Lab Practical Test Questions – Flashcards
Unlock all answers in this set
Unlock answersWhat is liquid broth media used for? |
enrich sample cultivate organisms present in low numbers |
What is a general purpose media? |
Supports growth of broad range of organisms Tryptic soy agar (TSA) Nutrient Agar Not selective or differentiating |
What is enriched media? |
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 |
What is cultured on chocolate agar? |
Chocolate agar supports fastidious organisms N. gonorrheae and H. influenzae contains hemolyzed RBCs plus supplements |
What is selective media? |
Contains additives that enhance growth of some organisms while inhibiting others
used for contaminated samples |
What is differential media? |
aids in presumptive ID or grouping of organisms based on the appearance of colonies
|
What is SBA used for? |
demonstration of hemolytic properties (esp. streptococci) |
What are the types of hemolysis? |
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 |
What is MacConkey Agar used for? |
Differential AND Selective
|
What is Eosin-Methylene Blue (EMB) used for? |
|
What is Columbia CNA Agar used for? |
|
What is Mannitol Salt Agar (MSA) used for? |
|
What genera constitute most clinically isolated aerobic G+ cocci? |
Staphylococcus Streptococcus Enterococcus |
What are the characteristics of Staphylococcus? Aerobe vs. anaerobe Gram stain morphology catalase test salt growth? |
facultative anaerobe Gram + Cocci in irregular clusters catalase + grow in high salt media (MSA) |
What test is used to differentiate groups in the genus Staphylococcus? |
Coagulase test
Coagulase positive=S. aureus group
Coagulase negative=S. epidermidis and S. saprophyticus groups |
How is the S. epidermidis CoNS differentiated from S. saprophyticus CoNS? |
S. epidermidis is Novobiocin SUSCEPTIBLE
S. saprophyticus is Novobiocin RESISTANT |
What regions of the body are colonized by different Staph groups? |
S. aureus mainly on anterior nares
S. epidermidis mainly colonized skin |
What are the characteristics of S. aureus colonies |
mostly hemolytic mod. large golden yellow/white |
What is useful about the catalase reaction test? |
Streptococci are catalase - Staphylococci are catalase + |
What is the composition of the Staphylococcus latex agglutination test? |
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 |
How is novobiocin resistance/susceptibility testing performed? |
Streak colony on SBA place 5 ug novobiocin disc on media resistance indicated by zone of inhibition less than or equal to 16mm |
What are some general characteristics of family Enterobacteriaceae? |
Enteric organisms Gram - bacilli grow well on MAC Biochemically active ferments glucose and other sugars (sometime with gas) catalase + Oxidase - reduce nitrate->nitrite |
What genera of family Enterobacteriaceae account for most extraintestinal infections? |
Eschericia (coli Klebsiella Proteus (mirabilis) Enterobacter Serratia |
What are the most common intestinal infections due to Enterobacteriaceae? |
E. coli
Salmonella Shigella
Y. enterocolitica (not covered) |
What toxins are produced by E. coli O157:H7? |
Stx1 Stx2
also known as verocytotoxins |
What are some biochemical characteristics of O157:H7? |
ferment lactose do NOT ferment sorbitol overnight SMAC colonies are colorless
Indole + (most E. coli are) |
What reaction is responsible for indole test? How do you know if it is indole +? |
metabolic degradation of tryptophan
Tryptophanase breaks down tryptophan->indole blue green color is + |
What is the MUG test and what is it used for? How do you know if a sample is MUG +? |
O157:H7 isolates are MUG- Other E. coli are MUG+ |
How is salmonella serotyping performed? |
Somatic O antigen (cell wall antigen-outer portion of LPS)
H antigen (flagella) |
What is used for serogrouping vs serotyping? how is it reported? |
|
What are the key biochemical reactions defining the Salmonella genus? |
Non-lactose fermenter (clear on MAC) Ferments glucose WITH production of H2S gas Agglutination of antisera for Salmonella O groups |
What does a TSI slant look like when inoculated with Salmonella? A urease tube? |
TSI: blackening and gas bubble in the deep (H2S production)
No change in slant color (non-fermenter)
Urease - (no pink color) |
What are the 4 serogroups of Shigella sp.? |
A-D |
What are the key biochemical reactions of Shigella? Lactose fermentation other Fermentation Motility Gas production Agglutination |
Non-lactose fermenter ferments GLUCOSE without gas non-motile biochemically inactive H2S negative Agglutinates with grouping antisera for Shigella |
What does a TSI slant look like when inoculated with Shigella?
