ASSESSMENT MICRO 2016 CLIN COR

1. What is the purpose of adding 0.025% to 0.050% sodium polyanethol sulfonate

(SPS) to nutrient broth media for collection of blood cultures?
a) It inhibits phagocytosis and complement
b) It promotes formation of a blood clot
c) It enhances growth of anaerobes
d) It functions as a preservative

Ratio:
Yellow top - for blood culture, contains 0.025%-0.050% SPS
SPS - inhibits coagulation, phagocytosis, complement activation and inactivates
aminoglycosides

2. Assuming that a clean-catch midstream urine was processed, which of the following
is indicative of urinary tract infection?
a) <1000 CFU/mL
b) >1000 CFU/mL
c) >10,000 CFU/mL
d) >100,000 CFU/mL or 1.0 x 105 CFU/mL

Ratio:
UTI - 90% cause by E.coli,
S. saprophyticus - leading agent of UTI among YOUNG FEMALES

3. Which of the following antimicrobial agents acts by inhibiting protein synthesis?

a) Penicillin
b) Gentamicin
c) Rifampin
d) Colistin

Ratio:
TARGET SITES
Cell wall synthesis Beta lactams, glycopeptides,
cycloserine, bacitracin
Protein synthesis Aminoglycosides, tetracycline,
chloramphenicol, erythromycin, fusidic
acid
Nucleic acid synthesis Sulfonamides, trimethoprim, quinolones,
rifampicin
Cell membrane function Polymyxins

Beta-lactams neutralized by beta-lactamase - common among H. influenzae

ESBL, ampC, carbapenemase
Tests to detect beta-lactamase - CHROMOGENIC CEPHALOSPORIN ASSAY,
NITROCEFIN ASSAY
MODIFIED HODGE TEST - confirmatory test for carbapenem resistance
TETRACYCLINE - inhibits bone growth in children
Chloramphenicol - aplastic anemia
Sulfonamides - inhibit FOLIC ACID synthesis
Rifampin - ORANGE-RED body fluids if high amounts ingested

4. For demonstration of Campylobacter jejuni, which of the following reagent is

substituted as counterstain instead of safranin?
a) Malachite green
b) Methylene blue
c) Tergitol
d) Carbol fuschin

Ratio: GRAM STAIN

Reagents GRAM (+) GRAM (-)
Primary stain Crystal violet violet violet
Mordant Grams Iodine violet violet
Decolorizer Acetone alcohol Violet (due to colorless
(95% ETOH) peptidoglycan
layer)
Secondary stain Safranin violet Red/pink
For C. jejuni, safranin is substituted by CARBOL FUSCHIN as counterstain
MTB - Gram (+), Gram ghost/neutral (do not take up crystal violet or safranin
counterstain)
Decolorizing stage - most critical step, stage where differentiation occurs
Gram positive = with THICK peptidoglycan, retains crystal violet
Plasma membrane, periplasmic space, peptidoglycan (THICK)
Gram negative = with THIN peptidoglycan, decolorized
Plasma membrane, periplasmic space, peptidoglycan (THIN), periplasmic
space, OUTER MEMBRANE

Two types of AFB stains

CARBOL FUSCHIN
FLUOROCHROME STAINS (auramine-rhodamine and auramine-O) - more
sensitive than carbol fuschin, preferred method in detecting AFB

ACID-FAST STAIN (2x3 cm)

ZIEHL-NEELSEN KINYOUN (COLD) ACID-FAST NON-ACID
(HOT) FAST
Primary stain Carbol fuschin Carbol fuschin Red Red

Mordant Steam Tergitol (no Red Red

heating required
because phenol
content is higher)

Decolorizer ACID ALCOHOL ACID ALCOHOL Red (cell wall Colorless

(3% HCl in 95% (3% HCl in 95% has high lipid
ETOH) ETOH) content)

Secondary Malachite green or Malachite green Red Blue or green

stain methylene blue or methylene blue

Nocardia - PARTIALLY ACID FAST

N. asteroides - nocardiosis (pulmonary infections, etc.)
N. brasiliensis - actinomycetoma (localized granulomatous mass with sinus
tracts draining pus and sulfur granules)
Rhodococcus equi - granulomatous pneumonia
Tropheryma whippelii - WHIPPLES DISEASE

5. Which of the following gram(+) bacilli is capable of producing hydrogen sulfide

(H2S)?
a) Bacillus spp.
b) Corynebacterium spp.
c) Erysipelothrix spp.
d) Lactobacillus spp.

Ratio:

ERYSIPELOTHRIX RHUSIOPATHIAE
Produces HYDROGEN SULFIDE on triple-sugar iron (TSI) agar
Causes ERYSIPELOID - localized cutaneous infection of the HANDS and FINGERS
Common in wild and domestic animals
High risks - BUTCHERS, abattoir workers, fishermen, FISH HANDLERS ,etc.
PIPE CLEANER pattern of growth in GELATIN STAB CULTURES
GRAM-STAIN REACTION OF MEDICALLY IMPORTANT BACTERIA

AEROBIC ANAEROBIC
(OBLIGATE)
GRAM POSITIVE Staphylococcus, Streptococcus, Peptostreptococcus,
COCCI Micrococcus Finegoldia,
Anaerococcus,
Peptoniphilus
GRAM NEGATIVE Neisseria, Moraxella Veilonella,
COCCI Acidaminococcus,
Negativicoccus,
Megasphaera,
Anaeroglobus
GRAM POSITIVE Corynebacterium, Listeria, Erysipelothrix, Clostridium,
BACILLI Bacillus, Nocardia Actinomyces,
Propionibacterium,
Bifidobacterium,
Eggerthella

GRAM NEGATIVE Fermentative: Bacteroides,

BACILLI ENTEROBACTERIACEAE list provided Bilophila,
on separate table** Porphyromonas,
Prevotella,
Nonfermentative: Pseudomonas, Fusobacterium
Acinetobacter, Stenotrophomonas,
Burkholderia, Alcaligenes,
Achromobacter, Flavobacterium,
Chryseomonas, Acidovorax,
Brevundimonas, Ralstonia

Vibrio
Campylobacter
Aeromonas
Helicobacter

HACEK group: Haemophilus,

Aggregatibacter, Cardiobacterium,
Eikenella, Kingella

Miscellaneous: Legionella, Bordetella,

Pasteurella, Gardnerella,
Capnocytophaga, Calymmatobacterium*,
Francisella, Streptobacillus
Calymmatobacterium reclassified into enterobacteriaceae family
Spirochetes are GRAM NEGATIVE
Oxidase test: differentiates fermenters from nonfermenters
Fermenters - oxidase NEGATIVE except P. shigelloides
Nonfermenters - oxidase POSITIVE except A. baumanii and S. maltophila

ATMOSPHERIC REQUIREMENTS
Obligate aerobes Grows only in the presence of
oxygen
Obligate anaerobes Grows only in the absence of Separate table provided**
oxygen
Facultative anaerobes Grows in the presence or
absence of oxygen
Microaerophilic 5% O2, 10% CO2, and 85% N2

6. An identifying characteristic of S. aureus is

a) DNAse negative
 b) Coagulase negative
c) Mannitol fermentation positive
d) Growth inhibition in the presence of increased salt

Ratio:
STAPHYLOCCOCUS AUREUS
S. aureus is DNAse (+), coagulase (+), mannitol fermentation (+), halophile (7%
NaCl)
B-hemolytic on BAP
Most common cause of spinal epidural abscess and suppurative intracranial
phlebitis, septic arthritis is prepubertal children, BULLOUS IMPETIGO

Two types of coagulase:

Cell-bound (CLUMPING FACTOR) coagulase
detected by SLIDE COAGULASE TEST
(emulsified colonies + plasma) (+) clumping within 30 seconds
Free coagulase
detected by TUBE COAGULASE TEST
(emulsified colonies + plasma), incubate a 35C and observe after 4 and 24
hours, definitive, may be FALSE NEGATIVE if observed only after 24
hours since some strains produce FIBRINOLYSIN

