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Microbiology Lab Practical
Terms in this set (225)
What does MAC stand for?
Isolation of Gram - enterics
What is the purpose of MacConkey's Agar?
Differential and Selective
What is the category of MacConkey's Agar?
What is the differential agent of MacConkey's Agar?
Bile Salt & Crystal Violet
What are the selective agents of MacConkey's Agar?
What is the pH indicator for MacConkey's Agar?
What is the inoculation of MacConkey's Agar?
Lactose fermentation +
What is the reading criteria of MacConkey's Agar when it is purple?
Lactose fermentation -
What is the reading criteria of MacConkey's Agar when it has colorless growth?
Mannitol Salt Agar
What does MSA stand for?
Isolate and differentiate Halotolerant species
What is the purpose of Mannitol Salt Agar?
Differential and Selective
What is the category of Mannitol Salt Agar?
What is the differential agent in Mannitol Salt Agar?
What is the selective agent in Mannitol Salt Agar?
What is the pH indicator for Mannitol Salt Agar?
What is the inoculation of Mannitol Salt Agar?
Mannitol Fermentation +
What is the reading criteria of Mannitol Salt Agar when it is lemmon yellow?
Mannitol Fermentation -
What is the reading criteria of Mannitol Salt Agar when it is pink?
What is the reading criteria of Mannitol Salt Agar when there is NO growth?
In the Mannitol Salt Agar, Staphylococcus Epidermidis is a __________________ __________________.
Sulfide Indole Motility
What does SIM stand for?
Screening for H2S, Indole production, and motility
What is the purpose of the Sulfide Indole Motility test?
What is the category of Sulfide Indole Motility test?
What is the reagent used to test for indole in the Sulfide Indole Motility test?
What is the reagent used to test for H2S production in the Sulfide Indole Motility test?
What is the inoculation method of Sulfide Indole Motility?
What is the reading criteria of the Sulfide Indole Motility test when there is black precipitate?
What is the reading criteria of the Sulfide Indole Motility test when there it is red?
What is the reading criteria of the Sulfide Indole Motility test when there is cloudiness throughout?
What does UB stand for?
Production of exoenzyme urease
What is the purpose of Urea Broth?
What is the category of Urea Broth?
What is the subtrate of Urea Broth?
What is the pH indicator of Urea Broth?
Dip & Swirl
What is the inoculation of Urea Broth?
What is the reading criteria of Urea Broth when it is orange?
What is the reading criteria of Urea Broth when it is Fuchsia?
What does BA stand for?
Test for hemolytic activity
What is the purpose of the blood agar?
Destruction of red blood cells
What does hemolytic mean?
What is the category of Blood Agar?
What is the inoculation of Blood Agar?
Autoclaved, cooled; blood then added
What is the preparation of Blood Agar?
What is the reading criteria of Blood Agar when there is a clear zone viewed from the bottom?
What is the reading criteria of Blood Agar when there is green/brown discoloration from the top?
What is the reading criteria of Blood Agar when there is no color change?
Trypticase Soy Agar
What does TSA stand for?
Growth of wide range of bacteria
What is the purpose of Trypticase Soy Agar?
What is the category of Trypticase Soy Agar?
What is the inoculation technique of Trypticase Soy Agar?
Bile Esculin Agar
What does BESC stand for?
Isolation of Enterococcus
What is the purpose of Bile Esculin Agar?
Selective and Differential
What is the category of Bile Esculin Agar?
What is the selective agent of Bile Esculin Agar?
What is the differential agent of Bile Esculin Agar?
What is the inoculation of Bile Esculin Agar?
What is the reading criteria of Bile Esculin Agar when there is black?
What is the reading criteria of Bile Esculin Agar when there is NOT black?
Eosin Methylene Blue Agar
What does EMB stand for?
Isolation of Gram - enterics
What is the purpose of Eosin Methylene Blue Agar?
