Outline 616
X Aerobic Gram-Positive Bacteria
>• Aerobic Gram-Negative Bacteria
>• Mycobacteria
>• Anaerobic Bacteria
>• Chlamydia, Rickettsia, and Mycoplasma
>• Spirochetes
>• Antimicrobial Agents and Antimicrobial Susceptibility
>• Procedures and Biochemical Identification of Bacteria
Review Questions 666
Answers & Rationales 705
References 752




A. Staphylococci and Similar Microorganisms
1. Staphylococcus aureus
a. Approximately 30% of the population carries S. aureus as resident flora,
primarily in the anterior nares.
b. Isolated from abscesses, wound infections, and carbuncles
c. Causes food poisoning (via enterotoxin), pneumonia, osteomyelitis,
endocarditis, wounds, staphylococcal scalded skin syndrome, etc.
d. Produces six types of enterotoxin and toxic shock syndrome toxin-1
e. Identifying characteristics
1) Gram-positive cocci arranged in clusters
2) Colonies are opaque and smooth. S. aureus grows well on most media
and is usually beta-hemolytic on sheep blood agar (SBA).
3) Catalase and coagulase positive
4) Latex agglutination assay detects clumping factor and protein A on the
surface of 5. aureus.
5) Negative for ability to metabolize the substrate pyrrolidonyl-
oc-naphthylamide (PYR) and ornithine
6) Staphylococci can tolerate the high salt concentration (7.5%) of
mannitol salt agar (MSA).
a) S. aureus ferments mannitol and produces yellow colonies on MSA.
b) Most coagulase-negative Staphylococci do not ferment mannitol and
therefore produce red colonies.
7) Penicillin resistance is due to beta-lactamase production. Methicillin-
resistant 5. aureus (MRSA) is resistant to (3-lactam antibiotics because
of production of altered penicillin-binding proteins. Rare strains of
vancomycin-intermediate S. aureus (VISA) and vancomycin-resistant
S. aureus (VRSA) have been reported. Vancomycin resistance is due to
the Van A operon that alters the target of vancomycin in the cell wall.
VISA occurs following oveiproduction of the target.
2. Coagulase-negative Staphylococci
a. Coagulase-negative Staphylococci are very common skin flora and are
mostly nonpathogenic. However, they can cause disease in
immunosuppressed and neutropenic patients. This group of bacteria causes
urinary tract infections (UTIs) and is associated with infections of
catheters and shunts.
b. Gram-positive cocci arranged in clusters
c. Colonies appear white to gray on blood agar and nonhemolytic.
d. Catalase positive and coagulase negative
e. Commonly encountered species
1) Staphylococcus epidermidis—Most common species of coagulase-
negative Staphylococci, novobiocin susceptible


2) S. saprophyticus—Significant only in UTIs, novobiocin resistant
3) S. lugdunensis—Frequent cause of endocarditis, ferments manitol,
PYR positive, and typically clumps in plasma (slide coagulase) because
of the presence of clumping factor
3. Micrococcus
a. Micrococcus spp. are considered normal flora of the skin and mucous
membranes; they rarely cause infections.
b. On Gram stain, arranged in tetrads and appear larger than Staphylococcus
spp., see Table 6-1 •.
c. Colonies often appear yellow and nonhemolytic on SBA.
B. Streptococcaceae and Similar Microorganisms
1 . General Characteristics
a. Catalase-negative, gram-positive cocci arranged in pairs and chains
b. Can be alpha- or beta-hemolytic, or nonhemolytic on SBA
c. Lancefield grouping is based on a cell wall antigen.
2. Group A Streptococcus (S. pyogenes)
a. Infections are spread by respiratory secretions, and some children may
carry the bacteria in the respiratory tract without illness. However,
S. pyogenes is always considered pathogenic.
b. Infections caused include strep throat (pharyngitis), impetigo, cellulitis,
scarlet fever, pneumonia, otitis media (middle ear infections), and
necrotizing fasciitis.
c. Sequelae include rheumatic fever and post-streptococcal
d. Susceptible to bacitracin (A disk) and PYR positive, but often identified
by serological latex agglutination test
e. Colonies are pinpoint (<1 mm), translucent, and will show a large zone of
f. Virulence factors include
1) Cell wall M protein inhibits phagocytosis, and antibodies to M protein
are protective.


Micrococci Staphylococci
Acid production from glucose under anaerobic conditions o +

Modified oxidasetest + 0

Bacitracin (0.04 unit disk) S R

O = negative, + = positive, R = resistant, S = sensitive

Group B Streptococcus (S. 25% of all females carry the bacteria as normal vaginal flora c. UTIs. Important cause of infections in OB/GYN patients. Isolation of group D streptococci in blood cultures is an indicator of colon cancer. 6) Hyaluronidase hydrolyzes hyaluronic acid. causes the rash seen in scarlet fever. STSS. Streptococcus dysgalactiae subsp. Unlike S. c. Some strains may be nonhemolytic.618 • CHAPTER 6: BACTERIOLOGY 2) Streptococcal pyrogenic exotoxin (Spe A. respiratory tract. Normal flora of the oral cavity. and gastrointestinal (GI) tract mucosa . 4. equisimilis a. neonates acquire infections during birth. S. With early onset infections. b. Strains that produce a hyaluronic acid capsule will not produce hyaluronidase. endocarditis. and abdominal abscesses. e. Group D Streptococcus a. skin infections. round. negative for growth in 6. Normal fecal and oral flora b. CAMP test and hippurate hydrolysis positive. formerly referred to as erythrogenic toxin. resulting in sepsis and meningitis. Colonies are medium-size (>1 mm). and show small zones of beta-hemolysis. flat. agalactiae is resistant to bacitracin. 5. an interstitial barrier. 5) Streptolysin O and streptolysin S lyse erythrocytes. and Spe F). Additionally. Bile-esculin positive. agalactiae) a. platelets. and convex. d. These bacteria are associated with wound infections. rarely beta-hemolytic e. osteomyelitis. d. PYR negative f. These isolates typically express Lancefield group C or G antigens. Colonies are gray to white.5% NaCl. pyogenes and includes pharyngitis. necrotizing fasciitis. and acute rheumatic fever. However. 4) Hyaluronic acid capsule inhibits phagocytosis. Spe B. Viridans streptococci a. creamy. glomerulonephritis. isolates are often identified by serological latex agglutination kits. and pyelonephritis in immunosuppressed patients. S. agalactiae can cause postpartum fever. translucent. and neutrophils. The clinical spectrum of disease resembles 5. PYR negative 6. 3. Many other beta-hemolytic streptococci are also resistant to bacitracin. 3) Streptokinase dissolves clots. and wound infections. Spe C. pneumonia. Alpha-hemolytic or nonhemolytic. as well as endocarditis. facilitating spread of the infection. Normal flora of the gastrointestinal tract of humans and animals b. These toxins act as superantigens interacting with macrophages and T helper cells to stimulate the massive release of cytokines and are associated with Streptococcal toxic shock syndrome (STSS). pyogenes.

2) Umbilicated. Colony morphology 1) Mucoid strains produce a large polysaccharide capsule. 5. also causes wound infections and brain abscesses c. AEROBIC GRAM-POSITIVE BACTERIA • 619 b. Alpha-hemolytic. May enter the blood after dental procedures d. depressed centers caused by autolytic enzymes 3) After 48 hours. salivarius group. Viridans Streptococcus spp. and bile-esculin negative 10. does not grow on bile-esculin medium 7. Gemella spp. ventriculitis. 8. 5. mutans group. pneumoniae is an important cause of community-acquired bacterial pneumonia. S. Leuconostoc a. faecium. and osteomyelitis. including endocarditis. a pediatric vaccine is available that has reduced the number of childhood meningitis cases b. c. however. Vancomycin-resistant enterococci (VRE): Resistance is due to altered peptidoglycan cross-link target. Sputum samples are often rust-colored from blood. bovis group. PYR and leucine aminopeptidase (LAP) positive. Most commonly encountered species are E. Identifying characteristics 1) Bile-esculin positive 2) Positive for growth in 6. brain abscesses. include S. and alpha- hemolytic d. have been associated with a number of infections. Optochin (O or P disks) will inhibit growth (zone of inhibition). . have been linked to osteomyelitis. e. Leuconostoc spp. mitis group. Gemella a. or most commonly nonhemolytic c. The vast majority of VRE are E. Normal upper respiratory tract flora but can cause: 1) Lobar pneumonia in the elderly and alcoholics 2) Otitis media in infants and children 3) Meningitis. faecalis and E. S. b. 9. some strains nonhemolytic f. Major cause of bacterial endocarditis in people with damaged heart valves. postsurgical endophthalmitis. Optochin resistant and insoluble in bile. sanguis group. at 48 hours e. and S. pneumoniae is bile (10% sodium deoxycholate) soluble. beta-. b. colonies become nonviable. f. and bacteremia in neonates. Enterococcus a. and S. Streptococcus pneumoniae a. meningitis. Grows on SBA with 5-10% CO2.5% NaCl 3) PYR positive 4) Express Lancefield group D antigen 5) Can be alpha-. Gram-positive diplococci that are lancet or bullet shaped. lung abscesses. D-Ala-D-Ala to D-Ala-D-Lac or D-Ala-D-Ser. faecium.

Meningitis is a common outcome of infection. b) Nontoxigenic C. monocytogenes demonstrates both umbrella motility in semisolid media at room temperature and end-over-end (tumbling) motility in a wet mount. Causes spontaneous abortion and meningitis in animals (e. C.g. and infections of the central nervous system (CNS). Found in the environment (soil and water). diphtheriae strains are infected with a bacteriophage that contains the gene for the diphtheria toxin. G. pregnant women. c. L. nitrate and catalase positive. monocytogenes grows on most media. and G. elegans. are vancomycin resistant. d. These species are normal flora of the oral cavity and have been associated with endocarditis. 1) Diphtheria is characterized by a pseudomembrane formed by dead cells and exudate at the back of the throat. and irnmunosuppressed patients. and catalase negative. require vitamin B6 (pyridoxal or pyridoxamine) for growth b. diphtheriae strains lack the bacteriophage gene and do not produce the diphtheria toxin. esculin. ophthalmic infections. Formerly referred to as nutritionally variant streptococci. CAMP test. Species include A. sheep) b. 11. Corynebacterium diphtheriae causes diphtheria. and is normal flora of the vagina and intestines in humans c. monocytogenes causes a variety of infections in neonates. and catalase positive 2. Aerobic Non-Spore-Forming Gram-Positive Bacilli 1. c) Urease and pyrazidamidase negative.620 • CHAPTER 6: BACTERIOLOGY b. 2) Bacterial toxin damages major organs. Listeria monocytogenes a. adiacens. resulting in a high death rate. Abiotrophia and Granulicatella a. and PYR. can be very pleomorphic b) Staining with methylene blue will reveal metachromatic granules. which are red to purple intracellular granules. and nonmotile .. colonies are small and white with a narrow zone of beta-hemolysis. The Leuconostoc spp. 3) Found only in humans 4) Identifying characteristics a) Gram stain: Diphtheroid morphology arranged in "picket fences" or "Chinese letters". 2) Closely resembles group B streptococci on SBA 3) L. 4) Hippurate hydrolysis. a) Toxigenic C. defectiva. Identifying characteristics 1) L. Corynebacterium a. LAP.

Corynebacterium pseudotuberculosis also exhibits this phenomenon. d. rhusiopathiae primarily infects animals. iv. although A. e) Media i. bernardiae. white. pyogenes. diphtheriae will grow on SB A as small. haemolyticum has been associated with pharyngitis and wound and tissue infections. Six species of Arcanobacterium have been named. sheep. and A. form small beta- hemolytic colonies on SBA and are catalase negative and nonmotile. E. b. three are clinically significant: A. pyogenes is found on mucous membranes of cattle. 4. ii. Loeffler agar is a nonselective medium that supports growth and enhances pleomorphism and the formation of metachromatic granules. produce small. dry colonies. Erysipelothrix rhusiopathiae a. white to gray colonies. C. Most Corynebacterium spp. wound. Corynebacterium jeikeium 1) C. This bacterium produces phospholipase D. form brown to black colonies with halos from hydrolysis of tellurite. A. 2) Pyrazidamidase positive 3) Resistant to most antimicrobial agents c. and swine and is linked to abscess formation. jeikeium is an important cause of nosocomial infections and produces infections after prosthetic device implants and infections in immunocompromised patients. . c. Cystine-tellurite: Corynebacterium spp. and soft tissue infections in cattle and humans. A. Corynebacterium urealyticum 1) Cause UTIs 2) Is rapid urease positive and grows very slowly 3. A. form black colonies from hydrolysis of tellurite. Tinsdale's agar: Corynebacterium spp. Human infections often result in cellulitis (erysipeloid lesions that can resemble erysipelas caused by Streptococcus pyogenes} but may also present as bacteremia or endocarditis. which inhibits the activity of the Staphylococcus aureus beta-lysin. b. b. The clinically significant Arcanobacterium spp. iii. haemolyticum can be identified by the CAMP inhibition test. Most strains are nonhemolytic. The natural habitat of these organisms has not been confirmed. Humans generally become infected through contact with infected animals (occupational exposure) or rarely by consuming infected meat. Arcanobacterium a. AEROBIC GRAM-POSITIVE BACTERIA • 621 d) The Elek test uses antitoxin to detect toxin production. haemolyticum.

brasiliensis and N. Spores can be central or terminally located. Three clinical forms of anthrax 1) Cutaneous anthrax: Most common form worldwide. It produces large. catalase positive. cereus (food poisoning and wounds) are pathogenic species. Bacterial spores can survive adverse conditions for prolonged periods of time and are frequent contaminants of laboratory cultures. appear fungal-like 2) Partially acid-fast. Aerobic Spore-Forming Gram-Positive Bacilli 1 . in 2001.S. flat colonies. B. branching gram-positive bacilli in chains that produce a beading arrangement. with square ends (boxcar morphology) appearing in chains and singly. nonmotile 3) Requires up to 6 weeks for growth 4) Exudate contains masses of filamentous organisms with pus that resemble sulfur granules. . follows ingestion of spores c. a. b. d. B. Generally found in immunocompromised patients with chronic pulmonary disorders b. other species include N. anthracis (anthrax) and B. e. are nonpathogenic. asteroides is the most clinically relevant species. c.622 • CHAPTERS: BACTERIOLOGY c. N. D. Identifying characteristics 1) Nonmotile. otitidiscaviarum. General Characteristics of Bacillus spp. d. Identifying characteristics 1) Pleomorphic. pleomorphic gram-positive bacilli 2) Catalase negative 3) Hydrogen sulfide positive 5. and only genus identification is necessary.S." spread by inhalation of spores from sheep's wool 3) Gastrointestinal anthrax: Rarest form. g. f. Bacillus spp. Nocardia asteroides a. Cultures form large. Causes anthrax. b. form straight bacilli. Most Bacillus spp. c. a zoonosis that is rare in the U. nonhemolytic colonies with filamentous projections. characterized by necrotic skin lesions called black eschars 2) Pulmonary anthrax: "Wool-sorter's disease. Bacillus anthracis a. sometimes referred to as "Medusa-head" colonies. anthracis is considered a potential bioterrorism agent and was used as such in a series of attacks in the U. 2. The majority of the species will grow on SBA and phenylethyl alcohol (PEA) agar and are catalase positive.

e. flat. and nonmotile g. B.. Preliminary testing by sentinel laboratories: Typical colony morphology. gonococcal arthritis. Neisseria and Similar Microorganisms 1. 2) In females it causes urethral infections and cervicitis. N. Neisseria gonorrhoeae a.g. Kingella. and clinical material must be free of lubricants and spermicides 1) N. gram-positive bacilli with spores. and blood. resulting in gonococcal ophthalmia neonatorum. N. therefore. Includes the genera Neisseria. fastidious. It may also cause opportunistic eye. 1) In males it causes acute urethritis. which is a severe conjunctivitis leading to blindness. cereus is an important cause of food poisoning and occasionally wounds. bone. Many species are normal flora of the upper respiratory tract of animals. antimicrobial eye drops (e. endometritis. Infections can be asymptomatic or produce cervical discharge. The Neisseriaceae are oxidase positive. gonorrhoeae is not normal flora. To prevent newborn conjunctivitis. AEROBIC GRAM-NEGATIVE BACTERIA A. N. gonorrhoeae can also cause pelvic inflammatory disease (PID). cereus and B. anthracis typically does not grow on PEA agar at 24 hours. d. b. beta-hemolytic colonies with irregular edges d. c. gonorrhoeae. Bacillus cereus a. gonorrhoeae can be isolated from the urethra. c. gonorrhoeae often appears intracellular in neutrophils. skin lesions. and dysuria. oropharynx. acute pain. gonorrhoeae is fastidious. Must culture immediately. joints. catalase positive. On direct Gram stain. Motile and resistant to 10 ug of penicillin II. media must be at room temperature before plating. . N. It is fastidious and does not survive long outside the host. salpingitis. Eikenella. They cannot tolerate cold. Family Neisseriaceae a. and grow best in 5-10% CO2 at 37°C. Humans are the only host for N. fever. Colony morphology: Large. subtilis are also common laboratory contaminants. which is characterized by a pus- containing urethral discharge and dysuria and can also cause prostatitis and epididymitis. AEROBIC GRAM-NEGATIVE BACTERIA • 623 e. requiring enriched media such as chocolate. 2. anal canal. and Simmonsiella b. and peritonitis. b. Neonates may be infected during vaginal delivery. cervix. Kidney bean-shaped. c. B. Confirmatory testing is performed by a reference laboratory. and brain infections. erythromycin) are administered to all infants at birth. It does not grow on SBA. f. B. gram-negative diplococci or coccobacilli d. 3.

Many strains are positive for beta-lactamase production. Normal flora Neisseria a. Martin-Lewis. N. K. sicca. K. c. lactamica. catalase. and glistening gray or tan. 3. mucosa. The ability of K. dentifricans is most often associated with endocarditis. gonococci cannot be incubated for prolonged times. meningococcemia. Neisseria meningitidis a. subflava. oralis has been isolated from the mouth of adults. DNase. kingae colonizes the throat of children. gonorrhoeae. smooth. They will not grow on MacConkey agar. Many Neisseria spp. 6. Resembles Neisseria and is normal flora of the upper respiratory tract . Identifying characteristics 1) Superoxol. N. Spread by respiratory droplets and may be normal flora of the nasopharynx b. 4) Because of autolysis. endocarditis. whereas the habitat of K. Causes meningococcal meningitis. N. sucrose. In rare cases. chocolate. Moraxella catarrhalis a. and nitrate negative h. kingae is best isolated from joints and bones in blood culture media. leading to dissemi- nated intravascular coagulation. Identifying characteristics 1) Catalase. denitrificans to reduce nitrates is a key test for its differentiation from N. New York City. d. Many species are not fastidious and will grow on most nutrient agars. meninigitidis will grow on SBA incubated in increased CO2 and produce bluish-gray colonies. and glucose positive 2) Maltose. elongata. e.624 • CHAPTER 6: BACTERIOLOGY 2) Selective media include modified Thayer-Martin. flavescens. f. smooth. and gray to white on chocolate agar. Colonies are flat. Kingella will grow on sheep blood. and other infections. Colonies are flat. these organisms can cause meningitis. Specimens: Cerebrospinal fluid (CSF). and modified Thayer- Martin agars. and N. N. blood. glucose. cinerea. K. sputum. and GC-Lect agars. Infections in adults are generally limited to those who are immunocompromised. are normal flora of the upper respiratory tract. and Waterhouse-Friderichsen syndrome c. and maltose positive 2) DNase and nitrate negative 4. g. denitri- ficans is unknown. oxidase. N. kingae shows a predilection for infections of the bones and joints in children. lactose. K. c. K. N. N. 3) The bacteria require increased CO2 with a humidified atmosphere. b. Species include N. b. oxidase. Member of the family Moraxellaceae b. 5. polysaccharea. and nasopharyngeal swabs d. Kingella a.

and lactose nonfermenters are colorless. 8) Selenite broth: The broth is an enhancement medium for stool cultures. 2) Eosin-methylene blue (EMB) agar: Colonies of lactose fermenters have a dark center. 6) Bismuth sulfite agar: Salmonella Typhi produces black colonies. Most medically important family of gram-negative bacilli b. are oxidase negative. DNase. Diseases include UTIs. Will grow on most nutrient agars e. 3) Hektoen enteric (HE) agar: Lactose and/or sucrose fermenters form yellow/orange colonies. 5) Salmonella-Shigella (SS) agar: Lactose fermenters produce red colonies. Enterobacteriaceae 1. Salmonella. sinusitis. and butyrate esterase positive 2) Asaccharolytic. whereas gram-positive and . with the exception of Plesiomonas. AEROBIC GRAM-NEGATIVE BACTERIA E 62S c. Shigella. Salmonella colonies are colorless with black centers. and Shigella colonies are green. c. Identifying characteristics 1) Catalase. lactose-fermenting colonies are yellow-orange. pneumonia. Causes otitis media. 7) Brilliant green agar: Proteus and Salmonella species produce red/pink colonies. gastroenteritis. Salmonella growth is enhanced. are nitrate and catalase positive and. g. food poisoning. and Yersinia are not normal GI flora. and respiratory tract infections d. E. and lactose-negative colonies are colorless. Enterobacteriaceae are facultative anaerobes. and meningitis e. coli has a dark center and usually shows a green metallic sheen. wound infections. and Shigella have clear colonies. The family exhibits four serological characteristics 1) O (somatic) antigen—A cell wall antigen (heat stable) 2) K (envelope) antigen—Capsular antigen (heat labile) 3) H (flagellar) antigen—Flagellar antigen (heat labile) 4) Vi antigen—Capsular antigen of Salmonella Typhi (heat labile) f. Major cause of nosocomial infections d. Enteric media 1) MacConkey (MAC) agar: Lactose-positive colonies are pink/red. and Shigella colonies are colorless. oxidase. Salmonella produce red colonies with black centers (H2S). nitrate. They ferment glucose. Salmonella colonies are green with black centers (H2S positive). peritonitis. septicemia. 4) Xylose-lysine-desoxycholate (XLD) agar: Colonies of lactose and/or sucrose fermenters are yellow. General family characteristics a. whereas Shigella and most lactose fermenters will not grow. all carbohydrate tests are negative B. Most species are normal flora of the GI tract.

methyl red (MR). meningitis in newborns. Many strains of Stx-producing E. coli (EPEC) causes a watery diarrhea. off-white.626 • CHAPTER 5: BACTERIOLOGY coliform (normal GI flora) bacteria are inhibited. Outbreaks are known to occur in daycare centers and nursing homes. several other serogroups have been reported that produce Stx-1 and/or Stx-2. opaque. and diarrhea 2) Infections are most severe in children and the elderly. coli strains rapidly ferment sorbitol. endocarditis. c) Enteropathogenic E. 2) E. 2-4 mm in diameter. Shigella 1) Causes shigellosis. coli O157:H7 does not metabolize sorbitol. gastroenteritis. leading to kidney failure in young children. and urease negative 4) Enterohemorrhagic E. coli O157:H7 is thought to cause over 80% of all cases of HUS in the U. coli is normal GI flora and a very common clinical isolate. 5) Other strains of E. Confirmatory testing is by detection of Stx-1 and Stx-2. coli (EHEC) causes hemorrhagic colitis and hemolytic uremic syndrome (HUS). appendicitis. and o-nitrophenyl-p-D- galactopyranoside (ONPG) positive f) Voges-Proskauer (VP). 2. characterized by abdominal pain. and usually beta-hemolytic. Escherichia 1) E. citrate. 3) Identifying characteristics a) Triple sugar iron (TSI): Acid over acid (A/A) and H2S negative b) MacConkey agar: Pink/red colonies c) On SBA. coli belong to the serogroup O157:H7. coli causing human intestinal infections a) Enterotoxigenic E. coli (ETEC) produces severe epidemic diarrhea. peritonitis. It is no longer commonly used. coli (EIEC) causes bloody diarrhea by invading the intestinal epithelium. most other E. E. b) Enteroinvasive E. mainly from drinking contaminated water. b. coli causes UTIs. a form of bacterial dysentery. etc. producing pink colonies on SMAC. c) Growth on sorbitol-MacConkey (SMAC) agar: Sorbitol replaces lactose in the medium. gallbladder infections. sprouts. colonies are shiny. a) Acquired by eating undercooked hamburger or other contaminated foods such as apple cider. Important genera a.S. fever. and food poisoning. coli O157:H7 colonies appear colorless on SMAC. E. motility. However. basil. b) The principal virulence factor is Shiga toxin (Stx) 1 and 2. E. d) EMB agar: Green metallic sheen colonies with dark centers e) Indole. .

c) Klebsiella spp. b) S.S. are H2S and MR negative and nonmotile. citrate. motility. the Klebsiella are indole negative. Many infections are nosocomial. 6) Four serogroups based on O antigens a) S. They are found in soil. a sexually transmitted disease b) Identification i. AEROBIC GRAM-NEGATIVE BACTERIA • 627 3) Highly pathogenic. In clinical specimens Wright or Giemsa stained. 7) Identifying characteristics a) TSI: Alkaline over acid (K/A) b) H9S. and a neurotoxin that may result in paralysis. and lactose positive 4) Klebsiella (Calymmatobacterium) granulomatis a) The causative agent of granuloma inguinale. Except for K. d) 5. sonnei is mannitol and ONPG positive. sonnei (serogroup D) produces a mild diarrhea. 2) The most common species isolated is K. boydii (serogroup C) produces a mild diarrhea. c) S. dysenteriae (serogroup A) produces an enterotoxin.flexneri (serogroup B) produces a mild diarrhea. . flexneri from S. It is difficult to biochemically distinguish S. citrate. boydi. dysenteriae is mannitol and ONPG negative. 5. and lactose negative c) MR positive c. boydii is mannitol positive and ONPG negative. urease. pneumoniae. oxytoca and K. which affects the large intestines. ornithinolytica. Less than 50 bacteria can cause disease 4) Causes food poisoning by direct fecal contamination from infected humans 5) Incubation period is between 1 and 7 days. 3) Identifying characteristics a) TSI: A/A with gas b) On MAC agar. It is mannitol positive and ONPG negative. Donovan bodies may be seen. and diabetics and alcoholics are prone to infections. d. Does not Gram stain or grow on laboratory media ii. and dairy products. It is a delayed lactose fermenter. S. Klebsiella 1) Klebsiella spp. typically cause UTIs and pneumonia. S. d) VP. the pink colonies are very mucoid because of capsule production. It is the most common cause of shigellosis in the U. water. Enterobacter 1) The genus includes about 12 species. Donovan bodies are intracellular pleomorphic bipolar staining bacterial cells. VP.

