Chapter 16

. An isolate with the appropriate colony and a. Perform an Elek test to determine
microscopic morphology may be suspected to whether the organism produces
be Bacillus anthracis if it is: exotoxin
a. β-Hemolytic on SBA b. Subculture the organism to cystine-
b. Nonmotile tellurite agar and examine for black
c. Catalase-negative colonies
d. Gram-negative, non–spore forming c. Prepare a methylene blue stain and
examine for metachromatic granules
2. An aerobic, gram-positive, spore-forming d. . Gram stain the isolate and observe for
bacillus was isolated from raw vegetables that its pleomorphic morphology
were associated with an outbreak of
gastroenteritis. The organism produced β- 9. Respiratory diphtheria is common in the
hemolysis, was catalase-positive, and was United States, and the natural hosts for C.
motile. The most likely organism is: diphtheriae are humans and birds.
a. Bacillus anthracis a. True
b. Nocardia asteroides b. False
c. Bacillus cereus
d. Tsukamurella spp. 10. True infections with nondiphtheria
Corynebacterium spp., such as C. jeikeium or C.
3. Bacillus cereus is most noted for causing: striatum, are often in immunocompromised
a. Food poisoning patients or patients who have had:
b. Meningitis a. Insertion of hardware or prosthetic devices
c. Sexually transmitted disease b. Coronary artery bypass surgery
d. Urinary tract infections c. Vitamin B12 deficiency
d. A lengthy hospital stay
4. Which forms of infection are caused by
Bacillus anthracis? 11. A _________ test can help to differentiate
a. Injectional which clinically significant Corynebacterium spp.
b. Inhalation recovered from urine samples?
c. Gastrointestinal a. Gelatin hydrolysis; C. ulcerans
d. Cutaneous b. Reverse CAMP; C. pseudotuberculosis
e. All of the above c. Alkaline phosphatase; C. amycolatum
d. Urease; C. urealyticum
5. The functionality of lethal factor requires the
presence of what other protein from Bacillus 12. A newborn girl becomes febrile and will not
anthracis to form an active toxin? feed about an hour after birth. A gram-positive
a. Edema factor rod is recovered from blood cultures from the
b. D-glutamic acid newborn. The isolate has the characteristics
c. cAMP listed. What is the most likely identity of the
d. Protective antigen isolate?Weakly β-hemolytic on SBA Gram-
positive bacilli, no spores observed Catalase-
6. Describe the appearance of spore-forming positiveHydrogen-sulfide-negative Motile at
bacteria seen with the spore stain. room temperature
- In the spore stain, vegetative cells are red, a. Erysipelothrix rhusiopathiae
but the spores stain green b. Listeria monocytogenes
c. Corynebacterium ureilyticum
7. Corynebacterium species often appear as d. Gardnerella vaginalis
______________ on Gram stain.
a. Pleomorphic gram-positive club-shaped 13. A commercial fisherman who had red sores
bacilli that appear in palisades or in V and L on his hands was seen by his physician. Biopsy
formations and culture of one of the lesions was
b. Branching gram-positive bacilli that appear as performed. The culture grew an organism with
fine, intertwining, delicate filaments the characteristics listed. What is the most likely
c. Short, thin gram-positive bacilli that appear in organism?Nonhemolytic on SBA
chains Gram-positive bacilli, no spores observed
d. Large square-ended, gram-positive or gram- Catalase-negativeHydrogen-sulfide-production-
variable bacilli in chains where the ends of the positiveGrowth in gelatin resembled a test-tube
single cells fit snugly together brush
a. Rhodococcus equi
8. The biochemical tests performed on a gram- b. Listeria monocytogenes
positive bacillus were consistent with c. Lactobacillus acidophilus
Corynebacterium diphtheriae. As a definitive d. Erysipelothrix rhusiopathiae
test, the laboratory scientist should now:
14. A 42-year-old man from Guatemala cuts his diarrhea,arthralgia,abdominalpain,malabsorptio
bare feet on thorns while walking. A n, and weight loss of 10 lb over the last month.
subcutaneous abscess develops, and when the A duodenal biopsy is performed, but no
patient is seen by a physician, his foot is infectious agent is recovered on culture media.
swollen. When the wound is pressed by the However, on Gram stain, a gram-positive
physician, purulence is expressed along rod is observed in macrophages in the biopsy
with some soft, white granules. A filamentous tissue. The organism is identified by 16S rRNA
organism that is partially acid-fast is recovered gene sequencing. What is the most
from the granules. This is most likely identity of this organism?
likely an __________ mycetoma caused by a. Bacillus anthracis
____________. b. Nocardia asteroides
a. Actinomycotic, Pseudallescheria boydii c. Tropheryma whipplei
b. Actinomycotic, Nocardia brasiliensis d. Erysipelothrix rhusiopathiae
c. Eumycotic, Madurella mycetomatis
d. Eumycotic, Tsukamurella paurametabola Chapter 17
15. What other organisms can give similar 1. Most Neisseria species are:
clinical and laboratory findings as Listeria a. Oxidase-positive, gram-positive diplococci
monocytogenes? How are these organisms b. Oxidase-positive, gram-negative diplococci
differentiated from L. monocytogenes? c. Oxidase-negative, gram-negative diplococci
- Streptococcus agalactiae (group B d. Oxidase-negative, gram-positive diplococci
Streptococcus) and Enterococcus spp. can
produce clinical laboratory findings similar to 2. Identify the virulence factor of the
those for L. monocytogenes. Initial pathogenic Neisseria species that is responsible
differentiation between L. monocytogenes and for the initial attachment of the organism to
similar microorganisms can be made by the host tissues.
Gram stain, catalase test, and esculin a. Pili
hydrolysis b. Endotoxin
c. Cell membrane proteins
16. A 17-year-old boy presented to an d. Peptidoglycan
emergency department with a history of
multiple episodes of febrile pharyngitis followed 3. Asymptomatic gonococcal infections in
in 10 to 14 days with extensive desquamation of women may result in:
his hands and feet. The reoccurrences have a. Pelvic inflammatory disease
followed several courses of antimicrobial b. Ectopic pregnancy
therapy including amoxicillin and c. Fitz-Hugh–Curtis syndrome
cephalosporins. Rapid group A streptococci d. All of the above
screens and cultures have been consistently
negative for Streptococcus pyogenes. A 4. What is the optimal specimen to collect for
specimen with a request for an alternative the diagnosis of gonorrhea by culture in male
agent was submitted to a reference laboratory. patients?
The results listed were observed. The patient a. Pharyngeal swab
was subsequently treated with erythromycin, b. Rectal swab
and he recovered. What was the etiologic c. Urethral swab
agent? d. Urine
SBA: dry, wrinkled, slightly hemolytic colony, 5. True or False: Direct Gram stain is more
which at 48 hours is a dark spot sunken in the useful in detecting genital
agar Catalase-negative Nitrate-negative gonorrhea in female patients than in male
Reverse-CAMP-test-positive patients.
a. Corynebacterium diphtheriae
b. Arcanobacterium haemolyticum 6. Identify a selective medium that allows the
c. Listeria monocytogenes isolation of N. gonorrhoeae and N.meningitidis.
d. Rhodococcus equi a. SBA
b. CHOC agar
17. A sample is sent to the laboratory from a c. Modified Thayer-Martin
female patient suspected to have bacterial d. All of the above
vaginosis. Which of the following would
be an appropriate medium for this specimen? 7. Identify the test that can be used for
a. Loeffler medium definitive identification of both N. gonorrhoeae
b. HBT agar and N. meningitidis.
c. CBTA a. Gram stain
d. BCYE b. Catalase
c. Oxidase
18. A 57-year-old man from New York City d. Carbohydrate utilization
presents to the emergency department with
8. A limitation of nucleic acid amplification tests
for diagnosing gonorrhea is that the tests: Chapter 18
a. Are rapid and sensitive 1. Are adults immune to Bordetella. pertussis
b. Require viable organisms in the specimen infection? Explain.
c. Are not approved for use in children - No, adults are not immune; however,
d. Require strict transport conditions infections in adults are normally mild or
asymptomatic. Adults serve as reservoirs
9. What are the current recommendations for for disease in children and adolescents
treatment of genital gonorrhea?
- Uncomplicated genital infections with
gonorrhea are treated with ceftriaxone (IM) or 2. What are the clinical samples of choice for
cefixime (PO) or an injectable cephalosporin. the diagnosis of B. pertussis infection?
Azithromycin or doxycycline is also - Nasopharyngeal aspirates and calcium
recommended because patients with alginate swabs are the clinical samples of
gonorrhea are often coinfected with Chlamydia choice for the diagnosis of B. pertussis
trachomatis infection