Selective and Differential
What is the category of Eosin Methylene Blue Agar?
Eosin and Methylene Blue
What is the Selective Agent of Eosin Methylene Blue Agar?
What is the differential agent of Eosin Methylene Blue Agar?
What is the inoculation of Eosin Methylene Blue Agar?
What is the reading for Eosin Methylene Blue Agar when there is metal green sheen, black, or pink mucoid?
What is the reading criteria for Eosin Methylene Blue Agar when it is NOT metal green sheen, black, or pink mucoid?
Lauryl Sulfate Broth
What does LSB stand for?
Detect or ID coliforms in food
What is the purpose of Lauryl Sulfate Broth?
Selective and Differential
What is the category of Lauryl Sulfate Broth?
Sodium Lauryl Sulfate
What is the selective agent in Lauryl Sulfate Broth?
What is the differential agent in Lauryl Sulfate Broth?
What is the selective property of Lauryl Sulfate Broth?
Small test tube that detects gas production
What is a Durham Tube?
Dip & Swirl
What is the inoculation technique of Lauryl Sulfate Broth?
What is the reading criteria of Lauryl Sulfate Broth when there is a gas bubble in the Durham tube?
What is the reading criteria of Lauryl Sulfate Broth when there is NO gas bubble in the Durham tube?
Pseudomonas Isolation Agar
What does PA stand for?
Isolation of Pseudomonas species
What is the purpose of the Pseudomonas Isolation Agar?
What is the category of Pseudomonas Isolation Agar?
What is the inoculation of Pseudomonas Isolation Agar?
What is the reading criteria of Pseudomonas Isolation Agar when there is growth on the streaked line?
What is the reading criteria of Pseudomonas Isolation Agar when there is no growth?
Glucose, Adonitol, Lactose, Arabinose, Soribitol
Which tests are associated with sugar fermentation in the EnteroPluri Test?
When reading GLU, ADN, LAC, ARAB, and SORB, what does the color RED represent?
When reading GLU, ADN, LAC, ARAB, and SORB, what does the color YELLOW represent?
Loss of Carbon Dioxide
What is Decarboxylation?
Complete separation of wax
What is gas (GAS) production from the GLU test?
Which color indicates decarboxylation?
Lysine and Ornithine Tests
Which tests are associated with Amino Acid Decarboxylase?
What is the end product of Lysine?
What is the end product of Ornithine?
In the EnteroPluri test, Hydrogen Sulfide (H2S) production is what color?
In the EnteroPluri test, Indole (IND) production is what color?
What reagent is used in IND test in the EnteroPluri test?
What does VP stand for in the EnteroPluri test?
What does the color red indicate in the Voges Proskauer test?
Alpha-naphthol and KOH
What are the reagents in the Voges Proskauer (VP) test?
Phenylalanine Test (PA)
Which test in the EnteroPluri is associated with Amino Acid Deaminase?
Removal of amine to form pyruvic acid
What is deamination?
Black to Smoky Gray
In the Phenylalanine (PA) Test in the EnteroPluri, what color indicates deamination?
In Dulcitol Fermentation (DUL), what does the color green represent?
In Dulcitol (DUL) Fermentation, what does the color yellow represent?
Urease has been used to produce CO2 and ammonia
What does the color fuchsia indicate in the Urea (UREA) hydrolysis?
Citrate has been utilized as a carbon source
What does blue indicate in the Citrate Utilization test?
Catalase and Coagulase
What are the two rapid screening tests for Staphylococcus?
Which rapid screening test differentiates Staphylococcus from Streptococcus?
What is the reagent for Catalase test?
What does bubbling indicate in the Catalase test?
What does it mean to be Catalase (+)?
What does it mean to be Catalase (-)?
Which rapid screening test differentiates Staphylococcus aureus from other Staph species?
What reagent is used in the Coagulase test?
What does clumping once represent in the Coagulase test?