2) There are many animal reservoirs. are occasional clinical isolates that have been linked to respiratory tract infections and wounds and isolated from blood. and E. sakazakii produces a yellow pigment that aids in its presumptive identification.628 • CHAPTERS: BACTERIOLOGY 2) E. e. The majority of human cases of salmonellosis are due to serotypes belonging to the species S. 3) Identifying characteristics a) DNase. 6) Salmonella isolated from stool cultures form lactose-negative and H2S-positive colonies on enteric media. d) E. Most serotypes are pathogenic to humans and cause moderate to severe gastroenteritis. f. Serratio. aerogenes is the second most common species isolated. . Typhi. especially chicken 4) Salmonella Typhi causes typhoid fever. e) E. which is enhanced with room temperature incubation. with over 2400 serotypes.S. and indole negative b) VP and citrate positive c) All species except E. MR. and lactose negative c) Colonies on HE agar are green with black centers. Humans are the only reservoir for 5. cloacae is the most common. and lipase positive. urease. 5) Most human infections in the U. aerogenes is arginine negative and lysine positive. motility. and citrate positive b) Indole. unique among the enterics b) VP and citrate positive c) ONPG positive but a delayed lactose fermenter d) Some strains produce a red pigment. taylorae are lactose positive. enterica. the most severe form of salmonellosis. 5. which includes the serotype S. cloacae is arginine positive and lysine negative. bongori. Enterobacter spp. 1) Causes opportunistic infections in patients undergoing chemotherapy and immunosuppressed patients 2) 5. Typhi. E. 3) Identifying characteristics a) H2S. 7) Identifying characteristics a) H2S. marcescens is the most common clinical isolate. gelatinase. Salmonella 1) The genus now contains two species. enterica and S. are caused by serotypes Salmonella Enteritidis and Salmonella Typhimurium. 3) Transmitted through contaminated water and undercooked food. which is characterized by a septicemia followed by a GI tract infection.

mimbilis are the most common isolates. Proteus vulgaris and P. Aeromonas spp. 5) Presumptive identification of Y. E. Edwardsiella 1) E. or lymph nodes with the following characteristics: pinpoint colonies on SBA after 24 hours incubation. but A/A on TSI. Y. Isolates are ONPG positive but delayed lactose fermenter and sucrose positive. mirabilis and many strains of P. tarda is indole positive and citrate negative. neutral red. 4) All species except Y.g. and ONPG negative 3) Unlike Salmonella. Therefore. rodents) are natural reservoirs. pestis is based on isolation of the bacterium from respiratory tract. AEROBIC GRAM-NEGATIVE BACTERIA m 629 Proteus 1) Four species are recognized. will also grow on this medium and form colonies with pink centers. enterocolitica. tarda is the most common isolate.S. pestis is nonmotile at both temperatures. b) Y. Colonies of Yersinia will ferment mannitol and absorb the dye. 2) Small coccobacilli 3) Cefsulodin-irgasan-novobiocin (CIN) medium is a selective and differential medium for isolation of Y. Yersinia 1) Three pathogenic species of Yersinia a) Y. Confirmatory testing is performed by a regional reference laboratory. 2) Identifying characteristics a) P. d) ONPG and therefore lactose negative e) P. and the bacteria are transmitted by fleas. pestis is considered a potential bioterrorism agent. pestis causes plague. mimbilis are typically H2S positive. c) Y. Growth may be better at 28°C than 35°C. c) All species are urease. it is acquired by drinking contaminated water or by eating contaminated meat. tryptophan deaminase (TDA). Y. vulgaris exhibit swarming motility on SBA. pseudotuberculosis is a rare cause of lymphadenitis in children. enterocolitica causes enterocolitis in humans. colorless on MAC at 18 hours. 2) Resembles Salmonella. oxidase and urease negative. vulgaris and P. producing clear colonies with a pink center. it is endemic to the southwestern U. blood. and catalase positive. b) P. P. and phenylalanine deaminase (PDA) positive. pestis are nonmotile at 37°C but motile at 25°C. .. H2S positive. gram-negative bacilli. vulgaris is indole positive. mirabilis is indole negative. Small animals (e.

Morganella 1) M. I-VIII 2) H. opaque. and/or NAD (V factor). morganii is the only species.e. Members of this group are fastidious (i. k. 2) C.. especially H. The widespread use of the Haemophilus influenzae type b (Hib) vaccine has greatly reduced childhood meningitis and other invasive diseases caused by this serotype. Citrobacter 1) C. Therefore. and tan on chocolate agar h. and citrate positive and VP negative C. Haemophilus will grow near the S. Haemophilus spp. and Kingelld) group. e. coli on MAC but can be differentiated because of being H. i. Nonmotile.freundii is the most common species isolated. and eight biotypes. round. which is released from hemoglobin.S (+) and indole (—). a-f. Clinically relevant species a. which is a heat-labile compound. catalase and oxidase positive 2. Pleomorphic gram-negative coccobacilli ranging from very small to filamentous d. TDA. The most common isolate is probably P. Colony morphology: Smooth. rettgeri. aureus colonies. c. Grows at 35-37°C with 5-10% CO2 and is susceptible to drying and temperature changes g. are considered members of the HACEK (Haemophilus. Cardiobacterium hominis. 2) Indole positive VP and citrate negative 3) PDA and TDA positive 1. Haemophilus influenzae 1) Six capsular serotypes. aphrophilus. can grow around colonies of S. aureus releases NAD. b. Eikenella corrodens. Chocolate agar is routinely used for cultures. Satellitism: Haemophilus spp. do not grow on SBA because of NADase in the agar (NADase inactivates NAD) but will grow on horse or rabbit blood agar. freundii resembles E. Haemophilus spp. indole. Growth requirements include hemin (X factor). aureus growing on an SBA plate. which contains no NADase. forming tiny clear pinpoint colonies.630 • CHAPTER 6: BACTERIOLOGY j. Most species are normal upper respiratory tract flora. influenzae type b was a major cause of meningitis in children. flat. f. 2) PDA. see Table 6-2 •. S. and four have been isolated from humans. Actinobacillus actinomycetemcomitans. require complex nutrients for growth) and important causes of endocarditis. Haemophilis and Similar Organisms 1 . . Providencia 1) There are five species of Providencia. General characteristics a..

a sexually transmitted disease 2) Produces chancroids and buboes (swollen lymph nodes) 3) Chocolate agar with vancomycin is used to inhibit normal flora and contaminants. b. and pneumonia. + *Aminolevulinic acid (ALA) is converted to porphyrin. including blood. + = positive. Haemophilus ducreyi 1) Causes genital ulcers. followed by invasive disease known as Brazilian purpuric fever 2) It resembles H. - H. Otitis media with effusion (middle ear infections) and sinusitis are often caused by nontypeable strains. haemolyticus + + + - H. and ornithine decarboxylase negative. . CSF. - H. parainfluenzae . Haemophilus influenzae biogroup aegyptius 1) Causes a conjunctivitis. aegyptius + + . . 4) H. influenzae isolates should be tested for beta-lactamase. 6) Specific detection of Hib capsular antigen is by latex agglutination. Haemophilus aegyptius 1) Causes pink eye. 5) This species can be isolated from a variety of specimens. those lacking a capsule. + H. AEROBIC GRAM-NEGATIVE BACTERIA • 631 TABLE 6-2 IDENTIFICATION OF HAEMOPHILUS SPP Requires (3-Hemolysis on X Factor V Factor Horse Blood ALA* H. a very contagious conjunctivitis 2) Similar to H. . . including acute sinusitis. sputum. influenzae with the exception of being sucrose positive c.= negative 3) Serotypes other than b are frequent cause of respiratory tract infections. urease positive. influenzae + + - H. influenzae biotype III in that it is indole negative. ducreyi + . . + . + . aphrophilus . and eye swabs. chronic bronchitis. d. paraphrophilus . + H.

General characteristics of NFB 1) Most species are obligate aerobes. It is a common clinical isolate that can infect humans. a yellow pigment that fluoresces. Growth on cetrimide agar enhances fluorescein production. fluoresceins and P. It causes eye (in contact lens wearers) and ear infections and is responsible for "swimmer's ear. food. c. Important pathogen in immunocompromised individuals g. and plants. and swimming pools. 4) TSI: K/no change 5) Grow on SBA but varied growth on MAC 2. 5) Oxidative-fermentative glucose test: This test determines if glucose is broken down oxidatively or fermentatively. Oxidative bacteria . and a few are normal flora of humans b. aeruginosa is resistant to a number of disinfectants and has been responsible for serious nosocomial infections. Found in water. Lower respiratory tract infections in patients with cystic fibrosis (CF) e. Pyocyanin mixes with fluorescein to produce a blue-green color. Nonfermentative Gram-Negative Bacilli 1. plants. animals. one is overlayed with mineral oil to produce anaerobic conditions. Two tubes are used. b. and fish. d. P.632 • CHAPTER 6: BACTERIOLOGY D. irregular colonies with a grapelike odor and metallic sheen on SBA b) (3-hemolytic colonies with a feathery edge on SBA c) Mucoid colonies when isolated from patients with CF d) Pigment: Only P. 3) Most are oxidase positive. All members of the fluorescent group produce fluorescein (pyoverdin). 2) They do not form spores and do not metabolize carbohydrates under anaerobic conditions (fermentation). The other tube contains no mineral oil (aerobic). Causes burn wound infections f. h. P. c. aeruginosa is the most important NFB." which is an external otitis. P. putida. Pseudomonas aeruginosa a. It is especially associated with hospital environments and equipment. Approximately 20% of all gram-negative bacilli isolates are nonfermentative gram-negative bacilli (NFB). Identifying characteristics 1) Oxidase positive 2) Motile 3) Lactose negative 4) Colony morphology a) Large. aeruginosa is a member of the Pseudomonas fluorescent group along with P. soil. whirlpools. aeruginosa produces pyocyanin. General characteristics a. a blue pigment.

Colony morphology: Colorless or yellow on nutrient agar d. Burkholderia cepacia a. Stenotrophomonas maltophilia a. B. UTIs. mules. Acinetobacter spp. coccobacillus found as normal flora of the GI and respiratory tracts b. reduces nitrate to nitrite with gas. maltophila is one of the only nonfermentative. It is one of two clinically encountered NFB that is lysine decarboxylase positive. Identifying characteristics 1) Oxidase negative 2) S. B. pseudomallei is found in soil. Enhanced growth on P. B. baumanii) and UTIs. aeruginosa c. 4) Resistant to most antimicrobials 4. cepacia causes nosocomial infections and is also an important respiratory tract pathogen in patients with CF. Oxidative/fermentative bacteria produce acid in both tubes. Acinetobacter a. Very resistant to antimicrobial agents 3. a disease of humans and animals endemic to Southeast Asia and northern Australia b. and oxidizes glucose. aeruginosa. Burkholderia mallei a. B. Acquired as transient flora from hospitals b. . and infections are acquired through the skin or by inhalation. pseudomallei grows on MacConkey agar. b. The bacterium is also considered a potential bioterrorism agent. particularly among horses. whereas the closed tube is green. including glucose and lactose. 6. Causes melioidosis. Oxidase and lactose positive e. Isolates tend to be multidrug resistant. c. mallei grows on MacConkey agar. AEROBIC GRAM-NEGATIVE BACTERIA • 633 produce acid (yellow color) in the open tube. reduces nitrate to nitrite without gas. c. is oxidase positive and motile. 7. and oxidizes several sugars. are important causes of nosocomial infections (most often A. 5. i. second most common cause to P. B. and donkeys b. a highly contagious disease of livestock. Burkholderia pseudomallei a. Causes pneumonia. mallei can be transmitted to humans by animal contact. Causes glanders. cepacia (PC) agar that inhibits P. Obligate aerobic. gram-negative bacillus that is oxidase negative and maltose positive 3) One of two clinically encountered NFB that is lysine decarboxylase positive. wound infections c. is oxidase variable and nonmotile.

such as A. B. 2. also known as undulant fever b. rabbits. Some species. whereas others are asacchrolytic. beavers. and modified Thayer-Martin agars. Humans may acquire the infection by skinning animals or eating undercooked game. d. eyes. produce acid from glucose (saccharolytic group). c. Isolated from blood and bone marrow h. Brucella has been considered a potential bioterrorism agent. canis. e. Humans usually acquire the infection by drinking contaminated milk or from slaughter house exposure. Francisella a. mis. tularensis causes tularemia and is a potential agent of bioterrorism. F. and squirrels. Isolation in automated blood culture monitoring systems is recommended. Four species infect humans: B. Biosafety level 3 organism g. Miscellaneous Gram-Negative Bacilli 1. Acinetobacter spp. Facultative intracellular parasite f. buffered charcoal yeast extract (BCYE). 3) Colony morphology: Small and grayish 4) Agglutination and direct fluorescent antibody tests are used to confirm the identification. The bacteria are carried by wild animals. d. g. Identifying characteristics 1) They are fastidious organisms but will grow on Brucella. d. tularensis causes skin ulcers at the site of inoculation and can cause infections of the lymph nodes. lungs. and require 10% CO2 in humidified air and 3^ weeks for growth. and GI system. The incubation period is 1-3 weeks. and B. b. Identifying characteristics 1) Faintly staining coccobacilli 2) The medium of choice is glucose-cystine blood agar. B. Causes brucellosis. Intracellular bacteria that resist phagocytosis e. Normal gastrointestinal flora of animals c. f. F. melitenis causes the most severe infections. grow on most media and may resemble enterics on MAC and EMB agars. abortus. baumanii. Identifying characteristics 1) Nonmotile and oxidase negative 2) Nitrate negative and catalase positive E. melitenis. including deer. or from animal bites and the bite of deerflies or ticks.634 • CHAPTER 6: BACTERIOLOGY c. Brucella a. B. 2) Strict aerobe 3) Oxidase and catalase positive . Biosafety level 3 is required when handling the organism or suspect specimens.

Identifying characteristics 1)5. whereas all other species are urease positive. they appear like mercury droplets and are beta-hemolytic. Actinobacillus a. 3) Most species will grow on MAC agar except B. Causes cellulitis but can progress into osteomyelitis. Pasteurella spp. Actinobacillus spp. B. bronchiseptica causes kennel cough in dogs and is an infrequent cause of respiratory infections in humans. f. Infections are caused by animal bites. pertussis causes pertussis. is generally done serologically. poorly stained coccobacilli. and pneumonia c. Media are often made selective by adding cephalexin. grow well on SBA and chocolate agar but will not grow on MacConkey agar. Most species are catalase and glucose positive. pertussis colonies are small and smooth. meningitis. Other species 1) B. A. b. pertussis is urease negative. actinomycetemcomitans is normal oral flora of humans. 2) Gram stain shows minute. A. c. are found mostly as oral flora of animals. e. Three stages of pertussis (whooping cough) 1) Catarrhal: General flulike symptoms 2) Paroxysmal: Repetitive coughing episodes 3) Convalescent: Recovery phase d. Actinobacillus spp. Phage and dye sensitivity tests are used for identification to the species level. B. Cultures are needed for definitive identification. P. multocida causes most human infections. pertussis grows on Bordet-Gengou (potato infusion) and Regan-Lowe agars (charcoal-horse blood agar). AEROBIC GRAM-NEGATIVE BACTERIA 4) Serology can be used for presumptive diagnosis of brucellosis. c. Confirmation of identification of isolates as Brucella sp. which can result in cellulitis. actinomycetemcomitans is associated with endocarditis and also causes gum disease. 2) B. Humans acquire the bacteria from animal bites (cats and dogs) or by inhalation of dried animal feces. b. parapertussis causes mild respiratory infections in humans. d. single or in pairs. pertussis. b. . joint infections. are normal respiratory/GI flora of animals. It inhabits the mucous membranes of the respiratory tract of humans. Bordetella a. They produce colonies that show starlike centers. Pasteurella a. 4) B.

are sometimes used for cultures for the diagnosis of the pneumonic form of the disease. which can be asymptomatic or mild to severe pneumonia. catalase. is the severe form of legionellosis. and most biochemical tests are negative. and nitrate positive 3) Nonmotile. appeal' as thin. They are resistant to commonly used concentrations of chlorine. Requires hemin (factor X) for growth. poorly staining gram- negative bacilli. Most human infections are caused by L. urine antigen test. pneumophila. Normal flora of the mouth and upper respiratory tract b. a medium more nutritious than SBA. pneumophila serogroup 1. Produces a bleachlike odor Legionella a. expectorated sputum. Legionella spp. characterized by flulike symptoms. whirlpools. and air conditioning chillers. Pontiac fever is a mild form. Identifying characteristics 1) Grows well on nonselective agars but not MAC 2) Oxidase. unless 5-10% CO2 is present e.1% basic fuchsin as the counter stain instead of safranin. c. 5) Other identifying tests: Direct fluorescent antibody test. However. 2) Legionella spp. require L-cysteine for growth. Chromobacterium violaceum is found in water and soil. bronchial wash. are aquatic organisms that may be found in various water systems. etc. It is better to use 0.636 • CHAPTERS: BACTERIOLOGY d. some species will grow on Brucella blood agar. including humidifiers. e. Causes legionellosis. including L. a primary pneumonia. Identifying characteristics 1) On a Gram stain. Specimens from the lower respiratory tract. gram-negative coccobacilli that may show bipolar staining 4) Very susceptible to penicillin Eikenella corrodens a. Legionella spp. . d. Legionnaires disease. First discovered in 1976 as the cause of pneumonia in people attending an American Legion convention in Philadelphia b. Approximately 50% of the strains corrode or pit the agar surface. and nucleic acid probes Chromobacterium a. The urine antigen test is the most common laboratory assay used for the diagnosis of legionellosis. pleomorphic. indole. lung biopsy. d. 4) Most species will autofluoresce when exposed to ultraviolet light. 3) They are asacchrolytic. f. They can produce tiny colonies on chocolate agar. g. Causes abscesses of oral cavity and human bite wound infections c. They will grow on BCYE agar but not on SBA.

G. pairs. S. synovial fluid. hominis appears as short chains. Presence of clue cells. b. Causes wound infections acquired from contaminated soil or water 9. Found as normal oral flora in rats and other rodents b. It is oxidase positive. Ingestion of contaminated food or water results in Haverhill fever. facultative. Oxidase negative. Gardnerella are very small gram-variable coccobacilli. catalase negative. In low numbers. . UTIs. gram-negative. 12.. PID. c. and abscess material. Gardnerella vaginalis a. They differ from Lactobacillus spp. moniliformis is a nonmotile. d. c. Produces a purple or violet pigment on nutrient agar c. d. Approximately 50-60% of women who do not meet the criteria for BV are positive for G. Mainly associated with endocarditis c. vaginalis probably does not cause B V. Found as normal flora in humans in the upper respiratory tract and possibly the gastrointestinal and genital tracts b. Streptobacillus moniliformis a. but its presence is indicative of the condition. G. Amsel and Nugent scoring systems are used to diagnose BV. although growth is enhanced in media containing yeast extract. epithelial cells with numerous bacteria attached. vaginalis is associated with bacterial vaginosis (BV). Cultures alone are too sensitive. 10. and weakly indole positive. Catalase negative f. which are large gram-positive bacilli. C. is suggestive of BV. e. Bartonella quintana 1) Agent of trench fever 2) Also causes growth of neoplastic blood vessels in various parts of the body (bacillary angiomatosis) and other infections such as endocarditis 3) Spread by human lice c. e. Bartonella a. vaginalis is considered normal vaginal flora. B. gram-negative pleomorphic bacillus. curved bacilli b. Cardiobacterium hominis a. AEROBIC GRAM-NEGATIVE BACTERIA • 637 b. The bacteria are best isolated from blood. 11. Infections following animal bites results in a disease called rat-bite fever. G. In Gram stains. or rosettes of irregularly staining bacilli with bulbous ends. hominis requires CO2 for initial isolation and can be recovered on SBA. and postpartum sepsis and may infect the newborn. C. henselae: Causes cat-scratch disease and also bacillary peliosis hepatitis and bacillary angiomatosis. It grows on media enriched with SBA (15% is optimal) incubated in a CO7 incubator. d. vaginalis.

particularly in Japan and Taiwan g. causing septicemia after ingestion of undercooked seafood. Most are indole positive. cholerae Ol serological group causes cholera. Vibrio a. Vibrio cholerae 1) V. cholerae Ol is subdivided into three serotypes: Inaba. Vibrio vulnificus 1) Highly virulent. General characteristics a. and South America. b. Serogroup O139 produces severe disease similar to V. cholerae and V. and mucinase. motility. pili. 2) Serogroups non-Ol generally cause a mild choleralike illness. Ogawa.CHAPTER 6: BACTERIOLOGY F. 2. 4) V. The genus contains about 12 species that are inhabitants of marine water. cholerae Ol. Some species cause GI disease. c. d. and Hikojima. resulting in a massive release of water. wound infections. an enterotoxin). 3) V. Thiosulfate citrate bile salt sucrose agar (TCBS) is a selective and differential (based on sucrose fermentation) medium that supports the growth of most species and is particularly useful for isolating V. cholerae can also cause bacteremia. whereas V. characterized by severe watery diarrhea with flecks of mucus sometimes referred to as "rice-water" stool. Vibrio parahaemolyticus 1) Causes a mild to moderate choleralike diarrhea disease 2) Acquired by eating raw shellfish 3) Important cause of food poisoning in Asia. e. cholerae include cholera toxin (choleragen. All species are halophilic (salt loving) except V cholerae and V. 7) Symptoms seen in cholera are caused by an enterotoxin that alters ion transport of intestinal mucosa. and all are oxidase positive except V. Africa. notably raw oysters 2) Causes a rapidly progressive wound infection after exposure to marine water . parahaemolyticus is sucrose negative. Vibrio and Similar Microorganisms 1 . mimicus. V. Most laboratories use biochemical testing to presumptively identify species and then confirm with serology based on somatic O antigens. cholerae is sucrose positive and will produce yellow colonies on TCBS agar. 6) Important virulence mechanisms of V. parahaemolyticus. V. b. and otitis media. It is endemic to Southeast Asia. 5) Cholera infections are acquired by ingestion of undercooked seafood or contaminated drinking water. f. 8) In addition to causing cholera. metschnikovii. cholerae Ol has two biotypes: classical and El Tor.

Treatment is required only in immunosuppressed patients or other severe cases. citrate. A. causing gastroenteritis. Vibrio alginolyticus 1) Very common in marine environment 2) Suspected causes of otitis media and wound infections 3. diarrhea. Plesiomonas shigelloides a. they will show darting motility. 6) On wet mount. 4) C. jejuni and C. Clinically important species include A. however. indole. Acquired by eating undercooked seafood b. . d.jejuni causes most infections in this genus. 8) C.g. caviae. wound infections may require antimicrobial therapy. c. AEROBIC GRAM-NEGATIVE BACTERIA • 639 h. Infects humans and fish c. jejuni and C. however. e. and A. Found in fresh and salt water b. hydrophilia. Causes cellulitis and diarrhea d. A. veronii biovar sobria. P. Campylobacter a. Part of many routine stool culture work-ups e. coli. and septic arthritis. c. shigelloides was recently moved to the family Enterobacteriaceae. shigelloides is primarily associated with a self-limiting gastroenteritis. Infection is acquired by eating undercooked contaminated poultry or other meat products. VP. and ONPG positive. and most human isolates are catalase and oxidase positive. 7) Resistance to cephalothin and sensitivity to nalidixic acid has been used in the past for identification of C. b. fetus is a rare cause of extraintestinal infections and does not grow at 42°C. Based on DNA homology. Identifying characteristics 1) Curved bacilli that may appear S-shaped or spiral on Gram stain 2) Most species are microaerophilic. 5. because of variability in the sensitivity pattern. veronii biovar veronii.. P. Campylobacter spp. despite being oxidase positive. are a major cause of food poisoning.jejuni grows at 42°C but will grow slowly at 37°C. 3) A number of selective media (e. hydrophila is typically beta-hemolytic and oxidase. Aeromonas a. Generally cause a self-limiting infection not usually requiring treatment. A. f. 5) They do not oxidize or ferment carbohydrates. disk identification tests are no longer recommended. 4. charcoal cefoperzone deoxycholate agar and Campy-colistin vancomycin amphotericin B) are available for the isolation of C. C. coli from stool specimens.

and catalase positive c. 5. MYCOBACTERIA A. pylori colonization include fecal antigen detection. Mycobacteria are slender. d. b. CSF.. H. mild treatments. rapid urease. b. Oxidase. and Skirrow's agars incubated microaerophilically. such as 2% NaOH with W-acetyl-L-cysteine (NALC). 9. The primary stain in the acid-fast stain is carbol fuchsin. Urine: 3 to 5 different morning voids c. nonmotile. Centers for Disease Control and Prevention acid-fast smear evaluation and reporting criteria. Lower respiratory tract: Sputum and bronchial washings. It is necessary to decontaminate samples containing normal flora before culturing. non-spore-forming. Selective media for enteric Campylobacter are not recommended. NaOH increases the pH to a level that is antibacterial. usually 3 to 5 samples. Blood and bone marrow d. 7. Cause tuberculosis (TB) and other diseases 2. pylori causes peptic and duodenal ulcers and has been linked to stomach cancer. obligate aerobes.g. Mycobacteria are acid-fast and are referred to as acid-fast bacilli (AFB). etc. such as sputum that contains bacteria from the oral cavity.640 • CHAPTER 6: BACTERIOLOGY 6. need to be decontaminated. and sputum must also be digested. see Table 6-3 B. 3. Brucella. b. are effective.) do not require decontamination. General Characteristics 1. . blood. NALC is a mucolytic agent that liquefies mucus in respiratory specimens. The microorganism can be isolated from gastric biopsy on SBA. Specimens a. Digestion and decontamination of sputum samples a. The Ziehl-Neelsen stain requires heating during the staining step. III. Other methods to determine H. Mycobacteria resist Gram staining because of lipids in their cell wall that prevent penetration of crystal violet and safranin. urea breath test. whereas the Kinyoun's stain does not. Therefore. 14 of which are pathogenic to humans. releasing mycobacteria. are collected early in the morning on different days. 6. There are about 50 species of Mycobacterium. 4. Specimens from normally sterile sites (e. c. Only specimens containing normal flora. Helicobacter pylori a. The mycobacteria are slightly more resistant to acids and alkalis than contaminating bacteria making up the normal flora. Tissue and body fluids 8. and demonstration of urease activity in stomach biopsy material.

Mycobacterium growth index tube (MGIT®) by Becton Dickinson (Franklin Lakes. Lowenstein-Jensen (LJ) contains egg components for growth and malachite green to inhibit growth of normal flora. repeat 1-2/300 fields 1+ 1-9/100 fields 2+ 1-9/10 fields 3+ 1 -9/field 4+ >9/field 10. b. Lowenstein-Jensen-Gruft is made selective by the addition of penicillin and naladixic acid. Antimicrobials can be added to make the media selective for the mycobacteria. Runyon groups a. Solid media a. Most common pathogens are slow growers. b. These media generally exhibit growth several days before egg-based media. c. Liquid media a. and the fluorchrome will fluoresce when exposed to ultraviolet light. the mycobacteria are placed into groups according to their growth rate and photoreactivity. b. avium complex (MAC). Growth rate: Rapid growers produce colonies on solid media within 1 week. Middlebrook medium is agar based and contains 2% glycerol to support the growth of M. MYCOBACTERIA • 641 TABLE 6-3 REPORTING CRITERIA FOR AFB ON DIRECT SMEAR Report Fuchsin Stain ( x 1000) No AFB seen Doubtful. tuberculosis complex. Growth of mycobacteria releases radioactive CO9 that is detected by the instrument. BACTEC 460TB system contains a radioactively labeled substrate that can be metabolized by mycobacteria. Middlebrook 7H9 broth is often used to maintain stock cultures and prepare isolates for biochemical tests. c. . NJ) contains a modified 7H9 broth. The large amount of oxygen in the broth quenches the fluorescence of a fluorchrome. and weakly pathogenic species are rapid growers. As mycobacteria grow. Except for members of the M. It is also supplemented with RNA. 12. they consume the oxygen. 1 1 .