10. True or False: Military recruits and college 3. What transport media are appropriate for
students have an increased risk of exposure to maximum recovery of B. pertussis?
N. meningitidis. - Amies, 1% casein hydrolysate, and Regan-
Lowe with cephalexin transport media are the
11. Entrance of N. meningitidis into the most appropriate for themaximum recovery of
bloodstream may lead to: B. pertussis
a. Meningococcemia
b. Meningitis 4. Which method is preferred for the detection
c. Waterhouse-Friderichsen syndrome of Bordetella in nasopharyngeal smears?
d. All of the above - Direct fluorescent antibody testing is the
preferred method for the detection of
12. True or False: N. meningitidis can be Bordetella in nasopharyngeal smears.
recovered from CSF, blood, naspopharyngeal The PCR assay is the most recent accepted
swabs, and joint fluids. methodology for the rapid detection of B.
pertussis.
13. Which organism is an opportunistic
pathogen associated with otitis media and 5. Describe the colony morphology of B.
sinusitis in children? pertussis on charcoal agar supplemented with
a. N. lactamica horse blood after 72 hours incubation.
b. N. meningitidis - On charcoal–horse blood, young colonies of
c. N. sicca B. pertussis are smooth, glistening, and silver,
d. M. catarrhalis resembling mercury droplets.
Colonies turn whitish-gray as they age
14. Describe the colony morphology of M.
catarrhalis, and explain how it is identified in 6. How many immunizations are required for
the laboratory. protection against pertussis in children?
- Moraxella catarrhalis will grow on SBA and - Six doses of vaccination are required for
CHOC agar,producing smooth, opaque, gray to protection against pertussis, four in the initial
white colonies. The term hockey puck has been infant series and two boosters(before the age
used to describe the colony because it remains of 6 years and in adolescence). However,
intact when pushed across the platewith a immunity is not lifelong.
loop. Older colonies may have a wagon wheel
appearance. Identification tests will show that 7. Compare the diseases caused by B. pertussis
M. catarrhalis is oxidase- and catalase- and B. parapertussis.
positive. The organism is asaccharolytic, and it - Both species cause pertussis in children;
may be differentiated from Neisseria spp. by however, pertussis caused by B. parapertussis
positive DNase or butyrate esterase tends to be milder.
reactions.Several of the commercial multitest
systems for the identification of Neisseria 8. Is serology a good method to identify and
species also identify M. catarrhali respond to pertussis outbreaks in real time?
- No, serologic testing is not widely available,
15. The ONPG test is used to distinguish but it can be used as a retrospective
between N. meningitidisand: epidemiologic tool.
a. N. gonorrhoeae
b. M. catarrhalis 9. The antimicrobial agent of choice for the
c. N. lactamica treatment of pertussis is:
d. N. mucosa a. Ampicillin
b. Amoxicillin bacteria and algae, and ability to multiply
c. Erythromycin within free-living protozoanare factors
d. Penicillin contributing to human infections caused by
Legionella spp.
10. B. bronchiseptica is considered normal oral
biota in: 8. What presumptive identification methods are
a. Humans currently used to identify Legionella spp. in
b. Cats culture?
c. Cows - Presumptive identification methods include
d. Rats Gram staining a suspicious colony growing only
on BCYE medium and finding thin, faintly-
1. What three bacterial agents are the main staining gram-negative rods that may show
causes of communityacquired, atypical size variation, from 2 to 20 µm in length. Also,
pneumonia? testing for L-cysteine by subculturing to BCYE
- Mycoplasma pneumoniae, Chlamydophila and SBA should be performed. Smears
(Chlamydia) pneumoniae, and Legionella prepared from colonies that require L-cysteine
pneumophila are the three most common for growth are tested with DFA conjugates
causes of community-acquired atypical
pneumonia
9. What are common phenotypic characteristics
2. What risk factors contribute to the more of Legionella?
severe form of legionellosis? - Legionella spp. are pleomorphic, weakly
- Travel, age (older adults), smoking, alcohol staining, gramnegative rods that are
consumption,and an immunocompromised approximately 1 to 2 µm by 0.5 µm
state are risk factors that contribute to severe in size but can elongate. On BCYE medium,
infections caused by Legionella spp. colonies appear grayish-white or blue-green,
convex, and glistening, measuring
3. What environmental factors contribute to approximately 2 to 4 mm in diameter
infection caused by Legionella spp.? Young colonies have a ground glass
- Crowded conditions and warm, humid, appearance and are lightgray and granular,
environmental sources contribute to infections whereas the periphery appears pink and/or
by Legionella light blue, or with bottle green bands.
Illumination with a
4. What are the advantages and disadvantages long-wave ultraviolet light (366 nm) can show
of DFA testing for Legionella spp.? differencesin colony autofluorescence
-Advantages of DFA testing include rapid,
moderately sensitive, and specific results. 10. Besides respiratory tract specimens, what
Disadvantages are having trained clinical specimen is
personnel for rapid tests being required on all useful for the sensitive detection of Legionella
shifts, highcost, potential false-positives antigen?
