Lab Practical Test Questions – Flashcards

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question
What is liquid broth media used for?
answer

enrich sample

cultivate organisms present in low numbers

question
What is a general purpose media?
answer

Supports growth of broad range of organisms

Tryptic soy agar (TSA)

Nutrient Agar

Not selective or differentiating

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What is enriched media?
answer

Contains specific nutrients for the growth of particular species

 

SBA-TSA with 5% sheep RBC added (used for general isolation of organisms directly from specimens)

 

Chocolate agar- hemolyzed RBCs and supplements

question
What is cultured on chocolate agar?
answer

Chocolate agar supports fastidious organisms

N. gonorrheae and H. influenzae

contains hemolyzed RBCs plus supplements

question
What is selective media?
answer

Contains additives that enhance growth of some organisms while inhibiting others

 

used for contaminated samples

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What is differential media?
answer

aids in presumptive ID or grouping of organisms based on the appearance of colonies

 

 

question
What is SBA used for?
answer
demonstration of hemolytic properties (esp. streptococci)
question
What are the types of hemolysis?
answer

beta hemolysis: complete clearing of SBA

Streptolysn O: oxygen labile

Streptolysin S: oxygen stable

 

alpha hemolysis: incomplete lysing of RBCs leads to

greenish discoloration around colonies

 

gamma: non-hemolytic

question
What is MacConkey Agar used for?
answer

Differential AND Selective 

  • Selective: inhibits gram positive organisms
  • Differential: lactose plus pH color changing indicator
  • Lactase fermentation turns colonies magenta
  • non-lactose fermenters stay clear
question
What is Eosin-Methylene Blue (EMB) used for?
answer
  • Selective-eosin and methylene dyes inhibit Gram + organisms
  • Differential: lactose fermenters turn dark due to acid production
  • E. coli colonies turn green w/sheen because of high amounts of acid produced
  • Lactose non-fermentors are colorless
question
What is Columbia CNA Agar used for?
answer
  • Selective only: selects for G+ bacteria
  • colistin and nalidixic acid inhibit G - bacteria
question
What is Mannitol Salt Agar (MSA) used for?
answer
  • Screens for S. aureus
  • Differential: mannitol fermentation produces yellow discoloration (S. aureus)
  • Selective: high salt concentration selects for organisms that grow in presence of salt (S. aureus)
question
What genera constitute most clinically isolated aerobic G+ cocci?
answer

Staphylococcus

Streptococcus

Enterococcus

question

What are the characteristics of Staphylococcus?

Aerobe vs. anaerobe

Gram stain

morphology

catalase test

salt growth?

answer

facultative anaerobe

Gram +

Cocci in irregular clusters

catalase +

grow in high salt media (MSA)

question
What test is used to differentiate groups in the genus Staphylococcus?
answer

Coagulase test

 

Coagulase positive=S. aureus group

 

Coagulase negative=S. epidermidis and S. saprophyticus groups

question
How is the S. epidermidis CoNS differentiated from S. saprophyticus CoNS?
answer

S. epidermidis is Novobiocin SUSCEPTIBLE

 

S. saprophyticus is Novobiocin RESISTANT

question
What regions of the body are colonized by different Staph groups?
answer

S. aureus mainly on anterior nares

 

S. epidermidis mainly colonized skin

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What are the characteristics of S. aureus colonies
answer

mostly hemolytic

mod. large

golden yellow/white

question
What is useful about the catalase reaction test?
answer

Streptococci are catalase -

Staphylococci are catalase +

question
What is the composition of the Staphylococcus latex agglutination test?
answer

Latex beads coated with fibrinogen and Ab to Protein A (component of S. aureus cell wall)

 

Fibrinogen detects S. aureus coagulase

Ab detect protein A in S. aureus

question
How is novobiocin resistance/susceptibility testing performed?
answer

Streak colony on SBA

place 5 ug novobiocin disc on media

resistance indicated by zone of inhibition less than or equal to 16mm

question
What are some general characteristics of family Enterobacteriaceae?
answer