Catalase: All mycobacteria typically produce catalase./V-naphthylenediamine dihy- drochloride) are added. A 1:1 mixture of 30% hydrogen peroxide and 10% Tween 80 is added. Nitrate 1) In the test for nitrate reductase. This is not the same formulation used for gram-negative bacilli. d. and . 2) Semiquantitative catalase: A 1 mL aliquot of a mixture of hydrogen peroxide and Tween 80 is added to a 2-week-old culture deep of mycobacteria. 13. tuberculosis is one of the few species positive for the accumulation of niacin. Biochemical tests for the identification of the mycobacteria a. Niacin accumulates in the medium and is detected by reacting with a cyanogen halide. b. Group 4 contains the rapidly growing mycobacteria. d. 2) Scotochromogens produce yellow to orange pigment in the light and in the dark. some species produce an excess amount that is excreted from the cell. there are different forms of catalase that can be differentiated in the laboratory. tuberculosis complex. Members of the M. Members of Runyon group 1 are slow growers and photochromogens. 2) M. c. Although all mycobacteria produce niacin. however. the suspension is observed for bubbles. 2) M. . 2) M. and most other Mycobacterium spp. are negative. sulfanilamide. Runyon groups are no longer commonly used. bovis. NaNO3 is added to a heavy suspension of mycobacteria. tuberculosis and M. kansasii are nitrate reductase positive. Group 2 members are slow growers and scotochromogens. The suspension is incubated for 2 hours at 35°C. and then nitrate reagents (HC1. Growth on MacConkey agar 1) MacConkey agar without crystal violet is inoculated with a 7-day broth culture of the test organism. Mycobacteria that are slow growers and nonchromogens belong to group 3. chelonei are the only mycobacteria able to grow on MacConkey agar in 5 days. after 5 minutes. After 5 minutes. the height of the column of bubbles is measured. 1) Heat-sensitive catalase: A suspension of a Mycobacterium sp. including M.642 • CHAPTER 6: BACTERIOLOGY c. tuberculosis and M. is heated at 68°C for 20 minutes. are negative. Because of variation within the species. Niacin 1) Niacin (nicotinic acid) is a precursor in the synthesis of NAD. Photoreactivity 1) Photochromogens produce yellow to orange pigment only when exposed to light. fortuitum and M. Members of the M. tuberculosis complex produce columns of bubbles <45 mm. Formation of a pink color is a positive reaction. 3) Nonchromogens (nonpigmented) do not produce pigment.

MYCOBACTERIA • 643 e. etc. Susceptibility to thiophene-2-carboxylic acid hydrazide (T2H): differentiates M. 3) Treatment requires long-term combination therapy. 2) The bacteria can spread to the lymphatic system. Direct nucleic acid amplification: Commercial amplification assays approved for use on respiratory specimens are available for detection of M. Mycobacterium tuberculosis a. Clinically Important Mycobacterium 1 . gordonae. It is approved for use on culture isolates. the patient is typically asymptomatic. rifampin. AccuProbe® (Gen-Probe Inc. M. 4) Primary TB may not lead to active TB in people with healthy immune systems. because of alteration in the cell-mediated immune response. can be triggered by poor nutrition. Strip assays: Probes are immobilized onto a membrane strip and bind biotinylated-polymerase chain reaction products. d. b. Together the cells form granulomatous lesions called tubercles. 14. The lesions can calcify. which can be seen on chest X-rays. tuberculosis. which can last up to 24 months. bovis (susceptible) from most other species (resistant). and pyrazinamide. c. . The assay is highly specific and takes less than 2 hours. Only a few bacteria are necessary to cause disease. ethambutol. kansasii. which are eventually surrounded by fibroid cells. dust.): This commercial assay has probes for Mycobacterium tuberculosis complex. 3) Macrophages phagocytize the bacteria and form multinucleated cells. M. avium. The target is a region of the 16s rRNA gene. c. e. Reactivation or secondary tuberculosis 1) Occurs in people who have had latent TB 2) Reactivation. These assays are also commercially available for culture isolates. a. a chronic primarily lower respiratory tract disease b. or hormonal factors associated with pregnancy and diabetes. Mycobacterium avium complex. intracellularae." While the bacteria are contained within the granulomas. First-line drugs include isoniazid. at which point they are called "ghon complexes. B. Nucleic acid assays: Because of their slowly growing nature. Primary tuberculosis 1) Infection begins in the middle or lower areas of the lungs. rapid and specific nucleic acid assays are becoming more common in clinical laboratories for the identification of the mycobacteria. and M. Causes tuberculosis. CNS. and heart. Spread by person-to-person contact via infected droplets. alcoholism. M. This stage of the disease is called a latent infection.

in addition to isoniazid and rifampicin. intracellulare are difficult to distinguish and are referred to as M. Mycobacterium scrofulaceum causes cervical lymphadenitis and other types of infections predominantly in children. Acid-fast stain often shows ropelike formations (cording) from broth culture. Agent of Hansen disease (leprosy) b. MAC may cause disseminated disease in immunosuppressed patients. M. M. avium and M. defined as simultaneous resistance to isoniazid and rifampin. Purified protein derivative (PPD): Skin test that determines exposure to M. Cannot be grown on artificial media c. lymphadenitis. except in patients with severe immunosuppression. 3. It is a slow grower and is nitrate and catalase positive. and granular in 14-21 days at 37°C. MAC is a slowly growing nonchromogen. g. heaped. Mycobacterium leprae a. filtered. 4. tuberculosis 1) Antigen is composed of heat-killed. was first discovered in 1991. j. indicates a poor prognosis for recovery. kansasii causes pulmonary infections and is the most commonly isolated photochromogen in the U. Extremely-drug-resistant M. avium complex (MAC). b. if present. kansasii a. Multidrug-resistant M. kanamycin. M. p-nitro-a-acetylamino-(3- hydroxypropiophenone (NAP) susceptible 2. 5. h. It is a slowly growing scotochromogen. producing lung infections. and at least one of three injectable second-line drugs (capreomycin. b. c. tuberculosis (MDR-TB). dry. It causes a lung disease that resembles classic TB and rarely disseminates. tuberculosis. tuberculosis (XDR-TB) is defined as resistance to any fluoroquinolone.644 • CHAPTER 6: BACTERIOLOGY f. avium complex a. and intestinal infections. ammonium sulfate precipitated protein from M. Diagnosis is based on characteristic skin lesions and visualizing AFB in lesions. k. such as patients with human immunodeficiency virus infection. . M. Niacin and nitrate positive. c. and amikacin). and. Colonies on LJ medium appear nonpigmented (tan or buff). 2) Injected intradermally and is examined at 48 hours for swelling (induration) 3) A positive skin test indicates previous exposure to the bacteria but not necessarily active disease. It has been isolated from tap water around the world.S. i.

Obligate anaerobe: Bacterium that cannot use oxygen for metabolism and oxygen is lethal to the microorganism b. Obligate aerobe: Bacterium that requires oxygen at concentrations found in room air. Surgical specimens 3. Mycobacterium ulcerans. General Characteristics 1. 9. chelonae. about 20% e. andM. Anaerobic bacteria (i. and M. Mycobacterium gordonae is found in fresh water. including tap water. Suspect anaerobic bacteria in the following situations: a. Anaerobic body sites. Centers for Disease Control and Prevention (CDC) anaerobic blood agar: For general growth of all anaerobes . ANAEROBIC BACTERIA • 645 6. Vitamin K is added to enhance the growth of Prevotella and Porphyromonas. M. M. bovis is a nonchromogen like M. haemophilium have all been implicated in skin infections. It is a slowly growing scotochromogen often isolated as a contaminant. osteomyelitis. Their predilection for surface areas and extremities of the body is related to their optimal growth temperature range of 30-32°C. 2..e. abscessus. M. Foul odor (from gas production) and necrotic tissue b. IV. 2. fortuitum. 7. they are weakly virulent. abscesses. may cause abscesses. Mycobacterium bovis is responsible for a zoonosis. M. albeit more slowly. M. obligate anaerobes) comprise most normal flora of the mucous membranes. and wounds c. Anaerobic Media 1. wound and lung infections. 8. haemophilium requires ferric ammonium citrate or hemin for growth and can be grown on chocolate agar. Definitions a. Capnophile: Bacterium that requires increased concentration of CO2 B. and hemin is an added enrichment for Bacteroides and Prevotella. and is rarely pathogenic. tuberculosis. but it is nitrate and niacin negative and sensitive to T2H. however. ANAEROBIC BACTERIA A. Facultative anaerobe: Bacterium that will use oxygen if it is present and can grow. producing pulmonary infections primarily in cattle and occasionally in humans. Media contain supplements that enhance anaerobic growth. These species are among the rapidly growing mycobacteria. without oxygen d. Microaerophile: Bacterium that requires oxygen at concentrations of 5-10% f. marinum. Aerotolerant anaerobe: Bacterium that cannot use oxygen but can grow in its presence c.

is added. pleomorphic gram-negative coccobacilli with bipolar staining 2. thin. a catalyst.646 • CHAPTER 6: BACTERIOLOGY 3. releasing glycerol. 10. and to a chocolate agar plate incubated under conditions of increased CO2. Bacteroides and Prevotella: Pale. An oxidation-reduction indicator (Eh) must be used to determine if anaerobic conditions have been met. such as thioglycolate and cooked (or chopped) meat. Commonly used systems include anaerobic GasPak jars and bags and anaerobic hoods. including Clostridium and Bacteroides. 9. Solid media must be placed in anaerobic conditions in order for obligate anaerobes to grow.g. Aerotolerance testing: Before attempting to identify a possible anaerobic bacterium. Lipase cleaves lipids. Methylene blue is the most commonly used oxidation-reduction indicator. a. 8. Enterobacteriaceae) 6. it first must be demonstrated to be an obligate anaerobe. can be used to grow anaerobic bacteria. In the presence of palladium. filamentous gram-negative bacilli with tapered ends arranged end to end . laked blood enhances pigment formation 5. When anaerobic conditions are achieved. Gram Stain Morphology 1. anaerobic. Broths with reducing agents. Columbia-colistin-naladixic agar with 5% sheep blood: Inhibits gram- negative organisms and is used to grow most gram-positive anaerobes and facultative anaerobes 7. Lecithinase activity produces an opaque zone from the cleavage of lecithin releasing insoluble fats (diglyceride). which floats to the top of the medium producing a blue-green sheen (mother-of-pearl) on the agar surface. which is incubated anaerobically. indicating reduction. Phenylethyl alcohol (PEA) agar: Enriched and selective medium used to grow most anaerobes. Isolates growing only on the plate incubated anaerobically are obligate anaerobes. the following reaction occurs: 2H2 + O2 -> 2H2O2 b. C. The indicator is pink in the presence of oxygen and colorless when reduced. A colony is inoculated to an anaerobic blood agar plate. Fusobacterium: Long. Kanamycin-vancomycin laked sheep blood (KVLB) agar: Enriched selective medium for isolation of slowly growing anaerobes such as Prevotella and Bacteroides. Sometimes resazurin. Bacteroides bile esculin (BBE) agar: Selective and differential medium used to culture and presumptively identify Bacteroides fragilis 4. an oxidation-reduction indicator. inhibits the growth of facultative.. Egg yolk agar is used to detect proteolytic enzymes (lipase and lecithinase) produced by Clostridium. gram-negative bacilli (e. the methylene blue indicator will turn from blue (oxidized) to white.

Identifying characteristics 1) Brick red fluorescence under UV light . Causes infections of the head. fragilis is resistant to penicillin. b. kanamycin. Clostridium: Large gram-positive bacilli. nose. neck. bile-esculin positive. urease. lipase negative. Porphyromonas a. ANAEROBIC BACTERIA • 847 3. Pigmented saccharolytic gram-negative bacilli b. Identifying characteristics 1) Nonmotile gram-negative bacilli with rounded ends and may be pleomorphic 2) Nonhemolytic on anaerobic blood agar 3) Biochemistry: Growth in 20% bile. central. Causes head. Actinomyces: Branching gram-positive bacilli 4. and GI and urogenital tracts c. lecithinase negative. and urogenital tract d. B. 2. Antimicrobial susceptibility disks can also be used to help identify anaerobes. nose. fragilis is a nonpigmented bacillus responsible for most anaerobic infections. spore location (terminal. Biochemical Reactions 1. Important anaerobic biochemical tests include catalase. Bacteroides fragilis group a. nitrate. 2) Biochemistry: Ferments glucose and many other carbohydrates and is inhibited by 20% bile 3) Susceptible to rifampin and resistant to kanamycin 3. Asaccharolytic or weak fermenters. Normal flora of the oropharynx. E. Identifying characteristics 1) Young colonies appear tan and exhibit brick-red fluorescence under ultraviolet (UV) light. Major normal flora of the colon c. It may take up to 3 weeks to see brown to black pigment. and many isolates are becoming more resistant to antimicrobial agents. neck. Normal flora of the oropharynx. or disease d. catalase positive. Anaerobic Gram-Negative Bacilli 1. especially after surgery. and lower respiratory tract infections d. and vancomycin and susceptible to rifampin. and gelatinase negative 4) Produces brown to black colonies on BBE agar 5) B. gram-negative bacilli b. pigmented colonies. and indole. trauma. Prevotella melaninogenica a. oral cavity. 2. or subterminal) is important in species identification D. Older colonies are brown to black. A polysaccharide capsule is an important virulence mechanism. Causes infections by gaining entry into normally sterile body sites. and GI and urogenital tracts c.

648 • CHAPTERS: BACTERIOLOGY 2) Porphyromonas spp. post-abortion sepsis. water. C. Catalase negative e. Asaccharolytic or weak fermenters. gram- negative bacilli b. penicillin. Anaerobic Gram-Positive Spore-Forming Bacilli 1. abdominal infections. will not grow on KVLB agar and are inhibited by bile. It causes gas gangrene (myonecrosis). Normal flora of the upper respiratory and GI tracts c. C. Identifying characteristics 1) Colony morphology: Opalescent with speckles 2) Indole and lipase positive. nonmotile species include C. Diabetics and patients with circulatory disorders are more prone to infection. C. e. perfringens is normal flora of the GI tract and can spread from this site following trauma. F. Most species are anaerobic. nucleatum (causes serious pulmonary infections) and F. Produce a variety of exotoxins 2. However. resistant to vancomycin F. Some species are normal GI flora of humans and animals. some species appear gram-negative d. and others are found in soil. . Clostridium perfringens a. necrophorum causes more serious infections. and rifampin. 4. f. b. nonpigmented colonies. resulting in a mild to moderate diarrhea without vomiting c. nitrate and catalase negative 3) Relatively biochemically inactive 4) Inhibited by kanamycin and colistin. and dental infections in addition to brain abscesses. Major cause of food poisoning (from meats and gravy). nucleatum is the more common isolate. c. they are resistant to kanamycin. ramosum. perfringens is also normal flora of the female genital tract and can cause post-abortion infections. C. Fusobacterium spp. blood. inocuum. sinus. b. cause pulmonary. Fusobacterium a. perfringens. Large gram-positive bacilli. and enterocolitis. and dust. General characteristics of the Clostridium a. and C. a few are aerotolerant. Two important species are F. d. d. e. perfringens is the most important pathogen in the genus. Many infections are associated with metastatic conditions. but F. Bacteria are acquired through puncture wound or by ingestion. and arthritis). necrophorum (lung and liver abscesses. Most Clostridium species are motile. vancomycin.

C. Identifying characteristics 1) Produces a double zone of beta-hemolysis on SBA incubated anaerobically 2) C. indole and motility positive. diagnosis made by signs and symptoms and toxin detection 4. g. The antibody will neutralize lecithinase. which may include gunshots. Causes tetanus b. Bacteria and spores gain entry into the host by puncture wounds contaminated with soil. a neurotoxin that affects the anterior horn cells of the spinal cord. A to E. or by wounds. C. ANAEROBIC BACTERIA • 649 f. Identifying characteristics 1) Gram-positive bacilli with round/terminal spores that resemble drumsticks 2) Gelatinase. pertussis. This test is not performed much today. Botulism is usually acquired by ingestions of spoiled. maltose. . resulting in acute (flaccid) paralysis and death. C. Clostridium botulinum a. c. Clostridium tetani a. home-canned foods in which the spores are not destroyed. Treatment and prevention: Antitoxin and vaccine (DPT: diphtheria. and fructose fermentation 4) Spores are subterminal but difficult to induce. The isolate is inoculated onto both halves of the plate. d. tetani produces tetanospasmin. Contractions begin with the neck and jaw ("lockjaw") and progress to a backward arching of the back muscles. In this assay. The bacteria produce little necrosis. Causes botulism b. 3. Streptococcus agalactiae (group B streptococci) is substituted for Staphylococcus aureus in the standard CAMP test. and tetanus trivalent vaccine) booster every 5 years e. lecithinase and lipase negative 3) Generally not cultured. d. resulting in involuntary muscle contractions. based on the quantities and types of exotoxins produced. 3) Positive for lecithinase and glucose. preventing the opaque zone from forming. c. burns. perfringens produces lecithinase. 5) Nonmotile 6) Nagler test: Antilecithinase antibody is swabbed onto half of an egg yolk agar plate. Infant botulism is the most common type of botulism. Bacteria are ingested and grow in the infant GI tract and can cause a rapidly fatal infection. lactose. which will produce an opaque zone on the half of the plate without the antibody. Botulism toxin is a neurotoxin that binds to the synapse of nerve fibers. This organism is divided into five types. petfringens exhibits a positive (enhanced hemolysis) reverse CAMP test. perfringens secretes enzymes and exotoxins that cause severe tissue damage. or animal bites.

difficile is weakly fructose positive. difficile should only be performed on watery or unformed stools. Causes antibiotic-associated pseudomembranous colitis and diarrhea b. Cycloserine-cefoxitin-fructose agar (CCFA) is used for isolating C. The toxins can be detected using various immunologic methods. and by examining cell monolayers for cytopathic effect after the addition of stool filtrates. Normal flora of animal and human mucous membranes b. including enzyme immunoassay. glucose. indicates colon cancer when isolated in blood cultures. are infrequently associated with infections. C. difficile can be normal flora. High carriage rate in the intestines of patients who have received broad- spectrum antimicrobial agents that have eliminated the normal intestinal flora d. stool cultures can sometimes be too sensitive. is a member of the histotoxic group and is occasionally linked to gas gangrene. a. b. Clostridium difficile a. the colonies are yellow. anaerobic) conditions. Identifying characteristics 1) Lipase. 5. C. normal flora of the gastrointestinal tract. It is also important to test isolates for toxin production. Identifying characteristics 1) Because C. Produces enterotoxin A and/or cytotoxin B e. Other Clostridium spp. f. difficile is normal GI flora in a small percentage of the population. is the most common pathogen. Anaerobic Non-Spore-Forming Gram-Positive Bacilli 1 . In reduced (i. ismelii. Infections can be diagnosed by detecting either toxin in the stool.650 • CHAPTER 6: BACTERIOLOGY e. c. The product will also fluoresce yellow-green. 2) Lecithinase. This is a characteristic associated with the Streptococcus bovis group as well. which causes abdomen and chest infections and pelvic actinomycosis in women with intrauterine devices. C. A. Despite being fructose positive. the pH indicator turns yellow at a pH of about 5. and motility positive 2) Spores are oval/subterminal and resemble tennis rackets. G.e. and positive for motility and glucose and fructose fermentation 3) Spores are oval and subterminal. 2) Gram-positive bacilli with a beaded appearance. c. Anaerobic Actinomyces a. lipase. 6. C. lecithinase.3. Cultures for C. along with C. perfringens. septicum.. Identifying characteristics 1) Exudate contains sulfur granules—dense clumps of bacteria. and as many as 30% of hospitalized patients may carry the bacteria. often filamentous 3) Colony morphology: Smooth to molar toothlike morphology . and indol negative. difficile from stool specimens. septicum.

and P. This organism helps to maintain an acidic environment in the vagina. Catalase and indole positive 3. Associated with BV. Rarely pathogenic e. Lactobacillus is normal flora of the GI and female genital tracts. Veillonella b. niger 1) Catalase positive 2) Produces olive-green colonies that become black b. the only species is P. Anaerobic gram-positive cocci a. Species include P. Peptococcus. does not ferment any carbohydrates 3) Inhibited by kanamycin and colistin but resistant to vancomycin . 3. female genital tract. Curved bacilli c. acnes and P. asaccharolyticus was renamed Peptoniphilus asaccharolytica. Rarely pathogenic c. catalase and indole negative d. propionicus. inhibited by sodium polyanethol sulfonate (SPS) 2) P. Catalase negative and nonmotile bacilli 5. and abdominal infections b. Normal flora of the skin. gram-negative cocci 2) Reduces nitrate to nitrite. Anaerobic gram-negative cocci a. the risk of BV increases. General characteristics of anaerobic cocci a. PID. b. and respiratory tract b. Motile. Propionibacterium a. Normal flora of the intestines. Anaerobic Gram-Positive and Gram-Negative Cocci 1. b. Often called anaerobic diphtheroids c. Mobiluncus a. Lactobacilli are generally aerotolerant anaerobes that will form small alpha-hemolytic colonies on SB A. Lactobacillus a. If the population of lactobacilli decreases. Inhibited by vancomycin 4. Associated with polymicrobial liver and brain abscesses and wound infections 2. Peptostreptococcus 1) P. Identifying characteristics 1) Small. d. Bifidobacterium: Mostly nonpathogenic normal oral and intestinal flora 6. Eubacterium: Mostly nonpathogenic normal oral and intestinal flora H. mouth. and GI tract d. ANAEROBIC BACTERIA • 651 2. oral cavity. anaerobius. magnus was renamed Finegoldia magna.



A. Chlamydia and Chlamydophila
1. Obligate intracellular parasites
2. Cannot produce ATP; require ATP from host cell
3. Contain both DNA and RNA and are susceptible to antimicrobial agents
4. Diagnosis
a. Cytological methods: Detect chlamydia inclusions in epithelial cells
b. Cell cultures are required to grow the bacteria.
c. Nucleic acid amplification tests (NAATs) are the most common
diagnostic method.
d. Serology: Antibody to lipopolysaccharide and outer membrane protein
5. Three important species
a. Chlamydia trachomatis
1) Causes lymphogranuloma venereum, trachoma, urethritis,
conjunctivitis, and infant pneumonia
2) Trachoma is the leading cause of blindness in the world.
3) No animal vectors; it is spread by human-to-human contact.
4) Diagnosis: Cell cultures, direct fluorescent antibody tests, enzyme
immunoassays, NAATs, and serologic procedures
b. Chlamydophila (formerly Chlamydia) pneumoniae
1) Mild respiratory tract infections producing flulike symptoms, may also
cause Guillain-Barre syndrome
2) No animal vectors; it is spread by human-to-human contact.
3) Diagnosis is often made by using fluorescence-labeled C. pneumoniae
c. Chlarnydiophila (formerly Chlamydia) psittaci
1) Causes psittacosis (ornithosis) or parrot fever, a disease of parrots,
parakeets, cockatiels, and other birds such as turkeys and chickens
2) Humans get infections by the inhalation of bird fecal dust; infections
are uncommon in the U.S.
3) Incubation period 1-2 weeks: Chills, fever, malaise, can progress to
pneumonia, which can be fatal
4) Occupational hazard to farmers, pet shop employees, and bird owners
5) Diagnosis by serology
B. Rickettsia and Similar Genera
1. Rickettsia and Ehrlichia are obligate intracellular parasites requiring
nucleotides and other metabolic building blocks from host cells.
2. Infections are generally spread by insect vectors (ticks, mites, and lice).


3. Diagnosis is often made by clinical symptoms, patient history, and serology.
Immunohistology and polymerase chain reaction assays are also available.
4. Weil-Felix serologic test utilizes Proteus antigens. This assay is nonspecific
and is not used much today.
5. The bacteria can be grown in embryonated eggs and tissue cells. However,
cultures require a biosafety level 3 laboratory and, for safety concerns,
cultures are not recommended.
6. Clinically important species
a. R. rickettsii causes Rocky Mountain spotted fever (RMSF) and is the most
important species in the U.S. It is a member of the spotted fever group and
is carried by ticks. RMSF is a very serious disease; death rates are
approximately 25%.
b. R. prowazekii causes typhus, also called epidemic or louse-borne typhus;
it is carried by human lice. Brill-Zinsser disease is a reactivation of the
original infection.
c. R. typhi causes endemic or murine typhus. It is transmitted by fleas.
d. Coxiella burnetii causes Q fever. It is transmitted by inhalation, contact
with fomites, and ingestion of contaminated milk.
e. Ehrlichia chaffeensis causes ehrlichiosis or human monocytic
ehrlichiosis. It is transmitted by ticks and is endemic to the U.S.
f. Anaplasma (formerly Ehrlichia) phagocytophilum causes human
granulocytic anaplasmosis.
C. Mycoplasma and Ureaplasma
1. Smallest free-living organisms, about the size of a large virus and beyond the
resolution of light microscopes
2. They lack a cell wall, making them pleomorphic and resistant to all antibiotics
that inhibit cell wall synthesis (e.g., beta-lactams).
3. They contain both RNA and DNA and can self-replicate.
4. Infections can be diagnosed by serology.
5. Many species of Mycoplasma and Ureaplasma grow on special laboratory
media, including SP4 and A8 agars and Shepard's 10 B broth.
U. urealyticum produces a strong alkaline pH because of the activity of
urease. Some species will also grow on chocolate agar. Some species
produce fried egg colony morphology.
6. Clinically important species
a. Mycoplasma pneumoniae
1) Causes tracheobronchitis and community-acquired primary atypical
(walking) pneumonia, resulting in a dry, nonproductive cough
2) Spread by direct respiratory contact
3) Mostly seen in teenagers and young adults; lacks a seasonal distribution
4) M. pneumoniae produces hydrogen peroxide, which causes lysis of red
blood cells in vitro.


b. M. hominis
1) Opportunistic pathogen linked to PID in sexually active adults
2) May cause infant meningitis and postpartum fever
c. Ureaplasma urealyticum
1) Causes nongonococcal urethritis and may cause other genital tract
2) Requires urea

A. Genera Causing Human Disease: Treponema, Leptospira, andBorrelia
1. Spirochetes are long, slender, helically curved bacilli that cannot usually be
seen on Gram stain.
2. Special stains such as silver and Giemsa will stain Spirochetes, silver for all
spirochetes and Giemsa only for Borrelia.
3. Spirochetes can be observed by darkfield or phase-contrast microscopy.
B. Treponema pallidum subsp. pallidum
1. Causes syphilis
2. Transmitted by sexual contact, direct blood transmission, or transplacentally
(congenital syphilis)
3. Stages of syphilis
a. Primary: Chancre at the site of inoculation
b. Secondary: Skin rash and lesions on oral mucosa
c. Latent: Absence of clinical symptoms
d. Tertiary: CNS disorders (neurosyphilis), aneurysms, and skin, liver,
and bone disorders
4. T. pallidum cannot be cultured in the laboratory. The bacteria exhibit
corkscrew motility seen by darkfield microscopy on material taken from
5. Generally diagnosed by serology
a. Nontreponemal antigen tests include the Venereal Disease Research
Laboratory (VDRL) and rapid plasma reagin (RPR) tests, which detect
antibodies to cardiolipin-lecithin-cholesterol and are nonspecific. These
antibodies are sometimes referred to as reagin. Biologic false positives
are caused by Lyme disease, various viruses, autoimmune disorders
(e.g., systemic lupus erythematosus), and pregnancy.
b. Treponemal antigen tests include the fluorescent treponemal antibody
absoiption (FTA-ABS) test and the Treponema pallidum particulate
antigen (TP-PA) test, which are specific and confirmatory.
6. Other clinically important species include T. pallidum subsp. pertenue (yaws),
T. pallidum subsp. endemicum (endemic syphilis), and T. carateum (pinta).

