = Optimal temperature range for bacterial growth. pH range 6.0-8.0 = Optimal pH range for bacterial growth. 50% humidity = Optimal humidity level for bacterial growth. 90% water = Optimal water content for bacterial growth.</p> Signup and view all the answers

    Match the type of environment with the type of bacteria that grows in it:

    <p>Aerobic environment = Bacteria that require oxygen. Anaerobic environment = Bacteria that grow in the absence of oxygen. Microaerophilic environment = Bacteria that require a 5% oxygen environment. Capnophilic environment = Bacteria that grow in the presence of 10-15% CO2.</p> Signup and view all the answers

    Match the type of media with its purpose:

    <p>Enrichment media = Selectively isolates microorganisms that require specific nutrients. Selective media = Differentiates between various microorganisms. Supportive media = Supports the growth of microorganisms. Differential media = Differentiates between various microorganisms.</p> Signup and view all the answers

    Match the type of loop with its usage:

    <p>Calibrated loop = Used for quantitative urine cultures. Sterile loop = Used for inoculating culture plates. Candle loop = Used for anaerobic bacteria. BA loop = Used for throat swabs.</p> Signup and view all the answers

    Match the following types of media with their characteristics:

    <p>Selective Media = Used to selectively isolate microorganisms that require specific nutrients Supportive Media = Used to support the growth of all microorganisms Differential Media = Used to differentiate between various microorganisms Enrichment Media = Used to enrich the growth of microorganisms that require specific nutrients</p> Signup and view all the answers

    Match the following growth environments with their characteristics:

    <p>Aerobic Environment = Requires oxygen to grow Anaerobic Environment = Requires absence of oxygen to grow Microaerophilic Environment = Requires low oxygen levels to grow Capnophilic Environment = Requires high CO2 levels to grow</p> Signup and view all the answers

    Match the following incubation conditions with their characteristics:

    <p>Incubation Temperature = Between 30-40°C for bacterial growth pH Range = Between 6.0 and 8.0 for bacterial growth Humidity Level = Between 50-60% for bacterial growth Water Requirement = Between 90-95% for bacterial growth</p> Signup and view all the answers

    Match the following specialized growth environments with their characteristics:

    <p>Candle Jar = Creates an environment with low oxygen levels Anaerobic Jar = Creates an environment with absence of oxygen Amies Transport Media = Used to transport bacteria while maintaining their viability Cary Blair Transport Medium = Used to transport bacteria while maintaining their viability</p> Signup and view all the answers

    Match the following growth curves with their characteristics:

    <p>Lag Phase = Initial phase of slow growth Log Phase = Phase of rapid growth Stationary Phase = Phase of no growth Death Phase = Phase of decline in growth</p> Signup and view all the answers

    Match the following nutrient requirements with their characteristics:

    <p>Facultative Anaerobes = Can grow in the presence or absence of oxygen Obligate Anaerobes = Can only grow in the absence of oxygen Aerotolerant Anaerobes = Can tolerate oxygen but do not use it Microaerophilic Anaerobes = Can only grow in low oxygen levels</p> Signup and view all the answers

    Match the following types of bacteria with their characteristics:

    <p>Staphylococcus = Forms clusters coli = Forms rods Staph aureus = Forms clusters and has a thick peptidoglycan layer E. coli = Forms rods and is commonly found in the gut</p> Signup and view all the answers

    Match the following agar types with their characteristics:

    <p>MacConkey's Agar = Used for selective isolation of Gram-negative bacteria Blood Agar = Used for selective isolation of fastidious bacteria Chocolate Agar = Used for selective isolation of fastidious bacteria MacConkey's Agar with Crystal Violet = Used for selective isolation of Gram-positive bacteria</p> Signup and view all the answers

    Study Notes

    Anaerobes and Microbiology

    • Anaerobes examples: Clostridium, Propionibacterium, Bifidobacterium, Bacteroides, Fusobacterium
    • Anaerobes are inoculated into anaerobic blood agar (BAK) and Pedro bottles (yellow top blood culture bottle) for 5 days