because of cross-reactivity, and requirement a. Blood
for culture confirmation when a test is b. Stool
negative c. Urine
d. CSF
5. What is the culture medium of choice for the
recovery of Legionella spp.? 1. Describe the appearance of H. influenzae
- Buffered charcoal yeast extract (BCYE) with the X and V strip test.
agar is the preferred medium for the recovery - H. influenzae will grow only where both X and
of Legionella spp. V factors are present; therefore, the bacteria
will grow between the two strips where the
6. What is the best nonrespiratory specimen for two factors have diffused and around the XV
rapid detection of Legionella? strip
- Urine for the urine antigen test is the best
nonrespiratory specimen for the detection of 2. The porphyrin test for H. influenzae would be
Legionella spp __________ because the organism __________
biosynthesize heme. The fluorescence result of
7. What factors of Legionella can contribute to the test would be __________.
the colonization of human-made water a. Negative; cannot; negative
supplies? b. Positive; cannot; positive
- Chlorine tolerance below 2 to 3 mg/L, ability c. Negative; can; positive
to grow at 20° to 43° C and survive for varying d. Positive; can; negative
periods at 40° to 60° C, capability to adhere to
components of piped water systems, ability to
survive in the presence of environmental
3. Infections caused by β-lactamase–positive H. c. C. hominis
influenzae should be treated with which of the d. Capnocytophaga spp.
following?
a. Ampicillin 8. An isolate from a wound infection is oxidase-,
b. Penicillin catalase-, ornithine decarboxylase-, indole-,
c. Either of the above and urease-positive. After 48 hours of
d. None of the above incubation at 37°C, growth on SBA was
described as mucoid colonies exhibiting a
4. H. ducreyi is one of the most fastidious greenish brown halo. A MAC agar plate shows
species of Haemophilus. Describe the optimal no growth. What organism is the mostly likely
growth conditions for the recovery of this cause of the infection?
organism. - The most likely identification is Pasteurella
- H. ducreyi is fastidious and requires enriched multocida. This microorganism is noted for
media for growth. GC agar supplemented with causing skin infections following cat and dog
1% hemoglobin, 5% fetal calf serum, 1% bites. P. multocida, the most common
IsoVitaleX, and 3 mg/L of vancomycin Pasteurella spp. isolated, is an oxidase-
is recommended. The plates need to be negative bacterium that grows on SBA but not
incubated in an atmosphere of increased on MacConkey agar.
humidity and CO2 (5% to 10%) and at a
temperature of about 32° C 9. Which microbiologic tests are most useful in
differentiating B. melitensis from H. influenzae?
5. Compare the pathogenesis of H. aegyptius - Brucella melitensis will grow on SBA and does
with H. influenza biogroup aegyptius. not require X or V factor, whereas H.
- Both H. aegyptius and H. influenzae biogroup influenzae will not grow on SBA and does
aegyptius are noted for causing conjunctivitis. require X and V factors
H. influenzae biogroup
aegyptius, however, is associated with a more
invasive disease known as Brazilian purpuric 10. A patient is complaining of a painful cervical
fever (BPF) characterized by conjunctivitis, high lymph node following a case of pharyngitis.
fever, vomiting, petechiae, purpura, Further investigation reveals that the patient
septicemia, and shock consumed a medium-cooked wild rabbit in a
restaurant in Germany 2 months earlier. What
6. A cervical culture for possible gonococcal is the most likely cause of the infection
infection is sent to the microbiology laboratory. - Francisella tularensis is the most likely
After 24 hours of incubation, the modified causative agent. The primary reservoirs for F.
Thayer-Martin plate has small opaque colonies tularensis are rabbits. Although some
that adhere slightly to the medium. Microscopic infections are acquired by ingestion, it is more
examination reveals gramnegative coccobacilli, common to find ulceroglandular infections
many with square ends. The organism ferments following direct contact with rabbits.
glucose and is superoxol- and catalase-negative.
The most Chapter 19
likely identification is: 1. What are the three general characteristics a
a. N. gonorrhoeae gram-negative bacillus must possess to belong
b. K. denitrificans to the family Enterobacteriaceae with a few
c. Moraxella catarrhalis exceptions?
d. H. ducreyi - Most members of the family
Enterobacteriaceae are able to ferment
glucose, are oxidase- negative (except for
7. A 52-year-old man who had recently received Plesiomonas shigelloides), and are able to
a kidney transplant was admitted to the hospital reduce nitrate to nitrite(except for
with a low-grade fever, a heart murmur, and Photorhabdus spp. and Xenorhabdus
neutropenia. He had a history of periodontal
disease and recently had two teeth extracted. 2. Match the Shigella spp. with the
Blood cultures were positive after 48 hours. The corresponding group antigen:
isolate grew on CHOC A, B, C, and D.
agar and SBA in 5% CO2. The colonies were a. S. sonnei (D)
nonhemolytic, slightly adhered to the surface of b. S. boydii ( C )
the media, and had a slight yellow appearance c. S. dysenteriae ( A )
when removed. The isolate was catalase-, d. S. flexneri ( B )
indole-, and oxidase-negative. Microscopic