Urease slant? |
Red in the slant (oxidation of peptones) Yellow in the deep (production of acid due to glucose fermentation)
Urease - |
What media are used to culture and isolate Shigella and Salmonella? |
enrichment broth (maximal recovery)
selective media (MAC, HK)
biochemical screening
serogrouping/(serotyping for Salmonella) |
What do Salmonella and Shigella look like on MAC? |
Gram - lactose negative=clear colonies on MAC (no acid) |
What is Hektoen Enteric Agar? What does it select for/differentiate? |
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 |
What do Salmonella and Shigella look like on HK agar? What about E. coli? |
Salmonella: clear/green with black center Shigella: clear green E. coli (non-O157:H7): yellow |
What are the contents of TSI agar? |
Glucose, lactose, and sucros phenol red pH indicator |
How do you read TSI? |
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 |
How do you read a Urease test? |
Positive:media turns hot pink if urea->ammonia+CO2 Negative: no color change, yellow |
Which Strep are beta, alpha, and gamma hemolytic? |
Beta: S. pyogenes (GAS) and S. agalactiae (GBS) Alpha: S. pneumoniae and "viridans" strep Gamma: S. bovis |
What are the key characteristics of Streptococci? Gram Colony morphology Catalase an/aerobic
|
Gram positive cocci, spherical/ovoid usually in pairs or chains Grey/white translucent colonies Catalase - Facultative anaerobes
|
How are Beta hemolytic strep grouped? |
Lancefield System Detects cell wall polysaccharides A,B,C,F,G Cell wall lipteichoic acids (D and Enterococci) |
How is Lancefield grouping performed? |
Extract antigens enzymatically by incubating for 10min-1hr
use latex agglutination to identify via homologous antisera |
What clinical syndromes are caused by Group A Strep? |
S. pyogenes pharyngitis, impetigo cellulitis/necrotizing fasciitis endocarditis, sepsis scarlet vever streptococcal TSS Sequelae: ARF, glomerulonephritis |
What is the PYR test? What is it used for? |
Determines the activity of prrolidonyl aminopeptidase enzyme
Presumptive ID of GAS and Enterococci |
How do you isolate GAS? |
Blood agar with sulfamethoxazole trimethoprom (STX) to inhibit normal flora
Select colonies with beta hemolysis |
What are the biochemical findings that indicate GAS? |
Catalase -
PYR + |
What species of Streptococcus is GBS? What clinical significant syndromes does it cause? |
LOS NICU Transmission from Mother to child during birth Sepsis, amnionitis, UTI in pregnant women |
How is GBS isolated and identified? |
Colony morphology on SBA: narrow B hemolysis (some are non-hemolytic)
Lancefield grouping (positive for B antigen)
CAMP +
|
How does the CAMP test work? What does it identify? |
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 |
What species comprise the alpha hemolytic strep? |
Streptococcus pneumoniae "viridans" strep |
What are the biochemical characteristics of S. pneumo? |
Optochin sensitive Bile sensitive |
What syndromes does S. pneumoniae cause? |
**Community acquires pneumonia Otitis media, sinusitis meningitis, peritonitis, arthritis Oropharyngeal carriage common |
What is the colony morphology of S. pneumoniae? |
Large zone of alpha hemolysis colony encapsulation gives "mucoid" wet appearance (can decrease over time) Prolonged incubation can cause center of colonies to collapse |
What is the quellung reaction? |
Anticapsular antibodies applied to S. pneumo cause capsule to swell and become more conspicuous under microscopy |
What special cellular morphology identifies S. pneumo vs. other strep species? |
Pairs of coccobacilli non-staining halo indicates capsule |
What does the Optochin test allow you to do? |
Presumptive ID of S. pneumo
Disk with optochin placed on media plated with colony
Zone of inhibition >14mm = presumptive ID of S. pneumo |
How is the bile solubility test performed? |
Addition of 10% Na desoxycholate (bile salt)
activates lytic enzymes in S. pneumo and causes colonies to disappear |
What are the biochemical characteristics of viridans sp. strep? |
alpha or gamma hemolytic bile esculin and optochin - |
what are the biochemical characteristics of Serogroup C and G strep? Group D? |
C and G: beta hemolytic, bile esculin and optochin -
D: gamna hemolytic, Bile Esculin + optochin - |
What characteristics describe Enterococci?
Gram/morphology hemolysis catalase lancefield group pyr bile esculin salt tolerance |
Gram + cocci (single, pair or short chain) Gamma hemolytic Catalase - Lancefield group D (but not GDS) facultative anaerobe PYR+ Bile Esculin and Salt tolerant |