    Fluids

    • Joint fluids are usually viscous, so a gram stain is made straight from the original sample and directly planted to the agar
    • Non-viscous fluids are centrifuged, and a cytospin gram is made
    • Looking for microorganisms like Staphylococcus aureus, Group A Streptococci, Enterobacteriaceae family, and anaerobic bacteria

    Stool

    • Inoculated into BA plate, MacConkey's, HE (Hektoen enteric agar), XLD (Xylose-Lysine-Desoxycholate), and SS Agar (Salmonella/Shigella)
    • Looking for Salmonella species, Shigella species, Campylobacter species, E. coli O157:H7, Yersinia species, and Vibrio species

    Media

    • Selective media: inhibits the growth of some organisms while enhancing the growth of others
    • Examples of selective media: Salmonella-Shigella (SS), Mannitol salt agar, Lim Broth, and MacConkey's
    • Differential media: have indicators that some bacteria will use (pink) and others won't (colorless)
    • Examples of differential media: MacConkey's

    Lab Techniques

    • Order of inoculation: supportive, enrichment, differential, and selective
    • Setting up cultures: inoculating from swabs or other sources, using a sterilized inoculation loop or sterile swab
    • Streaking plates: to isolate and identify bacteria, using a three-phase streaking pattern (T-Streak)
    • Quality control tests: testing growth performance, stability, and labelling media with type, lot number, and expiry date

    Microbiology Specimens

    • Collecting specimens: stool, sputum, urine, throat swabs, blood cultures, and wound swabs
    • Patient instructions: providing clear instructions for collecting and handling specimens
    • Amies transport media: used for transporting specimens, maintaining microorganisms without overgrowth
    • Cary Blair transport medium: used for transporting stool and rectal swabs

    Growth and Incubation

    • Growth curve: lag phase, log phase, stationary phase, and death phase
    • Incubation: plates are incubated for 18-24 hours before colonies appear, and no longer than 48 hours

    Streaking Plates

    • Streaking plates are used to isolate a single species of bacteria from a mixed population.
    • The three-phase streaking pattern is known as the T-Streak.
    • The process involves dragging a sterile loop or swab across the surface of the agar in a zigzag motion, covering approximately 30% of the plate.
    • The loop is then re-sterilized, and the plate is turned 90 degrees.

    Streaking Procedure

    • Step 1: Drag the sterile loop or swab across the surface of the agar in a zigzag motion, covering approximately 30% of the plate.
    • Step 2: Re-sterilize the loop, and turn the plate 90 degrees.
    • Step 3: Repeat the process, being cautious not to touch the previously streaked sectors.
    • The final section will have the least amount of growth and many isolated colonies.

    Bacterial Physiology

    • Aerobic bacteria require 21% oxygen.
    • Anaerobic bacteria grow in the absence of oxygen.
    • Facultative anaerobes can grow in the presence or absence of oxygen.
    • Microaerophilic bacteria require 5% oxygen.
    • Capnophilic bacteria grow in the presence of 5-10% CO2.
    • Bacteria require moisture, with 80% of bacteria being H2O, and 70% humidity is required in the incubator.
    • Most bacteria that affect humans require an isotonic environment (0.9% saline).
    • The ideal pH range is 7.2-7.6, and the ideal temperature is 35-37°C.

    Anaerobic Organisms

    • Anaerobic organisms are placed in an anaerobic jar with a catalyst.
    • The catalyst pouch is broken, and water is poured inside to activate the catalyst, which depletes O2 from the jar.
    • An indicator, such as Methylene Blue, is used to detect anaerobic conditions.

    Blood Cultures

    • Blood cultures are used to determine the presence and extent of infection, identify the type of organism responsible, and determine the best antibiotic to use.
    • Blood cultures are ordered for patients with suspected bacteremia or fever of unknown origin.
    • Blood is collected using the butterfly method, and a blood culture adapter is used.
    • The aerobic bottle is filled first to prevent air from the tubing of the butterfly from entering the anaerobic bottle.