Leptospira 1. Diagnosis 1) Serologic tests are sensitive in diagnosing Lyme disease. Humans are the only known reservoir for this species. kidney. Stages of Lyme disease 1) Early localized (stage I): A rash at the bite site (erythema migrans) produces a characteristic "bull's eye" pattern in many patients. b. burgdorferi is the most common tickborne disease in the U. Borrelia 1 . and headache. c. or by darkfield microscopy. also known as Lyme borreliosis b. malaise. B. Patients present with fatigue. Diagnosis is based on observing bacteria in the peripheral blood-stream via the Giemsa or silver stains. 5. and headaches. cause endemic relapsing fever transmitted by arthropods such as ticks. d. 3) Late stage (stage III): This stage is characterized by chronic arthritis and acrodermatitis that can continue for years. Causes Lyme disease. Borrelia spp. and cattle 3. 2. bacteria are transmitted by body lice. L. interrogans causes leptospirosis (Weil's disease). chills. Zoonosis of rodents. or with silver stain . and CNS involvement. Borrelia recurrentis a. or heart (palpitations. it can be difficult to diagnose infections by staining. d. arthralgia. 4. SPIROCHETES • 655 C. 2. It is transmitted by the deer tick (Ixodes damninii). Direct examination via darkfield microscopy. Diagnosis of leptospirosis a. are difficult to culture. 2) Difficult to culture and too few bacteria to detect by direct microscopy D. paralysis). myalgia. and serological tests are insensitive. c. The infection can produce fever. Other Borrelia spp. Borrelia burgdorferi a. carditis). Due to low bacterial numbers. B. Humans acquire the infection by contact with contaminated animal urine. liver.S. muscle pain. Western immunoblotting is considered the most accurate method for antibody detection. which is characterized by recurrent high fever. recurrentis causes epidemic relapsing fever. dogs. CNS (meningitis. 2) Early disseminated (stage II): Bacteria enter the blood stream (producing flulike symptoms) and then can go to the bones (arthritis).

Beta-lactamase inhibitors can be given with a beta-lactam antibiotic to provide effective treatment. Tetracyclines inhibit protein synthesis at the 308 ribosomal subunit (e.. Beta-Iactamase inhibitors: Bacteria can exhibit resistance to the beta-lactam antibiotics by producing an enzyme (beta-lactamase) that cleaves the beta- lactam ring. monobactams. Clavulanic acid. dosage should be monitored using peak and trough values in peripheral blood. tobramycin and netilmicin). bacteristatic compound inhibits bacterial growth) 4. Most cases are diagnosed by serology. Because of potential toxicity. and tazobactam are examples of beta-lactamase inhibitors.. 3. c. inactivating the antibiotic.g.CHAPTER 6: BACTERIOLOGY b. Microorganisms can be recovered in cultures. An antibiotic is a molecule produced by microorganisms that inhibits the growth of other microorganisms. Increased resistance has limited their use.. Blood is the most sensitive specimen during early infections.g. They have no activity against obligate anaerobes. cephalosporins. Aminoglycosides inhibit protein synthesis at the 30S ribosomal subunit and are active against gram-negative and gram-positive bacteria (e. Media include Ellinghausen-McCullough-Johnson-Harris and Fletcher's.. 2. .g.g. although methods vary in sensitivity. bactericidal compound kills bacteria) 3. Cidal: Kills microorganisms (e. ANTIMICROBIAL AGENTS AND ANTIMICROBIAL SUSCEPTIBILITY TESTING A. broad spectrum (third generation). Spectrum of Action 1. expanded spectrum (second generation). and extended spectrum (fourth generation). and carbapenems). 2. Classes of Antimicrobial Agents and Their Mode of Action 1 . Definitions 1. gentamicin. Tobramycin is bactericidal whereas the others are bacteristatic. Urine should be cultured after the second week. sulbactam.g. Narrow-spectrum antimicrobial agent: Limited range of action 2. Synergy: When two or more antimicrobials are used and the combined effect is greater than what would be expected for the simple additive effect of the agents B. Beta-lactam antibiotics inhibit cell wall synthesis (e. Broad-spectrum antimicrobial agent: Active against a wide range of bacteria C. Antibiotics can also be synthetic. 4. Static: Inhibits the growth of microorganisms (e. penicillins. doxycycline and minocycline). They are active against gram-positive and gram-negative bacteria and Mycoplasma and Chlamydia. The class cephalosporin contains a large number of agents categorized as narrow spectrum (first generation). VII..

Bacteria are inoculated onto the agar plates. 6. aureus have been isolated. In the case of fastidious microorganisms (e. Glycopeptides inhibit cell wall formation by inhibiting peptidoglycan synthesis. 4 mm thick in Petri dish at a pH 7.g. 10. this value is the minimal inhibitory concentration (MIC). Also referred to as Kirby-Bauer sensitivity test b. polymixin B and polymixin E). 8. is required. For Haemophilus influenzae. 2. erythromycin and clarithromycin). Macrolides inhibit protein synthesis (e. Haemophilus test medium (HTM) is used. and yeast extract. Quinolones inhibit DNA activity by inactivating DNA gyrase.g. Nitrofurantoin inhibits bacterial enzymes. HTM is Mueller-Hinton base supplemented with hematin. Streptococcus pneumoniae). nitrofurantoin is used to treat UTIs. and vancomycin-resistant S. ciprofloxacin and levofloxacin). they are used to treat infections caused by gram-negative bacteria (e. ANTIMICROBIAL AGENTS AND ANTIMICROBIAL SUSCEPTIBILITY TESTING • 657 5. Sulfonamides inhibit folic acid synthesis by forming nonfunctional analogs of folic acid. 1 1 . The smallest concentration that inhibits growth of the bacteria is recorded.. vancomycin is the only glycopeptide approved for use in the U..g. aplastic anemia (bone marrow suppression). In these assays. . vancomycin-intermediate S.g.S. MHA with 5% sheep red blood cells is used. Antimicrobial Susceptibility Testing 1. The sample taken from the tube with the lowest concentration of antimicrobial agent showing no growth is representative of the MBC.2-7. This can be determined by first performing a broth dilution test and then subculturing the tubes without visible growth to media without antimicrobial agents. NAD. Polymyxins disrupt plasma membranes.. Disk diffusion a. aureus.9% of the bacteria in the original inoculum. c. Chloramphenicol inhibits protein synthesis by binding to the 50S ribosomal subunit. Dilution tests a. d. It is broad spectrum and used to treat serious gram-negative infections such as meningitis. Broth dilutions: Dilutions of the antimicrobial agents are prepared in broth. Risk of bone marrow toxicity. Standardization 1) Mueller-Hinton agar (MHA).4. D. limits use to serious infections. Vancomycin-resistant enterococci. bacteria are exposed to different concentrations of antimicrobial agents. Newer agents are known as fluoroquinolones (e. 7. 9. The minimum bactericidal concentration (MBC) of an antimicrobial agent is defined as the lowest concentration of an antimicrobial agent that kills at least 99. The assays are generally performed in microtiter plates. b. Agar dilutions: Dilutions of the antimicrobial agents are prepared in agar.

3) Cefoxitin is a more powerful inducer of oxacillin resistance and can be used in disk diffusion assays. 2) The bacteria are lawned onto a Mueller-Hinton agar plate and the Etest strips are placed on top of the agar.5 turbidity standard 3) MHA plates are incubated for 18 hours at 35°C in ambient air. 10s colony forming units/mL. 2) Because populations of MRSA are often heteroresistant (some cells sensitive and much fewer resistant). The procedure is the same as for routine disk diffusion except that interpretive criteria are changed: for 5. Both HTM and MHA with sheep red blood cells are incubated in 5-7% CO2 for 18-20 hours.g. Quality control organisms vary depending on the susceptibility test used. 3) After incubation. b. 4) In broth dilution tests with oxacillin. Gradient diffusion a.5 turbidity standard is prepared. the bacteria produce an elliptical zone of inhibition around the strip. . or resistant (R).S. Detection of MRSA 1) Methicillin is no longer available in the U. Some Enterobacteriaceae can produce an extended-spectrum beta-lactamase (ESBL). and the results are reported as sensitive (S). Procedure 1) A bacterial suspension equal to a McFarland #0. c. Miscellaneous assays a. The MIC is read from a scale on the strip where the zone of inhibition crosses the strip. The zone sizes are compared to standard interpretation charts. testing procedures should be modified to be sensitive for the detection of MRSA. 3. zones of <19 mm are reported as oxacillin resistant and results >20 mm are reported as sensitive. which is equal to a McFarland #0. d. These enzymes inactivate the extended spectrum cephalosporins such as ceftriaxone and cefotaxime. Each strip contains a different antimicrobial agent.658 • CHAPTER 6: BACTERIOLOGY 2) Bacterial inoculum. so when referring to MRSA.. After incubation. Etest® (AB Biodisk) provides quantitative antimicrobial susceptibility testing results. the diameters of the zones of inhibition are measured. intermediate (I). e. Several methods are available for detecting the presence of beta-lactamase. it is actually oxacillin or nafcillin resistance that is being discussed. Beta-lactamase is an enzyme that confers resistance to penicillin and some of the semisynthetic penicillins (e. aureus.. 4. S aureus isolates with MICs <2 ug/mL are considered sensitive and results >4 ug/mL are resistant. ampicillin).

Even though clindamycin and erythromycin are in different classes. However. d) Bacteria are in highest numbers in the blood just before fever spikes. Cerebrospinal fluid 1) CSF surrounds the brain and spinal cord and carries nutrients and waste. Blood 1) Blood is normally sterile. before antimicrobial therapy. an anticoagulant that also inhibits complement and inactivates neutrophils. 2) Definitions and characteristics a) Bacteremia: Bacteria in the blood b) Septicemia: Bacteria increasing in numbers in the blood causing harm to the patient c) When drawing blood cultures. indicating the presence of inducible resistance to clindamycin. two bottles are inoculated: one for aerobes and one for obligate anaerobes. PROCEDURES AND BIOCHEMICAL IDENTIFICATION OF BACTERIA A. It is important to collect several specimens at different times for greatest potential of bacterial yield (sensitivity). VIII. The clinical specimen and the suspected pathogens will determine the selection of the primary plating media. The media used will vary among different laboratories because of local pathogens and personal preference of the laboratorians. 3) Cultures a) Blood culture systems utilize bottles containing liquid media. Clinical specimens a. the plate is examined for a flattening of the zone of inhibition around the clindamycin disk. b. A 15-jag erythromycin disk is placed 15 to 20 mm from a 2-ug clindamycin disk. Plating Procedures 1 . A plate is inoculated as for a disk diffusion assay. General information a. avoid skin contamination and collect sample. After incubation. the mechanisms of resistance are similar. The volume of blood collected probably has the greatest effect on isolation of bacteria. 2. resembling the letter D. d) Blood culture bottles often contain sodium polyanethol sulfonate (SPS). a number of hospitals have stopped using anaerobic bottles. SPS has been shown to inhibit the growth of some bacteria. The D-zone test is used to detect the presence of inducible clindamycin resistance by erythromycin. b) Generally. it is normally sterile. because of the reported decrease in the incidence of anaerobic bacteremias. b. PROCEDURES AND BIOCHEMICAL IDENTIFICATION OF BACTERIA • 659 b. . if possible. c) Most aerobic bottles contain 5-10% CO2.

group C streptococci. . 2) Alpha-hemolytic streptococci viridans group. pyogenes (group A Streptococcus) is the most important pathogen isolated in throat cultures. group G streptococci. d) Pseudomonas aeruginosa causes nosocomal and severe pneumonia in patients with CF. e) Haemophilus influenzae causes infection in infants. Staphylococcus aureus. The incidence of infections has decreased dramatically since routine use of the Hib vaccine. c. and Arcanobacterium spp. Sputum 1) Used to diagnose lower respiratory tract infections (e. usually secondary to another infection or predisposing factor.CHAPTER 6: BACTERIOLOGY 2) Meningitis is an inflammation of the meninges. b) Klebsiella pneumoniae is associated with nosocomial pneumonia and pneumonia in alcoholics. Typically. 4) The most common isolates found in CSF are Neisseria meningitidis. and chocolate agars. pneumonia) 2) The lower respiratory tract is normally sterile. This does not pertain to neutropenic or atypical pneumonia samples. group B streptococci. which often have nonpurulent sputum. 3) Encephalitis is an inflammation of the brain. and Listeria monocytogenes. whereas polymorphonuclear cells (PMNs) indicate a quality specimen. and it is the most common cause of pneumonia in geriatric patients. MAC. Acceptable specimens are cultured on SBA. sputum from the lungs acquires normal flora passing through the oral cavity.. However. 3) Culture on SBA and other media as needed by special request d. MAC.. and chocolate agars. Screening for other pathogens may occur upon request. children. E. Streptococcus agalactiae. are also clinically significant. 5) Diagnoses are made by a direct Gram stain and culturing on SBA. c) Staphylococcus aureus causes community-acquired and nosocomial pneumonia.. 5) Common significant sputum isolates a) Streptococcus pneumoniae is an important cause of community- acquired pneumonia. and the immunosuppressed. Streptococcus pneumoniae. Throat 1) S. Neisseria spp. A general rale for an acceptable specimen might be <10 squamous epithelial cells and >25 PMNs/low power field. squamous epithelial cells are an indication of contamination with oral flora.g. Corynebacterium spp. and coagulase negative staphylococci make up the majority of the normal oral flora. coli. 3) A direct Gram stain is performed to determine the quality of the specimen. 4) Several methods are used to determine specimen acceptability.

Legionella spp. E. gonorrhoeae and C. coli. but it may not indicate a UTI. Lactobacillus spp. SBA.. 3) Types of genital tract infections a) Cervicitis and urethritis usually caused by N. 3) Plating protocols vary widely but in general include selective and differential media for the isolation and screening of specific pathogens. chocolate. and MAC). most likely Mobiluncus. Stool 1) Feces contain many species of anaerobic and facultative anaerobic normal flora. gonorrhoeae or C.g. 5) Common significant urine isolates include E. Earlier and late in life. c) PID is a complication of infection caused by N. 4) Urine specimens are generally plated onto SBA and MAC or EMB... There is a corresponding decrease in lactobacilli. g) Mycoplasma pneumoniae causes primary atypical pneumonia. is due to overgrowth of some species of normal vaginal flora. or nonspecific vaginitis. Yersinia enterocolitica. and Vibrio spp. coli (e. Urine 1) Urine is normally sterile. Staphylococcus aureus.. 2) Bacteria causing gastroenteritis include Shigella spp.. PROCEDURES AND BIOCHEMICAL IDENTIFICATION OF BACTERIA • 661 f) Legionella pneumophila primarily infects middle-aged males. trachomatis involving the endometrium or fallopian tubes. Klebsiella spp. Gardnerella vaginalis is considered normal vaginal flora and may only be an indicator ofBV. g.. 2) Bacteriuria is bacteria in the urine. Pseudomonas aeruginosa. are the predominant flora during childbearing years. 3) Calibrated loops are used to determine colony counts on media.. 4) Plating protocols for N. Mycoplasma will not grow on routinely used media. modified Thayer-Martin). Genital tract 1) Laboratorians commonly look for Neisseria gonorrhoeae and Chlamydia trachomatis. which is mostly seen in young adults. gonorrhoeae include using specific selective media (e.. trachomatis b) BV. . 2) The cervix is typically a sterile site. The vagina contains normal flora that changes with age. Staphylococ- cus saprophyticus. Proteus spp. e.e. f. Salmonella. staphylococci and corynebacteria predominate. Enterobacter spp. Campylobacter jejuni. and yeast. will not grow on routinely used media (i. Enterococcus spp. d) Prostatitis is usually caused by enterics. O157:H7).g. Clostridium difficile (must test for cytotoxin).

If catalase is present. pyogenes and 5. aureus. water and oxygen (bubbles) will form. bedsores. Newer tests are based on latex agglutination and detect protein A in the cell wall and have higher sensitivity and specificity for S. A colony is placed on filter paper with the substrate pyrrolidonyl-a-naphthylamide (PYR). Catalasetest a. 3. but it is incubated at 37°C for up to 24 hours. Wounds/abscesses 1) Superficial skin infections: Staphylococcus aureus and Streptococcus pyogenes 2) Folliculitis (hair follicle infection): 5. This test detects the enzyme L-pyrrolidonyl arylamidase. 2. Biochemical Identification of Bacteria 1. both of which are positive. 5. Several drops of H2O2 are added to a bacterial smear on a microscope slide. aureus. However. Staphylococci are positive and streptococci are negative. b. Some strains produce staphylokinase. The human and animal pathogen S. which can dissolve the clot. The PYR test can also be used to differentiate 5. S. aureus is positive. intermedius and 5. Clumping factor (slide coagulase): Formerly slide coagulase tests used rabbit plasma. Coagulase test a. schleiferi. etc. h. Catalase is an enzyme that produces water and oxygen from hydrogen peroxide (H2O2). aureus 5) Erysipelis: 5. lugdunensis and S. Streptococcus pyogenes and Enterococcus spp. A red color after the addition of /?-dimethylaminocinnamaldehyde (DMACA) is a positive PYR test. The tube coagulase test uses rabbit plasma like the slide method.: 5. are typically positive. b. lugdunensis and 5. aureus (negative) from S. b.662 • CHAPTER 6: BACTERIOLOGY 5) Molecular techniques are commonly used for detecting both N. Tests must be checked at 4 hours for clot formation. pyogenes and less commonly Erysipelothrix rhusiopathiae 6) Deep and surgical wounds and abscesses: Anaerobes from normal body sites B. schleiferi can also produce positive results. aureus 4) Impetigo: S. aureus and Pseudomonas aeruginosa 3) Boils. trachomatis. PYRtest a. Clumping indicates a positive reaction and identification of S. gonorrhoeae and C. producing a false negative result. . hyicus are animal pathogens that are also positive.

7. using DMACA. c. The color of uninoculated medium is reddish-orange. Bile solubility test a. Colonies should be taken from nonselective. nondifferential media. yellow is acid. A positive urease test is indicated by a bright pink color. lactose. A spot indole test. The hippurate hydrolysis test detects the bacterial enzyme hippuricase. Media with a high concentration of glucose can inhibit oxidase activity. Triple sugar iron agar (TSI) a. or Kovac's reagent. the test can be used to differ- entiate Campylobacter jejuni (positive) from most other Campylobacter spp. The Ehrlich method is more sensitive but requires an extraction step with xylene. b. In the presence of the bile at 37°C. eosin methylene blue and MAC) should not be tested because of the risk of a false positive result. Indole is detected by an aldehyde indicator (Ehrlich's reagent). c. not Enterobacteriaceae . Hippurate hydrolysis test a. b. The indole test detects the bacterial enzyme tryptophanase. Colonies of Streptococcus pneumoniae are soluble in sodium deoxycholate (bile). Urease test a. Several drops of oxidase reagent (tetramethyl-p- phenylenediamine dihydrochloride) are placed on filter paper containing bacterial colonies or directly on plate colonies. 6. Urease breaks down urea to form ammonia (NH3). Group B streptococci are hippurate positive. The presence of a blue to blue-green color is positive. and red is alkaline. and indole.g. 9. PROCEDURES AND BIOCHEMICAL IDENTIFICATION OF BACTERIA 4. c. Indoletest a. which hydrolyzes hippurate to glycine and benzoic acid. and disappear from the agar surface. in addition to H2S and gas production.. and sucrose fermentation. A positive hippurate will give a purple color after the addition of ninhydrin. b. TSI will show the pattern of glucose. ammonia. whereas most other beta- hemolytic streptococci are negative. Alkaline slant/alkaline deep (K/K): Nonfermenter. 8. The oxidase test detects cytochrome oxidase that is used in the electron transport system. b. Oxidasetest a. has been shown to be more sensitive in detecting indole activity. Organisms are inoculated onto a urea agar slant and incubated at 37°C for 18-24 hours. the colonies autolyse within 30 minutes. A positive oxidase test is indicated by a purple color within 10 to 15 seconds. yielding a red color. c. Tryptophan is broken down by tryptophanase into pyruvic acid. In addition. b. yielding a bright pink color. Phenol red is the pH indicator. 5. b. Colonies from media containing dyes (e.

one end of the strip is held in placed by the cap. c. 10. methyl red. it is yellow at an acid pH. ammonia. glucose fermenter e. Alpha-naphthol followed by 40% KOH is used to detect acetoin. If H2S gas is produced. see above. . Red is negative. Examples include tryptophan (tryptophan deaminase). Yellow is a positive reaction. The color of positive and negative reactions depends on the pH indicator used. Citrate: This test determines if citrate is used as a sole carbon source. Indole: Bacteria positive for indole produce tryptophanase. Carbohydrate fermentation test a. and yellow is negative. g. After the slant is inoculated. IMViC (indole. indicating glucose fermentation. which breaks down tryptophan to pyruvic acid. Deaminase reaction: Detects the ability of an organism to remove the amino group from specific amino acids c. b. Blue is positive and green is negative. Methyl red (MR): MR is a pH indicator. it reacts with the lead acetate. an enzyme that cleaves ONPG and lactose. production of H2S gas: Test systems detect enzymes that produce hydrogen sulfide (H2S) from sulfur-containing molecules in the medium. b. 12. b. 11. Decarboxylation reaction: Detects the ability of bacteria to remove the carboxyl group from a specific amino acid d. ONPG (0-nitrophenyl-(3-D-galactopyranoside) a. Black deep. This test detects the presence of pi-galactosidase. This test is useful in detecting delayed (late) lactose fermenters that lack. Bacteria are usually MR or VP positive. Lead acetate is a more sensitive method to detect H2S gas. beta-galactoside permease. forming a black color (lead sulfide). indicating the ability to ferment lactose. and ornithine (ornithine dihydrolase). Amino acid degradation test a. and indole. Voges-Proskauer. and glucose fermenter f. Red is positive. Acid slant/acid deep (A/A): Lactose and/or sucrose fermenter. A positive test detects bacterial enzymes that break down various amino acids. Voges-Proskauer (VP): A positive VP reaction detects the metabolism of glucose to acetyl-methyl-carbinol (acetoin). lysine (lysine decarboxylase). 13. H2S reacts with iron salts in the medium to form a black precipitate composed of ferrous sulfide. A positive test detects the ability of bacteria to produce organic acids by the fermentation of various carbohydrates. d. Lead acetate is added to filter paper strips. or are deficient in. Alkaline slant/acid deep (K/A): Nonlactose and nonsucrose fermenter. A pink color is a positive reaction. and citrate) a.CHAPTER 6: BACTERIOLOGY d.

C. Inc. a pink color is positive for reduction of NO3 to NO2. Enterotube II (Becton. Multitest Systems 1. . 2. A positive test determines the ability of an organism to reduce nitrate (NO3) to nitrite (NO2) and nitrogen gas (N2). After the addition of the reagents (N. MIDI. API (Analytical Profile Index. A colorless reaction requires the addition of zinc dust to confirm a negative result. Development of a pink color after adding zinc indicates a true negative. Inc. bioMerieux Clinical Diagnostics) b. formerly Dade Behring) D. a positive result for nitrate reduction. Dickinson and Company) c. Vitek (bioMerieux Clinical Diagnostics) e. Nitrate reduction test a. have identification and minimal inhibitory concentration combination plates. A^-dimethyl-a-naphthylamine and sulfanilic acid).) identifies the fatty acid composition of the bacterial cell wall as determined by gas chromatography. 14. PROCEDURES AND BIOCHEMICAL IDENTIFICATION OF BACTERIA • 665 b. b. Most biochemical testing is performed using multitest methodologies. such as the Vitek and Microscan. Micro-ID (Remel) d. Tubed media are inoculated and overlayed with sterile mineral oil to produce an anaerobic environment. The fatty acids in mycobacteria have a larger molecular weight and are identified via high-performance liquid chromatography in the Sherlock Mycobacteria Identification System. Multitest systems a. Microscan (Siemens Healthcare Diagnostics. 2. Remaining colorless after adding the addition of zinc indicates that NO3 was completely reduced to N2. Semiautomated systems. The Sherlock Microbial Identification System (MIDI. Identification Systems 1. Positive and negative reactions depend on the pH indicators used.

Streptococcus pyogenes tively identified using a(an) D. ONPGdisk aureus is responsible for causing C. Corynebacterium B. A test for the hydrolysis of esculin in the ally variant streptococci. Select the best answer or completion statement in each case. Enterotoxin produced by Staphylococcus B. Staphylococcus D. Staphylococcus aureus 6. Enterocolitis C. Abiotrophia A. Streptococcus pyogenes can be presump- C. preview questions l_j^ O JL. Staphylococcal pneumonia B. Scalded skin syndrome 666 . SPSdisk A. Impetigo D. Exfoliatin produced by Staphylococcus characterized by the presence of a aureus is responsible for causing pseudomembrane in the throat and the A. Scalded skin syndrome A. Vitamin B ]2 D. formerly known as nutrition- 1. will not grow on presence of bile is especially useful in routine blood or chocolate agars because identifying species of the genus they are deficient in A. The organism associated with a disease 5. PYRdisk 3. Enterocolitis production of an exotoxin that is absorbed B. Corynebacterium diphtheriae A. Optochin disk B. Thiophene-2-carboxylic hydrazide 2. JLV LJ C_x JL JLV-/JNI O Each of the questions or incomplete statements that follows is comprised of four suggested responses. Aerobic Gram-Positive Bacteria 4. Enterococcus C. Carbuncles D. Toxic shock syndrome into the bloodstream with a lethal effect is C. Pyridoxal C. Hemin B. Arcanobacterium haemolyticum D. Abiotrophia.

B. is a cause of A. Carbuncles D. S. Listeria monocytogenes 8. licheniformis D. In the CAMP test. Central nervous system pathology D. Streptococcus agalactiae 11. Propionibacterium acnes characteristically produce C. S. a recog- nized pathogen. pyogenes D. post-traumatic infections 10. bovis B. Abiotrophia sp. B. C. Erysipelothrix infections in humans characteristically produce A. Pathology in the lower respiratory 14. equinus C. Micrococcus isolate on blood agar produced small. S. nonmotile. Loeffler's serum medium is recommended indicates the presumptive identification of for the cultivation of A. subtilis incubation. blood culture on a 23-year-old pregnant and modified oxidase positive is best woman who presented with fever and identified as a member of the genus flulike symptoms in her ninth month. Staphylococcus etiologic agent in this case? A. S. circulans perpendicular to a streak of beta-lysin. Impetigo C. Streptococcus pneumoniae B. and endocarditis? beta-hemolytic Streptococcus is placed A. After C. The formation of abscesses in visceral in opportunistic infections such as organs bacteremia. a zone of increased lysis in the shape of an arrowhead is noted. Which of the following is the most tract commonly isolated species of Bacillus D. The A. Streptococcus agalactiae A. Urinary tract infections . Staphylococcus saprophyticus. A gram-positive coccus that is catalase 12. Leuconostoc sp. a single streak of a of the eye. B. Pediococcus Which of the following is the most likely D. Color Plate 26 • shows the Gram stain of a positive. agalactiae A. The etiologic agent most commonly D. Serous effusions associated with septicemia and meningitis of newborns is 9. Streptococcus bovis group of the organism C. Lactococcus translucent beta-hemolytic colonies. B. Streptococcus agalactiae A. B. Nocardia asteroides infections in humans B. Otitis media D. Pathology at the point of entrance B. lysostaphin resistant. cereus producing Staphylococcus aureus. Septic shock 13. this 15. B. REVIEW QUESTIONS • 667 7. Draining cutaneous sinuses C. D. B. Streptococcus pyogenes B. Furuncles B. Corynebacterium diphtheriae C. Streptococcus pyogenes C.