morphology indicated gram-negative fusiform 3. Which of the following test results is most
bacilli. The most probable identification is: helpful in categorizing an isolate as a member
a. A. aphrophilus of the tribe Proteeae?
b. K. kingae a. Positive Voges-Proskauer
b. Positive urea d. Enterobacter cloacae
c. Positive phenylalanine deaminase
d. Positive lactose fermentation 10. An enteric organism that is acquired by
eating improperly prepared and cooked or
4. The causative agent of plague is: preserved contaminated food and produces
a. Yersinia pestis dysentery is:
b. Klebsiella rhinoscleromatis a. P. vulgaris
c. Citrobacter freundii b. Y. enterocolitica
d. Serratia marcescens c. S. marcescens
d. Shigella spp.
5. A 47-year-old patient who had just returned
from Mexico was admitted to the hospital with Chapter 20
a 3-day history of vomiting and diarrhea, 1. A gram-negative bacillus isolated from a stool
without fever, and no fecal leukocytes were specimen produces clear colonies on
found in the stool. When he was admitted to MacConkey agar and yellow colonies on
the hospital, a stool culture grew an organism thiosulfate citrate bile salt sucrose medium. The
identified as Escherichia coli. Which of the isolate is subcultured to a sheep blood agar
following strains is the most likely cause of the plate with an O/129 disk. The isolate is sensitive
infection? to O/129 and is oxidase-positive. You should
a. EPEC suspect:
b. ETEC a. Vibrio parahaemolyticus
c. EHEC b. Vibrio cholerae
d. EIEC c. Plesiomonas
d. Aeromonas
6. A gram-negative, oxidase-negative
coccobacillus was isolated from the 2. Which of the following Vibrio spp. would you
cerebrospinal fluid of an infant in the newborn expect to be most likely isolated from a blood
nursery. The organism produced dark pink culture?
colonies on MAC agar and had the following a. V. cholerae
biochemical results: triple sugar iron, acid over b. V. parahaemolyticus
acid with gas; phenylalanine deaminase- c. V. vulnificus
negative; sulfide-indolemotility agar, H2S- d. V. alginolyticus
negative, indole-positive, and motile;
ureasenegative; and citrate-negative. The most 3. Which of the following genera is typically
probable identity of this microaerophilic?
organism is: a. Helicobacter
a. Escherichia coli b. Aeromonas
b. Enterobacter aerogenes c. Plesiomonas
c. Klebsiella pneumoniae d. Vibrio
d. Serratia marcescens
4. Campylobacter jejuni is most noted for
7. What organism is often associated with lobar causing:
pneumonia in elderly hospitalized patients? a. Wounds
a. Shigella spp. b. Septicemia
b. Proteus vulgaris c. Gastric ulcers
c. Escherichia coli d. Gastroenteritis
d. Klebsiella pneumoniae
5. Which of the following is a risk factor for
8. The most common cause of community- acquiring V. alginolyticus infection?
acquired UTIs is: a. Farming
a. Klebsiella pneumoniae b. Hunting
b. Escherichia coli c. Fishing or swimming in ocean water
c. Providencia stuartii d. Drinking unpasteurized milk
d. Citrobacter freundii
6. An oxidase-positive, indole-positive, β-
hemolytic, gram-negative bacillus resistant to
O/129, cannot grow in 6% NaCl broth, and
9. Which organism is an opportunistic pathogen is a non–lactose fermenter on MacConkey agar
that causes wound and urinary tract infections is isolated from an adult stool culture. You
and may cause the production of should suspect:
kidney stones? a. Aeromonas hydrophila
a. Yersinia enterocolitica b. Aeromonas caviae
b. Citrobacter freundii c. Plesiomonas shigelloides
c. Proteus mirabilis d. Vibrio parahaemolyticus
 d. Clostridium botulinum
7. Darting motility is a characteristic of: e. Actinomyces spp
a. Aeromonas .
b. Campylobacter 2. An organism that can live and grow in
c. V. cholerae O1 reduced concentrations
d. V. cholerae non-O1 of oxygen but prefers an anaerobic
environment is known as a(n):
8. Which of the following tests is most helpful in a. Capnophile
differentiating C. jejuni from the other b. Obligate anaerobe
Campylobacter spp.? c. Facultative anaerobe
a. Nitrate reduction d. Aerotolerant anaerobe
b. Urease activity
c. Hippurate hydrolysis 3. Some anaerobes are particularly susceptible
d. Susceptibility to nalidixic acid to oxygen because
they lack the enzyme:
9. When attempting to recover enteric a. Amylase
Campylobacter spp., which specimen, media, b. β-Lactamase
and incubation conditions should be used? c. Superoxide dismutase
- Appropriate specimens for the isolation of d. Glucose-6-phosphate dehydrogenase
enteric campylobacters are stool samples and
rectal swabs; stool samples are preferred. Two 4. Endogenous anaerobes least likely to be
categories of media are available for isolation, involved in cases of
blood-based and charcoal-based media. A bacteremia are:
commonlyused blood-based medium is a. Bacteroides
CAMPY-BAP. This is a Brucella agar–based b. Eubacterium
medium, with 10% sheep red blood cells and a c. Fusobacterium
combination of antimicrobials. Charcoal d. Peptostreptococcus
cefoperazone deoxycholate agar is an
alternative. The addition of antimicrobial 5. Which of the following specimens would be
agents and incubation of the plates at 42° C unacceptable for anaerobic culture?
inhibits normal fecal biota. Because the a. Aspirated pus
campylobacters requireoxygen at a b. Cerebrospinal fluid
concentration less than room air, they must c. Tissue from biopsy
beincubated in a microaerophilic atmosphere. d. Urethral swab