    Growth Curve

    • The lag phase is the adjustment period to a new medium and environment, during which the bacteria prepare for growth.
    • The log phase is the rapid growth period, during which the number of bacteria doubles in proportion to time.
    • The stationary phase is when nutrients are used up, and there is a balance between cell growth and cell death.
    • The death phase is when bacteria stop multiplying.

    Culture Plates

    • Culture plates are incubated for 18-24 hours before colonies appear.
    • Most plates are incubated for no longer than 48 hours.
    • Different body sites require different inoculation procedures, such as throat swabs for BA plates and urine for BA and MacConkey's plates.

    Media

    • Media can be solid, semi-solid, or liquid.
    • Different types of media include Blood, MacConkey's, and Chocolate.
    • The order of inoculation is supportive, enrichment, differential, and selective.
    • Dehydrated media is weighed out and dissolved in distilled water, then sterilized and poured into plates.
    • Media is stored in the fridge at 2-8°C, and labels must include the type of media, lot number, and expiry date.

    Streaking Plates

    • Streaking plates are used to isolate a single species of bacteria from a mixed population.
    • The three-phase streaking pattern is known as the T-Streak.
    • The process involves dragging a sterile loop or swab across the surface of the agar in a zigzag motion, covering approximately 30% of the plate.
    • The loop is then re-sterilized, and the plate is turned 90 degrees.

    Streaking Procedure

    • Step 1: Drag the sterile loop or swab across the surface of the agar in a zigzag motion, covering approximately 30% of the plate.
    • Step 2: Re-sterilize the loop, and turn the plate 90 degrees.
    • Step 3: Repeat the process, being cautious not to touch the previously streaked sectors.
    • The final section will have the least amount of growth and many isolated colonies.

    Bacterial Physiology

    • Aerobic bacteria require 21% oxygen.
    • Anaerobic bacteria grow in the absence of oxygen.
    • Facultative anaerobes can grow in the presence or absence of oxygen.
    • Microaerophilic bacteria require 5% oxygen.
    • Capnophilic bacteria grow in the presence of 5-10% CO2.
    • Bacteria require moisture, with 80% of bacteria being H2O, and 70% humidity is required in the incubator.
    • Most bacteria that affect humans require an isotonic environment (0.9% saline).
    • The ideal pH range is 7.2-7.6, and the ideal temperature is 35-37°C.

    Anaerobic Organisms

    • Anaerobic organisms are placed in an anaerobic jar with a catalyst.
    • The catalyst pouch is broken, and water is poured inside to activate the catalyst, which depletes O2 from the jar.
    • An indicator, such as Methylene Blue, is used to detect anaerobic conditions.

    Blood Cultures

    • Blood cultures are used to determine the presence and extent of infection, identify the type of organism responsible, and determine the best antibiotic to use.
    • Blood cultures are ordered for patients with suspected bacteremia or fever of unknown origin.
    • Blood is collected using the butterfly method, and a blood culture adapter is used.
    • The aerobic bottle is filled first to prevent air from the tubing of the butterfly from entering the anaerobic bottle.

    Growth Curve

    • The lag phase is the adjustment period to a new medium and environment, during which the bacteria prepare for growth.
    • The log phase is the rapid growth period, during which the number of bacteria doubles in proportion to time.
    • The stationary phase is when nutrients are used up, and there is a balance between cell growth and cell death.
    • The death phase is when bacteria stop multiplying.

    Culture Plates

    • Culture plates are incubated for 18-24 hours before colonies appear.
    • Most plates are incubated for no longer than 48 hours.
    • Different body sites require different inoculation procedures, such as throat swabs for BA plates and urine for BA and MacConkey's plates.

    Media

    • Media can be solid, semi-solid, or liquid.
    • Different types of media include Blood, MacConkey's, and Chocolate.
    • The order of inoculation is supportive, enrichment, differential, and selective.
    • Dehydrated media is weighed out and dissolved in distilled water, then sterilized and poured into plates.
    • Media is stored in the fridge at 2-8°C, and labels must include the type of media, lot number, and expiry date.

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