Virulence factors
Protein A - found in the cell wall, has ANTI-PHAGOCYTIC properties, can bind
the Fc fragment of IgG
TSST-1 - responsible for TOXIC SHOCK SYNDROME, occurs 1 to 6 hours after
ingestion of enterotoxin
Enterotoxins A to E - responsible for STAPHYLOCOCCAL FOOD POISONING
Epidermolytic toxins A and B - SCALDED SKIN SYNDROME and erythema
Panton-Valentin leukocidin - responsible for NECROTIZING SKIN and soft
tissue infections
90% of staphylococci are resistant to penicillin due to production of beta-
lactamase

mecA gene which encodes for PENICILLIN-BINDING PROTEIN 2a (PBP2a) with

LOW AFFINITY to beta-lactams
responsible for resistance of MRSA to METHICILLIN and S. pneumoniae to
PENICILLIN

7. What is the composition of the acid-alcohol decolorizer used in acid-fast staining?

a) 3% HCl in 95% ethyl alcohol
b) 95% HCl in 3% ethyl alcohol
c) 3% H2SO4 in 95% ethyl alcohol
d) 95% H2SO4 in 3% ethyl alcohol

8. The beta-hemolysis produced by Group A Streptococcus seen on the surface of a

sheep blood agar plate is primarily the result of streptolysin
a) H
b) M
c) O
d) S

Ratio:
Streptococci
Catalase (-), gram (+) cocci in chains; facultatively anaerobic
Groups A, B, C and G are universally susceptible to PENICILLIN, susceptibility
testing is NOT required
Hemolysis Lancefield Species
Group
Beta (COMPLETE A S. pyogenes
hemolysis) B S. agalactiae

Alpha (PARTIAL hemolysis,
GREENING of the agar) or
gamma (NO HEMOLYSIS)
Alpha
C S. dysagalactiae
D Enterococcus spp. (Enterococcus
faecium and Enterococcus faecalis)
D Enterococcus spp.
D Streptococcus bovis complex
NONE Viridans group (Streptococcus mutans,
Streptococcus sanguis,
Streptococcus mitis, Streptococcus
salivarius, and Streptococcus anginosus.)
NONE S. pneumoniae

Nutritionally variant streptococci - requires THIOL or PYRIDOXAL for growth;

reassigned into genus ABIOTROPHIA or GRANULICATELLA; satellites around
colonies of S. aureus

GROUP A (S. pyogenes)

Clinical manifestations
Pharyngitis - most common clinical manifestation
Scarlet fever
Skin lesions: cellulitis, ERYSIPELAS, pyoderma
Acute rheumatic fever, acute glomerulonephritis, necrotizing fasciitis
Virulence factors:
M PROTEIN - ANTIPHAGOCYTIC
Lipoteichoic acid - permits bacterial ADHERENCE to respiratory epithelium
Enzymes
SLO: oxygen-labile, antigenic, subsurface hemolysis on BAP
SLS: oxygen-stable, nonantigenic, surface hemolysis on BAP
Streptokinase: dissolves fibrin
DNase
NADase
Pyrogenic toxins - produced by isolates of S. pyogenes infected with
bacteriophage
Superantigens - responsible for necrotizing fasciitis or toxic shock-like
syndrome

GROUP B
Clinical manifestations
Neonatal sepsis, pneumonia, meningitis

9. In the CAMP test, a single streak of beta-hemolytic Streptococcus is placed

perpendicular to a streak of beta-lysin producing S.aureus. After incubation, a zone
of increased lysis in the shape of an arrowhead is noted. This indicates the
presumptive identification of
a) S. agalactiae
b) A. pneumoniae
c) L. monocytogenes
d) E. faecalis

Ratio:
CAMP = Cristie, Atkins, Munch-Petersen
(+) CAMP test = S. agalactiae (catalase +), L. monocytogenes (catalase -)
Reverse CAMP test = C. perfringes
Group B strep colonies appear as YELLOW to ORANGE in GRANADA AGAR

10. Which of the following is able to hydrolyze sodium hippurate to benzoic acid and
glycine?
a) S. agalactiae
b) S. pneumoniae
c) L. monocytogenes
d) E. faecalis

Ratio: = S. agalactiae (+) CAMP test, Hippurate test

C. jenuni and L. pneumophilia = (+) Hippurate test

11. The inclusion bodies found in the cytoplasm of Corynebacterium diphtheriae are
called?
a) Much' granules - MTB, for mycolic acid
b) Babes-Ernst granules
c) Bipolar bodies - Yersinia pestis
d) Guarnieri bodies - smallpox
Ratio:
CORYNEBACTERIUM DIPHTHERIAE
Gram (+) rods with CLUBBED appearance, catalase (+)
Part of the normal flora of skin and mucous membrane
Causes grayish-back PSEUDOMEMBRANE in the oropharynx Bullous neck,
spreading into larynx causing respiratory obstruction
Only C. diphtheriae infected with bacteriophage are toxigenic
ELEK IMMUNODIFFUSION TEST - IN VITRO test for the detection of exotoxin
Preferred media:
CYSTINE-TELLURITE - gray or black colonies after 48 hours
TINSDALE MEDIUM

12. Which of the following is(are) used to stain spores?

a) Schaeffer and Fultons
b) Wirtz-Conklin
c) Dorner's
d) All of the above

Ratio:
SPORE-FORMING BACTERIA
Bacillus spp. (AEROBIC), Clostridium spp. (ANAEROBIC)

BACILLUS SPP.
All species are MOTILE except B. anthracis

B. anthracis - agent of ANTHRAX

Four forms: CUTANEOUS (Black eschar), INHALATION WOOLSORTERS
DISEASE), intestinal, injectional (NEWEST form)
Virulence factor: glutamyl POLYPEPTIDE capsule (antiphagocytic)
Exploited as agent of BIOTERRORISM
MEDUSA- HEAD colonies on BAP
BEATEN-EGG WHITE consistency when touched with an inoculated loop

B. cereus
Produces heat-stable toxic that causes FOOD POISONING (esp. FRIED RICE),
poisoning occurs 1 to 6 hours following ingestion of contaminated food

13. Which of the following bacillus species has round and terminally located spores?
a) Clostridium perfringes
b) Clostridium septicum
c) Clostridium tetani
d) Clostridium botulinium

Ratio:
Endospore = resting stage for bacteria exposed to unfavorable conditions
Composition = calcium dipicolinate
(+) Spore
Bacillus spp. = sporulates aerobically, catalase (+)
Clostridium spp. = sporulates anaerobically, catalase (-)
Spores are oval and subterminal EXCEPT C. perfringes (round and
terminal = tackhead drumstick, lollipop appearance)
14. Colonies that are slimy in appearance denotes the presence of what cellular
structure?
a) Mesosomes
b) Plasmids
c) Cell Wall
d) Capsule

Ratio:
Capsule = antiphagocytic, antigenic test (Quellung reaction)
Colonies are slimy
Composition = polysaccharide (HNS, B.fragilis), polypeptide (B. anthracis)
Stains: India Ink, Hiss

15. Which of the following corresponds to flagellar antigen?

a) K antigen
b) O antigen
c) H antigen
d) Vi antigen

Ratio:
K is CAPSULAR antigen, O is heat-stable SOMATIC antigen, H is heat-labile
FLAGELLAR antigen
Flagellar arrangements
Atrichous, Monotrichous (Vibrio spp.) (Amphitrichous), (Lophotrichous)
Peritrichous - Enterobacteriaceae EXCEPT Kleibsiella and Shigella (nonmotile)
Periplasmic flagella/Axial filament/Endoflagella - locomotive structure of spirochetes
(TBL), corkscrew like motility

Motility is best seen at 25C (37C is inhibitory for Yersinia and Listeria)

16. Which genus of bacteria produce a gliding motility on hanging drop preparation
technique?
a) Listeria spp.
b) Capnocytophaga spp.
c) Campylobacter spp.
d) Helicobacter spp.
Ratio:
Test for motility: Hanging drop preparation, SIM
Tumbling motility = Listeria spp.
Darting: Campylobacter
Gliding: Capnocytophaga
Stains for flagella contains tannic acid (ppt and coat flagella) = Leifsons, Grayson,
Fischer-Conn, Cesares-Gil, Van Emersons