Lactobacillus the upper respiratory tract is D. Staphyloccus aureus most other bacteria by B. 37°C could most likely be recovered from a D. Precipitates of diphtheria toxin and D. Pitting of the agar medium surround- B. The etiologic agent of the disease 24. Hodge test C. Heat shocking the culture C. The sheep blood agar plate is primarily the name of this test procedure is the result of streptolysin A. Xylose-lysine-desoxycholate B. Streptococci obtain all their energy from Corynebacterium diphtheriae are the fermentation of sugars to characterized by the observance of A. H B. when present. Streptococcus pyogenes C. Streptococcus agalactiae B. Streptococci are unable to synthesize the C. O D. Succinic acid B. Staphylococcus aureus. When an infection due to Streptomyces is erysipelas is suspected. CHAPTER 6: BACTERIOLOGY 16. Valeric acid precipitate in the surrounding agar 22. Opalescent colonies with a white D. Erysipelothrix 20. A common member of the normal flora of C. D-test A. S 18. Corynebacterium jeikeium B. M C. isolates can be separated from A. Incubating the culture at greater than 19. Hyaluronidase 17. Phenylethyl alcohol aerobic gram-positive bacilli? C. Bismuth sulfite tic used to differentiate which of the B. The beta-hemolysis produced by group A vitro means for detecting toxigenic strains Streptococcus seen on the surface of a of Corynebacterium diphtheriae. Drying the specimen before inoculat- stool sample if the primary plating ing the culture media medium included 25. Catalase D. Nocardia A. Lactobacillus C. The production of H2S is one characteris- A. Formic acid A. Naglertest D. Corynebacterium D. Thiosulfate citrate bile salts sucrose A. Streptobacillus moniliformis A. Liquefaction of the agar surrounding B. Staphylococcus epidermidis D. colonies of 21. Incubating the culture at 25°C D. Elektest B. Lactic acid the colonies on the medium C. Viridans streptococcus . Kinase ing the colonies C. Lipase antitoxin formed in agar gels are an in 23. Black colonies on the culture medium enzyme surrounded by brown halos A. On Tinsdale agar.

Enterococcus faecium pathogenic for swine. Corynebacterium urealyticum 31. and cattle is C. This organism when grown 32. appropriate when describing B. Alpha-hemolytic on the culture isolate. Rhodococcus equi A. The large. Nocardia being catalase positive C. which are A. Streptococcus pyogenes . Which one of the following is not the culture isolate. and the diagnosis of B. gram-positive D. An optochin test should be performed B. D. B. A child presented in August at the 29. Nocardia being an obligate anaerobe lesions were cultured. PYR negative A. 33. Enterococcusfaecalis naphthylamide) test is demonstrated by D. Viridans streptococci mised hosts. Inhibited by the presence of bile in filamentous growth away from the stab culture media near the top of the medium. Listeria monocytogenes A. Virulent strains are encapsulated. Growth in a 48-hour semisolid agar stab 30. Which of the B. Arcanobacterium hemolyticum primarily diplococci. itchy A. An Elek test should be performed on Streptococcus pneumoniae ? the culture isolate. Lancet-shaped. Streptococcus pyogenes also known to cause disease in compro. A. Staphylococcus saprophyticus D. The partial-acid fast staining reaction impetigo was made. D. REVIEW QUESTIONS • 669 26. C. A negative PYR (L-pyrolidonyl-a- C. Enterococcus faecalis 27. Beta-hemolytic B. Rhodococcus sp. This observa. A hippurate hydrolysis test should be diplococcus performed on the culture isolate. A Gram stain of a sputum specimen from on culture media produces pale pink a patient with a suspected case of lobar colonies that help to presumptively pneumonia reveals many white blood cells identify it as and many gram-positive cocci. given these findings? D. A PYR test should be performed on 28. Nocardia can be differentiated from pediatric clinic with a superficial skin Actinomyces based on infection of the neck. A former species of Corynebacterium B. Enterococcus faecium is characteristically culture at room temperature reveals lateral A. horses. Gardnerella vaginalis ate. Corynebacterium diphtheriae D. One of the etiologic of Actinomyces agents of this clinical condition is C. Erysipelothrix rhusiopathiae cases of nocardiosis B. The production of sulfur granules in A. D. Actinomyces naeslundii following statements would be appropri- C. Bile-resistant C. Able to grow in the presence of high tion is most characteristic of which concentrations of salt organism? C.

Corynebacteriumjeikeium D. B. is most commonly tic of Listeria monocytogenes ? associated with which of the following A. Streptococcus agalactiae B. Scarlet fever increased salt 41. Subacute bacterial endocarditis 37. Which of the following is not characteris. Growth inhibition in presence of D. Staphylococcus aureus 38. Otitis media C. Mannitol fermentation postive C. Which of the following is not associated with Staphylococcus aureus? 43. An identifying characteristic of 40. Streptococcus pyogenes isolation from clinical specimens? 44. Rust-colored sputum in cases of lobar A. Diphtheria C. Deoxyribonuclease production A. B. CAMP test postive clinical conditions? B. Motile C. Streptococcus bovis group D. thuringiensis . Enterococcusfaecalis 42. cereus B. Streptococcus pneumoniae of staphylococci that would help in their D. streptococcus. Resistance to novobiocin A. Staphylococcus saprophyticus 39. Growth at 55°C coccus (> 100. DNase negative A.670 • CHAPTER 6: BACTERIOLOGY 34. Bile resistance grew a catalase-positive gram-positive B. Staphylococcus aureus B. Catalase negative A. Coiynebacterium diphtheriae A. which would C. B. Esculin hydrolysis positive B. Clumping factor production the following possible etiologic agents? C. Streptococcus pneumoniae C. Coagulase negative B. Which of the following organisms is able commonly submitted to the clinical to hydrolyze sodium hippurate to benzoic laboratory for the detection of acid and glycine? A. Listeria monocytogenes D. Streptococcus viridans A. Streptococcus sanguis. Hemolysin production B. Relapsing fever D. Cutaneous anthrax B. B. Cultures of the posterior pharynx are most 35. Which of the following species of Bacillus B. A urine culture from a 23-year-old female A. Endotoxin production pneumonia is characteristic of which of B. Enterococcus faecalis is nonmotile? C. subtilis C. Which one of the following diseases Staphylococcus aureus is involves erythrogenic toxin? A. a viridans 36. Streptococcuspneumoniae C. anthracis D. Pharyngitis D. High salt tolerance most likely be D. Streptococcus pyogenes D. Impetigo D. Which of the following is a characteristic C.000 cfu/mL).

Streptococcus agalactiae 47. Corynebacterium C. which of healthcare workers? stained weakly acid-fast. Staphylococcus epidermidis A. The description of "Medusa head" C. Staphylococcus capitis compromised patients is C. Streptococcus bovis group Gram stain best describes D. Staphylococcus aureus B. filamentous. A bone marrow transplant patient on A. B. Corynebacterium ulcerans colonies on solid agar is most characteris- D. Streptococcus agalactiae was detected by MRI. Mycobacterium tuberculosis D. Streptococcus mutans B. Streptococcus agalaciae B. Staphylococcus saprophyticus neurologic involvement. Rhodococcus equi A. Actinomyces israelii C. Nocardia asteroides D. artificial heart valves and cerebrospinal theriticum fluid shunts is 48. Colonies of Listeria monocytogenes on a A. Leuconostoc D. The pulmonary form of anthrax is known as 51. Cystine-tellurite blood agar plates are 50. Corynebacterium diphtheriae D. Corynebacterium urealyticum cause bacteremia in hospitalized imrnuno. Streptococcus pyogenes C. A brain abscess D. Propionibacterium acnes . Listeria 46. Bacillus anthracis immunosuppressive therapy developed a B. Staphylococcus saprophyticus C. Bacillus anthmcis 52. Streptococcuspneumoniae A. Valley fever sheep blood agar plate most closely B. Enterococcusfaecalis pulmonary abscess with symptoms of C. The most common etiologic agent of B. An aerobic gram-positive rod known to A. Listeria monocytogenes insertion of prosthetic devices such as D. Corynebacterium urealyticum tic of 49. Bacillus cereus etiologic agent in this case would be B. Group D streptococci D. Farmers' lung A. and aspirated 54. Woolsorters disease B. Walking pneumonia resemble colonies of C. REVIEW QUESTIONS • 671 45. Which of the following is most likely to be material grew an aerobic. Which of the following is catalase recommended for the isolation of negative? A. Corynebacterium pseudodiph. Pleomorphic gram-positive bacilli in a C. Bacillus anthracis D. isolated in cultures from the anterior nares branching gram-positive organism. Bacillus B. Corynebacterium diphtheriae A. The most likely A. Bacillus subtilis infections associated with the surgical C. Corynebacterium jeikeium 53.

Streptococcus agalactiae etiologic agent in cases of C.1. Staphylococcus saprophyticus produced a black pigment on bile-esculin 60. A. The child's epidermis 58.672 CHAPTER 6: BACTERIOLOGY 55. A catalase-negative gram-positive coccus B. Abiotrophia sp. The most likely bacterial pustule" is cause of such symptoms would be food A. The bacterium D. Cojjnebacterium ulcerans A. Enterococcusfaecalis 62. Impetigo C. Streptococcus pneumonias B. Bacillus anthracis 59. Staphylococcus aureus C. Which of the following is associated with presumptive test for the identification of infections in humans often linked to deli A. Streptococcus pneumoniae C. Streptococcus pyogenes associated with C. The D. An infant was hospitalized with a severe. Group B streptococci D. Group D streptococci . likely identification of this bacterium? B. Streptococcus mutatis A. Bacillus subtilis D. Listeria monocytogenes 56. Erysipelothrix rhusiopathiae B. Streptococcus pneumoniae A. Bacillus cereus has been implicated as the B. Streptococcus pyogenes products? C. The novobiocin susceptibility test is used was loose. Enterococcus faecalis D. Food poisoning D. and large areas of skin could for the identification of be peeled off. Streptococcus pneumoniae B. Toxic shock syndrome illness was characterized by rapid onset of 63. Listeria monocytogenes 64. Ethylhydrocupreine HC1 susceptibility is a 6. Leuconostoc desoxycholate is characteristic of D. Viridans streptococci meats and improperly pasteurized dairy B. Erysipelothrix rhusiopathiae A. Streptococcus agalactiae A. Corynebacterium diphtheriae is isolated from a urine sample of a C. What is the most A. Listeria monocytogenes C. Corynebacterium diphtheriae most consistent with a clinical syndrome B. D. Staphylococcus saprophyticus B. Streptococcus pyogenes D. The condition described is A. Abiotrophia sp. The etiologic agent of the majority of agar and formed acid from glucose in the adult joint infections is presence of 6. Family members attending a picnic became ill about 2 hours after eating. Bacillus anthracis poisoning caused by B. Solubility in the presence of sodium C. Bacillus subtilis D. Enterococcusfaecium C. The causative agent of "malignant violent vomiting. Streptococcus agalactiae A. Leuconostoc sp. Erysipelothrix rhusiopathiae hospitalized patient. Bacillus anthracis 65.5% NaCl. Staphylococcus aureus tender erythema. C. Tellurite reduction is used for the presumptive identification of D. Staphylococcus aureus B. Pelvic inflammatory disease 57.

The enterotoxins of both Vibrio cholerae several of the nebulizers. The color D. Pseudomonas aeruginosa C. Pseudomonas A. Penetration of the bowel mucosa negative. K. The organism produces disease most commonly in the pediatric population. pneumoniae 71. K. Cytochrome oxidase A. A. The organism is a delayed lactose Klebsiella are fermenter. Blood B. An environmental sampling study of C. nonfermentative (at 48 hours). oxytoca cultures of a yellow. K. REVIEW QUESTIONS • 673 Aerobic Gram-Negative Bacteria 70. Colonies ofNeisseria sp. Salmonella enterica a redox reagent is applied. citrate negative. Urine tory condition in the large intestine C. The majority of clinical isolates of D. C. C. Shigella dysenteriae change is indicative of the activity of the bacterial enzyme 73. 67. aerogenes respiratory therapy equipment produced D. The characteristics of being lactose B. Proteus vulgaris B. Which of the following is not true of 66. Nasopharyngeal exudates C. A fermentative gram-negative bacillus that A. Urease on MacConkey agar is C. D. turn color when C. Cerebrospinal fluid with bloody diarrhea. and D. urease negative. Stenotrophomonas maltophilia D. Yersinia enterocolitica . Phenylalanine deaminase B. Alcaligenes which gives rise to excessive fluid D. Stimulation of colicin production lysine decarboxylase negative. Beta-galactosidase is oxidase positive. Chryseobacterium by what mechanism? B. Aeromonas hydrophila D. gram-negative bacillus from 68. ozaenae B. the Shigella sonnei? optimal specimen to be collected for the A. The organism produces an inflamma- B. Moraxella secretion by the cells of the small intestine 72. K. Stimulation of adenylate cyclase. Yersiniapestis 69. motile. and grows well B. In suspected cases of brucellosis. Elaboration of a dermonecrotizing nonmotile best describe which organism? toxin A. A. Large numbers of organisms must be isolation of the etiologic agent is ingested to produce disease. which would Ol and noninvasive (toxigenic) strains of most likely be species of Escherichia coli produce serious diarrhea A.

Aeromonas hydrophila A. Chromobacterium C. Escherichia coli B. Moraxella catarrhalis C. Escherichia coli C. Kingella D. The most likely identification of this etiologic agent would be A. Campylobacter D. Stenotrophomonas maltophilia D. Vibrio follows: C. Citrobacter (diversus) koseri D. Bartonella bacilliformis C. culture media supplemented with either glucose nonfermenting organism is cysteine or cystine? A.674 • CHAPTER 6: BACTERIOLOGY 74. The somatic antigen ONPG positive C. Pseudomonas aeruginosa D. acid butt. The causative agent of melioidosis is The organism was identified as A. Shigella gas. Located on the flagellum Voges-Proskauer negative D. Salmonella Citrate negative 75. Test reactions in other media were as B. Klebsiella pneumoniae C. Enterobacter cloacae B. Actinobacillus lignieresii B. Providencia stuartii 77. Capnocytophaga B. On TSI agar specifically for the isolation of it produced an acid slant. Plesiomonas . Francisella tularensis D. The antigen used to group Shigella 76. Which microorganism will grow only on 79. A fastidious gram-negative bacillus was isolated from a case of periodontal disease. Burkholderia cepacia A. Heat labile B. Burkholderia pseudomallei B. Kingella kingae 80. The K antigen of the family Indole positive Enterobacteriaceae is Urease negative A. Fecal cultures are inoculated on 78. A culture of a decubitus ulcer grew a thiosulfate-citrate-bile salts-sucrose gram-negative facultative bacillus. An example of an oxidase-positive. and A. which upon darkfield examination was noted to have gliding motility.

Syphilis reactions shown in Color Plate 27 •? D. Which of the following organisms would B. The etiologic agent of whooping cough is 84. The bacteria survive long periods small. Pseudomonas dermatitis D. which differentiates B. Campylobacter spp. Francisella tularensis of the following is not characteristic of D. V. V. Gastroenteritis A. Acinetobacter sp. C. Endocarditis B. Neisseria meningitidis an enteric pathogen because it produces a 83. gram-negative bacillus. Citrobacterfreundii B. which is caused by a D. Serotype d 89. Produces disease in humans and ized by genital ulcers and tender inguinal domestic animals lymphadenopathy. The sexually acquired disease character. Brucella suis it from the other salmonellae. Asymptomatic infections are common D. Citrate negative in females. Q fever A. Osteomyelitis septicemia caused by B. Bacterial vaginosis most likely produce the biochemical C. 85. is known as outside the host's body. Pseudomonas putida C. Chancroid 91. Burkholderia cepacia A. REVIEW QUESTIONS • 675 81. Which of the following diseases is most with rapidly progressing wound infections likely to be acquired from a hot tub or seen in patients with underlying liver whirlpool? disease is A. A. Bordetella pertussis biochemical pattern. are associated most 82. An organism occasionally misidentified as D. Serotype c D. Serotype a B. Appendicitis C. V. Severe disseminated intravascular frequently with cases of coagulation often complicates cases of A. Burkholderia pseudomallei B. Serotype b C. A. C. C. Typhi? 90. Shewanella putrefaciens D. Which C. Acinetobacter cellulitis C. Salmonella Typhi exhibits a characteristic A. Salmonella . Neisseria gonorrhoeae 88. Trachoma A. parahaemolyticus 87. Agglutination in Vi grouping serum gram-negative bacilli. Moraxella sp. alginolyticus B. Haemophilus ducreyi S. Erysipelas B. vulnificus D. A Gram stain of the organism reveals B. The species of Vibrio closely associated 86. An important characteristic of Neisseria A. Large amounts of H2S are produced in gonorrhoeae or the infection it produces is TSI agar. V. cholerae C. D. Proteus mirabilis C. Providencia rettgeri D. The Haemophilus influenzas vaccine large amount of H2S is protects against which serotype? A. Lysine decarboxylase positive B.

gergoviae D. Lactose infect reptiles and fish as well as humans C. Stenotrophomonas maltophilia D. Chryseobacterium B. Pseudomonas aeruginosa C. Swine fever rarely isolated in the U. Oxidase negative A. tarda . Erythromycin eye drops are routinely culture? administered to infants to prevent A. Nasopharyngeal swab A.676 CHAPTER 6: BACTERIOLOGY 92. E. Haemophilus influenzae D. Chromobacterium A. A. Which of the following nonfermenters is A. Enterobacter C. Chlamydia trachomatis 103. A straight gram-negative bacillus C. Haemophilus aegyptius C. The single species in the genus Hafiiia is B. Requires cysteine istic of B. Hippurate hydrolysis positive B. A causative agent of the form of conjunc. Pseudomonas aeruginosa 101. A. Aeromonas tivitis known as pinkeye is B. Yersinia pestis D. Hemolytic uremic syndrome is a compli- D. 99. co//O157:H7 Neisseria meningitidis but can be B. Rift Valley fever B. Burkholderia cepacia the most sensitive specimen to submit for 94.S. Stool C. Neisseria gonorrhoeae cation after infection with 95. Neisseria lactamica closely resembles A. coli C. alvei C. Moraxella lacunata D. Legionella pneumophila is the etiologic D. Is oxidase negative A. Bartonella spp. Nonmotile 97. Actinobacter spp. Identify the fermentative agent that may B. Ferments glucose B. Klebsiella ozaenae A.? B. In suspected cases of brucellosis. Acinetobacter baumannii morphology is a distinguishing character. Bone marrow infections by B. Glucose when they are exposed to contaminated D. Maltose 1. Does not grow on MacConkey agar C. E. A gram-negative. Yersinia enterocolitica A. Salmonella Typhi differentiated from it by its ability to C. Sputum B. Campylobacterjejuniis D.02. Sucrose soil or water. "kidney bean" cellular 98. Burkholderia mallei 100. Pontiac fever A. Vibrio cholerae Ol metabolize D. San Joaquin Valley fever C. 96. what is D. agent of both Legionnaires disease and 93. Neisseria meningitidis C. ruckeri D.

Which of the following has a negative 109. watery diarrhea A. Vibrio supplemented media for isolation. Hafnia B. They are fastidious and require D. agglutination will occur in which grouping sera? A. Plesiomonasshigelloid. Oxidase negative A. C. Generally coccobacillary in morphology an infection caused by C. Vibrio cholerae D. An example of a halophilic micro. B. A. Adversly affected by fatty acids in D. Infection may occur via abrasions of the B. and genitals. A B. O antigens 111. REVIEW QUESTIONS • 677 104. most characteristic of infection caused by organism is A. C. Campylobacter jejuni A. Enterohemorrhagic Escherichia coli D. Helicobacterpylori A. Shigella dysenteriae C. Morganella morganii B. Explosive watery diarrhea with severe abdominal pain after eating raw shellfish is 108. Yersinia enterocolitica 112. Salmonella Typhi B. After heating the same suspension. An unheated suspension of Salmonella Typhi typically produces agglutination of Vi C. Vibrio parahemolyticus D. conjunctiva. Campylobacterjejuni in the clinical laboratory is based on B. Which of the following statements is not oxidase test? true of Brucella? A. Helicobacter pylori B. D .? 107. Phage and dye sensitivity tests are used clinical specimens for identification to the species level. The method of serogrouping Shigella used A. Chryseobacterium oral mucosa. Bacteriocins C. The symptom of diffuse. Yersinia pestis 113. The risk of accidental laboratory 105. Infections associated with use of B. Aeromonas A. K antigens D. Ferments glucose and maltose 110. Commonly susceptible to most that produces a relatively clear stool antimicrobials containing mucus flecks is suggestive of B. Shigella dysenteriae medical devices C. Which of the following is true of infection is no greater than with any Neisseria gonorrhoeae ? other organism. Vibrio parahaemolyticus D. B C. H antigens D. G! D. Rapid growth on sheep blood agar C. Which of the the following does not describe Acinetobacter sp. ONPG positive gastritis or peptic ulcers is most com- monly associated with 106. Infection of the gastric mucosa leading to D. Shigella sonnet C.

Lack of motility is characteristic of B. Beta-hemolytic on sheep blood agar 115. Its positive phenylalanine deaminase D. A positive Voges-Proskauer reaction is C. sputorum A. Specimens may be cold enriched to infections of Vibrio cholerae? enhance recovery of the organism A. C. C. Person-to-person transmission is A. Intracellular pathogens B. Its negative oxidase reaction 121. Green colonies with black centers on reaction Hektoen enteric agar C. by C. Colorless colonies on xylose-lysine- A. O4 A. laris for Escherichia coli? D. Patients can present with a self-limited 118. Which of the following reactions is typical C. C. Enterobacter aerogenes 122. Shigella sonnet is differentiated from B. Ol 124. Enterobacter cloacae C. Hydrogen sulfide positive 116. Mucinase production B. Is the causative agent of granuloma inguinale D. Farm animals are important animal C. Is often sexually transmitted B. Escherichia coli A. septic arthritis. Its ability to demonstrate motility at A. Morganella morganii 111. coli D. Toxigenic C. B. Klebsiella oxytoca D. O2 istic of Klebsiella (Calymmatobacteriuni) C. The classic toxigenic strains of which nonpneumonic febrile illness serogroup are implicated in epidemic D. The species of Campylobacter noted to 119. Urease negative 22°C B.878 • CHAPTER 6: BACTERIOLOGY 114. A positive ONPG desoxycholate agar B. Colorless colonies on MacConkey agar other Shigella sp. jaundice with hepatomegaly. C. Oxidase positive A. Yersinia pestis is characteristically D. O3 granulomatis? D. In cases of legionellosis cholerae? A. Nonmotile reseroirs D. Which of the following is not true D. Proteus vulgaris B. Motile at 20-25°C characteristic of D. The etiologic agents of relapsing fever meningitis. Is isolated on chocolate agar C. Small spiral organisms and thrombophlebitis in debilitated C. Adherent to enterocytes common B. Can be diagnosed by detection of Donovan bodies in clinical specimens . Providencia stuartii regarding virulent strains of Vibrio 123. A. Which of the following is not a character- B. Primarily a cause of endogenous patients is human infections A. Providencia rettgeri C. fetus 120. are produce septicemia. Brucella spp.

Serratia marcescens isolated on chocolate agar incubated in CO2. Salmonella Choleraesuis B. Shewanella putrefaciens human pathogen that is known to cause A. an inhabitant of butt on TSI agar after incubating 18 the upper respiratory tract of humans. An identifying B. Pneumonia C. Escherichia coli cerebrospinal fluid from a 1-year-old girl B. Moraxella catarrhalis 129. Shigella boydii D. Pseudomonas aeruginosa B. Positive catalase 130. Stenotrophomonas maltophillia B. REVIEW QUESTIONS • 679 125. Escherichia coli is applied? B. From contaminated sites is made C. Isolation ofNeisseria gonorrhoeae 132. Cardiobacterium hominis. Stenotrophomonas maltophilia D. Indole negative produced by D. A positive indole reaction is one will turn a dark purple when tetram- characteristic of ethyl-p-phenylenediaminehydrochloride A. Pyelonephritis D. Klebsiella oxytoca suspected of having meningitis. Positive oxidase D. Citrobacterfreundii A. Sheep blood agar also incubated 126. has hours? been recovered as the etiologic agent from A. which 128. Inability to grow on sheep blood agar A. Yersinia enterocolitica would produce a yellow slant and a yellow 134. Color Plate 28 • shows the Gram stain of A. Shigella sonnei B. Haemophilus influenzae D. A positive DNase would be seen with 131. small tan colonies were D. Proteus mirabilis A. Neisseria meningitidis 127'. Pyocyanin is characteristically C. Serratia marcescens C. Vibrio vulnificus is a well-established C. A negative citrate reaction is in CO2 had no growth. Enterobacteraerogenes B. Bordetella parapertussis C. Which of the characteristic of following organisms should be suspected? A. Is not affected if clinical specimen is worldwide refrigerated before culturing 133. Of the following microorganisms. Escherichia coli cases of endocarditis. Proteus mirabilis hours of growth. Gastroenteritis D. Which of the following is true concerning A. Is enhanced by cold enrichment Campy lobacter jejuni ? B. Isolated best at 24°C C. A leading cause of bacterial diarrhea D. Catalase negative CO2 B. Wound infections . Requires incubation under increased A. Proteus mirabilis characteristic of the organism is C. Hydrogen sulfide positive easier by the use of CIN agar D. Acinetobacter baumannii C. After 24 C. Pseudomonasfluorescens 135. Serratia marcescens C. Salmonella Typhimurium A. Which one of the following organisms D. Brucella canis B.