If the Coagulase test is Coagulase +, which Staphylococcus species is it?
Semiquantitative Method using the volumetric loop
What inoculation method is used in the Urine Lab?
What is the number of CFUs needed to be considered a countable plate?
Colony Forming Units
What does CFU stand for?
What is the CFU reading of a UTI?
OCD = CFUs/loop volume
What is the formula for original cell density (OCD)?
Neutrophils, Lymphocytes, Monocytes, Eosinophils, Basophils
What is the order of leukocytes in normal blood?
Which leukocyte is associated with Bacteria?
Which leukocyte is associated with Viruses?
Which leukocyte is nonspecific and includes many types of infections?
Which leukocyte is associated with fungi, protozoa, helminthic infection, and allergies
Which leukocyte is associated with allergies?
Methylene Blue Reductase Test
What test is used to test for milk quality?
Oxidation & Reduction
What is the principle of the Methylene Blue Reductase Test?
What is the quality of milk when it changes from blue to white in under 2 hours?
What is the quality of milk when it changes from blue to white after 6 hours?
BESC and LSB
Which tests indicate fecal contamination from the food lab?
Which test indicates handling contamination from the food lab?
Which test indicates spoilage contamination from the food lab?
Live bacteria absorb new DNA and express new gene traits
What is transformation?
GFP and AMPr gene
Which two genes are carried on the plasmid?
Green fluorescent protein
In the transformation experiment, green fluorescent protein gene causes the expression of ______ _________ ___________.
In the transformation experiment, the AMPr gene causes production of ____________.
Ampicillin is a _______ __________ used to isolate transformed cells.
Transduction and Conjugation
What are two methods (other than Transformation) that bacteria can acquire new genes?
Prove bacteria were alive and adding plasmid did not harm
What was the purpose of the plasmid (-) and plasmid (+) on the TSA plate?
Penicillinase diffuses into medium and white bacteria that weren't transformed are able to survive
What causes the "halo effect" in the plasmid (+) TSA+AMP plate?
Screening of HIV
What is the clinical significance of indirect ELISA?
What is indirect ELISA used to detect?
What is direct ELISA used to detect?
What is another diagnostic test used to antibodies?
Which blood types can O- receive?
Which blood types can O+ receive?
Which blood types can A- receive?
A+, A-, O+, O-
Which blood types can A+ receive?
Which blood types can B- receive?
B+, B-, O+, O-
Which blood types can B+ receive?
A-, B-, AB-, O-
Which blood types can AB- receive?
Which blood types can AB+ receive?
In the Ultraviolet Radiation lab, UV light is a _________.
What forms as a result of UV radiation?
Covalently bonded thymine bases that are adjacent on the DNA strand
What are thymine dimers?
Which enzyme is involved in light repair?
Endonuclease, DNA polymerase, DNA Ligase
Which enzymes are involved in dark repair?
Which inoculation technique was used in the UV radiation experiment?
1.0mL in 9.0mL = 10^-1
0.1mL in 9.9mL = 10^-2
What are the numbers for serial dilution?
In serial dilutions, you _________ exponents are you move from one tube to the next and dilution factors are always ____________.
OCD = CFUs/Final Dilution Factor
What is the formula for Original Cell Density in the water contamination lab?
5.6 x 10^6
What is the original cell density when:
CFU = 56
Final Dilution Factor = 10^-5
Demonstrate how quickly a pathogen can spread through contact with body fluid
What was the purpose of the body fluid lab?
What is the formula for exponential spread of pathogens?
What was the inoculation method used in the urine lab?
What are zones of inhibition measured with?
Bacteria are resistant and antibiotic should not be prescribed
If any of the colonies grow within the zone of inhibition, what does this mean?
Which antibiotic is associated with cell wall synthesis?
Which antibiotic is associated with nucleic acid synthesis?
Which antibiotic is associated with protein synthesis?
Which antibiotic is associated with folic acid synthesis?