Enteric organisms

Gram - bacilli

grow well on MAC

Biochemically active

ferments glucose and other sugars (sometime with gas)

catalase +

Oxidase -

reduce nitrate->nitrite

question
What genera of family Enterobacteriaceae account for most extraintestinal infections?
answer

Eschericia (coli

Klebsiella

Proteus (mirabilis)

Enterobacter

Serratia

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What are the most common intestinal infections due to Enterobacteriaceae?
answer

E. coli 

 

Salmonella

Shigella

 

Y. enterocolitica (not covered)

question
What toxins are produced by E. coli O157:H7?
answer

Stx1

Stx2

 

also known as verocytotoxins

question
What are some biochemical characteristics of O157:H7?
answer

ferment lactose

do NOT ferment sorbitol overnight

SMAC colonies are colorless

 

Indole + (most E. coli are)

question

What reaction is responsible for indole test?

How do you know if it is indole +?

answer

metabolic degradation of tryptophan

 

Tryptophanase breaks down tryptophan->indole

blue green color is +

question
What is the MUG test and what is it used for? How do you know if a sample is MUG +?
answer
  • Used to differentiate other E. coli from O157:H7
  • beta-D-glucuronide enzyme in E. coli degrades MUG and produces fluorescence when incubated overnight

 

O157:H7 isolates are MUG-

Other E. coli are MUG+

question
How is salmonella serotyping performed?
answer

Somatic O antigen (cell wall antigen-outer portion of LPS)

 

H antigen (flagella)

question
What is used for serogrouping vs serotyping? how is it reported?
answer
  • Serogrouped based on O antigens
  • Serotyped based on flagella H antigens
  • reported by genus and serotype to state for tracking outbreaks
question
What are the key biochemical reactions defining the Salmonella genus?
answer

Non-lactose fermenter (clear on MAC)

Ferments glucose WITH production of H2S gas

Agglutination of antisera for Salmonella O groups

question
What does a TSI slant look like when inoculated with Salmonella? A urease tube?
answer

TSI: blackening and gas bubble in the deep (H2S  production)

 

No change in slant color (non-fermenter)

 

Urease - (no pink color)

question
What are the 4 serogroups of Shigella sp.?
answer
A-D
question

What are the key biochemical reactions of Shigella?

Lactose fermentation

other Fermentation

Motility

Gas production

Agglutination

answer

Non-lactose fermenter

ferments GLUCOSE without gas

non-motile

biochemically inactive

H2S negative

Agglutinates with grouping antisera for Shigella

question

What does a TSI slant look like when inoculated with Shigella?

 

Urease slant?

answer

Red in the slant (oxidation of peptones)

Yellow in the deep (production of acid due to glucose fermentation)

 

Urease -

question
What media are used to culture and isolate Shigella and Salmonella?
answer

enrichment broth (maximal recovery)

 

selective media (MAC, HK)

 

biochemical screening

 

serogrouping/(serotyping for Salmonella)

question
What do Salmonella and Shigella look like on MAC?
answer

Gram -

lactose negative=clear colonies on MAC (no acid)

question
What is Hektoen Enteric Agar? What does it select for/differentiate?
answer

Selects for enteric pathogens

 

contains lactose, sucrose and salicin + pH indicator

Fermenters produce yellow colonies

non-fermenters are green/transparent

 

Na thiosulfate and ferric ammonium citrate indicate H2S production through black center of colony

question

What do Salmonella and Shigella look like on HK agar?

What about E. coli?

answer

Salmonella: clear/green with black center

Shigella: clear green

E. coli (non-O157:H7): yellow

question
What are the contents of TSI agar?
answer

Glucose, lactose, and sucros

phenol red pH indicator

question
How do you read TSI?
answer

Reaction in the deep: anaerobic fermentation (yellow if acid production due to fermentation)

Reaction on slant: yellow if glucose and lactose/sucrose fermenter, reverts to red if oxidation of peptones causes pH increase

Blackening: H2S production

Gas production: lifting or cracking of media

question
How do you read a Urease test?
answer

Positive:media turns hot pink if urea->ammonia+CO

Negative: no color change, yellow

question
Which Strep are beta, alpha, and gamma hemolytic?
answer

Beta: S. pyogenes (GAS) and S. agalactiae (GBS)

Alpha: S. pneumoniae and "viridans" strep

Gamma: S. bovis

question

What are the key characteristics of Streptococci?