Motile 140. Stains easily on the routine Gram stain tic of Eikenella corrodens? D. Klebsiella pneumoniae B. Burkholderia cepacia organism was oxidase negative. Which of the following non-lactose. and motile.? A.• CHAPTER 6: BACTERIOLOGY 136. The B. cloacae C. Pseudomonas aeruginosa coccobacilli 145. It is a thin. Buffered charcoal yeast extract 137. Citrobacterfreundii A. Pseudomonas putida D. sakazakii D. Able to grow on MacConkey agar B. Pseudomonas alcaligenes A. Metabolizes a number of carbohydrates 146. Klebsiella pneumoniae B. E. Small. taylorae 138. E. E. Yersinia enterocolitica D. Stool respiratory tract of humans. Bordet-Gengou C. Which of the following organisms is 144. aerogenes B. Which of the following organisms 142.? A. 141. Which of the following is the optimal B. Sensitive to penicillin from environmental sources D. Regan-Lowe B. gram-negative bacillus. Production of a yellow pigment is fermenting organisms does not produce characteristic of which of the following fluorescein? Enterobacter sp. Pseudomonasfluorescens C. Modified Skirrow's medium D. Proteus mirabilis C. Plesiomonas shigelloides most likely identification of this agent would be 139. Grow well on sheep blood agar C. Blood D. Which of the following is not characteris- C. E. 143. Nasopharyngeal swab . Does not autofluoresce A. Acinetobacter baumannii is characteristi- D. The D. Bronchial washings recovered from infections. Obligate parasites B. lactose C. B. A gram-negative bacillus was recovered unable to grow on MacConkey agar? from the urine of a child with a history of A. Pseudomonas aeruginosa B. Providendia stuartii C. gram-negative D. Which of the following is not an appropri- produce a positive phenylalanine ate medium for primary isolation of deaminase reaction? Bordetella pertussis ? A. Many are found as normal flora in the cally human respiratory tract. Legionella pneumophila ? C. Escherichia coli A. Colonies sometimes pit the surface of clinical specimen for the recovery of the agar medium. D. Legionella pneumophila B. It is often found in pure culture when C. pleomorphic. Kingella denitrificans negative. A. urease positive. Oxidase positive A. Which of the the following is not true of Haemophilus spp. Infections are most often acquired C. It is found in the mouth and upper A. Bordetella bronchiseptica recurrent urinary tract infections.

pneumophila A. Able to grow on Thayer-Martin agar D. L. canis likely organism to be isolated in these C. Which of the following is not produced by Serratia species? 150. Anterior nares swab B. Kingella denitrificans can be differenti- A. Enterobacteriaceae D. Klebsiella pneumoniae 148. Blood C. Which of the following species of 152. P. Haemophilus ducreyi is the causative D. REVIEW QUESTIONS • 681 147. povidone iodine. Requires X factor ability to produce extracellular hydrolytic D. Which of the following Legionella spp. multocida cases is D. Lymphogranuloma venereum family? A. Glucose positive agent of A. avium of microbial contamination. Gelatinase it is C. DNase ated from Neisseria gonorrhoeae because B. The optimal specimen for the recovery D. gormanii most commonly associated with eating D. L. Chancroid 151. Serratia spp. L. Contaminated potato salad tic of Haemophilus influenzae biogroup D. Oxidase positive 155. Lipase A. Foodborne outbreaks of brucellosis are C. longbeachae 153. Requires V factor Enterobacteriaceae because of their C. stomatis A. Imported cheese 149. and other infections following cat bites? disinfectants have been recalled because A. Legionellaceae 156. P. Aminolevulinic acid positive enzymes. are unique in the family B. Whooping cough C. is C. NADase B. Bordetella bronchiseptica B. Trachoma B. Plesiomonas was recently moved to which B. Vibrionaceae of Bordetella pertussis is A. The most B. micdadei B. Raw shellfish B. Able to reduce nitrates C. P. tincture of Pasteurella is associated with human iodine prep pads and swabs. Pseudomonas aeruginosa positive for hippurate hydrolysis? D. L. Nasopharyngeal swab . Expectorated sputum D. Serratia marcescens A. P. Improperly cooked hamburger aegyptius? A. In the past. Indole negative 154. Brucelleaceae C. Which of the following is not characteris- C.

have had their microorganisms? vacations interrupted by a case of A. Campylobacterjejuni C. Yersinia enterocolitica B. Swimmer's ear. Campylobacter jejuni C. Eikenella A. positive? B. Gardnerella C. Pasteurella multocida vaginalis not recommended? A. Buffered charcoal yeast extract agar is the 168. Which of the following is the most 163. Vibrio cholerae C. Epiglottitis "seagull wings" is most characteristic of B. Yersinia enterocolitica 162. Neisseria lactamica D. Bordetella bronchiseptica C. Aeromonas hydrophila D. a morphology that resembles A. Mesenteric lymphadenitis B. Vibrio parahemolyticus manifest commonly as which of the following clinical conditions? 165. Listeria D. Rhodococcus equi A. Salmonella clinically significant. Neisseria gonorrhoeae 161. Acinetobacter baurnannii "traveler's diarrhea. Plesiomonas shigelloides D. Capnocytophaga B. Pseudomonas aeruginosa 158.S. On Gram stain. produces acid recommended medium for the recovery of from glucose but not from maltose. Chlamydia trachomatis A. Pseudomembranous colitis C. Brucella canis associated with which etiologic agent? C. Neisseria gonorrhoeae C. Which of the Neisseria spp. Neisseria meningitidis D. D. Cefsulodin-irgasan-novobiocin (CIN) agar is recommended for the recovery of 166." which is commonly B. Acinetobacter baurnannii B. It is unsafe to grow this bacterium. Yersinia pseudotuberculosis is known to D. B. i CHAPTER 6: BACTERIOLOGY 157. C. Neisseria sicca . Which of the following is not urease results take too long to be relevant. Why are cultures for Gardnerella D. Neisseria meningitidis B. A number of vacationers who have recovery of which of the following traveled outside the U. Blood cultures are recommended for the 164. Hafnia alvei lactose. Yersinia pseudotuberculosis 160. Legionella pneumophila A. Haemophilus influenzae D. Aeromonas hydrophila D. Neisseria gonorrhoeae D. a form of external otitis common indicator of bacterial vaginosis? is commonly caused by A. Hepatitis A. The bacteria grow so slowly that 167. Isolation of the bacteria may not be A. Artificial media are not available. Escherichia coli C. Cardiobacterium hominis said to resemble kidney beans? C. The flattened adjacent sides of the cellular A. Helicobacterpylori C. or sucrose? B. Yersinia enterocolitica B. Proteus mirabilis 159. Proteus mirabilis D. A. Brucella suis appearance of which microorganism are B.

Morganella morganii A. Haemophilus aegyptius showed many white blood cells and D. Klebsiella 178. Catalase positive A. Violet-colored colonies are typically A. Serratia marcescens isolated in stool specimens and can biochemically be confused with A. Oxidase positive C. It grows on sheep blood agar. Neisseria meningitidis their normal flora. Chromobacterium violaceum C. It is the most virulent of the species in the genus. Which of the Neisseria spp. Neisseria sicca C. Enterohemorraghic E. ONPG negative B. Yersinia enterocolitica . Yersinia pestis numerous gram-negative bacilli. Proteus vulgar is B. B. The most likely of etiologic agent in this case would be A. Serratia hydrophila? 174. Salmonella B. Growth on sheep blood agar 173. Salmonella description regarding Aeromonas D. Incubation at 22°C C. Chryseobacterium meningosepticum D. Edwardsiella tarda is occasionally D. Pus was aspirated from an empyema. Growth on typtic soy agar without A. Moraxella catarrhalis A. Most common human infections occur A. Salmonella Enteritidis C. Lack of motility is a characteristic of D. D. Vibrio cholerae D. Hafiiia alvei production is enhanced by B. Serratia liquefaciens D. Proteus vulgaris B. Neisseria lactamica B. Humans harbor the organism as part of C. Burkholderia mallei A Gram stain of the aspirated material C. and joints. D. Bartonella bacilliformis 176. The culture grew many colonies producing a 111. Pigment A. Legionellapneumophila C. Salmonella Typhimurium C. Enterobacter blood B. Pseudomonas aeruginosa D. REVIEW QUESTIONS • 683 169. A positive phenylalanine deaminase 111. Serratia marcescens 172. hemolytic disease known as Oroya fever is A. Which of the following is not true about from glucose and maltose but not sucrose Pasteurella multocida? or lactose? A. coli C. Incubation at 42°C D. A positive gelatin reaction is characteristic soluble green pigment. Beta-hemolytic produced by B. produces acid 175. Pseudomonas aeruginosa 179. bones. 170. Neisseria gonorrhoeas in soft tissues. Chromobacterium violaceum B. Which of the following is not a correct C. Some strains of Serratia marcescens reaction is characteristic of produce a red-colored pigment. The causative agent of the septicemic. B.

Buccal cavity C. When an epidemiologic survey for the agar is due to the inclusion of which one of the following? detection of upper respiratory tract A. Burkholderia cepacia C. Chronic carriers. Which of the following is an ocasional D. NAD A. Bile salts earners ofNeisseria meningitidis or Bordetella pertussis is being conducted. Cefsulodin-irgasan-novobiocin agar C. Haemophilus influenzas D. Salmonella Typhi 181. Klebsiella pneumoniae intensive care units is commonly a factor D. Colonies that are said to resemble for the cultivation ofNeisseria gonorrheae? A. Chocolate agar "droplets of mercury" are characteristic of A. Proteus vulgaris in outbreaks of infections with which of the following microorganisms? 183. fiz's'-sodium metasulfate C. Yersinia pestis cause of respiratory tract infections and is rapidly urea positive? 187. Glanders 182. Modified Thayer-Martin agar D. are typically associated with the B. Bordetella pertussis B. Campylobacterfetus cattle transmissible to humans. The porphyrin test determines an organ. Bordetella pertussis D. The characteristic growth pattern known A. 186. Enterobacter aerogenes 188. Pontiac fever cases of cystitis is A. such as D. Thiol B. Association with faucet aerators and B. Brucella abortus diseases. One of the most common etiologic agents C. Hemin dissemination of C. Campylobacter jejuni C. The selective nature of Hektoen enteric 185. Salmonella spp. Yersinia pestis 190. Campylobacterjejuni D. Which of the following is not appropriate 184. Brucellosis B. Escherichia coli humidifiers used with ventilators in C. Nasopharyngeal D. Bromthymol blue the optimal type of specimen to be D. B. Anterior nares B. BurkhoIderia pseudomallei C. CHAPTER 6: BACTERIOLOGY 180. Serratia marcescens C. Escherichia coli A. Campylobacter jejuni D. Martin-Lewis agar B. Pseudomonas aeruginosa A. especially for those diseases of C. Yersinia pestis \. Throat . Meliodosis of community-acquired uncomplicated D. Bordetella bronchiseptica associated with the transmission of some B. Milk has classically been the primary food A. Klebsiella pneumoniae as "satelliting" is associated with B. persons who remain ism's requirement for infected with an organism for long A. B. NaCl obtained for culture is A. Cystiene periods.

Campylobacterjejuni used. The most common C. which B. Neisseria gonorrhoeae D. Tinsdale agar A. When performing the oxidase test. Calcium alginate or Dacron swabs are C. DFA test results are definitive and do comitans not need to be confirmed by culture. which 194. Thiosulfate-citrate-bile salt-sucrose agar that had been immersed in a contaminated B. Chiyseobacterium meningosepticum 192. Yersinia enterocolitica the reagent. Chiyseobacterium meningosepticum recommended over cotton-tipped D. A. Actinobacillus actinomycetem- B. A nichrome wire loop should be used ulcers and is the most frequent cause of to acquire inoculum for testing. Burkholdia cepacia C. B. 196. Vibrio . D. B. 197. Charcoal yeast extract agar cleaning solution. Salmonella D. Neisseria 199. Helicobacter pylori D. Which of the following is not true about of the following is an important cause of the laboratory diagnosis of pertussis? lower respiratory tract infections in A. Urinary tract infections A. The organism that is linked to peptic B. Pneumonia 195. Pseudomonas aeruginosa of the following would not be appropriate? C. Salmonella Typhimurium within 10 seconds after application of D. 200. but it is most noted for causing ynx is optimal for detection of the A. Haemophilus C. Serratia marcescens swabs for specimen collection. B. REVIEW QUESTIONS • 685 191. Bordetella pertussis A. New York City agar was developed for the 193. gastritis is C. The reagent used is o-nitrophenyl-(3. Haemophilus influenzae C. patients with cystic fibrosis? mended medium. Escherichia coli D-galactopyranoside. Abscesses of the oral cavity causative agent. Besides Pseudomonas aeruginosa. Postsurgical wound infections associated with the transmission of D. Eikenella corrodens is an opportunistic D. Reptiles kept as pets are sometimes C. For the selective isolation of Vibrio spp. Helicobacter C. Campylobacter B. 198. Colonies from sheep blood agar can be A. Mannitol salt agar bacterial etiologic agent in such cases is D. A positive colony turns dark purple C. Francisella tularensis A. Material collected from the nasophar- pathogen. A young man developed keratitis the recommended agar is associated with the use of contact lenses A. Moraxella D. Regan-Lowe medium is the recom. Campylobacter fetus B. The porphyrin test is most useful for the isolation of identification of which of the following? A. Campylobacter B.

Slow growing likely etiologic agent in this case would be B. M. That component is for digestion and decontamination of the A. bovis C. marinum D. The species of Mycobacterium that would be most commonly associated with 206. Gimenez stain D. Mycobacterium bovis large institutions such as hospitals is B. Teichoic acid C. 6%NaOH C. the generally recommended method bacteria lack. Mycobacterium leprae B. When clinical specimens are processed for 201. M. orange-pigmented. kansasii B. chelonei A. Mycobacterium fortuitum. M. Mycobacterium avium-intracellulare C. M. Mycobacterium fortuitum D. Nonchromogenic complex D. M. Lipids sample is B. M. Kinyoun stain 204. avium complex B. Wright's stain acid-fast bacillus was isolated from a 209. Which of the following mycobacteria has an optimal growth temperature 30-32°C? A. Dieterle stain C. The etiologic agent of Hansen disease is contamination of the hot water system in A. M. Obligate aerobe A. grows on reported as MacConkey agar in 5 days. ulcerans 207. tuberculosis C. M. a rapidly smear of expectorated sputum should be growing Mycobacterium. xenopi . M. D. Murein A. M. 4+ A. Trisodium phosphate 202. NALC-NaOH D. Mycobacterium scrofulaceum 210. xenopi per field (X 800 to X 1000) in a fuchsin 203. HC1 D. bovis 208. haemophilum C. The most A. A slowly growing. Mycobacteria can be examined by using the B. Sterols B. Mycobacterium chelonei respiratory route C. ulcerans D. 1 + species of Mycobacterium is able to B. Mycobacteria have a large amount of a the recovery of Mycobacterium tuberculo- component in their cell wall that other sis. 3+ period on MacConkey agar? D. Which other A. 2+ demonstrate growth within the same time C. Mycobacterium fortuitum A. M.686 • CHAPTER 6: BACTERIOLOGY Mycobacteria 205. Which of the following is not characteris- cervical lymph node of a child with tic of Mycobacterium kansasii? symptoms of cervical adenitis. Mycobacterium tuberculosis C. Infections usually acquired by B. The finding of five to six acid-fast bacilli D. M.

bovis B. avium-intmcellularae complex 221. xenopi . Which one of the following tests would be A.fortuitum D. is C. Bactericidal for normal flora B. bovis D. In the decontamination and digestion 217. Which of the following is associated with A. Elek test D. M. tuberculosis D. M. M. appears as buff-colored colonies after ria from sputa samples. Which of the following Mycobacterium is A. M. M. Which of the following is a fluorescent D. M. M. asiaticum D. Which of the following Mycobacterium considered as primary antimycobacterial produces an orange pigment and is most therapy? commonly recovered from water? A. Fluorescein isothiocyanate syndrome? D. M. The most common photochromogenic B. bovis A. M. Growth stimulant D. PPDtest which of the following mycobacteria? 214. M. Ziehl-Neelsen A. M. Liquefies mucus C. kansasii 212. bovis C. M. Rifampin C. M. M. gordonae C. REVIEW QUESTIONS • B87 211. Which of the following Mycobacterium procedure for the isolation of mycobacte. M. Neutralizes pH to prevent damage to 218. M. M. kansasii A.fortuitum appropriate in the diagnosis of a mycobac. M. xenopi stain for mycobacteria? 215. CAMP test 219.S. kansasii C. M. xenopi B. M. tuberculosis C. M. M. tuberculosis C. M. Susceptibility to thiophene-2-carboxylic C. marinum Mycobacterium isolated in the U. avium-intracellularae complex livestock and causes a zoonosis? B. ulcerans D. M. what is the role of exposure to light and is niacin positive? NALC? A. scrofulaceum B. M. Which one of the following drugs is not 216. intracellulare B. Auromine-rhodamine most noted for being associated with B. M. Pyrazinamide D. Kanamycin B. M. B. Isoniazid A. Calcofluor white patients with acquired immunodeficiency C. M. Naglertest acid hydrazide (T2H) is characteristic of D. marinum A. The Mycobacterium that is the etiologic mycobacteria agent of "swimming pool granuloma" is 213. kansasii terial infection? C. gordonae 220. kansasii B. bovis A.

Clostridium septicum resembles 221. Which of the following is not considered a A. Botulism D. It is a gram-positive. and was negative for A. 42°C likely presumptive identification of this isolate would be 223. Which of the following specimens is grew an anaerobic gram-negative bacillus routinely decontaminated when trying to that was inhibited by bile. The characteristic colony morphology of C. should be incubated at plate shows colonies surrounded by an A. Clostridium perfringens be routinely used to culture B. Usually grows in peripheral limbs of A. The potentially lethal intoxication type of C. A molar tooth A. Leptospirosis . Lowenstein-Jensen-Gruft D. Clostridium perfringens Actinomyces israelii on solid agar D. CHAPTER 6: BACTERIOLOGY 222. The most D. Clostridium botulinum 231.? black pigment. Which of the following media would not A. Cerebrospinal fluid A. Easily transmitted from person to person Clostridium tetani ? D. Porphyromonas gingivalis A. Fusobacterium nucleatum A. Bacteroides fragilis group 225. B. Skin cultures for the recovery of Mycobac. B. Anthrax C. Bacteroidesfragilis acting on the central nervous system. Brucellosis D. It produces rapid tissue necrosis. Causes Hansen disease D. Chocolate agar sent to the laboratory for bacteriologic examination. Bacteroides ureolyticus Mycobacterium leprae? C. Ground glass C. improperly canned food is caused by D. produced a recover Mycobacterium spp. and lactose fermentation. The culture of this sample 224. A fried egg B. An anaerobically incubated blood agar terium spp. A cervical mucosal abscess specimen was D. "Medusa head" zoonotic disease? B. spore-forming Anaerobic Bacteria bacillus. Prevotella melaninogenica B.? C. infected patients B. MiddlebrookVHll 229. Sputum indole production and positive for glucose. 35°C (double zone of hemolysis). Disease is caused by an exotoxin A. Lowenstein-Jensen C. Which of the following is not true of B. Which one of the following is not true of C. 226. Difficult to grow in vitro 230. Pleural fluid sucrose. Microorganisms in soil contaminate food poisoning often associated with puncture wounds. 22°C inner zone of complete red cell lysis and B. Prevotella melaninogenica B. Lung biopsy isolate would most likely be D. Clostridium tetani Mycobacterium spp. 30°C an outer zone of incomplete cell lysis C. This C. 228.

Eubacterium lentum B. A puncture wound with a contami- seen? nated household item A. were found to be resistant to C. A scratch wound caused by a cat only under anaerobic conditions and was B. Assays to detect toxin in stool penicillin. Clostridium perfringens C. Propionibacterium acnes . gram-negative. Lactobacillus 234. Peptostreptococcus cular weakness and constipation. Growth of this organism was D. D. Mobiluncus obligately anaerobic gram-positive cocci? 239. Fusobacterium nucleatum C. The C. Veillonella firmed by the demonstration of toxin in 235. Ingestion of preformed toxin found B. gram-positive. Bacteroides fragilis B. primarily used for forming bacteria A. The anaerobic. spore-forming colitis (Clostridium difficile associated bacteria disease) is often made by 233. Ingestion of spores that germinated not acid-fast. Bacteroides fragilis in a contaminated jar of pureed C. Acid-fast stain of fecal material not inhibited by bile. curved. gram-negative. What is the most 238. REVIEW QUESTIONS • 689 232. Culturing blood specimens wound. Capnocytophaga department with symptoms of neuromus- B. Serology bacilli. Bifidobacterium D. Clostridium septicum vegetables D. Color Plate 29 • shows the filamentous the child's stool. non-spore. Which of the following is described as D. gram-negative. An infant was seen in the emergency A. Growth on kanamycin-vancomycin laked of the colon? blood agar incubated anaerobically is A. Aerobic. Anaerobic. Propionibacterium diagnosis of infant botulism was con- D. Actinomyces C. The child most likely gram-positive rod recovered from an contracted this disease by aspirate of a closed chest abscess. The diagnosis of pseudomembranous D. Peptostreptococcus anaerobius forming bacteria 237. B. likely identification of this isolate? motile bacilli associated with bacterial A. Aerobic. non-spore. non-spore. recovered from an abdominal B. Actinomyces israelii D. gram-positive. Anaerobic. Bacteroides fragilis group vaginosis belong to the genus B. What is the most likely in the intestine presumptive identification of the isolate C. Clostridium septicum A. Obligately anaerobic. Bifidobacterium dentium forming bacteria C. What is the predominant indigenous flora 236. gram-negative A. It grew A.

A. A. gram-positive? C. anaerobic bacilli isolated resazurin appears pink. Of the seven toxogenic types. Eubacterium lentum C. Veillonella recovered from cases of gas gangrene? 244. D. Bifidobacterium dentium A. Which Clostridium sp. F. The gram-negative. types A. C. F. bifermentans characteristic of Clostridium botulinum? B. F. Propionibacterium acnes D. Bifidobacterium dentium A. C. difficile toxin B B. E. perfringens A. A. tetani D. necrophorum 242. non. Infant botulism is the most common C. Bacteroides urealyticus B. Septicemia caused by which of the anaerobic bacillus frequently implicated following is generally associated with in serious clinical infections such as brain an underlying malignancy? and lung abscesses is A. varium B. Clostridium C. The majority of the gram-positive. gram-positive. C. Eubacterium lentum C. Suttonella wadsworthensis . C. Bifidobacterium dentium B. Which of the following clostridia has a 246. Lactobacillus catenaforme D. Identify the Fusobacterium sp. Erythrogenic toxin A. Presence of oxygen 241. Fusobacterium D. Peptostreptococcus anaerobius 248. perfringens C. Fusobacterium nucleatum D. C. What does this from clinical material will be indicate? A. Which of the following organisms is not neuro toxin. B. C. difficile B. F. 245. sordellii clinical form. is most commonly D. Botulinum toxin anaerobic bacillus? B. Acid environment B. non-spore-forming. Propionibacterium acnes botulism. C. 247. C. Propionibacterium 249. Alkaline environment C. Capnocytophagia ochracea B. Which one of the following is a bacterial exotoxin known? non-spore-forming. D. Which of the following is the most potent 243. Clostridium septicum B. Motility D. mortiferum D. difficile B. perfringens alpha-toxin C. C. and F are associated with human C. considered terminal spore that causes the cell to to be the most frequent isolate recovered swell? from clinical infections. CHAPTER 6: BACTERIOLOGY 240. Eubacterium limosum C. Oval spores are located terminally. Eubacterium lentum D. nucleatum C. Which of the following statements is not A. A tube of semisolid medium that contains spore-forming. botulinum A. C. Pathogenicity is related to a potent 250.

therefore. Prevotella A. Clostrdium appropriate for the identification of C. Porphyromonas gingivalis 252. Lactobacillus Clostridium difficile? D. Spores are readily seen in laboratory media. It is proteolytic toxin produced by C. It is characteristically nonmotile found in the normal flora of the colon D. Clinically significant clostridia are C. Fluorescence test is the likely cause of the infection? A. Botulism is caused by ingesting be presumptively identified by its Gram preformed toxin and can be prevented stain morphology. Bacteroides fragilis group presence of oxygen. 256. Which of the following D. SPS sensitivity test B. tetani spores will form in the A. D. . Bacillus 258. Which of the following statements is true D. C. There are five serologic types. Cytotoxin assay on a sheep blood agar plate after 48 hours D. It is lecithinase positive B. difficile. no growth is seen on the sheep blood A. Spores are terminally located. Which of the following tests is most are seen in the direct Gram stain of a appropriate for the presumptive identifica- tissue biopsy. Which of the following statements is not 255. Alpha-toxin is produced by all strains. D. Gram-positive bacilli with central spores 251. Fluorescence test of incubation on a vaginal culture. Which of the following tests is most B. Naglertest chocolate agar plate incubated in C. Cytotoxin assay 254. SPS sensitivity test D. and inhibition by bile by boiling food prior to eating. Clostridium tion of Clostridium perfringens? C. Fusobacterium nucleatum required to cause tetanus. B. Bacteroides appropriate for the presumptive identifica- B. Which of the following tests is most A. Cytotoxin assay increased CO2. Lactobacillus A. It produces terminal spores and in the soil. Small alpha-hemolytic colonies are seen C. Reverse CAMP test C. After 24 hours of incuba- tion of Prevotella melaninogenica? tion. Naglertest 253. and a 1-jag kanamycin disk? D. Porphyromonas B. This describes which of the following? 259. Which is a correct statement regarding true of clostridia? Clostridium tetani? A. Which anaerobic. B. Eubacterium lentum anaerobiosis in a wound is not C. C. SPS sensitivity test agar plate incubated aerobically and the B. B. Pseudomembranous colitis is due to a A. REVIEW QUESTIONS • 691 251. gram-negative rod can C. Esculin hydrolysis regarding Clostridium perfringens ? A.

Colistindisk A. Bacteroides fragilis Bacteroides fragilis ? B. Polysaccharide capsule 269. A curved appearance on Gram stain is D. SPSdisk tion pneumonia? B. Porphyromonas sp. Veillonella sp. D. Which of the following would tion of Peptostreptococcus anaerobius? not be a likely causative agent in aspira- A. Exotoxins D. Clostridium perfringens B. pneumonia. On direct Gram stain. Bacteroides gracilis C. A Gram stain reveals gram-postive D. Protease activity anaerobic agents? D. Which of the following is an important A. Eubacterium sp. Bromcreosol purple B. 268. and Veillonella sp. seen. Peptostreptococcus sp. Among the most antimicrobial- sensitive anaerobic bacteria . Clostridium histolyticum C. Vancomycin disk C. Fusobacterium nucleatum is employed. Methylene blue 262. Pumlent material from a cerebral abscess was submitted to the laboratory for smear C. and cocci. On the basis of the organisms A. Clostridium septicum chambers. Actinomyces israelii been achieved in anaerobic jars or B. B. Aspiration of vomitus can lead to appropriate for the presumptive identifica. Which of the following is not true of A. Endotoxin C. what is the most likely B. Fingoldia magna virulence factor of Bacteroides fragilis? B. tion of this organism? 263. What is the most likely identifica- Peptostreptococcus sp. gram. 261. Prevotella melaninogenica characteristic of which of the following? 266. Peptococcus niger A. and Clostridium sp. Anaerobic gram-negative bacillus D. Kanamycin disk B. Plates medium surrounding the colony is positive incubated aerobically exhibited no growth for at 24 hours. Mobiluncus sp. small olive-green to black colonies are C. Which of the following is an important D. After 72 hours of anaerobic incubation. Veillonella parvula C. such as D. Lipase production presumptive identification of the etiologic C. and Nocardia sp. Lipase and lecithinase negative C. Protease cause of food poisoning? 264. Bacteroides ureolyticus A. Commonly associated with intra- abdominal infections D. Fusobacterium sp. Peptostreptococcus anaerobius B. Egg yolk agar showing a precipitate in the bacilli with pointed ends were seen. Methyl red and culture. Phenol red positive cocci in chains and gram-negative 267. Lecithinase production seen on the smear. Which of the following tests is most 265. To ensure that anaerobic conditions have A. CHAPTERS: BACTERIOLOGY 260. Starch hydrolysis A. D. Propionibacterium acnes A. an oxygen-sensitive indicator C.

which D. humans 277. and Mycoplasma 276. Lactobacillus sp. Not able to survive extracellularly A. Chlamydia trachomatis C. Rickettsia. Mycoplasma hominis D. Easily stained using the Gram stain B. Peptococcus sp. The recommended medium for the D. A rash appears first on the extremities C. Ureaplasma urealyticum . Amphibians B. Mycoplasma hominis D. Corneal scrapings are collected and 273. SP4agar C. Rickettsia prowazekii recovery of Mycoplasma pneumoniae 279. It is an obligate intracellular parasite. Chlamydophila (Chlamydia) psittaci vaginal flora that helps resist the onset of infections in humans most commonly bacterial vaginosis? result after exposure to infected A. Ureaplasma urealyticum C. Chlamydia trachomatis for the diagnosis of infection caused by B. Arthropods C. Which one of the following microorgan. REVIEW QUESTIONS 270. D. Fletcher semisolid medium A. Ehrlichia chaffeensis D. The etiologic agent of primary atypical examined microscopically using a direct pneumonia is fluorescent test to detect inclusion bodies A. A skin disease found predominantly in and then on the trunk. A. A. It is transmitted from animals to A. Ureaplasma urealyticum 278. Mycoplasma pneumoniae B. Zoonosis in birds and parrot fever in may be acute or chronic. Peptostreptococcus sp. Is the etiologic agent of Q fever. Which of the following is not true of Coxiella burnetii? 271. B. Mobiluncus sp. Which of the following is true about 272. Charcoal yeast extract medium nected with human genital infections? B. Mycoplasma pneumoniae D. Rat-bite fever humans by inhalation. Mammalians Chlamydia. Resistant to penicillin free media? B. tropical areas D. the following? B. mycoplasmas? isms cannot be cultivated on artificial cell. Avians D. B. Mycoplasma pneumoniae media D. Chlamydia trachomatis C. Mycoplasma hominis 274. Middlebrook B. Chlamydia trachomatis causes which of A. Mycoplasma genitalium C. Inclusion conjunctivitis C. Chlamydiophila psittaci A. C. Which of the following from clinical specimens is Mycoplasmataceae has not been con- A. Which bacterium is part of the normal 275. Grow on routine nonselective culture C.