17. M protein is the primary antiphagocytic factor of what organism?

a) Streptococcus pyogenes
b) Haemophilus influenzae
c) Neisseria spp.
d) Staphylococcus aureus

Ratio: M protein is homologous to myosin light chains of the heart = ARF

18. Which of the following statements is NOT true of endotoxin?

a) Released by dead organisms
b) Usually made up of lipid
c) Heat labile
d) Not easily neutralized by antitoxin

Ratio:
Comparison Exotoxin Endotoxin
Source Gram (+) and Gram (-) Gram (-) only
Release Metabolic products Released by DEAD
released by LIVING cells organisms upon lysis
Heat labile except Heat stable
staphylococcal enterotoxin
Immunologic Can be converted into Cannot be converted into
toxoid toxoid
Easily neutralized by Neutralized by antitoxin
antitoxin
Toxicity High low
Lethal dose Smaller Larger
1. Tetanospasmin - tetanus
lock jaw, spasmodic
contractions
2. Botulinium toxin -
botulism canned-good
poisoning, most potent
exotoxin, flaccid paralysis

19. Which of the following test is used to detect the presence of endotoxin in body fluids
and surgical instruments?
a) Limulus lysate test
b) ELEK test
c) CAMP tests
d) PYR test

Ratio:
Reagent = blood from horseshoe crab (Limulus polyphemus)
In the presence of endotoxin, amoebocytes will release lysate (+) clumping

20. Which of the following genus of bacteria requires X and/or V factors?

a) Staphyloccocus spp.
b) Streptococcus spp.
c) Vibrio spp.
d) Haemophilus spp.

Ratio:
HAEMOPHILUS SPP.
Oxidase (+), facultatively anaerobic, small, gram (-) pleomorphic rods or coccobacilli
Fastidious, requires X (HEMIN)and V (NAD) factors for growth
X factor: heat-lysed (CHOCOLATIZED) red cells from HORSE BLOOD,
human blood, rabbit blood, HEAT STABLE
V factor: YEAST EXTRACT, suspension of S. aureus, HEAT LABILE
Major virulence factor: POLYSACCHARIDE CAPSULE, six serotypes (a to f),
vaccine is available for H. influenzae type b
Part of normal flora, may cause conjunctivitis, otitis media, meningitis, endocarditis
Haemophilus aphrophilus reclassified as AGGREGATIBACTER APHROPHILUS
V factor dependent Haemophilus strains satellites around colonies of V factor
producing S. aureus.
PORPHYRIN TEST - identify which species DO NOT REQUIRE X factor,
determines organisms ability to convert -aminolevulinic acid into porphobilinogen
and porphyrins.
(+) X-factor independent strains: development of red color upon addition of
KOVACS reagent or red fluorescence under a WOODS LAMP
SCHOOLS OF FISH (bacilli in pairs or rows) - gram stain reaction of H. ducreyi
from chancroid lesions
Some H. influenzae strains produce -lactamase

DIFFERENTIAL CHARACTERISTICS OF MEDICALLY IMPORTNT HAEMOPHILUS

SPECIES
Porphyrin X Factor V factor Hemolysis
 Dependent Dependent
H. influenzae - + + -
H. haemolyticus - + + +
H. + - + +
parahaemolyticus
H. ducreyi - + - -
H. parainfluenzae + - +

HACEK - bacteria associated with SUBACUTE BACTERIAL ENDOCARDITIS

Haemophilus spp.
H. influenzae, H. parainfluenzae
Aggregatibacter
A. aphrophilus (Haemophilus), A. actinomycetemcomitans (Actinobacillus)
Cardiobacterium hominis
Eikenella corrodens
Kingella kingae

21. Sulfide Indole Motility and Transport agars are examples of?
a) Liquid medium
b) Semi-solid medium
c) Solid medium, liquefiable
d) Solid medium, nonliquefiable

Ratio: Semi-solid - contains 0.5% agar, C= 2-3% agar D = chopped meat (anaerobes),
rice grain (fungi) M. canis, M. audinii (fluorescence under Woods light)

22. The recommended cell culture medium for the recovery of Chlamydia spp. from
clinical specimens is
a) McCoy cells
b) Charcoal yeast extract medium
c) Fletcher semisolid media
d) WI-38 cells

Ratio:
CHLAMYDIAS, RICKETTSIAE, MYCOPLASMAS
Difference from other bacteria: SMALLER, different cell wall structure, most
OBLIGATE INTRACELLULAR (except mycoplasma)

CHLAMYDIA
Infects primarily COLUMNAR EPITHELIAL CELLS
Unlike most bacteria, they are ENERGY PARASITES, unable to produce their
own ATP and thus cannot replicate outside host cells.
Two forms:
Elementary body - INFECTIOUS FORM, capable of extracellular survival
Reticulate body - ACTIVE FORM, intracellular
Three human pathogens:
Chlamydia trachomatis - two BIOVARS LYMPHOGRANULOMA VENEREUM
(Biovar LGV) and TRACHOMA (Biovar trachoma), most common cause of STD
in USA, primary infectious cause of BLINDNESS in trachoma-endemic regions
Chlamydophila (formerly Chlamydia) psittaci - PNEUMONIA associated with
exposure to BIRDS psittacosis, PARROT FEVER, ornithosis
Chlamydophila (formerly Chlamydia) pneumoniae - acute respiratory tract
disease, formerly considered as a strain (TWAR) of C. psittaci

RICKETSSIAE***Separate table provided

Have a gram-negative bacterial cell wall
Frequently transmitted by ARTHROPODS
Rickettsiae genera pathogenic to humans:
Rickettsia -
 Orientia
Ehrlichia
Anaplasma
Neorickettsia
Neoehrlichia
Coxiella
Bartonella**

MYCOPLASMA
Smallest free-living organisms
Unique, NO CELL WALL
Require CHOLESTEROL and related STEROLS for membrane synthesis
Human pathogens:
Mycoplasma pneumoniae - PRIMARY ATYPICAL PNEUMONIA, aerosol
transmission
Genital mycoplasmas (Ureaplasma urealyticum, Mycoplasma homonis,
M.genitalium), recovered from urethral, vaginal or endocervical swab specimens

Bartonella may be cultured in cell-free medium, not considered a Rickettsiae

Etiologic Disease Transmission

agent
SPOTTED FEVERS
R. rickettsii Rocky Mountain Spotted Fever Tick bite
R. conorii Boutonneuse fever Tick bite
TYPHUS FEVERS
R. prowazekii Epidemic typhus Feces of human body
louse
R. prowazekii BrillZinsser disease Recrudescence of latent
infection
R. prowazekii Flying squirrel typhus Recrudescence of latent
infection
R. typhi Murine typhus Flea feces
TRANSITIONAL GROUP RICKETTSIAL FEVERS
R. akari Rickettsialpox Mite bite
R. australis Queensland tick typhus Tick bite
R. felis Flea-borne spotted fever Presumably flea bite or
feces
SCRUB TYPHUS
Orientia Scrub typhus Chigger bite
tsutsugamushi
EHRLICHIOSES
E. chaffeensis Human monocytotropic ehrlichiosis Tick bite
E. ewingii Ehrlichiosis ewingii Tick bite
Anaplasma Human granulocytotropic Tick bite
phagocytophilum anaplasmosis
Neorickettsia Sennetsu rickettsiosis Unknown
sennetsu
COXIELLOSIS
C. burnetii Q fever Inhalation of aerosols from
infected animals, possibly
ingestion of animal
products
BARTONELLOSES
B. bacilliformis Oroya fever, verruga peruana Sandfly bite
B. henselae Cat scratch disease, bacillary Kitten scratch or bite, cat
angiomatosis flea
and peliosis, endocarditis,
osteomyelitis,
 neuroretinitis, lymphadenitis
B. clarridgeiae Cat-scratchlike disease Presumed cat scratch or
bite
B. quintana Trench fever, endocarditis, bacillary Feces of Pediculus louse
angiomatosis

23. Organisms that have a cell wall containing large amount of lipids belong to the
genus
a) Chlamydia spp.
b) Mycobacterium spp.
c) Mycoplasma spp.
d) Leptospira spp.

