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What is the primary purpose of inoculating a PEDS bottle in fluid culture?
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?
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?
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?
What type of media is required for Campylobacter species?
What is the purpose of using a gram stain in fluid culture?
What is the purpose of using a gram stain in fluid culture?
Which of the following is NOT an example of anaerobic bacteria?
Which of the following is NOT an example of anaerobic bacteria?
What is the primary purpose of using a BA plate in fluid culture?
What is the primary purpose of using a BA plate in fluid culture?
What type of agar is used for stool cultures?
What type of agar is used for stool cultures?
What is the primary purpose of using differential media in a microbiology lab?
What is the primary purpose of using differential media in a microbiology lab?
What is the correct order of inoculation in a microbiology lab?
What is the correct order of inoculation in a microbiology lab?
What is the purpose of streaking plates in a microbiology lab?
What is the purpose of streaking plates in a microbiology lab?
What is the most common type of media used in a microbiology lab?
What is the most common type of media used in a microbiology lab?
What is the purpose of the three-phase streaking pattern, known as the T-Streak?
What is the purpose of the three-phase streaking pattern, known as the T-Streak?
What is the correct way to inoculate a culture plate from a swab or other source?
What is the correct way to inoculate a culture plate from a swab or other source?
What is the purpose of using a sterile loop or swab in inoculating a culture plate?
What is the purpose of using a sterile loop or swab in inoculating a culture plate?
What is the purpose of Step 1 in the streaking process?
What is the purpose of Step 1 in the streaking process?
What is the primary purpose of performing quality control tests on prepared media?
What is the primary purpose of performing quality control tests on prepared media?
What information should be included on the label of a prepared medium before storage?
What information should be included on the label of a prepared medium before storage?
What is the primary function of enrichment media?
What is the primary function of enrichment media?
Which type of media is used to differentiate between various microorganisms?
Which type of media is used to differentiate between various microorganisms?
What is the purpose of performing stability tests on prepared media?
What is the purpose of performing stability tests on prepared media?
Which of the following is an example of selective media?
Which of the following is an example of selective media?
What is the primary purpose of using supportive media?
What is the primary purpose of using supportive media?
Which of the following media is used for sputum and stool cultures?
Which of the following media is used for sputum and stool cultures?
What type of environment is created in a Microaerophilic Jar?
What type of environment is created in a Microaerophilic Jar?
What is the primary purpose of the Lag Phase in the growth curve of bacteria?
What is the primary purpose of the Lag Phase in the growth curve of bacteria?
What type of specimen has priority when inoculating plates?
What type of specimen has priority when inoculating plates?
What is the purpose of using MacConkey's plate in urine cultures?
What is the purpose of using MacConkey's plate in urine cultures?
What is the primary purpose of incubating culture plates for 18-24 hours?
What is the primary purpose of incubating culture plates for 18-24 hours?
What type of agar is used for throat swabs?
What type of agar is used for throat swabs?
What is the primary purpose of using a calibrated loop in urine cultures?
What is the primary purpose of using a calibrated loop in urine cultures?
What is the primary purpose of using a Candle Jar?
What is the primary purpose of using a Candle Jar?
What is the primary purpose of using Amies Transport Media with charcoal?
What is the primary purpose of using Amies Transport Media with charcoal?
What type of specimen requires a sterile container for collection?
What type of specimen requires a sterile container for collection?
What is the purpose of using Cary Blair Transport Medium?
What is the purpose of using Cary Blair Transport Medium?
Which of the following specimens typically has normal flora?
Which of the following specimens typically has normal flora?
What should be written on the requisition during specimen collection?
What should be written on the requisition during specimen collection?
What should be avoided when collecting a specimen for C&S?
What should be avoided when collecting a specimen for C&S?
What is the purpose of using Amies Transport Media?
What is the purpose of using Amies Transport Media?
Which of the following specimens is considered sterile?
Which of the following specimens is considered sterile?
Facultative anaerobes can only grow in the presence of oxygen.
Facultative anaerobes can only grow in the presence of oxygen.
Bacteria require 50% humidity to grow.
Bacteria require 50% humidity to grow.
The anaerobic jar is used to create an environment with 5% oxygen.
The anaerobic jar is used to create an environment with 5% oxygen.
The pH range for bacterial growth is between 6.0 and 8.0.
The pH range for bacterial growth is between 6.0 and 8.0.
Bacteria require 90% water to grow.
Bacteria require 90% water to grow.
The incubation temperature for bacterial growth is between 30-40°C.
The incubation temperature for bacterial growth is between 30-40°C.
Capnophilic bacteria grow in the presence of 10-15% CO2.
Capnophilic bacteria grow in the presence of 10-15% CO2.
Methylene Blue is used as an indicator to detect aerobic environments.
Methylene Blue is used as an indicator to detect aerobic environments.
The primary purpose of using enrichment media is to selectively isolate microorganisms that require specific nutrients.