Rickettsia akari D. What is the reference method for detection C. Sao Paulo typhus "fried egg" are characteristic of D. The causative agent of endemic or murine B. Human infection with the causative agent C. The bite of a body louse A. Tissue culture caused by recrudescence of an initial B. Inhalation of infectious material 289. Rickettsia prowazekii of Q fever is acquired by D. Which of the following is not true about 285. Rickettsia conorii 283. Presence of Mycoplasma genitalium C. 286. The bite of the arthropod Phlebotomus B. D. Rickettsia akari A. an C.694 • CHAPTER 6: BACTERIOLOGY 280. Colonies said to have the appearance of a C. Rickettsia rickettsii reservoirs. DNA-amplification techniques A. After overnight incubation. Presence of Ureaplasma urealyticum . For nonspecific staining ofRickettsia the C. Rabbits 281. Culture on modified Thayer-Martin agar B. Presence of Mycoplasma hominis D. Gram stain 284. Catalasetest A. Brill-Zinsser disease D. prowazekii is associated with D. Gram stain alkaline reaction is noted without turbidity. The bite of a mite (chigger) Mycoplasma pneumoniae isolates is the C. Common agent of lower respiratory B. Qfever 282. Research has found an association A. Rickettsia conorii tract infection C. Tsutsugamushi disease A. Gimenez stain 290. Lysis of red blood cells recommended stain is D. The mild type of typhus fever that is A. A genital specimen is inoculated into 10 B B. Raccoons of Chlamydia trachomatis in cases of D. A screening test for the identification of B. pH change due to molecules in the clinical specimen B. Kinyoun stain What is the most likely explanation? A. Rocky Mountain spotted fever is transmit- Chlamydophila (Chlamydia) ted by the bite of a tick infected with pneumoniae? A. Humans become infected from animal D. Mycoplasma hominis A. Bats with artherosclerosis. Nonculture El A methods attack of epidemic typhus is known as C. Ehrlichia chaffeensis 288. Acid-fast stain D. Rickettsia typhi A. Tetracycline and erythromycin are typhus infection caused by Rickettsia effective treatments. B. Mycoplasma genitalium typhus is C. Rickettsia prowazekii B. Ureaplasma urealyticum B. Transmission of the sylvatic form of C. Gomori silver stain broth. Squirrels sexually transmitted disease? 287.

1 um wide. Lyme disease patient is adequately treated. A. a spirochete A. is the etiologic agent of C. Ticks . which is commonly transmitted by A. This D. Motility is via axial filaments. A helicoidal. Borrelia burgdorferi 298. Syphilis serves as the antigen. Fleas B. Leptonema dark field or phase optics. Sporangium motile organism was approximately 12 um long. Cytoplasmic membrance B. to the morphology of B. During the first week of leptospirosis. and had 299. Borrelia recurrentis closely resemble which bacterial C. Inactivated Treponema pallidum D. Culturing of blood transmitted by Ixodes dammini in the B.S. C. D.1-0. Rat-bite fever C. Treponema pallidum subsp. Examination of cerebrospinal fluid B. Lyme disease D. pertenue A. C. Treponema pallidum subsp. They are visualized best using B. Spirochetes are gram positive. The test is usually positive in 292. Borrelia burgdorferi. Q fever associated with A. Which of the following is not correct semicircular hooked ends. A. pallidum structure? D. most reliable way to detect the presence of the causative agent is by the direct 297. The etiologic agent of epidemic relapsing fever is Borrelia recurrentis. flexible organism was C. Those associated with human D. The axial fibrils of spirochetes most B. Leptospirosis B. Relapsing fever 293. False-positive tests are more frequent following diseases? than with the FTA-ABS test.. Pilus demonstrated in a blood smear. Serious congenital infections are D. Culturing of urine northeastern U. Lice C. Leptospira D.5 um in diameter and 5-30 um in length 295. The description regarding spirochetes? of this organism corresponds most closely A. Mosquitoes D. The antibody titer will decline if the B. Relapsing fever C. REVIEW QUESTIONS • 695 Spirochetes 296. Which of the following is not true of the 291. the secondary syphilis. Detection of antibody against cardiolipin VDRL test? is useful for the diagnosis of which of the A. Flagellum 294. approximately 0. Treponema disease are 0. Examination of blood A. Borrelia C.

Aminoglycosides transmitted disease. Penicillin . When testing the antimicrobial suscepti. Which of the following organisms would B. and yeast extract C. Resistance to clindamycin can be induced C. Inhibition of nucleic acid function A.696 • CHAPTER 6: BACTERIOLOGY 300. The minimum bactericidal concentration B. A suspension of the test organism for use B. When questioned by her acid that act by inhibiting bacteria via physician. Staphylococcus aureus false-positive caused by her liver D. Escherichia coli treponemal antibody-absorbed assay B. Gentamicin D. #2. 97% medium is C. she denied sexual contact with inhibition of folic acid synthesis are any partner symptomatic for a sexually A. the recommended B. Penicillin is active against bacteria by Antimicrobial Agents and Antimicrobial A. Sulfonamides A.0 McFarland standard defined as the lowest concentration D. at least of the original inoculum. Ampicillin D. Treat her with penicillin 304. A positive VDRL test for syphilis was 303. Test her serum using a fluorescent A. 99. Which of the B. Mueller-Hinton base supplemented in vitro by with 5% sheep blood A. Macrolides step for her physician? D.0 McFarland standard (MBC) of an antimicrobial agent is C. Inhibition of protein synthesis at the Susceptibility Testing 30S ribosomal subunit 301. #3.5% by disk-agar diffusion. Proteus mirabilis D. Identify her sexual contacts for not routinely undergo antimicrobial serologic testing susceptibiity testing? C. Chocolate agar D. Streptococcus pyogenes disease 305.9% A. Mueller-Hinton base supplemented B. Erythromycin with hematin. 95. Penicillins following would be the appropriate next C. The chemotherapeutic agents structurally reported on a young woman known to similar to the vitamin p-aminobenzoic have hepatitis. NAD. Reassure her that it was a biologic C. bility of Haemophilus influenzae strains A.5 McFarland standard 306. #1. Charcoal yeast extract agar 307. Reduction of dihydrofolic acid in broth dilution and disk diffusion testing C. 100% B.0 McFarland standard of that antimicrobial agent that kills 302. #0. Inhibition or peptidoglycan synthesis is adjusted to match the turbidity of a D.

High-level resistance D.4-7. REVIEW QUESTIONS • 697 308. Sensitivity 315. Additivism C. 7.2-7. Which drug known to be active against C. Erythromycin C. Metronidazole C. Tolerance therapeutic agent in cases of disease caused by anaerobic bacteria? A. Charcoal yeast extract agar B. Chloramphenicol B.4 organism's growth. MiddlebrookVHIOagar B.8 B. Rif ampin C. The term that denotes a situation in which 314. pneumoniae? Haemophilus influenzas are resistant to A. An example of a bactericidal antibiotic is A. 317. Erythromycin C. Tobramycin . for isolates of A.6-7.6 A. The pH of the agar used for the Kirby. Trimethoprim sulfamethoxazole D. The phenomenon of bacterial resistance to Bauer test should be the bactericidal activity of penicillins and A. tion is recommended. 7. Rifampin D. 7. Serratia marcescens B. The extended-spectrum beta-lactamases the effect of two antimicrobial agents confer resistance to together is greater than the sum of the A. Chocolate agar A. Synergism used for in vitro susceptibility testing of 309. is known as C. before initiation of ous bacteria is antimicrobial therapy. Intrinsic resistance 312. 7. Tetracycline D. Mueller-Hinton base supplemented with 310. Rapid testing for beta-lactamase produc- routine susceptibility testing of nonfastidi. Streptococcus pyogenes 311. Penicillin with 5% lysed horse blood D. MacConkey agar A. The agar recommended by the Clinical 1% hemoglobin and 1% IsoVitaleX and Laboratory Standards Institute for 316. Haemophilus influenzas C. Antagonism D. Mueller-Hinton agar C. Mueller-Hinton base supplemented C. Amoxicillin effects of either drug alone is B. Which of the following media should be D. Trimethoprim 313. Chloramphenicol B. Inducible resistance parasitic infections has importance as a D.2 cephalosporins. Ceftriaxone A. with only inhibition of the B.0-7. Beta-lactamase-producing strains of 5. Erythromycin B. Staphylococcus epidermidis D. Trypticase soy agar D. Isoniazid B.

Which of the following body sites is not agents acts by inhibiting protein synthesis? normally colonized by large numbers of A. Naladixic acid cephalosporin method for the detection of D. Aminoglycoside 25833) B. Chloramphenicol is an important laboratory has been asked to begin antimicrobial agent for the treatment of performing in vitro antimicrobial meningitis as well as several other serious susceptibility testing of Mycobacterium infections. Significant gastrointestinal mani- C. Gentamicin 325. The supervisor of a microbiology 323. Vagina . Photosensitivity 319. Skin D. Blue B. 321. Yellow recommended for treatment of infections B. Aerobic microorganisms D. Macrolide A. BACTEC method festations D. Clavulanic acid is classified as a 324. Obligate anaerobic microorganisms Procedures and Biochemical Identification D. Green caused by C. Broth microdilution method using B. Rifampin B. limit its clinical usefulness. Escherichia coli (ATCC 25922) synthesis? A. Colon C. Ampicillin C. Kirby-Bauer method C. Schlichter method D. When using the rapid chromogenic C. Gentamicin normal flora organisms? B. These effects Which of the following methods would include be appropriate for this testing? A. chlorampheni- tuberculosis because of an increase in the col exhibits significant complications that reported resistance in his community. CHAPTER 6: BACTERIOLOGY 318. Obligate intracellular microorganisms of Bacteria 322. Methicillin A. Microaerophilic microorganisms C. Pseudomonas aeruginosa (ATCC 320. Staphylococcus epidermidis (ATCC D. Enterococcus faecalis (ATCC 29212) B. Red A. Bone marrow suppression and aplastic Mueller-Hinton broth anemia B. Metronidazole is most commonly a positive test is indicated by the color A. Which of the following antimicrobial 326. Clindamycin D. Vancomycin beta-lactamase production by an organism. Allergic reactions and anaphylaxis A. Beta-lactamase inhibitor weekly testing of antimicrobial disks? C. Unfortunately. Which of the following is not one of the A. Which of the following antimicrobial 27853) agents acts by inhibiting cell wall C. Lungs D. Beta-lactam standard control organisms used for the B.

Edwardsiella tarda D. Coagulase-negative Staphylococcus C. 5% and 10% C. Lead acetate C. if the organism is positive. Morganella morganii. Blood drawn for culture may be with the test organism. Sodium citrate impregnated with a solution of A. Two or three blood cultures are D. Collection of 10-20 mL per culture for B. Heparin D. Escherichia C. Contamination of abraded skin C. Antibiograms B. the normal 332. Puncture wounds adults is recommended B. Sodium desoxycholate B. Sheep blood agar when a body fluid or joint fluid that may B. I%and5% D. 10% and 15% 335. Volume of blood cultured is more deaminate phenylalanine can be assessed criticial than timing of culture by inoculating a phenylalanine agar slant D. Escherichia coli B. 5% alpha-naphthol B. Which of the following is not a correct A. 15% and 20% selective and differential? 331. a 330. Infectious aerosols statement regarding blood cultures? A. In capnophilic incubators. Following allowed to clot incubation. Laboratory professionals are at risk for C. Suggested quality control organisms for flora of the vagina is predominantly Simmons citrate agar slants are A. Sodium thiosulfate . During childbearing years. Klebsiellapneumoniae. Edwardsiella 328. 10% feme chloride B. The recommended anticoagulant for use A. Mannitol salt agar A. Fatty acid analysis disease transmission. Lactobacillus B. Person-to-person transmission recommended as optimum 334. Sodium polyethanolsulfonate (SPS) 336. The ability of a microorganism to C. Potassium tellurite D. The paper strip test for the demonstration C. Sodium EDTA of hydrogen sulfide production is D. Enterococcus A. The MIDI identification system is based on tarda A. Shigella sonnet. carbon dioxide green color develops with the addition of concentrations should be maintained A. Escherichia coli. 2% sulfanilamide between C. Which of the following media is both D. The majority of D. Chocolate agar clot is sent for microbiologic examination is C. Propionibacterium coli D. Colony pigment on ChromAgar 333. REVIEW QUESTIONS 327. Mueller-Hinton agar B. 2 N sodium carbonate A. Multiple biochemical tests cases of laboratory-related infections are associated with 329.

The Moeller test for the detection of wall decarboxylase activity is dependent upon B. 10% FeCl3 and alpha-naphthol C. What two reagents must be added A. An acid pH basal medium D. Continuous blood culture systems that strated by the development of a red color. In the catalase test. Kovac's reagent and zinc dust D. The medium used to determine whether an 344. Alpha-naphthylamine and sulfanilic not apply to the acridine orange stain? acid A. C. Which of the following is not true of blood cultures for the recovery of bacteria? A. Fluorescence methods hydroxide B. the formation of organism is oxidative or fermentative with bubbles is due to respect to its metabolic activities is A. Is more sensitive than the Gram stain C. Binds to the teichoic acid of the cell 338. CTA medium B. The oxidation of gluconate C. Which of the following would not be 340. B. Alpha-naphthol and 40% KOH 341. A deamination of tryptophan with low bacterial concentrations 339. 342. Alpha-naphthol and potassium A. . Enterococcusfaecalis B. In the nitrate test. Production of hydrogen gas (H2) B. Which of the following statements does D. An alkaline pH shift in the medium microscope B. OF medium C.700 CHAPTER 6: BACTERIOLOGY 337. Ninhydrin and acetone 343. Production of oxygen (O2) A. No more than three cultures should be drawn in 1 day. XLD medium 345. Colorimetry methods amyl alcohol D. Collect 5 mL of blood for optimal recovery of pathogen. Requires the use of a fluorescence A. Cultures should be drawn before the expected fever spike. D. Manometric methods C. p-Dimethylaminobenzaldehyde and C. Oxidation of hydrogen peroxide D. 14C detection methods B. Creatine and 1N HC1 D. Staphylococcus aureus to determine if the bacterium is VP B. reduction is demon. Cultures should be incubated aerobically and anaerobically. Streptococcus pyogenes A. Is recommended for fluid and exudates D. HE medium D. Breakdown of water C. Methyl red-Voges Proskauer (MRVP) appropriate for a positive control in the broth is inoculated and incubated for 48 PYR test? hours. detect changes in headspace pressure in following the addition of bottles are refered to as A. Staphylococcus lugdunensis positive? C.

and hematuria. the appropriate selective medium chronic hepatitis B. C. He complained of fever and headache. A curved gram- D. fluid- extracellular gram-negative diplococci. Klebsiella pneumoniae C. Urine cultures . C. dysuria. The most likely etiologic agent in this case is A. and myalgia. The source of the urgency. A 45-year-old man was seen in the 346. She had oysters eaten by the deceased patient was no history of prior urinary tract infection. B. popular seafood restaurant. Blood cultures B.2°F. was admitted to B. A 36-year-old man was seen in the emergency department. Proteus mirabilis 348. A 21-year-old sexually active woman came the bullous leg wound. Plesiomonas shigelloides Cultures yielded more than 105 colony. The most likely Laboratory test showed a white blood cell etiologic agent in this case would be count of 10 X 109/L. year-old female with joint pain was sent for nausea. He had returned 1 week previously from a 6-week visit to a village in India. Stool cultures D. Yersinia enterocolitica gram-negative rod. Potassium tellurite agar the intensive care unit for presumed sepsis. The Gram stain days earlier he had eaten raw oysters at a of this sample revealed many polymor. On admission phonuclear cells with intracellular and he was febrile and had hemorrhagic. REVIEW QUESTIONS • 701 Case Studies 349. filled bullous lesions on his left leg. Aeromonas hydrophila contained innumerable white cells. The urine sediment A. Mannitol salt agar temperature of 102. and he died. findings. What is the most critical laboratory test necessary to establish or eliminate the diagnosis? A. emergency department with fever. On the third day. Cefsulodin-irgasan-novobiocin negative rod was isolated from blood cultures drawn on admission and fluid from 347. Among the differential diagnoses was typhoid fever. The patient. A purulent aspirate of joint fluid from a 28. Morganella morganii D. the Gulf of Mexico. He reported that 2 microbiologic examination. and heavy alcohol for primary isolation would be consumption. Sputum cultures C. chills. to the university student health service with disseminated intravascular coagulation a 2-day history of urinary frequency with developed. Vibrio vulnificus forming units/mL of a lactose-fermenting D. Escherichia coli B. who had a A. The Given the specimen type and microscopic patient had a history of diabetes mellitus. Modified Thayer-Martin agar and treatment was begun.

febrile with a loss of appetite for the past 24 diarrhea. pinpoint biochemical reactions confirmed that the colonies grew on sheep blood agar that etiologic agent was showed faint zones of beta-hemoloysis. right lower lobe pneumonia who lived on The characteristic symptomatology and the a dairy farm. A complete blood count showed a was 100 mg/dL. The culture showed etiologic agent in this case is many gram-negative bacilli. A 32-year-old male was seen in the gram-negative diplococci. Kawasaki disease Cultures of blood and cerebrospinal fluid B. Streptococcus agalactiae C. Toxic shock syndrome many polymorphonuclear cells containing 353. Cultures of the blood and CSF neutrophils. Gram-positive rods were recovered from organism was positive for urease and the chest fluid drawn from a teenager with ONPG and negative for phenylalanine. Yersinia enterocolitica etiologic agent in this case? A. where a culture of vaginal daycare center. The CSF protein diarrhea. The white blood emergency department with symptoms of cell count was 25 X 109/L. A young woman complaining of symp- department by her parents. Streptobacillus moniliformis . C. The 351. vomiting. A. and B. but there was D. Pelvic inflammatory disease (CSF) were sent to the laboratory. She was admitted to the it was difficult to arouse her. Most recently the parents noted that gynecologist. A. Shigella dysenteriae demonstrated a positive CAMP test. Listeria monocytogenes indole negative. She had been toms of sudden onset of fever. Neisseria meningitidis no evidence of gas or H2S production. citrate negative. At 24 hours. She attended a hospital. and the Gram stain showed D. Vibrio parahaemolyticus Which of the following is the most likely D. with 88% lower right quadrant abdominal pain and polymorphonuclear cells. The most likely diagnosis tion she demonstrated a positive Brudzinski in this case would be sign indicative of meningeal irritation. and rash was seen by her hours. Listeria monocytogenes B. which were A. and a stool grew the same organism. The triple sugar iron C. He was admitted. A 3-year-old was brought to the emergency 352. and her childhood discharge grew many coagulase-positive immunizations were current. Haemophilus influenzae oxidase negative. Salmonella Paratyphi The isolate was catalase negative and B. Scalded skin syndrome was cloudy. The most likely culture was obtained. On examina.702 CHAPTER 6: BACTERIOLOGY 350. Her CSF C. Moraxella catarrhalis reaction was acid over acid. and the glucose was 15 leukocytosis with an increased number of mg/dL. staphylococci. Arcanobacterium pyogenes D.

A biopsy of one of the lesions A. A college student got a summer job community hospital in New England with working at a marina. A 7-year-old female became ill with an 1 hour after the transfusion began. An anemic patient was transfused with D. REVIEW QUESTIONS • 703 354. Vibrio vulnificus diarrhea. headache. which of the C. and swollen lymph received several lacerations on his right nodes. Franciscella tularensis B. Neisseria meningitidis C. In August. the patient healing and he sought the opinion of his was diagnosed as having physician. consistent with endotoxic shock. The racers hiked in the mountains and 358. Several international participants in an B. Her case history during the race was highly associated with revealed that she had eaten cheese that had illness. Pseudomonas aeruginosa became ill with symptoms of chills. Borellia recurrentis agent in this case would be B. She had bloody blood cells had been stored at 4°C for diarrhea and went on to develop hemolytic approximately 30 days before their use. She was febrile and complained Contact with contaminated water and soil of flulike symptoms. and chickens. swam in rivers. The most likely etiologic A. Relapsing fever likely be the etiologic agent in this case? A. headaches. A silver-stained biopsy of a skin forearm. Brucella canis A. Given the history. stiff outboard motor on a rental boat. the intestinal illness after visiting a petting patient developed fever and hypotension zoo featuring farm animals such as calves. Rabbit fever following microorganisms would most D. Vibrio cholerae nonOl D. Yersinia enterocolitica packed red blood cells. he neck. Plague condition. The red lambs. A woman. The most likely The organism most likely to be involved etiologic agent in this case is in this case is A. Leptospira interrogans C. Yersinia enterocolitica . On the basis at the time of the injury. but after several of the clinical syndrome and laboratory weeks he noted that the lesions were not detection of a causative agent. a patient presented at a 357. Listeriamonocytogenes D. Vibrio cholerae 01 C. uremic syndrome. who had recently returned from jungles. D. No medical treatment was sought lesion showed spirochetes. Lyme disease revealed a cutaneous granulomatous B. was admitted to the through flooded streams for 2 weeks. and eye infections. Eschericia coli 0157:H7 A. hospital. muscle aches. Shigella dysenteriae B. Approximately 359. Mycobacterium marinum 355. What is the most likely etiologic been made from unpasteurized milk while agent in this case? on vacation. While repairing the symptoms of a skin rash. Campylobacterfetus B. and slogged a vacation in Mexico. Bordetella pertussis C. Nocardia asteroides Eco-Challenge adventure race in Borneo C. Pseudomonas aeruginosa D. Staphylococcus aureus 356.

704 • CHAPTER 6: BACTERIOLOGY 360. Moraxella catarrhalis . diarrhea. A middle-aged man with a history of smoking and drinking for over 40 years developed shortness of breath. frontal headache. Bordetella pertussis B. and cough. Legionella pneumophila D. Which of the following is the most likely etiologic agent in this case? A. His sputum Gram stain showed numerous polymorphonuclear cells but rare microorganisms. and a diagnosis of atypical pneumonia was made. His medical history included a kidney transplant several years ago for which he remains on antirejection therapy. fever. which routinely misted the fresh greens. He worked in the produce section of a supermarket. An X-ray of his chest showed an infiltrate in the left lower lobe. Klebsiella pneumoniae C.

Scalded skin syndrome is a form of dermati- characterized by myocarditis and neuritis. These clinically signifi- sodium azide or 4% bile salts. are frequently not able to duce esculetin. Transmission is by contact with a toxin have been identified: exfoliation A and human carrier or with contaminated fomites. beginning about the face and trunk and subse- toxin. which causes the appearance of the clear. This tis produced by strains of Staphylococcus aureus disease most commonly affects children aged 1 to that elaborate exfoliative toxin. of 1-4% bile salts. foods containing preformed thermostable entero. and watery diarrhea due to the loss of electrolytes 705 . answers rationales Aerobic Gram-Positive Bacteria and fluids into the lumen. exfoliation B. large. This form of intoxication causes a perfuse quently spreading to the extremities. Nutritionally variant streptococci (NVS) are tive for enterococci by the addition of either now termed Abiotrophia. or vitamin B6-supplemented culture media is 2. The medium is made selec. which account for 5-6% of grow on this medium and hydrolyze esculin pro. tum granulosum. C. which reacts with an iron salt to be recovered because of insufficient quantities of form a black color in the agar. Organisms able to cant microorganisms. Strains of Corynebacterium diphtheriae infected by a lysogenic bacteriophage produce an extremely potent exotoxin. toxin may cause a rapidly fatal hypertoxic disease C. Infants and children are most com- cases result from the ingestion of contaminated monly affected with this form of dermatitis. Absorption of the 5. This potent toxin acts by disturb- ing the adhesive forces between cells of the stra- 3. B. the cases of endocarditis. vitamin B6 in the culture medium. In many cases. D. Two types of this 10 years. required for isolation. a streak of Staphylococcus. flaccid bullae and the skin to B. Staphylococcal enterocolitis food poisoning peel off. the causative agent may never be recovered from L patient specimens. Enterococcus and other group D strepto- cocci can be presumptively identified based on their ability to hydrolyze esculin in the presence 4. The routine use of a pyridoxal disk.

The characteristic presentation is cutaneous often initially confused with group A or group B spreading lesions of the fingers or hand that are streptococci because of its beta-hemolysis. Munch- longer recommended because groups C and G Petersen) who discovered this phenomenon!. Staphylococci and micrococci are both cata. so it is important to differentiate Micrococcus from Staphylococcus. are tion of this coagulase-negative Staphylococcus is soil saprophytes that may produce disease in its resistance to novobiocin. positive CAMP reaction.blood agar plates from clinical samples. hydrolyzes esculin. 10. and is Erysipeloid is usually the result of contact with oxidase negative. A. Streptococcus agalactiae isolates can be pre- tococci can be achieved through the PYR sumptively identified by the demonstration of a (L-pyrrolidonyl-(3-naphthlylamide) disk test. Nocardia spp. produces catalase. siopathiae in humans is primarily erysipeloid. whereas Staphylococci are negative. Microscopic examination of pus from suspected cases will 12. Listeria is an important animal and human pathogen that is known to cause abortion. Staphylococci group A streptococci. of Escherichia coli. the rate of false-positive CAMP reactions by lase positive gram-positive cocci. This gram-positive rod is actively A. Infection caused by Erysipelothrix rhu. Atkins. meningitis. humans either by the inhalation of contaminated material or through skin abrasions. demonstrate partially acid-fast. are more clinically significant. and anaerobic incubation increase A. because 7.708 • CHAPTERS: BACTERIOLOGY 6. CAMP is an acronym The use of a 0. E. When recovered on sheep an infected animal or contaminated animal prod.motile at room temperature (but not at 35°C). n. The organism has a predilec- B. produced by beta-lysin-producing strains of Staphylococcus aureus. humans.04-unit bacitracin disk is no for the scientists (Christie. Key to the identifica- manifestation of the disease. streptococci are also susceptible to this agent. Although generally con- fined to the skin. it is uct. gram-positive. which acts to enhance the zone of hemolysis color change within 5 minutes. Draining sinus and is often seen in large numbers adhering to tracts in the subcutaneous tissue are a common these cells on Gram stain. A. rhusiopathiae has been impli- cated in rare cases of endocarditis. Presumptive identification of group A strep. . Incubation of test plates should be earned out in ambient air. The recovery rate of coagulase-negative Staphylococcus saprophyticus from urinary tract infections in young females is second only to that 8. raised and erythematous. A. The organism seen in Color Plate 26m is branching filamentous or coccoid organism. and septicemia in 9. A Group B streptococci elaborate the CAMP fac- positive test result is interpreted as a bright red tor. Nocardiosis is characterized by mycetoma tion for the epithelial cells of the urogenital tract or chronic suppurative infection. D. Listeria monocytogenes. increased CO. Micrococci are modified oxidase positive.