Ratio = High lipid content makes them resistant to acetone alcohol

24. Which of the following enrichment medium is used to enhance the recovery of Vibrio
spp?
a) Tomato juice agar
b) Alkaline peptone water
c) Selenite broth
d) Tetrathionate broth

Ratio:

VIBRIO SPP.
General characteristics: Facultatively anaerobic, oxidase (+), short, curved
COMMA-SHAPED gram (-) bacilli, motile by means of POLAR flagella, ferments
CHO and reduce nitrates to nitrites
Vibrio vulnificus causes the most severe disease, wound infection and septicemia is
often fatal, usually associated with consumption of raw OYSTERS
V. cholerae O1 - produces CHOLERA TOXIN which causes EPIDEMIC CHOLERA
characterized by hypersecretion of water and electrolytes due to activation of
adenylate cyclase
V. parahemolyticus produces a cell-free hemolysis that is lethal to mice when
injected in high amounts (KANAGAWA PHENOMENON)
All requires NaCl for growth EXCEPT V. cholerae and V. mimicus
TCBS - selective media for Vibrio spp.
ALKALINE PEPTONE WATER - enrichment medium for Vibrio spp.
TSI reaction - A/A acid slant/acid butt NO GAS and H2S
LIA reaction - K/K alkaline slant/alkaline butt

DIFFERENTIAL CHARACTERISTICS OF VIBRIO SPECIES

Test V. cholerae V. V. V. V.
mimicus paraheamolyticus alginolyticu vulnificus
s
Indole + + V V +
Voges- V - - + -
Proskauer
Lysine + + + + +
decarboxylase
Ornithine + + V V V
decarboxylase
Arginine - - - - -
decarboxylase
Sucrose + + + + V
Lactose + V - - +
Maltose + + + + +
Salicin - - - - +
Oxidase + + + + +
Nitrate reduction + + + + +
All are oxidase positive and capable of reducing nitrate to nitrite
All are capable of lysine decarboxylase
NONE are capable of arginine decarboxylase
All ferments lactose and sucrose but only V. vulnificus ferments salicin

SELENITE BROTH - enrichment medium for Salmonella and Shigella

C and D is for Salmonella and Shigella

25. Which of the following blood sources is NOT acceptable to be used for blood agar
plates?
a) Sheep
b) Horse
c) Rabbit
d) Human

Ratio: Human blood contains nonspecific inhibitors citrate and glucose

BAP = 5% defibrinated sheep blood > horse > rabbit
Chocolate agar = Horse blood is preferred

26. Lowenstein-Jensen medium for cultivation of MTB contains what inhibitor?

a) Crystal violoet
b) Potassium tellurite
c) Sodium azide
d) Malachite green

Ratio: Lowenstein-Jensen = egg-based, MTB requires high protein, Malachite green

inhibits contaminating organism
Petragnani = contain the highest amount of malachite green, for heavily contaminated
samples

27. What antibiotic incorporated in the New York City agar prevents the growth of Gram
(+) bacteria?
a) Vancomycin
b) Trimethoprim lactate
c) Anisomycin
d) Amphotericin B

Thayer Martin Modified Martin Lewis New York City

Thayer Martin agar
Gram (+) inhibitor Vancomycin Vancomycin Vancomycin Vancomycin

Gram (-) inhibitor Colistin Colistin Colistin Colistin

EXCEPT N.
gonorrhea

Antifungal Nystatin Nystatin Anisomycin Amphotericin B

Inhibitor of NONE Trimethoprim Trimethoprim Trimethoprim
Proteus lactate lactate lactate
swarming

28. After a week on incubation, Vibrio cholerae produce yellow colonies on TCBS agar.
This indicates that
a) Mannose is fermented
b) Sucrose is fermented
c) Mannose is not fermented
d) Sucrose is not fermented
Ratio: TCBS = CHO (sucrose), pH indicator (Bromthymol blue)
SF = V. cholerae, V. alginolyticus (yellow colonies)
NSF = V. parahemolyticus, V. mimicus = green colonies
+/- = V. vulnificus

29. All of the following enterobacteria will give a positive ONPG test EXCEPT
a) Kleibsiella pneumoniae
b) Enterobacter aerogenes
c) Salmonella typhi
d) Shigella sonnei

Ratio:
ENTEROBACTERIACEAE
General characteristics
Non-spore-forming, oxidase negative, gram (-) bacilli, ferments GLUCOSE, reduce nitrates to
nitrites
Includes the following genera: Citrobacter, Edwardsiella, Enterobacter, Escherichia,
Ewingella, Hafnia, Klebsiella, Morganella, Proteus, Providencia, Salmonella, Serratia,
Shigella, Yersinia
Recent inclusion: Plesiomonas spp., very similar to PROTEUS - only oxidase (+) member
Virulent factors
ENDOTOXINS - responsible for most clinical manifestation
K1 antigen - found in E. coli causing neonatal meningitis
Capsule - antiphagocytic, found in K. pneumoniae
Vi antigen - inhibits intracellular killing, fond in S. typhi
Fimbriae - adherence to mucosal surfaces
Heat-labile (LT) enterotoxin - stimulates ADENYLATE CYCLASE causing secretory
diarrhea
Heat-stable (ST) enterotoxin - stimulates GUANYLATE CYCLASE
Shiga toxin - produced by E. coli O157:H7 and S. dysenteriae

ONPG - test for beta-galactosidase, present on rapid and late lactose fermenters
All enterobacteriaceae ferments glucose
Rapid fermenters (lactose permease and beta-galactosidase)
Late lactose fermenters (beta-galactosidase)

30. Lysine Iron Agar (LIA) is not sensitive to the H2S production of which organism?
a) Edwardsiella spp.
b) Arizona spp.
c) Citrobacter spp.
d) Proteus spp.

Ratio:
Proteus - exhibit SWARMING on BAP
LIA, TSI - media for biochemical testing
Composition = protein, CHO (glucose, lactose, sucrose 0.1%/1%/1%)
pH indicator = phenol red A-yellow K-red
H2S indicator = ferrous sulfate (+) blackening of the media
Slant = LF or SF fermentation LF/SF yellow/red
Butt = Glucose fermentation = GF/NGF yellow/red
Cracks = gas production, all entero EXCEPT shigella

TSI Reactions Typical organisms

A/AG H2S - LF/SF over GF EKE
K/AG H2S + NLF/NSF over GF Salmonella, Proteus
K/A H2S - NLF/NSF over GF Shigella
K/K H2S - NLF/NSF over NGF Pseudomonas alcaligenes
31. Enterobacter cloaceae will produce what reaction on TSI agar?
a) A/AG H2S (-)
b) K/AG H2S (+)
c) K/A H2S (-)
d) K/K H2S (-)

32. Which of the following LIA reactions indicates lysine deamination?

a) K/K H2S (+)
b) K/A H2S (-)
c) R/A H2S (-)
d) K/K H2S (-)

Ratio: LIA = test organisms ability to deaminate, decarboxylate lysine and produce H2S
Composition: glucose, lysine
pH indicator = Bromcresol purple A-yellow, K-purple
H2S indicator = Ferric ammonium citrate - SACEd not sensitive to H2S production of
proteus
Slant - lysine deamination
(+) - red (R) PPM deaminase (+)
(-) - purple (K)
Butt - lysine decarboxylation
Purple(K) --> Glucose fermentation --> (A)Yellow (-)--->LDC(+)--->Purple(K)
K/K H2S + lysine deam - lysine decar + H2S + Salmonella
K/A H2S - lysine deam - lysine decar - Shigella
R/A H2S lysine deam +, lysine decar + PPM

33. The pH indicator used in MacConkey Agar is

a) Bromcresol purple
b) Bromthymol blue
c) Phenol red
d) Neutral red

Ratio: MacConkey - Selective/Differential for Gram (-) Enteric Bacilli

Selective = crystal violet and bile salt - inhibits gram (+)
Differential = CHO lactose
pH indicator: neutral red (A-red, K-yellow)
LF = pink colonies
NLF = colorless colonies

34. Which of the following organisms will produce yellow colonies on Mannitol Salt
Agar?
a) Staphyloccocus lugdunensis
b) Staphylococcus aureus
c) Staphylococcus epidermidis
d) Staphyloccocus saprophyticus
Ratio: MSA = Selective/Differential for S.aureus
Selective = 7% NaCl
CHO = Mannitol
pH indicator = phenol red
MF = S. aureus (yellow colonies)
NMF = S. epidermidis (sensitive to novobiocin), S. saprophyticus (pink colonies)
(resistant to novobiocin)

35. Growth on EMB medium produced pink colonies with dark center characteristically
described as "fish-eye" colonies. What is the possible identity of the genus isolated?
a) Escherichia spp.
b) Klebsiella spp.
c) Enterobacter spp.
d) Shigella spp.