Which microbe is Gram-positive (although with age, some have the potential to become Gram-negative) aerobic, streptobacilli, produce endospores; large nonpathogenic bacillus used in cytological studies because its size makes it easy to study under the light microscope; produces casease, which breaks down casein-the protein that gives milk its white color?
Which microbe is a Gram-negative rod; an opportunist bacterial pathogen that can lead to different nosocomial infections such as respiratory, gastrointestinal, wound, blood, and central nervous infections; found in the human gastrointestinal tract and generally does not cause disease in a healthy individual?
Which microbe is Gram-positive coccus; an opportunist that is a common resident of the digestive tract; enterococcal infection is often associated with bacteremia which can lead to endocarditis; can also cause cystitis and wound infections; Vancomycin Resistant Enterococci (VRE) are one of most important nosocomial pathogens; individuals who are at risk are those who have urinary or intravascular catheterization or long-term hospitalization with broad-spectrum antimicrobials?
Which microbe is Gram-negative rod; belongs to the coliform group; common resident of the digestive tract - produces vitamins K and B-complex; "indicator bacterium", the presence of which indicates fecal contamination; opportunist: usual cause of Urinary Tract Infections, also Montezuma's revenge, food poisoning; strain 0157:H7 enterohemorrhagic that produces a toxin; Lab media: EMB & VRB -selective and differential medium; LSB - ID coliforms?
Which microbe is Gram-positive; aerobic; cocci found in pairs or clusters; appears yellow in culture; nonpathogenic; grows well at 25°C?
Which microbe is Gram-negative rod; lives in soil and water; organic matter decomposer, highly motile; swarming colonies on agar plate cultures; opportunist of the UG tract, nosocomial infections; resistant to ampicillin and cephalosporin but susceptible to Trimethoprim/Sulfa compounds?
Which microbe is Gram-negative rod; aerobe; motile; fruity odor and a blue-green pigment; opportunistic pathogen with a phagocyte-resistant slime layer; nosocomial; can infect any part of the body - skin, ear, eye, lungs, blood, heart valves, CNS, GI tract, bones and joints?
Which microbe is Gram-negative rod, peritrichous flagella; zoonotic especially in poultry and swine; Foodborne infections linked to raw meat, poultry, eggs and egg products, milk and dairy products; causing fever, headache, vomiting, diarrhea - may be life-threatening in the very young and old or immunocompromised?
Which microbe is Gram-negative motile rod; ubiquitous - soil and water; commonly found growing in bathrooms where it looks pink and slimy; nosocomial; opportunist in AIDS victims, pneumonia in alcoholics, burn and wound infections; resistant to antibiotics; Lab - at 25 C red pigmented, DNAse, lipase, gelatinase producer?
Which microbe is Gram-positive, cocci in grape-like clusters; salt tolerant?
Which microbe is wide-spread, assoc. with skin and mucus membranes; most resistant of the non-spore formers; drug resistance common, MRSA; VRSA; many virulence factors; food poisoning, toxic shock, scalded skin syndrome, pinkeye, boils, etc.; most common nosocomial bacterial infection; Lab: MSA - selective and differential; positive coagulase test?
Which microbe is on the skin; opportunistic; most strains produce a slime layer which may enable them to attach to certain hospital apparatuses used in invasive procedures; infections at the site of prosthetic implantation; multiple antibiotic resistance and debilitated patient =severe and frequently fatal infection; MSA - selective for the organism?
Which microbe is opportunistic; a leading cause of urinary tract infections; young women are more susceptible to colonization; hydrolyzes urea to produce ammonia allowing for its growth in an acidic urine environment; spread through sexual intercourse?
Agar, not gram reaction
Why is MacConkey's Agar purple?
Endospores are resistant to UV light
Why did B. megaterium survive exposure to UV light?
Light repair - more CFUs in areas exposed to UV light.