The primary purpose of using enrichment media is to selectively isolate microorganisms that require specific nutrients.
Chocolate agar is a type of selective media used for the growth of fastidious microorganisms.
Chocolate agar is a type of selective media used for the growth of fastidious microorganisms.
The lag phase of the bacterial growth curve is the period of rapid cell division.
The lag phase of the bacterial growth curve is the period of rapid cell division.
A microaerophilic environment is one that is completely devoid of oxygen.
A microaerophilic environment is one that is completely devoid of oxygen.
The correct order of inoculation is supportive, enrichment, differential, and then selective media.
The correct order of inoculation is supportive, enrichment, differential, and then selective media.
Incubation of culture plates at 37°C is necessary for the growth of all microorganisms.
Incubation of culture plates at 37°C is necessary for the growth of all microorganisms.
The streaking process is used to isolate microorganisms from a mixed culture.
The streaking process is used to isolate microorganisms from a mixed culture.
MacConkey's agar is a type of supportive media used for the growth of all microorganisms.
MacConkey's agar is a type of supportive media used for the growth of all microorganisms.
A Microaerophilic Jar creates a 10% oxygen environment.
A Microaerophilic Jar creates a 10% oxygen environment.
The Log phase of a growth curve is a period of slow growth for bacteria.
The Log phase of a growth curve is a period of slow growth for bacteria.
Blood cultures must be incubated at 37 degrees C.
Blood cultures must be incubated at 37 degrees C.
Blood cultures are collected using a sterile loop or swab.
Blood cultures are collected using a sterile loop or swab.
Candle Jar is used to create a carbon dioxide enriched environment.
Candle Jar is used to create a carbon dioxide enriched environment.
Patients weighing more than 27 kg must have 10 mL of blood drawn from two different venipunctures or sites within one hour.
Patients weighing more than 27 kg must have 10 mL of blood drawn from two different venipunctures or sites within one hour.
The Stationary phase of a growth curve is a period of rapid growth for bacteria.
The Stationary phase of a growth curve is a period of rapid growth for bacteria.
Each blood culture bottle can hold up to 20 mL of blood.
Each blood culture bottle can hold up to 20 mL of blood.
Culture plates are typically incubated for 48 hours or more before colonies appear.
Culture plates are typically incubated for 48 hours or more before colonies appear.
Blood cultures are collected in a microaerophilic jar with 20% O2.
Blood cultures are collected in a microaerophilic jar with 20% O2.
Throat swabs are inoculated onto MacConkey's plate.
Throat swabs are inoculated onto MacConkey's plate.
Aerobic bottles are filled before anaerobic bottles when collecting blood cultures.
Aerobic bottles are filled before anaerobic bottles when collecting blood cultures.
Urine cultures are inoculated using a calibrated loop of 1 ml.
Urine cultures are inoculated using a calibrated loop of 1 ml.
Blood cultures can be collected from a single venipuncture site.
Blood cultures can be collected from a single venipuncture site.
Surgical specimens have a low priority when inoculating plates.
Surgical specimens have a low priority when inoculating plates.
The total blood collection for a blood culture should be at least 20 mL.
The total blood collection for a blood culture should be at least 20 mL.
MacConkey Media is used to grow anaerobic bacteria.
MacConkey Media is used to grow anaerobic bacteria.
A medium with a pH value outside the specified range should be discarded.
A medium with a pH value outside the specified range should be discarded.
All media must be stored at room temperature.
All media must be stored at room temperature.
A representative sample of each lot/batch of medium should be incubated for 7-10 days during sterility testing.
A representative sample of each lot/batch of medium should be incubated for 7-10 days during sterility testing.
The thermometer used in preparing media should have a range of -20°C to 150°C.
The thermometer used in preparing media should have a range of -20°C to 150°C.
The label on a prepared medium should include only the type of media and lot number.
The label on a prepared medium should include only the type of media and lot number.
Agar is not a necessary component of MacConkey Media.
Agar is not a necessary component of MacConkey Media.
The purpose of pre-incubation is to check for contamination and quality control.
The purpose of pre-incubation is to check for contamination and quality control.
Match the following types of bacteria with their oxygen requirements:
Match the following types of bacteria with their oxygen requirements:
Match the following growth conditions with their required humidity levels:
Match the following growth conditions with their required humidity levels:
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Match the following specimen collection requirements with the corresponding volume:
Match the following growth conditions with the relevant information:
Match the following growth conditions with the relevant information:
Match the following growth environments with their respective characteristics:
Match the following growth environments with their respective characteristics:
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Match the following incubation conditions with the relevant information:
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Match the following growth environments with their respective characteristics:
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Match the following media preparation steps with their corresponding descriptions:
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Match the growth phase of bacteria with its description:
Match the type of environment with its characteristics:
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Match the type of specimen with the priority order for inoculation:
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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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