Ratio: EMB = Selective Differential for Gram (-) Enteric bacilli

CHO: lactose
pH indicator: Eosin and methylene blue
LF = pink to purple colonies
NLF = colorless colonies

E. Coli = pink to purple colonies with green metallic sheen

Kleibsiella = pink MUCOID colonies
Enterobacter = pink colonies with dark center fish eye colonies

36. How will the growth of E.coli on Hektoen Enteric Agar be described?
a) Yellow colonies without black center
b) Yellow colonies with black center
c) Green colonies without black center
d) Green colonies with black center

Ratio: HEA = Selective/Differential for Gram (-) Enteric bacilli

Selective = bile salt (inhibits gram +)
CHO: lactose, sucrose, salicin
pH indicator = bromthymol blue
H2S indicator = Ferric ammonium citrate SPACEd colonies have black center

37. Salmonella Shigella Agar is able to differentiate between Salmonella and Shigella by
identification of what bacterial property?
a) Lactose fermentation
b) Sucrose fermentation
c) Hydrogen sulfide production
d) Acetoin production

Ratio:
Colonial appearance
Salmonella = NLF, H2S (+)/colorless colonies with a black center
Shigella = NLF, H2S (-)/colorless colonies without a black center
Salmonella serotypes typhimurium and enteriditis - most common isolates
Salmonella serotype typhi - possess heat-labile capsular polysaccharide Vi
ANTIGEN
Four serogroups
A
B
C
D - S. serotype typhi

38. In which of the following culture media will Y. enterocolitica produce the
characteristic "bulls's eye" colonies?
a) Castaneda medium
b) Feeley-Gorman medium
c) Blood-Glucose-Cysteine medium
d) Cefsulodin-Irgasan-Novobiocin medium

Ratio:
CIN agar - selective and differential for Y. enterocolitica ,colonies appear as BULLS-
EYES WITH RED CENTERS and transparent borders
Molar tooth = Actinomyces
Fried egg = Mycoplasma
Medusa head = B. anthracis
Dew drop or satellite = H. influenzae
Star-shaped colonies = A. actinomycetemcomitans

39. Which of the following is the preferred medium for fluorescence microscopy of
streptococci obtained from genital specimens?
a) Granada medium
b) Cetrimide agar
c) MacConkey agar
d) Todd-Hewitt broth
40. After 24 hours of incubation, growth of colorless colonies were observed on sorbitol-
substituted Mac Conkey agar (SMAC). What is the most probable identification of
these organisms?
a) E. coli O157:H7
b) Salmonella arizonae
c) Shigella sonnei
d) All of the above

Ratio:
SMAC - differential medium for sorbitol (-) E. coli 0157:H7 associated with
HEMOLYTIC UREMIC SYNDROME
E. coli 0157:H7 - COLORLESS colonies, nonfermenter

41. An erratic growth on Mueller-Hinton agar may be observed if

a) Antibiotic disks are placed 24mm from disk center to center
b) The cation content of the medium is increased
c) pH of the agar is maintained between 7.2-7.4
d) Agar depth is maintained at 4mm

Ratio: MHA = standard medium for susceptibility testing

Agar depth = 4mm
pH = 7.2-7.4
24 mm from disk center to center

42. What is the equivalent bacterial concentration of 0.5 McFarland standard in terms of
CFU/mL?
a) 1.0 x 106 CFU/mL
b) 1.5 x 106 CFU/mL
c) 1.0 x 108 CFU/mL
d) 1.5 x 108 CFU/mL

43. The biologic indicator used in autoclave is?

a) Bacillus pumilis
b) Bacillus stearothermophilus
c) Bacillus subtilis var. niger
d) Bacillus subtilis var. globigii

Ratio: A - ionizing radiation, C - dry heat oven, D - Ethylene oxide

44. Blood samples for microbial culture must be

a) drawn before a fever spike
b) drawn after administration of antibiotics
c) collected from identical sites at least 1 hour apart
d) collected after red-top tubes in vacutainer system

45. Blood cultures for brucellosis must be held for how long before being reported as
negative?
a) 5-7 days
b) 3-4 weeks
c) 6-8 weeks
d) 10-12 weeks

Ratio: C = MTB

46. The most abundant normal flora of the upper respiratory tract is?
a) Streptococcus pneumoniae
b) Viridans streptococci
c) Group A streptococci
d) Group B streptococci

Ratio:
 Viridans strep, alpha hemolytic
Bacterial endocarditis - most common infection

47. An acceptable sputum for microbiological examination must have

a) <10 SEC/lpf and >25 PMN's/lpf
b) >10 SEC/lpf and <25 PMN's/lpf
c) <15 SEC/lpf and >25 PMN's/lpf
d) >15 SEC/lpf and <25 PMN's/lpf

48. Which CSF tube is acceptable for use in microbiology if four separate tubes were
used during spinal tap?
a) Tubes 1 and 2
b) Tubes 3 and 4
c) Tubes 1 and 3
d) Tubes 2 and 4

49. Which of the following does NOT describe S. aureus?

a) produces creamy white pinhead colonies with beta-hemolytic pattern on BAP
b) produces porcelain white colonies on Loeffler's serum slant
c) Catalase positive
d) Grows on phenylethylalcohol agar

Ratio:

50. A gram (+) coccus that is catalase (+), nonmotile, lysostaphin resistant, bacitracin
sensitive, furazolidine resistant, and modified oxidase positive is best identified as a
member of the genus
a) Micrococcus
b) Lactococcus
c) Veilonella
d) Staphyloccus
Ratio:
Staphylococcus spp. Micrococcus spp.
Lysostaphin susceptibility S R
Aerobic acid production + -
from glycerol
Anaerobic acid production +/- -
from glucose
Furazolidone S R
Bacitracin susceptibility R S
(0.04 U)
Modified oxidase - +

51. The reagent used in catalase test is

a) 3% H2O2
b) 30% H2O2
c) KOH and alpha-napthol
d) Tetramethylparaphenylenediamine dihydrochloride with DMSO
Ratio:
B - Superoxol test (screening test for Neisseria spp.)
C - Vogues-Proskauer - detect acetoin
D - Modified oxidase (micrococcus + ; staph -)

52. All of the following uses bromthymol blue as indicator EXCEPT

a) Mannitol Salt Agar - phenol red
b) Oxidative-Fermentative (OF) tube test
c) Hektoen Enteric Agar
d) Thiosulfate Citrate Bile Salt Sucrose Agar (TCBS)

54. Which of the following microorganisms cannot be cultivated on an artificial cell-free

media?
a) Francisella tularensis
 b) Helicobacter pylori
c) Actinobacillus lignieresii
d) Mycobacterium leprae
Ratio:
F. tularensis - requires cystine/cysteine

55. Fecal cultures are inoculated on thiosulfate citrate bile salt sucrose agar specifically
for the isolation of
a) Shigella spp.
b) Vibrio spp.
c) Campylobacter spp.
d) Salmonella spp.