Was there more DNA repair using S. marcescens with incubation in the light or the dark and why?
Ensure bacteria without plasmid do not produce penicillinase
What is the purpose of including ampicillin in the medium with the (-) plasmid treatment?
Isolate transformed cells
What is the purpose of including ampicillin in the medium with the (+) plasmid treatment?
What is an example of a disease that attacks the human immune system?
Ensure reagents are working
Why do you need to assay positive and negative controls as well as your experimental samples in the indirect ELISA?
1. Gram-negative rod
2. Facultative anaerobe
4. Glucose fermenting
What are the four qualifiers for enteric?
Isolate gram-Negative enterics that can be commonly found in fecal cultures
What is the value of using MacConkey Agar in fecal cultures?
What is the biological principle associated with the UREA test in the EnteroPluri?
What is the end product of the Citrate test in the EnteroPluri?
What are three ways to avoid sexually transmitted pathogens?
What is the quickest way for a pathogen to spread through any given population?
Only those who eat the food would be infected, causing sporadic clusters & uneven distribution
How would the dynamics be different if we would be investigating the outbreak associated with a foodborne pathogen?
72C for 15 seconds
At what temperature and for how long is refrigerated milk pasteurized?
Lower temperature and less time than autoclaving; milk will curdle
How is pasteurization different from autoclaving? Why isn't milk autoclaved?
Salmonella, Norovirus, Campylobacter
What are the most common pathogens associated with foodborne illness?
Infection is slow onset fever, but intoxication is rapid onset fever
What is the difference between food borne infection and intoxication?
E. coli = diarrhea, dysentery
O157:H7 = hemorrhagic colitis
What are the symptom of intestinal infection caused by E. coli and how is it different from O157:H7?
How does one treat water when a "boil water advisory" is issued?
Flush pipes and run with cold water for 5 minutes
Once the boil water advisory is over, what advice does the CDC give?
Vibrio cholerae, Giardia, Camplyobacter
Name three waterborne infectious agents other than E. coli.
Staphylococcus saprophyticus and Proteus vulgaris
Which alkalinophilic bacteria are usually associated with UTIs?
What is the most common pathogen associated with UTIs?
What test is done in the clinical setting that is similar to the Kirby-Bauer method?
Cheapest, allergies, pregnant, patient history
If mroe than one antimicrobial demonstrated an S result, which one would you choose and why?
What is the biochemical principle of the indole test?
What color indicates a positive reading in the Lysine and Ornithine test?
What color indicates a negative reading in the Lysine and Ornithine test?
What color indicates a positive reading in the Urea test?
Opportunistic pathogen that causes infections in hosts that are compromised
Why are staphylococci among the leading cause of nosocomial infections?
Exotoxin that destroys red blood cells to release nutrients and growth factors that increase staphylococcus aureus multiplication
What is the role of alpha toxin in the pathogenesis of Staphylococcus aureus?
1. Keep refrigerated or frozen.
2. Thaw in refrigerator or microwave.
3. Keep raw meat and poultry from other foods
What are some safe handling instructions?
Colonies inside pattern undergone DNA repair, but most cells were killed
What were the results from Serratia marcescens with lid off, exposed to UV radiation for 30 seconds and Incubation for 24 hours in the dark at 25⁰C?
Partial to slight repair
What were the results from S. marcescens uncovered for duration of 30 seconds to 3 minutes exposed to UV light & incubated in light?
More repair than light
What were the results from S. marcescens uncovered, exposed to UV light for duration of 30 seconds incubated in the dark?
What were the results from S. marcescens covered with lid, exposed to UV light for 3 minutes?
5 parallel lines
What was the inoculation of media for the food lab for plates?
Kirby-Bauer and the UV light exposure are both examples of _________ _________.
Spread Plate Method
What method was used in the water contamination and transformation experiment?
Increased pH, growth, and tissue damage resulting in formation of stones
What role does urease play in an UTI?
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