Gram

Colony morphology

Catalase

an/aerobic

 

answer

Gram positive cocci, spherical/ovoid

usually in pairs or chains

Grey/white translucent colonies

Catalase -

Facultative anaerobes

 

question
How are Beta hemolytic strep grouped?
answer

Lancefield System

Detects cell wall polysaccharides A,B,C,F,G

Cell wall lipteichoic acids (D and Enterococci)

question
How is Lancefield grouping performed?
answer

Extract antigens enzymatically by incubating for 10min-1hr

 

use latex agglutination to identify via homologous antisera

question
What clinical syndromes are caused by Group A Strep?
answer

S. pyogenes

pharyngitis, impetigo

cellulitis/necrotizing fasciitis

endocarditis, sepsis

scarlet vever

streptococcal TSS

Sequelae: ARF, glomerulonephritis

question
What is the PYR test? What is it used for?
answer

Determines the activity of prrolidonyl aminopeptidase enzyme

 

Presumptive ID of GAS and Enterococci

question
How do you isolate GAS?
answer

Blood agar with sulfamethoxazole trimethoprom (STX) to inhibit normal flora

 

Select colonies with beta hemolysis

question
What are the biochemical findings that indicate GAS?
answer

Catalase -

 

PYR +

question
What species of Streptococcus is GBS? What clinical significant syndromes does it cause?
answer

LOS NICU

Transmission from Mother to child during birth

Sepsis, amnionitis, UTI in pregnant women

question
How is GBS isolated and identified?
answer

Colony morphology on SBA: narrow B hemolysis (some are non-hemolytic)

 

Lancefield grouping (positive for B antigen)

 

CAMP +

 

question
How does the CAMP test work? What does it identify?
answer

Streak S. aureus down middle of SBA plate

Streak Streptococcus perpendiculart to it

 

Look for synergistic hemolysis between GBS and S. aureus

characterized by zone of clearer media around points of intersection 

question
What species comprise the alpha hemolytic strep?
answer

Streptococcus pneumoniae

"viridans" strep

question
What are the biochemical characteristics of S. pneumo?
answer

Optochin sensitive

Bile sensitive
Quellung reaction positive 

question

What syndromes does S. pneumoniae cause?

answer

**Community acquires pneumonia

Otitis media, sinusitis

meningitis, peritonitis, arthritis

Oropharyngeal carriage common

question
What is the colony morphology of S. pneumoniae?
answer

Large zone of alpha hemolysis

colony encapsulation gives "mucoid" wet appearance (can decrease over time)

Prolonged incubation can cause center of colonies to collapse

question
What is the quellung reaction?
answer
Anticapsular antibodies applied to S. pneumo cause capsule to swell and become more conspicuous under microscopy
question
What special cellular morphology identifies S. pneumo vs. other strep species?
answer

Pairs of coccobacilli

non-staining halo indicates capsule

question
What does the Optochin test allow you to do?
answer

Presumptive ID of S. pneumo

 

Disk with optochin placed on media plated with colony

 

Zone of inhibition >14mm = presumptive ID of S. pneumo

question
How is the bile solubility test performed?
answer

Addition of 10% Na desoxycholate (bile salt)

 

activates lytic enzymes in S. pneumo and causes colonies to disappear

question
What are the biochemical characteristics of viridans sp. strep?
answer

alpha or gamma hemolytic

bile esculin and optochin -

question
what are the biochemical characteristics of Serogroup C and G strep? Group D?
answer

C and G: beta hemolytic, bile esculin and optochin -

 

D: gamna hemolytic, Bile Esculin + optochin -

question

What characteristics describe Enterococci?

 

Gram/morphology

hemolysis

catalase

lancefield group

pyr

bile esculin

salt tolerance

answer

Gram + cocci (single, pair or short chain)

Gamma hemolytic

Catalase -

Lancefield group D (but not GDS)

facultative anaerobe

PYR+

Bile Esculin and Salt tolerant

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