56. Which microorganism will grow only on a culture media that has been supplemented
with either cysteine or cystine?
a) Bartonella bacilliformis
b) Kingella kingae
c) Actinobacillus lignieresii
d) Francisella tularensis

Ratio:
FRANCISELLA TULARENSIS
Very small, strictly AEROBIC, coccobacillary to pleomorphic rod-shaped, gram (-)
bacillus that requires CYSTINE or CYSTEINE for growth
Faint bipolar staining occurs with aniline dyes
Ubiquitous, primary reservoir is cottontail RABBIT
One of the Class A agents of bioterrorism
Agent of TULAREMIA
Grows well on Blood-Cysteine Agar and BCYE

57. Which of the following genera will produce yellow colonies on both open and closed
tubes in Oxidation-Fermentation (OF) tube test?
a) Pseudomonas spp.
b) Micrococcus spp.
c) Streptococcus spp.
d) Bordetella spp.

58. When methyl-green in the DNAse medium fades in color, which of the following
organisms may have been isolated?
a) Moraxella
b) Serratia
c) S. aureus
d) All of the above

59. A positive CAMP reaction is indicated by

a) development of a purple color upon addition of ninhydrin
b) formation of precipitin lines at 45 angle
c) an arrowhead zone of hemolysis
d) formation of black precipitate

60. Resistance to penicillin, sulfamethoxazole and penicillin, production of PYRase and

growth in 6.5 NaCl are characteristics of
a) Pneumococcus
b) Meningococcus
c) Gonococcus
d) Enterococcus

Ratio:
All Enterococci grow in the presence of 6.5% NaCl and hydrolyze ESCULIN in the
presence of bile

61. Sensitivity to 0.04U of Bacitracin (Taxo A) is one of the identifying features of

 a) Group A strep
b) Group B strep
c) Group C strep
d) Group D strep

Ratio:
More than 99% of Group A is susceptible to BACITRACIN, presumptive
identification
All isolates of Group A and more than 99% of Enterococcus are PYRase

62. A susceptibility test for scarlet fever in which an erythrogenic toxin is injected into the
arm of the patient and a positive reaction indicated by redness or erythema
a) Schultz-Charlton test
b) Schick's test
c) Francis test
d) Dick's test

63. Which of the following is NOT a characteristic of S. pneumoniae?

a) Solubility in bile
b) Resistance to ethyhyldrocupreine hydrochloride
c) Positive Quellung's reaction
d) Ability to ferment inulin

Ratio:
S. pneumoniae - possess POLYSACCHARIDE capsule (antiphagocytic)
Seen in the normal flora of upper respiratory tract
Produce ALPHA hemolytic colonies that are MUCOID or flattened with a depressed
center on BAP
All isolates are susceptible to OPTOCHIN (P disk) and BILE SOLUBLE

64. A positive butyrate esterase spot test is a characteristic of

a) Neisseria gonorrhea
b) Moraxella catarrhalis
c) Veillonella parvula
d) Acidaminococcus intestini

65. The microbiological test for acetoin production uses what reagents?
a) sodium citrate and bromthymol blue
b) alpha-napthol and KOH
c) Kovac's
d) methyl red

66. DIC and Waterhouse-Friedrichsen syndrome are some of the clinical conditions
caused by
a) Neisseria gonorrheae
b) Neisseria meningitidis
c) Neisseria subflava
d) Neisseria lactamica
Ratio:
NEISSERIA
Nonmotile, catalase (+) except N. elongata, oxidase (+), gram (-) diplococci
resembling KIDNEY BEANS or COFFEE BEANS
FASTIDIOUS - requires blood, serum, cholesterol and CARBON DIOXIDE (candle
jar, citric acid bicarbonate tablet) for growth
Saccharolytic
Inoculated tubes are incubated at 35C to 37C for 24 hours
Clinical manifestations
N. gonorrhea
N. meningitidis - meningococcocal meningitis , penicillin G drug of choice
Lipopolysaccharide-endotoxin - main virulence factor of N. meningitidis
Pathogenic strains of N. gonorrhea have PILI which permits ADHERENCE and is
ANTIPHAGOCYTIC
IgA protease - secreted by both N. gonorrhea and N. meningitidis
Culture media - ENRICHED CHOCOLATE AGAR **separate table provided
Cysteine trypticase acid (CTA) medium - detects acid production from
carbohydrates, standard method of identification

CTA REACTIONS/CARBOHYDRATE HYDROLYSIS

Glucose Maltose Lactose Sucrose Fructose
N. gonorrheae + - - - -
N. meningitidis + + - - -
N. lactamica + + + - -
M. catarrhalis - - - - -
DIFFERENTIATION OF NEISSERIA SPP. FROM MORAXELLA CATARRHALIS
CHO metabolism DNase Growth on BAP
Neisseria spp. Saccharolytic - -
M. catarrhalis Assacharolytic + +

67. Which genus under the Neisseriaceae family is able to grow on MacConkey agar
and is catalase positive?
a) Neisseria
b) Moraxella
c) Acinetobacter
d) Kingella

Ratio:
A. baumanii - PURPLISH color on MacConkeys agar, reclassified as Gram (-) bacilli

68. S. pneumoniae and Viridans strep will produce what type of hemolysis on BAP?
a) Alpha
b) Beta - S. pyogenes, S. agalactiae
c) Gamma
d) Alpha-prime

69. Which is NOT a characteristic of L. monocytogenes ?

a) produce a positive CAMP reaction
b) bile esculin hydrolysis positive
c) sodium hippurate hydrolysis positive
d) exhibit an "umbrella-like" growth on semi-solid medium

Ratio:
L. monocytogenes - ONLY listeria species that is pathogenic to humans, causes
LISTERIOSIS common among PREGNANT women, NEONATES and
immunocompromised
 Optimal growth is observed between 30C to 37C, but growth may occur as low
as 4C
Narrow zone of beta-hemolysis on BAP
Group B strep - catalase (+), L. monocytogenes - catalase (-)
CAMP (+), esculin hydrolysis (+)
TUMBLING motility at room temp on saline suspensions
UMBRELLA growth pattern on semi-solid media
Commonly found in dairy products, esp CHEESE, UNPASTEURIZED MILK,
vegetables, raw meat and poultry

70. This organisms grows optimally at 42C, exhibits darting motility in hanging drop
slide test, hydrolyzes sodium hippurate and best demonstrated on Gram-stain when
safranin is substituted by carbol fuschin
a) P. aeruginosa
b) C. jejuni
c) H. pylori
d) Proteus spp.

71. Which of the following organism is a rapid urease producer?

a) Morganella morgani
b) Serratia rubidea
c) Citrobacter diversus
d) Kleibsiella oxytoca

72. The end product of ornithine decarboxylation is

a) cadaverine
b) putresceine
c) citrulline
d) arginine

73. The primary reagent used in the phenylalanine decarboxylase test

a) Alpha-naphtol
b) PDAB
c) 10% KOH
d) 10% ferric chloride

74. Which of the following organisms have the ability to liquefy gelatin into amino acids?
a) Proteus vulgaris
b) Providencia stuartii
c) Morganella morganii
d) Citrobacter freundii

75. Positive DNAse, gelatin hydrolysis, and lipase tests and production of the red
pigment prodigiosin are identifying features of
a) Proteus spp.
b) Providencia spp.
c) Serratia spp.
d) Campylobacter spp.
76. Yersinia pestis
a) Is the most common cause of death associated with transfusion of
contaminated blood units
b) produces "Bull's eye" colonies on CIN medium
c) Causes mesenteric lymphadenitis
d) nonmotile both at room temp and 37C

77. A common agent of nosocomial infections due to its resistance to a number of

disinfectants including povidone iodine and the causative agent of "swimmer's ear",
whirlpool dermatitis is?
a) Pseudomonas aeruginosa
b) Burkholderia cepacia
c) Burkholderia mallei
d) Chryseobacterium spp.