10. What nonculture methods are used to 6. A gram-positive bacillus was isolated from a
diagnosis Helicobacter pylori infections? wound specimen and had the following
- The most commonly used nonculture method characteristics: double zone of β-hemolysis,
for the diagnosis of H. pylori is the noninvasive lecithinase-positive, lipase-negative, indole-
13C- or 14C-labeled ureambreath test. The negative. What is the most likely identification
patient receives an oral dose of labeled urea. of this organism?
Urease activity by H. pylori results in the a. Clostridium perfringens
formation of radioactive-labeled CO2, which is b. Clostridium ramosum
absorbed into the bloodstream and then c. Clostridium septicum
exhaled. Other nonculture methods include d. Clostridium tetani
microscopic examination of stained gastric
tissue, direct fecal antigen detection, 7. An anaerobic, pleomorphic, gram-negative
polymerase chain reaction assay, and bacilli was recovered from a liver abscess. The
determining urease activity of gastric biopsy special potency antimicrobial disks
material. demonstrated that the organism was
vancomycin-resistant, and colistin- and
kanamycin-sensitive. Other results were as
chapter 22 follows: chartreuse fluorescence, spot indole–
. Match the following infectious diseases with positive, and lipase-positive.
their associated What is the most likely identification of the
causative organism: organism?
___b__ Myonecrosis a. Fusobacterium mortiferum
___c__ Tetanus b. Fusobacterium necrophorum
___d__ Botulism c. Fusobacterium nucleatum
___a__ Pseudomembranous colitis d. Fusobacterium varium
___e__ Actinomycosis Indicate whether the following statements are
a. Clostridium difficile
b. Clostridium perfringens true or false:
c. Clostridium tetani
__f__ 8. Exogenous anaerobes more commonly
cause infectious diseases than endogenous 7. What are the stages of a Treponema pallidum
anaerobes. subsp. Pallidum infection? Is the final stage
__f__ 9. Clostridium spp. are especially easy to usually seen in developed countries?
identify in Gramstained smears of clinical - Primary, secondary, and tertiary are the three
specimens because they always appear as gram- stages ofsyphilis. In the United States, the
positive rods with terminal or subterminal tertiary stage of thedisease is not often seen
spores. because most patients are adequately treated
__t__ 10. Failure to isolate fusiform gram- with antimicrobial agents before the
negative organisms that were observed on a tertiary stage is reached.
Gram-stained smear of a clinical specimen could
be an indication that a problem exists with the 8. Where are most cases of leptospiroses
primary medium used for the isolation of contracted within the United States, and why is
anaerobes or the system being used for this important when considering the typical
anaerobic incubation of primary plates. incubation period of the infection?
__t__ 11. Large, dark colonies (>1 mm) growing - Most cases are contracted in Hawaii.
on a BBE plate at 24 hours can presumptively be Because the incubation period is typically 10 to
called a member of the Bacteroides fragilis 12 days, it is likely that visitors to Hawaii can
group. become infected but not show symptoms until
__t__ 12. A pleomorphic gram-positive bacillus they return home.
that is spot indole– and catalase-positive can be 9. Compare the difference(s) between
presumptively identified as Propionibacterium treponemal and nontreponemal tests for
acnes. syphilis.
- The treponemal tests for syphilis detect
Chapter 23 antibodies specific for treponemal antigens.
1. What are the general characteristics of The nontreponemal tests are nonspecific and
spirochetes? detect reaginic antibodies that develop against
- he spirochetes are slender, flexuous, helically lipids released from damaged cells
shaped, unicellular bacteria ranging from 0.1
to 0.5 µm wide and from 5 to 20 µm long, with 10. What is the recommended methodology for
one or more complete turns in the helix. They laboratory diagnosis of Lyme borreliosis?
differ from other bacteria in that they have a - The first tier is an immunofluorescent
flexible cell wall around which several fibrils antibody (IFA) orenzyme immunoassay (EIA)
are wound. screen. Positive or equivocal results are
confirmed with Western blot, which is the
second tier.
2. What risk factors are associated with Borrelia
spp. endemic relapsing fever infection?
- Geographic location, season, outdoor Chapter 24
exposure, and a history of tick bites are risk 1.What organisms should be considered as
factors for endemic relapsing fever. possible causes of neonatal conjunctivitis?
- The two most common causes of
3. Which tickborne species of Borrelia is neonatal conjunctivitis are Neisseria
associated with a skin rash or lesion? gonorrhoeae and Chlamydia trachomatis. The
- B. burgdorferi prophylactic use of erythromycin eye drops has
helped control this serious disease.
4. What is the significance on infectious disease
transmission of finding partially engorged ticks 2. What stains should be performed on the
attached to the skin? discharge or conjunctival scraping for
- Pathogen transmission is more probable the microscopic examination?
longer the vector is attached - Although the Giemsa stain is easy to perform
on eye scrapings, it is not very sensitive for
5. What is the test of choice for the laboratory detecting C. trachomatis andrequires
diagnosis of relapsing fever borreliosis? expertise. A direct fluorescent antibody stain is
- Peripheral blood smear stained with Giemsa more reliable. Alternatively, an antigen
stain is the test of choice for the laboratory detection assay or nucleicacid amplification
diagnosis of relapsing fever borreliosis. test could be performed