Ratio:
P. Aeruginosa - resistant to many antibiotics

78. After P. aeruginosa, what is the second most common agent implicated to infect
patients with cystic fibrosis
a) Pseudomonas putida
b) Burkholderia mallei
c) Burkholderia pseudomallei
d) Burkholderia cepacia

Ratio:
Burkholderia cepacia - NOSOCOMIAL pathogen associated with contaminated
equipment, medications, disinfectants, important pathogen in patient with CYSTIC
FIBROSIS and CGD
B. pseudomallei - melioidosis

79. Which gram-negative bacteria will be positive in the string test using 0.5% sodium
desoxycholate
a) Plesiomonas spp. - P. shigelloides
b) Aeromonas spp.
c) Vibrio spp.
d) Shewanella spp.

Ratio:
P. shigelloides - found in AQUATIC environments esp. in tropical countries,
implicated as a cause of GASTROENTERITIS following ingestion of uncooked
SHELLFISH
Aeromonas spp.- mainly found in aquatic environment, associated with use of
medicinal leeches to decrease vascular congestion

80. The enterotoxins of both Vibrio cholerae O1 and ETEC strains of E. coli produce
serious diarrhea by what mechanism?
a) Stimulation of adenylate cyclase giving rise to excessive fluid secretion
by the cells of the small intestine
b) Penetration of the bowel mucosa
c) Stimulation of colicin production
d) The elaboration of a dermonecrotizing toxin

81. Which Haemophilus spp. will be positive in D-ALA test?

a) H. ducreyi
b) H. parainfluenzae
c) H. influenzae
d) H. haemolyticus

82. Which organism is the causative agent of Brazilian purpuric fever, the invasive form
of pink-eye conjunctivitis?
 a) H. aegypticus
b) H. ducreyi
c) H. haemolyticus
d) H. aphrophilus

83. This microaerophilic bacteria is a strong urease producer and commonly implicated
in cases of peptic ulcers
a) Campylobacter coli
b) Campylobacter jejuni
c) Campylobacter fetus
d) Helicobacter pylori

Ratio:
Microaerophilic: 5% O2, 10% CO2, and 85% N2
CAMPYLOBACTER
SMALL, motile, non-spore-forming, curved (COMMA-SHAPED) or S-SHAPED gram
(-) bacilli
MICROAEROPHILIC: optimal growth in an atmosphere containing 5% to 10%
oxygen (the amount of oxygen in candle jar is too little and therefore must not be
used)
C. jejuni - one of the most common cause of BACTERIAL ENTERITIS, the most
recognized antecedent cause of Guillain-Barre syndrome
C. fetus subsp fetus - cause of septic thrombophlebitis, arthritis, peritonitis, abscess,
pericarditis
Selective media for Campylobacter spp.: charcoal-cefoperazone-deoxycholate agar,
SKIRROWS medium
Incubation at 42C - enhances selectivity for C. jejuni
C. jejuni - HIPPURATE POSITIVE

C. jejuni C. fetus subsp. fetus

Hippurate hydrolysis + -
Growth at 42C + -
Nalidixic acid S R

HELICOBACTER
SPIRAL-SHAPED or curved, gram (-), non-pore-forming bacilli, motile,
MICROAEROPHILIC
One of the GASTRIC helicobacters, lives in the epithelium of the STOMACH
Associated with 90% of DUODENAL ULCERS and nearly all GASTRIC ULCERS;
also associated with gastric lymphoma and gastric carcinoma.
UREA BREATH TEST - employed in diagnosis of H. pylori infection, patient ingests
labeled urea and the amount of exhaled labeled CO2 is measured
Hydrolyze UREA VERY RAPIDLY
Must be cultured under MICROAEROPHILIC atmosphere (5% to 10% CO2, 80% to
90% N2, and 5% to 10% O2)

84. Which Brucella species is inhibited by thionine but grows in the presence of basic
fuschin?
a) Brucella abortus
b) Brucella mellitensis
c) Brucella suis
d) Brucella canis

Ratio:
BRUCELLA
Small, gram (-) coccobacillus in pairs of short chains resembling the appearance of
SAND, nonmotile, STRICTLY AEROBIC, nonfermenter, INTRACELLULAR,
produces UREASE
Growth is enhanced by 5% to 10% CO2 and the addition of blood or serum
 Causative agent of BRUCELLOSIS characterized by diurnal or UNDULANT FEVER
Common risk factor - consumption of imported CHEESE from unpasteurized
goats milk
Recovered most often from blood or BONE MARROW
Preferential hosts:
B. melitensis - sheep and goat
B. abortus - cattle
B. suis - swine
B. canis - dogs
Recognized as CLASS A bioterrorism agent
Grows SLOWLY, cultures must be held for up to 14 DAYS before reporting as
negative

DIFFERENTIATION OF BRUCELLA SPP.

Species CO2 requirement H2S production DYE INHIBITION
Thionine Fuschin
B. abortus + + + -
B. melitensis - + - -
B. suis - + - +
B. canis - - - +
85. A biphasic medium (with agar and broth component) which is used to cultivate
Brucella spp. is?
a) Feeley-Gorman medium
b) 10B broth
c) Castaneda bottles
d) Shepard's medium

86. This bacterium can be acquired by inhalation of infectious aerosols. It is not airborne
and not transmitted from human to human. The first recorded outbreak was in
Philadelphia during the convention of the American Legion.
a) Francisella tularensis
b) Bordetella pertussis
c) Legionella pneumophila
d) Pasteurella multocida
Ratio:

DIETERLE SILVER STAIN - stain for Legionella spp.

Buffered Charcoal Yeast Extract (BCYE) agar - culture medium for Legionella spp.

BORDETELLA SPP.
STRICTLY AEROBIC, nonfermentative, catalase (+) minute coccobacilli
B. pertussis - agent of PERTUSSIS or Whooping cough
B. parapertussis - agent of PERTUSSIS-LIKE disease
B. bronchiseptica - respiratory disease in humans; KENNEL COUGH in dogs
NASOPHARYNGEAL ASPIRATE - specimen of choice
REGAN-LOWE - culture medium for Bordetella spp., contains CEPHALEXIN;
culture provides the MOST SPECIFIC diagnosis for whooping.
DROP OF MERCURY appearance of colonies

87. This bacteria is associated with wound infections due to animal bites or scratch.
a) Pasteurella spp.
b) Francisella spp.
c) Streptobacillus spp.
d) Calymmatobacterium spp.

PASTEURELLA SPP.
Facultatively anaerobic, nonmotile, gram (-) coccobacillus to filamentous rods.
Of the eight species that infect humans, P. multocida is the most important
 Found as commensals in the upper respiratory tract of fowl and mammals and are
frequently isolated from ANIMAL BITE or SCRATCH WOUNDS
Highly susceptible to PENICILLIN, as evidence by WIDE ZONE of inhibition, thus
the drug of choice for Pasteurellosis

88. The Human Blood Bilayer Tween (HBT) Medium is used for culturing what
bacterium?
a) Streptobacillus moniliformis
b) Capnocytophaga canimorsus
c) Gardnerella vaginalis
d) Eikenella corrodens
Ratio:
GARDNERELLA SPP.
Associated with BACTERIAL VAGINOSIS but is NOT THE CAUSE
Part of the anorectal flora of both sexes and children, part of the endogenous flora
of women of reproductive age
Diagnosis is made by examination of CLUE CELLS (epithelial cells covered with
bacteria on cell margins), the absence of lactobacilli, pH greater than 4.5 and a
FISHY AMINE ODOR of the vaginal secretions after addition of 10% KOH.
Susceptibility testing is not required, METRONIDAZOLE is the drug of choice

STREPTOBACILLUS MONILIFORMIS
Facultatively anaerobic, nonmotile pleomorphic gram (-) rod, occurring in long
chains or long filaments resembling STRING-OF-BEADS appearance
Agents of RAT-BITE or HAVERHILL DISEASE

89. The agent of Donovanosis or granuloma inguinale is

a) Chromobacterium violaceum
b) Cardiobacterium hominis
c) Eikenella corrodens
d) Calymmatobacterium granulomatis
Ratio:
Renamed as Kleibsiella granulomatis
Gram (-), nonmotile bacilli, ENCAPSULATED, pleomorphic, may be cultured in
YOLK SAC or FRESH EGG YOLK MEDIUM
Agent of GRANULOMA INGUINALE, a rare sexually transmitted disease
Safety pin appearance on stained smears due to presence of polar granules

90. This bacterium pits or corrodes the agar, produces bleachlike odor, and causes
polymicrobial abscesses of the oral cavity
a) Cardiobacterium hominis
b) Eikenella corrodens
c) Chromobactecterium violaceum
d) Chryseobacterium spp.