6. Name the four strains of the genus 3. For the infant described in the Case in Point,
Treponema that are pathogenic for humans. what other clinical conditions could be due to
- T. pallidum subsp. pallidum (syphilis), T. the causative organisms?
pallidum subsp. pertenue (yaws), T. carateum - The infant in the Case in Point was infected
(pinta), and T. pallidum subsp. endemicum with C. trachomatis. This organism can cause a
(endemic syphilis or bejel)
variety of infections in neonates, including develop within a phagosome. C. burnetii forms
pneumonia, pharyngeal and enteric infections spores,whereas Rickettsia spp. do not. In
addition, although C. burnetii is known to
infect more than 12 genera of ticks and other
4. What STD is caused by Chlamydia arthropods, it is generally not transmitted by
trachomatis serotypes L1, L2, L2a, L2b, and L3? arthropods
- .Lymphogranuloma venereum (LGV)
Chapter 25
5. How does lymphogranuloma venereum differ 1. From what source did the infant described in
from other STDs caused by C. trachomatis? the Case in Point likely acquire the infection?
- LGV differs from other diseases caused by C. - The infant in the Case in Point most likely
trachomatis because the serovars causing this acquired the infection when passing through
disease are more invasive. These serovars are the birth canal; however,studies have also
able to survive inside mononuclear cells and demonstrated neonatal infections following
are carried into the lymphoid tissue, where cesarean deliveries.
they produce a strong inflammatory response
2. Would routine prenatal culture of the mother
6. With what types of infections are have yielded this organism?
Chlamydophila pneumoniae associated? - Because of their fastidious nature,
- Chlamydophila pneumoniae is thought to be routine prenatal cultures would not have
a commoncause of pharyngitis and detected most mycoplasma. M. hominis will
pneumonia. This organism has been linked to grow on routine media, but it forms pinpoint
atherosclerosis and coronary heart disease colonies after 48 hours of incubation that could
be easily missed.
7. What is psittacosis or ornithosis?
- Psittacosis is an infection caused by 3. Why was the Gram stain negative?
Chlamydophila psittaci; it typically manifests as - The mollicutes lack a cell wall; therefore, they
pneumonia. Because of the highly contagious do not Gram stain
nature of C. psittaci, cultures are not
recommended. Diagnosis is best made by 4. How does primary atypical pneumonia
detecting antibodies tothe bacterial antigens caused by Mycoplasma pneumoniae differ from
pneumonia caused by Streptococcus
8. What is the most common laboratory method pneumoniae?
used to diagnose rickettsial diseases? Explain. - Primary atypical pneumonia is milder than
- Serologic assays are the most commonly used pneumococcal pneumonia and more often
testingmethods for the diagnosis of rickettsial seen in young adults compared with older
diseases. Unfortunately, antibodies are adults. In addition, unlike pneumococcal
generally detected in convalescent specimens pneumonia, primary atypical pneumonia does
too late to affect treatment. The Weil-Felix test not have a seasonal incidence. The numbers of
is a nonspecific agglutination assay those infected do not increase in the winter.
infrequently used in theUnited States that uses
antigens from various strains of Proteus. The 5. Name the four common species of mollicutes
immunofluorescent antibody test is the associated with the genitourinary tracts of
methodology most commonly used. humans.
- The four species of mollicutes associated with
9. What cells do the Ehrlichia and Anaplasma the urogenital tract of humans are
species typically infect Mycoplasma hominis, M. genitalium,
in humans? Ureaplasma urealyticum, and U. parvum
- . Ehrlichia spp. generally infect monocytes
and macrophages. 6. What special stain is used on suspected
They prevent phagolysosome formation and colonies of Mycoplasma?
survive withinphagosomes. The Anaplasma are - Dienes or methylene blue stains are used to
unusual in that they preferentially infect stain suspected mycoplasmal colonies
granulocytes. They also prevent
phagosomelysosome fusion and survive within 7. What culture media are used to isolate
phagosomes Mycoplasma pneumoniae, M. hominis, and
Ureaplasma urealyticum?
10. How does Coxiella burnetii differ from the - A7 and A8 are selective and differential media
Rickettsia spp.? for the isolation of M. hominis and U.
- Coxiella burnetii is an obligate intracellular urealyticum. In addition,Shepard’s 10B
parasite that develops within the arginine broth can also be used. M. hominis
phagolysosome of infected cells. Rickettsia and M. pneumoniae can be grown on SP4
spp. prevent phagolysosome formation and broth, if arginine is added for the latter. M.
hominis is the least fastidious of the mollicutes
and will grow on sheep blood and chocolate the harsh environment of the alveolar
agars. macrophage. This organism has evolved
effective mechanisms to survive most
8. Why is cold agglutinin titer not a useful macrophage defense activities and
diagnostic tool for M. pneumoniae? infection, once established, can lead to the
- The cold agglutinin test detects antibodies formation of lung granulomas. The pathogenic
that agglutinate red blood cells incubated at 4° mycobacteria are slow growers, which also
C. Historically, the test has been used to aid in lends to drug resistance. Antimicrobial agents
the diagnosis of primary atypical pneumonia are more active against rapidly growing
caused by M. pneumoniae. However, it is not bacteria. The American Thoracic Society,
very sensitive (only about 50% of the patients Centers for Disease Control and Prevention,
infected are positive) or specific and International Union Against Tuberculosis
9. What current serologic assays are available to and Lung Disease recommend a 6-month
demonstrate M. pneumoniae antibodies? regimen of isoniazid and rifampin,
- Complement fixation assays had been supplemented by pyrazinamide for the first
the primary serologic method to detect anti– 2 months, for the treatment of tuberculosis.
M. pneumoniae antibodies. However, because Multidrugresistant tuberculosis, defined as an
of technical problems, enzyme immunoassays isolate resistant to at least isoniazid and
and immunofluorescent antibody methods are rifampin, may be treated with a combination
now more commonly used of antimicrobial agents. The treatment
. regimen may include the administration of
10. Why are the mollicutes universally resistant antimicrobial agents such as isoniazid,
to penicillin? rifampin,pyrazinamide, and ethambutol.
- The mollicutes lack a cell wall and are Longterm treatment is necessary to ensure a
therefore inherently resistant to the β-lactams cure, and the use of
—penicillins and cephalosporins. multiple drugs maintains antibacterial activity
in case thebacteria become resistant to one of
Chapter 26 the drugs
1. Describe the current recommendations for
the identification of Mycobacterium 3. Compare and contrast the different levels of
tuberculosis in the clinical laboratory. mycobacterial laboratory testing, and cite the
- A number of mycobacterial species, reasons why smaller volume laboratories should
saprophytes and potential pathogens, may be consider not performing full identification and
isolated from humans. Historically, susceptibility testing on mycobacterial isolates.
mycobacteria have been identified by growth - Mycobacteriologic services have
characteristics and biochemical testing. More spread to many laboratories. Clinical
recently, molecular biology assays have been laboratory functions that contribute to the
developed. These assays include mycolic diagnosis and management of tuberculosis
acid analysis of bacterial cell walls and DNA have been divided
probe technology and DNA sequencing. into the following three major categories of
Genetic probe technology offers tremendous service offered.
promise in microbial identification at a variety • Level 1: Collection and transport of
of levels—family, genus, species, and specimens, preparation and examination of
subspecies. The most common probe smears for acid-fast bacilli
technology is the commercially available, • Level 2: Procedures of level 1, plus
single-stranded, acridinium ester–labeled DNA isolation and identification of M. tuberculosis
probe for the detection of rRNA (Gen-Probe, • Level 3: All procedures of level 2, plus
San Diego, CA). Probes specific for the M. identification of mycobacteria other than M.
tuberculosis complex (M. tuberculosis, M. tuberculosis The determination of drug
bovis, M. africanum, M. canettii, and M. susceptibility may be performed at level 2 and
microti), M. kansasii, and M. gordonae, and should be performed at level 3. A laboratory
separate probes for M. avium and M. may choose to develop or maintain the skills
intracellulare, are available. Laboratories defined under one of the above levels,
should perform identification according to the depending on the frequency with which
level of service for which they are qualified. All specimens are received for isolation of
isolates should be identified to the species mycobacteria,nature of the clinical community
level. being served, and availability of a specialized
referral service. All laboratories that
2. Explain the reason why mycobacterial perform clinical mycobacteriology should
infections have to be treated for 6 months or participate in recognized proficiency testing
longer and the need to use multiple drugs when programs, and levels of service should be
treating M. tuberculosis infections. established and limited by the quality of
- Slowly growing M. tuberculosis has the performance demonstrated in these
extraordinary ability to persist and replicate in examinations.
 - The mycobacteria are easily spread by the
4. Discuss the methods used to process clinical airborne route; therefore, it is important to
specimens for mycobacterial culture and the avoid aerosol-generating procedures. In
reasons why specimens need to be addition, mycobacterial laboratories should be
decontaminated and digested before culture. under negative air pressure, so that when
- Most clinical specimens contain an doors to the laboratory are opened, air flows
abundance of nonmycobacterial contaminants. inward. Specimen processing should be
Unless an attempt is made to inhibit these conducted in a biological safety cabinet;
usually fast-growing contaminants, they can laboratory scientists must wear laboratory
quickly overgrow the more slowly growing coats, eye protection,
mycobacteria. Organic debris (e.g., tissue, and a respirator (not a surgical mask). When
serum, other proteinaceous material) disposable inoculating loops and needles are
surrounding the organism in the specimen also not used, electric incinerators should be used
must be liquefied so that decontaminating instead of open flames for sterilizing
agents will kill undesirable microbes allowing metal loops and needles. When specimens are
surviving mycobacteria to can gain access to centrifuged, they must be in a screw-capped
the growth media. Mycobacteria are more tube in a secondary screwcapped container.
refractory to harsh chemicals; therefore
chemical digestion and decontamination 7. Which of the following is (are) fluorescent
procedures have been used with success to stain(s) used in the detection of the
enhance the recovery of acid-fast bacteria mycobacteria?
from clinical specimens. NaOH digests and a. Auramine-rhodamine
decontaminates specimens, whereas N- b. Kinyoun’s
acetylL-cysteine effectively digests the c. Ziehl-Neelsen
specimen but does not decontaminate. d. Both b and c