Ratio:
Eikenella corrodens - CORRODING bacilli, colonies pits or corrodes the agar

91. All the following descriptions fit B. anthracis EXCEPT

a) Colonies on BAP resemble "medusa-head"
b) Colonies have "beaten-egg" consistency when lifted by loop or needle
c) String of pearl" appearance on MHA with penicillin disk
d) Large sporulating bacilli in "Bamboo fishing" pole/rod arrangement
e) None of the above

92. The disease resulting from ingestion of live C. botulinium which then multiply in the
gut and produces toxins
a) Classical botulism
b) Canned-good poisoning
c) Floppy Baby syndrome
d) Wound botulism
93. Cycloserine-Cefoxitin-Fructose Agar (CCFA) is used for the cultivation of what
Clostridium species?
a) C. perfringes
b) C. botulinium
c) C. tetani
d) C. difficile

Ratio:
CLOSTRIDIUM SPP.
Obligate anaerobes
Infections are usually POLYMICROBIAL
C. perfringes
The species most frequently involved in GAS GANGRENE
DOUBLE-ZONE of hemolysis on BAP
Positive NAGLERS reaction on EGG YOLK AGAR
Boxcar appearance on Gram stain
Lipase negative, lecithinase positive, indole negative, REVERSE CAMP positive
C. tetani
Produces TETANOSPASMIN that causes TETANUS
Snowshoe or tennis racket appearance on Gram stain
Lecithinase negative, lipase positive, urease negative
C. botulinium
Produces BOLINIUM TOXIN associated with CANNED GOOD POISONING
C. difficile
The major cause of nosocomial/antibiotic-associated diarrhea
Primary pathogen responsible for PSEUDOMEMBRANOUS COLITIS
Colonies have horse-barn odor
Selective media: CCFA ; yellow, ground glass appearing colonies

OTHER MEDICALLY IMPORTANT ANAEROBES

Fusobacterium necrophorum - associates with Lemiers syndrome
Finegoldia magnus and Parvimonas micra - most commonly isolated anaerobic
gram-positive cocci
Veilonella spp. - most commonly isolated gram-negative cocci
Propionibacterium acnes - associated with cases of ENDOPHTHALMITIS after
cataract surgery and ventricular shunt infection in patients with hydrocephalus
Actinomyces spp. (esp. A. israelii) - involved in LUNG ABSCESS, brain abscess,
skin and soft tissue infections, mycetoma

94. Infection with this bacterium is associated with pseudomembrane formation in the
oropharynx "bull's neck" leading to obstructions
a) Clostridium difficile
b) Corynebacterium diptheriae
c) Corynebacterium jeikeium
d) Corynebacterium pseudodipthericum

95. The toxigenic strains of C. diphtheriae can be detected in vitro using

a) Animal inoculation test
b) Anton's test
c) ELEK's test
d) Schick's test

96. Considered as the most effective means of decontaminating and digesting sputum
specimens for the detection of MTB?
a) N-acetyl L-cysteine (NALC) and 2-4% NaOH
b) Trisodium phosphate and sbenzalkonium chloride (Zephiran)
c) Dithiotheritol and NaOH
d) QUATS

Ratio:
NALC-NaOH procedure - MOST COMMON method to liquefy, decontaminate and
concentrate specimens for detection of MTB
97. Which of the following culture media M. tuberculosis is NOT egg-based?
a) Lowenstein-Jensen
b) American Thoracic Society medium
c) Petragnani
d) Middlebrook 7H10 and 7H11

Ratio:
Two types of culture media for mycobacterial culture
Egg-based: Lowenstein-Jensen
Agar-based: Middlebrook 7H10, 7H11
Cultures are incubated at 37C, 5-10% CO 2, MTB colonies appear are colorless,
CAULIFLOWER-LIKE
Specimens from cutaneous sites - incubate at 30C, mycobacteria that causes skin
lesions grow better at LOWER TEMP
All cultures should be examined for at least 6 WEEKS
NAT - MOST RAPID method for diagnosis of pulmonary TB
MYCOBACTERIA
AEROBIC, NON-MOTILE, ACID-ALCOHOL fast, slightly curved to straight
bacilli
Cell wall contains MYCOLIC ACID

MYCOBACTERIA
CLASSIFICATIO MEMBERS
N
M. tuberculosis Mycobacterium tuberculosis
complex (MTBC) Mycobacterium bovis
M. bovis Bacille Calmette-Gurin
strain (BCG)
Mycobacterium caprae
Mycobacterium pinnipedii
Mycobacterium africanum
Mycobacterium microti
Mycobacterium canettii
Mycobacterium caprae
Nontuberculous Mycobacterium avium complex
Mycobacteria (MAC)
(NTM)
Mycobacterium kansasii
Mycobacterium xenopi
Mycobacterium genavense
Mycobacterium marinum
Mycobacterium heamophilum
Mycobacterium ulcerans
Mycobacterium leprae
COMMENTS
causes respiratory and disseminated
infection among immunocompromised
patients
YELLOW BACILLUS, cross-barred
bacilli shepherds crook in AFB stain
Birds nest colonies
SWIMMING POOL GRANULOMA/Fish
tank granuloma - chronic
granulomatous disease of skin and soft
tissue
Requires hemoglobin or hemin for
growth
Bairnsdale ulcer, Buruli ulcer
LEPROSY/HANSENS DISEASE
Has never been cultivated in vitro
Animal models - MOUSE FOOTPAD
and NINE-BANDED ARMADILLO
Cardinal signs of leprosy - skin lesions,
areas of cutaneous anesthesia and
enlarged peripheral nerves
Two types of leprosy - LEPROMATOUS
LEPROSY (DISSEMINATED) and
TUBERCULOID LEPROSY (LOCAL)
RUNYONS CLASSIFICATION OF NTMS

PHOTOCHROMOGEN pigmented only when

EXPOSED TO LIGHT
SCOTOCHROMOGEN PIGMENTED when grown
in dark and light
NONCHROMOGEN NOT PIGMENTED when
grown in dark or light
RAPID GROWER grows on solid media <7 M. fortuitum group
days M. chelonae/M. abscessus
group
M. smegmatis

Multidrug resistant tuberculosis (MDR-TB) - resistance to RIFAMPIN ALONE or any

TWO other primary antituberculous drugs
Primary drugs for MTB: RIPE Rifampin Isoniazid Pyrazinamide Ethambutol
Secondary drugs: Capreomycin, ethionamide, ethambutol (higher concentration),
amikacin, kanamycin, levofloxacin, ofloxacin, moxifloxacin, isoniazid (higher
concentration), p-aminosalicylic acid, rifabutin, streptomycin

98. Which of the following is FALSE of M. leprae?

a) highly contagious
b) cannot be cultured on cell-free medium
c) may be treated by sulfone dapsone
d) acid-fast stain will reveal the presence of Lepra cells

99. The most important test for differentiating MTB from M. bovis
a) Susceptibility to T2H
b) Arylsulfatase test
c) Niacin test
d) Tween 80 hydrolysis test

Ratio:
Tests to detect infection with MTBC
TUBERCULIN skin test
Skin test reagent: PURIFIED PROTEIN DERIVATIVE (PPD), contains
proteins of the bacilli (+) INDURATION after 48 to 72 hours
Preferred method of skin testing: MANTOUX test - INTRACUTANEOUS
injection of 0.1 mL PPD
False positive: infection with NTM, BCG immunization
Interferon-y release assay - based on detection of interferon-y released by T
cell in response to presence of MTB proteins, preferred test for persons who
received BCG vaccine

100. Which of the following bacteria requires sterol for growth?

a) Chlamydia psitacii
b) Chlamydia trachomatis
c) Mycoplasma pneumoniae
d) Ureaplasma urealyticum