5. With respect to laboratory technique in the 8. A nonpigmented mycobacterium is isolated

isolation and identification of mycobacteria, that reduces nitrate to nitrite and is niacin-
discuss some causes of false-negative positive. You should suspect:
and false-positive results. a. M. kansasii
- Because low levels of mycobacteria organism b. M. xenopi
may be present in clinical samples, meticulous c. M. tuberculosis
care must be practiced during the processing d. M. avium complex
of specimens for the demonstration and
isolation of mycobacterial organisms. False- 9. The causative agent of Hansen disease:
negative results may occur if processing a. Is highly contagious
protocols are not followed appropriately. b. Readily grows on most mycobacterial media
Prolonged decontamination can have a c. Grows best at core body temperature (37°C)
negative effect by d. None of the above
killing the mycobacteria. Improper
centrifugation force may also lead to false- 10. The skin test for tuberculosis:
negative results. If insufficient force is a. Detects antibodies to mycobacterial antigens
applied, the mycobacteria may remain in the b. Detects a cell-mediated immune response to
interface of the processed specimen and can mycobacterial antigens
be inadvertently discarded during processing. c. Uses the bacillus of Calmette-Guérin strain as
Care must be applied during culturing of the antigen source
the specimen. False-positive results may occur d. Both a and b
through carryover from the mouths of
containers used to transfer processing agents,
introduction of the organism from
environmental sources, such as water; or 1. Steps in gram staining
introduction of organisms from aerosols  Primary stain
produced when specimen containers are  Mordant
opened. When making slides, care must be  Decolorization
taken to prevent carryover from one side to  Secondary stain
another. Slides should never come into contact
with one another. Cross-contamination 2. In the gram staining procedure, this
between specimens may occur if instruments function as mordant.
are not properly maintained. -gram iodine

6. Describe important safety considerations for 3. Toxin produced by V. parahaemolyticus

laboratories attempting mycobacterial isolation -
and identification.
4. In the bacterial growth cycle, growth 17. Clinical manifestation seen in patient
ceases because nutrients are exhausted with this disease is “strawberry tongue”
or toxic metabolic product have -
accumulated in which phase?
- 18. The type of hemolytic colonies
produced by S. pneumoniae?
5. An individual who harbors an organism -
and is capable of spreading the
organism to others, but has no 19. Selective medium that allows the
symptoms or signs of disease is called? isolation of N. gonorrhea an N.
- Carrier meningitides
-Modified thyer Martin
6. Typical gram stain appearance of
campylobacter jejuni. 20. A sexually transmitted disease caused
- by c. trachomatis and is characterized
by suppurative inguinal adenitis.
7. Test is used to identify S.
saprophyticus? 21. The optimal specimen to collect for the
- diagnosis of gonorrhea by culture in
male patients.
8. Presumptive identification test for the
detection of H. pylori.
- -

9. Oxygen labile, antigenic toxin, involved 22. This test is the key step in the
in ASO titer test, responsible for sub preliminary identification of V. cholera
surface hemolysis from other vibrios.
- Streptolysin O -

10. Oxygen stable, non antigenic toxin, 23. A solution placed in the eyes of
responsible for surface hemolysis. newborns during vaginal delivery
- Streptolysin s through infected birth canal of
gonococcal infected pregnant woman.
11. This is performed when patients are not -
responding to treatment and assess
susceptibility patterns. IDENTIFY THE ORGANISM
- 24. It has been associated with invasive
infection such as necrotizing fasciitis
12. A multisystemic disease that occurs in -
menstruating women using using a
cloth or re- usable tampons. 25. Streptococcus organism that is not part
- of the lancefield group
-
13. A useful test to differentiate Moraxella
catarrhalis from saprophytic neisserian 26. Coagulase- negative, DNAse-negative,
in respiratory samples. non-hemolytic, novobiocin-resistant.
- -

14. Microbialogical specimens are usually 27. The ability to clot blood plasma is one
incubated for specific length of time of the most reliable laboratory criteria
that is how long? available for the identification this
-24 hours organism.
-
15. This culture media demonstrate the
“green metallic sheen” appearance of E. 28. Coagulase- negative, DNAse-negative,
Coli. novobiocin-sensitive
- -

16. This disease is caused by S. pneumoniae 29. The ONPG test is used to distinguish
that produces rusty or anchovy red between N. meningitides and ___.
sputum. -
-
30. An opportunistic pathogen associated
with otitis media and sinusitis in
children.
-

31. It is the causative agent of plague.

-

32. Known to be a biochemically inert

pathogen.
-

33. Known as the top water bacillus

-

34. Non pathogenic specie of leprospira

-

35. Stained bacterial smear

- Compound light microscope

36. Unstained bacterial cells” the cells are

small, and not detail is needed.
-darkfield microscope

37. Unstained live tissue when it is

desirable to see some intracellular
detail.
- Phase-contrast microscope

38. A sample that emits light when

illuminated with ultraviolet light.
- Fluorescence microscope

39. Intracellular detail of a cell that is 1um

long.
-electron microscope

40. Unstained live cells in which

intracellular structures are shown in
color.
- Differential interference contrast
microscope.