Accidents in lum streak. transmission of group A streptococci. other corynebacteria and some staphylo- gitis and septicemia in neonates. is transmitted by the mother before birth. may 19. and 18. Neonatal infection with group B B. The potassium tellu. Streptococcus agalactiae (group B Strep- brown halo are seen on this agar medium. The vegetative cells and spores of Bacillus both sides and without touching the strip. cereus are an precipitin line at a 45-degree angle to the inocu- enterotoxin and a pyogenic toxin. diphtheriae are presumptively identified when black colonies surrounded by a A. This medium is Corynebacterium diphtheriae. B.disease referred to as erysipeloid. Symptoms occur when nasopharyngeal infection spreads to the face. an Elek agar plate. traumatic introduction into a normally sterile site through the use of contaminated medical D. when inoculated. A pos- cereus are widely distributed in the environment.15% 16.tightness at the site of invasion. for the primary isolation of gram-negative aerobic bacilli. The organism. it is not a differential medium. riae from clinical samples. Colonies of C. The rare complication of an upper res- 15. Blood agar medium is supplemented with 0. Erysipelas results from person-to-person equipment are associated with infection. ingestion of contaminated foods. The Elek immunodiffusion test is recom- streptococci may occur either as an early-onset mended for detecting toxigenic strains of disease (at birth) or as a delayed-onset syndrome Corynebacterium diphtheriae. How- tococcus) is a principal cause of bacterial menin- ever. fler's serum medium. cocci will produce a similar reaction. rite in the medium is taken up by colonies of Corynebacterium. intravenous drug abuse. In the test. Phenylethyl alcohol agar (PEA) is a selective medium for the isolation of gram-positive cocci. piratory infection with Streptococcus pyogenes B. . due to the inhibition of swarming by PEA. ANSWERS & RATIONALES 707 13. not only inhibits particularly helpful when a specimen containing indigenous respiratory flora but differentiates gram-positive cocci is contaminated with a colonies of C. which is pressed onto the surface of 14. The agar slant. diphthe. Although this medium is Erysipelothrix rhusiopathiae causes a similar primarily designed for the recovery of C. theria antitoxin is impregnated on a sterile filter paper strip. diphtheriae. Erythema asso- lar morphology seen in methylene blue stains is ciated with this superficial cellulitis rapidly enhanced when the organism is grown on Loef. itive reaction by toxigenic strains produces a The virulence mechanisms of B. the birth canal. which is a part of the indigenous microbial flora of the vagina. which is inhibitory to most C. The formation of the characteristic is characterized by sensations of burning and Corynebacterium diphtheriae granules and cellu. usually as the baby passes through 17. Test and control strains are then inoculated perpendicular to the strip on B. nature resulting in cuts or abrasions contami- nated with soil or vegetation. phenylethyl alcohol. diph- that manifests itself weeks after birth. Tinsdale medium. causing them to appear black.Proteus spp. demonstrate growth of corynebacteria within 8 to 24 hours.spreads with an advancing elevated margin.

mycelia and were formerly in the genus Nocardia. A. Rhodococcus equi is found in soil and com- sheep blood agar plate inoculated with a group A monly produces disease among livestock. Some strains of Streptomyces grow better at 25°C than at 35°C. is the human infection produced most commonly by this agent. tion on culture media and its inability to ferment ence of oxygen. 25. tolysin S is stable in the presence of atmospheric This species is characterized by its pink pigmenta- oxygen. gram-positive bacillus that often appears are all catalase negative and grow on coventional as long filaments. Listeria monocytogenes is motile at room and O2. Therefore. streptococci obtain all their energy from the B. Viridans streptococci are the most common B. Of the two hemolysins secreted gram-positive bacilli can demonstrate primary by beta-hemolytic group A streptococcus. when the agar has been stabbed and subsurface hemolysis is revealed. Streptomyces are weak pathogens rarely asso- normal flora in upper respiratory cultures. Streptolysin O is inactivated in the pres. and it is best demonstrated carbohydrates. monocyto- genes is nonmotile at 35°C. Unlike other aerobic gram- media such as sheep blood agar. Most are part of positive bacilli. the beta-hemolysis seen on the surface of a D. somaliensis. which is most frequently caused by previously damaged heart valves. D.708 • CHAPTERS: BACTERIOLOGY 20. These streptococcus. . Whether growing aerobically or anaerobi. growth away from the stab is character- catalase negative. Strep. the soil. a skin disease of the hands usu- introduced in tissues or blood. throat.just below the agar surface. they grow better on istic of motility. When inoculated into a semisolid heme prosthetic group for this enzyme and are medium. B. this organism produces H2S. Motility is generally enhanced blood-containing media because of the catalase. giving the growth like activity of hemoglobin. Erysipelothrix is a nonmotile. 21. L. Streptolysin S is primarily responsible for 27. S. Streptococci are unable to synthesize the temperature. They ciated with disease. The bacteria normally inhabit are opportunistic pathogens with low virulence. the normal flora of human skin. Streptococci negative. The most common human infection is Subacute endocarditis is seen in patients with myectoma. cally. catalase- fermentation of sugars to lactic acid. which can be demonstrated in triple sugar iron tine but produce a wide variety of infections when agar. 22. Organisms that synthesize the enzyme cata- lase are able to protect themselves from the 26. ally associated with the handling of infected ani- mals. and intes. pattern an "umbrella" appearance. Erysipeloid. 24. D. killing effects of H2O2 by converting it to H2O D.

is the acid fastness of the filamentous bacilli or coccoid forms. There are approximately 80 types of pneumococci based on specific capsular antigens. Viridans streptococci do not produce the logic agent of lobar pneumonia.. have been developed for the recovery of this organism. The optochin disk test inhalation of contaminated fomites or a trau. colonies of S. It is the bul- lous form of impetigo for which S. there- lated. 32. lanceolate diplococcus.or red color. C. . 31. A diag. 34. Selective media and rapid identifi- cation tests are important for this widely recog- nized opportunistic pathogen. Micrococcus and often mucoid in appearance and bile soluble. E. pyo- pus and upon examination would reveal non. ANSWERS & RATIONALES 709 28.enzymes coagulase and DNase and its ability to tive and is usually nonhemolytic. forming anaerobic bacilli. a primary etio.can be performed to presumptively identify this matic incident with soil contamination. pro- nostic characteristic. pneumoniae undergo tion as well. in the PYR test do not produce a positive Fastidious in its growth requirements. Unlike Actinomyces spp. Streptococcus pneumoniae. Streptococcus pneumoniae is a leading cause of lobar pneumonia as well as other serious bacte- 29.33. is primarily caused by S. Nocardia spp. the organ. faecium is PYR posi.genes and Staphylococcus aureus. The Gram stain smear of clinical D. autolytic changes. non-spore. is an encapsu. gram-positive. Species of the genus Nocardia are ubiqui. other enterococcal strains is of importance because of their resistance to most clinically use- ful antimicrobial agents. The route of infection is direct B. are D. The thick crust acid-fast branching filaments.nosis when the characteristic morphology and duce exogenous forms of infection as a result of Gram reaction is observed. Optochin lyses pneumococci. such as mannitol salt agar.D. Identifying characteristics of Staphylococcus is characteristic of the clinically significant aureus include the production of the extracellular species of Enterococcus. Bacteriologic cultures of a typical impetigo catalase-positive aerobic organisms. depending on the species. form of impetigo. Enterococcus faecium is an important agent inoculation of the causative agents into abraded or of human infection.organism. rial infections. ducing a zone of inhibition around the disk.specimens can provide a rapid presumptive diag- tous in the soil and thus characteristically pro. aureus is the etiologic agent. which is most commonly seen. The ability to tolerate a high concentration of salt C. grow in the presence of high salt concentrations. pyogenes. which are convex and tococci and Enterococcus. although the reaction may be delayed. Differential and selective media. The PYR test is used predominantly ism on sheep blood agar produces characteristic for the presumptive identification of group A strep- alpha-hemolytic colonies. Lactococcus are known to produce a positive reac- Upon aging. "Sulfur lesion may yield either a pure culture of Strepto- granules" are characteristic of actinomycotic coccus pyogenes or a mixed culture of S. which are catalase-negative. fore. A. gram-positive. Their differentiation from otherwise compromised areas of the skin.enzyme pyroglutamyl aminopeptidase and. including vancomycin.

Strep- coccal virulence by overcoming the host's natu. Many factors contribute to staphylo. pendicular to a streak of Staphylococcus aureus. therapy are required to prevent sequelae (i. The use of ninhydrin to detect 39.41. The test is per. A sample of this actively growing cul- B. scarlet fever is rare in most developed 37. and they are sodium hippurate to benzoic acid and glycine. at the point where the zone of hemolysis pro- D. Group B streptococci (Streptococcus agalac. Cultures of the tonsillar fossae and posterior hemolysis when the isolate is grown on sheep pharynx are most commonly obtained in sus- blood agar. countries. fever and erysipelas in addition to wound infec- tions (e. monocytogenes is catalase positive. the addition of ninhydrin to the medium will reduce the glycine to produce a purple color. Unlike 5.D. can hydrolyze high as 55 °C for long periods. If not bile resistant. agalac- group A streptococci. C. glycine is a sensitive and rapid test of hippurate C. The coagulase. which produce beta. Listeria monocy to genes and Streptococcus agalactiae produce an extracellular factor ture should be examined using the hanging-drop known as the CAMP factor. producing staphylococci are most commonly producers of staphylolysins. glycine is produced. them to remain infectious in the environment longer than many other pathogenic bacteria.pected cases of streptococcal pharyngitis. aureus joins with that produced by action of an erythrogenic toxin produced by the beta-hemolytic test isolate..g. The physiology of staphylococci enables rheumatic fever and acute glomerulonephritis). A positive CAMP reaction is indicated by a zone of enhanced beta-hemolysis (arrowhead shape) 40. Most species are sensitive to novobiocin. Because of the rapid diag- tiae. Staphy- A. anthracis is nonmotile and can therefore be easily differentiated from formed by making a streak of the test isolate per- commonly encountered motile species. Rapid identifica- tion of this organism and prompt antimicrobial 38. lococci. unlike other streptococci. Their high salt toler- ance enables strains to grow in salt-preserved . The rash of scarlet fever is a result of the duced by S. necrotizing fasciitis). The production of hemolysins and the involve an erythrogenic toxin. B.tococcus pyogenes is most often associated with ral defenses. however. enzymes coagulase and DNase is associated with the virulence of staphylococci.710 • CHAPTERS: BACTERIOLOGY 35.e.technique for motility. Endotoxin is found in the cell wall cases of pharyngitis but is also the agent of scarlet of gram-negative bacteria. Bacillus anthracis from other species of Bacillus. The other diseases listed do not A. Motility is a key test for the differentiation of hydrolysis. nosis and treatment of group A streptococci infections. cannot resist temperatures as tiae). foods and causes cases of food poisoning.. L. Suspect Bacillus colonies are inoculated in a 36. broth medium and allowed to grow to a visible turbidity. Staphylococci are somewhat heat resistant and can survive dry conditions.

morphologically similar to all other members of Patients characteristically produce blood-tinged. pneumoniae forms alpha-hemolytic colonies These bacteria often stain irregularly and have a when grown on sheep blood agar. . 43.D. On tellurite-contain.ciated with dissemination and have a poor prog- entiation. the genus Corynebacterium. Clinical material sent to the laboratory for 49. Iden.and can often spread hematogenously throughout ing media. and it is often cultured ble S. Its cases in young females. The mode of infection is the inhalation viridans group of streptococci. and tested for their biochemical activity and toxin infection is acquired by traumatic inoculation or production. C. the recovery of Corynebacterium diphtheriae B. S. usually during the per- sanguis is one of several species that may lodge formance of his/her occupation (sheep shearing in an abnormal heart or on valves damaged by or processing of animal hair).is known to progress rapidly to a fatal form of tory tract. D. 47. They are all gram- rust-colored sputum in which the characteristic positive. Corynebacterium pseudodiphtheriticum is associated with cases of lobar pneumonia. multiple drug resistance allows it to remain in coagulase-negative staphylococci closely resem. A. istically resemble Chinese characters or palisades. septicemia. Prompt diagnosis previous infection. inhalation. In immunocompromised patients. Corynebacterium jeikeium is a low virulence as an etiologic agent of uncomplicated cystitis organism resistant to multiple antimicrobials. 46. and endocardi- tis in association with intravenous catheter use. woolsorters disease or the pulmonary form of 1 which most often is caused by a member of the anthrax. colonies of this pathogen will appear the body. ANSWERS & RATIONALES • 711 42. Suspicious colonies should be further nosis. non-spore-forming bacilli that character- gram-positive lanceolate diplococci can be found. pleomorphic club-shaped appearance. 48. Nocardia should be inoculated on cystine-tellurite agar asteroides can cause invasive pulmonary infection plates or Tinsdale medium. 44. These nonhemolytic. epidermidis on sheep blood agar. of septicemia. Lesions in the brain are commonly asso- dark-brown to black. Bacillus anthracis is the causative agent of mation of the lining membrane of the heart. hospital environments. from the skin of hospitalized patients. Streptococcus pneumoniae is most commonly D. A. which aids in their differ. Subacute bacterial endocarditis is an inflam. In com- tification of 5. wound infections. Viridans streptococci are and treatment of this disease is needed because it normal inhabitants of the human upper respira. The organism is ubiquitous in nature. Staphylococcus saprophyticus is recognized B. saprophyticus is facilitated by promised patients it has been implicated in cases demonstrating its resistance to novobiocin. Streptococcus of spores by the patient. 45.

of viridans streptococci typically remain intact when bile is applied. Of the genera listed. Viridans streptococci Biochemically L. which may be visually mistaken for beta-hemolytic streptococci. mucosal surfaces in humans. levels of 105 projections referred to as "Medusa head" forms. pneumoniae will be totally dissolved. Listeria monocytogenes is a small. 5L B. only Leuconostoc is catalase negative. gram-positive bacillus that char. This coagulase-negative Staphylococcus is seen frequently as a contami. or ethylhy- agar. such as technique has not been used. S epidermidis has sodium desoxycholate. organisms per gram of food can be found. aureus. translucent drocupreine HC1. is a presumptive test for the beta-hemolytic colonies. 53. Colonies on described. The susceptibility of alpha-hemolytic strep- room temperature. anterior nares are recommended when screening for earners in the hospital environment. membranes of humans. 55. S. aureus is carried . differentiation of Streptococcus pneumoniae from viridans streptococci. A. Leuconostoc is vancomycin is associated with urinary tract infections. pital personnel may harbor resistant strains of S.D. Staphylococcus epidermidis is a saprophytic microorganism found on the skin and mucous 56. human anthrax that occurs in any of three forms: cutaneous. Within a few hours. When grown on sheep blood tococcal isolates to optochin. The ingestion of food contaminated with Gram stain this organism appears as a large.712 • CHAPTER 6: BACTERIOLOGY 50. S. Colonies thetic devices. amounts of toxin results in a rapid onset (1-6 C. is a widely used pre- been implicated in serious human infections sumptive identification procedure. this organism produces small. pneumoniae characteristi- cally is susceptible and produces a zone of inhi- 52. Cultures of the ticemias in neonates. Hos- resistant and associated with infections in hospi. with a 6-mm disk. enterotoxin produced by Staphylococcus aureus spore-forming. Staphylococci colonize various skin and hours) of vomiting and diarrhea as a result of a neural response. bition greater than 14 mm. aureus multiplies rapidly in improp- agar plates are large and opaque with fingerlike erly stored food. S. as transient flora in the anterior nares. monocytogenes differs from are typically resistant to this agent and show no streptococci because it possesses the enzyme zone of inhibition or a zone of less than 10 mm catalase. Bacillus anthracis is the etiologic agent of 57. gram. and person-to-person contact is a sub- talized patients. It has also been linked to sep- stantial infection control concern. positive bacillus that is actively motile at D.solution of this reagent is applied to test colonies. sapro- phyticus is less likely found as normal flora and C. 5. When a 10% associated with the surgical insertion of pros. pulmonary. 5. On D. C. Solubility of Streptococcus pneumoniae nant in blood cultures when proper venipuncture colonies by surface-active agents. Entero- toxin is elaborated when the organism reaches stationary growth phase. Ingestion of small the most likely cause of the disease in the case acteristically grows in long chains. and gastrointestinal.

diphtheriae is differentiated from other cornybac. ANSWERS & RATIONALES • 718 58. This each account for a significant number of adult syndrome occurs primarily in infants and chil- infections. neither malignant nor a provided by the addition of the serum. B. however. are gram-positive. ologic agent of human cases of food poisoning. childhood infections. toxin produced by strains of Staphylococcus Bacterial arthritis can occur following infection aureus. resembling the flu salt concentration. gastrointestinal. Corynebacterium 63. Enterococcus faecalis and E. The organism can streptococcal group D antigen. spore-form- cus are resistant to novobiocin. Potassium pustule. 62. but corynebacteria are black eschar surrounded by a red raised ring. A. hydrolyze esculin. most commonly associated with cases of food poisoning following ingestion of reheated rice served at Asian restaurants. This organism is ing bacilli widely found in the environment. creating large flaccid bullae. On serum-cystine-sodium thiosulfate-tellurite medium (Tinsdale medium). which may prevent the death of the fetus. . B. Growth factors needed by C. adhesive forces between cells of the stratum tococcus pyogenes and Neisseria gonorrhoeae granulosum. Exfoliatin acts in humans to disrupt the in other parts of the body or bacteremia. 64. Scalded skin syndrome is the dermatitis C. tion. 61. Listeria monocytogenes is a cause of human the presence of bile.A. bovis group. frequently found in urine culture of young Bacillus cereus is of particular interest as an eti- women and may be misidentified as S. and cuta- its ability to produce black colonies surrounded neous. epider. and a zone This enterotoxin-producing microorganism is of 16 mm or less determines resistance.lesions seen in cutaneous anthrax in humans. therapy can be initiated. These bacteria also express and causing a low-grade fever. the primary infection is usually unrelated and Haemophilus influenzas predominate in to the areas where lesions appear. the mother's duce acid from glucose in the presence of high symptoms are usually mild. diphtheriae are The lesion is. B. 65. The ability to tol- be isolated from aborted fetuses as well as from erate high salt concentrations differentiates the the maternal placenta. Strep. Malignant pustule is the name given to by a brown-black halo after 48 hours of incuba. A 5-ug disk is used in the test. Bacillus spp. resistant. whereas Streptococcus pneumoniae dren. appropriate the S. Bacillus anthracis infects humans by three teria and other bacteria of the respiratory tract by routes: respiratory. Staphylococcus aureus is the predominant associated with the effects of the exfoliative pathogen involved in joint infections of adults. The disease produces a localized tellurite is inhibitory to many gram-positive and abscess on the skin. faecium grow in B. D. midis. In humans. When infection with this enterococci from the group D streptococci like etiologic agent is detected early. and pro- and bovine abortion. Most strains of Staphylococcus saprophyti. 59. which forms a characteristic gram-negative bacteria.

Klebsiella pneumoniae is the species most A. unable to decarboxy- 69. nosocomial infections of sev. Enterotoxins are produced in the intestinal tract and primarily cause diarrhea. and other extrain. which are small. 71. Chryseobacterium (formerly have a capsule and appear mucoid on cultures. These mended for optimal recovery of the agent. B. Chryseobacterium spp. ters and can be spread to family members. In . Shigella has a low infecting dose and has difficult. the type species of the resembles cholera toxin. The heat.Further biochemical testing would generally ride ions by the cells lining the small intestine. multiple blood cultures are recom. Members of the genus Klebsiella moist soil and water. Isolation of these organisms is A. 67. C. has been implicated in out- severe pneumonia. Diarrhea results non-lactose-fermenting. equipment. Flavobacterium) meningosepticum. labile enterotoxin of Escherichia coli. typically organism. a known This highly encapsulated organism can cause nosocomial pathogen. The etiologic agents of brucellosis are the brucellae.714 • CHAPTERS: BACTERIOLOGY Aerobic Gram-Negative Bacteria the presence of the enzyme and atmospheric oxygen. is a causative agent of bacillary dysentery. enzyme adenylate cyclase. Bone organisms are found in humans only at the time marrow cultures have been found to be positive of infection. Shigella dysenteriae. when cultures of blood failed to recover the Transmission is by the fecal-oral route. and urease negative. Adult human infections are rare. The stimulation of Differential and selective media for the recovery the enzyme adenylate cyclase by the toxin of enteric pathogens from stool samples would increases the production of cyclic AMP. late the amino acid lysine. infantile enteritis.breaks of meningitis in hospitals and is associated eral types. rapid gastrointestinal fluid loss. The genus Neisseria contains organisms that possess cytochrome oxidase activity. gram-negative bacilli. The reaction relies on the property of the molecule to substitute for oxygen as an electron acceptor.with the use of contaminated respiratory therapy testinal infections. gram- negative coccobacilli that are facultative intra. the molecule is oxidized to form 66. following stimulation of the secretion of chlo.72. which are generally been reported to cause outbreaks in daycare cen- job related. are ubiquitous in the frequently recovered from the vast majority of environment and are especially associated with clinical cases. show these organisms to be unable to use citrate as their sole carbon source. causing demonstrate Shigella species as H2S negative. they are not part of the normal flora. In suspected cases. which D. A. Colonies can be identified by the development of a dark purple color following the application of tetram- ethyl-p-phenylenediamine dihydrochloride. cellular parasites. by ingestion of contaminated foods or water. 70. 68. acts to stimulate the genus. A. nonmotile. these opportunistic microorganisms occur primarily in immunocompromised patients. indophenol blue.

78. Francisella tularensis requires cysteine or cystine for growth. C. except the recently added Plesiomonas shigelloides. Growth on MacConkey agar and a pos- cultivation of Legionella spp. acteristic of Escherichia coli. A highly selective medium. cholerae. from skin abscesses to abscess formation by the production of concentrically spreading in internal organs. The K (capsule) antigen surrounds the bacte. teriaceae. cystitis. Aeromonas hydrophila is typically found in ports the growth of F. Capnocytophaga spp. which are used to group members of the Enterobacteriaceae. 77. It is ubiquitous in nature and is found in homes A. pleural fluid. Species able to ferment sucrose. in systemic disease isolated from cerebrospinal B. ANSWERS & RATIONALES • 715 73. appendicitis. These heat-labile anti. The bacterium is found in The gliding motility is best observed during the soil and water in subtropical areas of Southeast log phase of growth and can be demonstrated by Asia and Australia.and hospitals. It is an opportunistic pathogen rial cell and masks the somatic antigens of the responsible for nosocomial infections. itive oxidase reaction are characterisic of this organism. thiosulfate-citrate. 75. Decubitus ulcers frequently contain normal intestinal flora. bile salt-sucrose (TCBS) is used for the isolation of Vibrio spp. Burkholderia pseudomallei is the causative fluid. organism associated with cases of neonatal B. The biochemical results are char- Aeromonas produce beta-hemolysis.meningitis. and in compromised hosts they have been implicated 76. On sheep blood agar. A positive oxidase reaction differenti- ates this organism from all of the Enterobac. are fermentative that contain K antibody can be used to demon. ity of humans. coli is associated with a variety of diseases. both require the addition of 5% sheep or rabbit blood. cell wall. such as V. monly encountered gram-negative species that is not a member of the family Enterobacteriaceae. tularensis. it is the predominant 74.80. Pseudomonasa aendgnosa is the most com- colonies. growth around primary colonies. E. Melioidosis exhibits several darkfield microscopy and on sheep blood agar forms. specticemia.gram-negative bacteria that inhabit the oral cav- strate the presence of the capsular antigens. produce yellow colonies. a medium gen- fresh water and has been implicated in human erally used by clinical laboratories for the infections. These organisms have been iden- tified as a cause of disease in the oral cavity. Buffered charcoal yeast extract also sup- A. many strains of B. ^^_ Non-sucrose-fermenting organisms produce green D. gens can be removed by heating a suspension of the culture at 100°C for 10-30 minutes. agent of melioidosis. 79. They . Glucose-cysteine with thi- amine and cystine heart media are commercially available for suspected cases of tularemia. and pulmonary secretions. Antisera A. and endocarditis.

C. Campylobacter jejuni rivals Salmonella as influenzae type b (Hib) vaccine beginning in the most common bacterial cause of diarrheal 1985 has significantly reduced the incidence of invasive H. D. and incubation under a microaerophilic atmo- sphere at 42°C are required for the recovery of A. Before the development of an effective vac- ual's swimsuit. and abdominal pain. gram-negative with contamination at the site with organisms in coccobacillus. Identification of Salmonella Typhi. The widespread use of Haemophilus B. In some cases 83. sule. meningitidis may produce only a transient bacteremia or meningitis or may go on to cause a rapidly fatal infection. the etiologic agent of typhoid fever. and possesses a capsular antigen (Vi). The incubation period following con- 82. Vibrio vulnificus is implicated in wound infections and septicemia. ments. tory system by N. 85. The Waterhouse-Friderichsen syndrome of round. Ingestion of contami.716 • CHAPTERS: BACTERIOLOGY 81. disease in humans. nated water or seafood is the typical mode of A. Pseudomonads are ubiquitous microorgan- acute adrenal insufficiency due to hemorrhage isms generally associated with moist environ- into the adrenal gland may result. Cases have been increasing as the popularity of health spas increases. a small. In cases of 86. The organism is found in brackish or salt water. Salmonella Typhi this organism from clinical samples. When not properly maintained. Painful genital lesions and painful water. This serotype is surrounded by a weakly immunogenic polyribitol phosphate cap. after which the patient notes the painful. patient's circulatory system several days before a stool culture will be positive. bloody diarrhea. nonindurated primary lesion on the exter- disseminated intravascular coagulation occurs in nal genitalia. is citrate negative. influenzae type b disease. Signs of regional lymphadenitis cases of fulminant meningococcemia—Neisseria appear in about one-half of the cases a few days meningitidis septicemia. Chancroid or soft chancre is caused by transmission. 87. bacterial meningitis in children 1-6 years of age was serotype B. produces only a small amount of hydrogen sul- fide. swelling of the inguinal lymph nodes characterize the disease. D. Wound infections are associated Haemophilus ducreyi. Special selective culture media 84. Campylobacter enterocolitis is characterized by fever. the pattern of dermatitis caused by these organ- isms matches the areas covered by the individ- B. Unlike other salmonellae. meningococcemia with intravascular coagulation. It is also important to note that the bacilli appear in the . cine. Invasion of the circula- after the appearance of the primary lesion. the strain of Haemophilus influenzae found whirlpools create a favorable environment for the to be implicated in the majority of cases of growth of these organisms. may be delayed if labora- tory professionals do not have a good apprecia- tion of its atypical characteristics. tact with an infected person ranges from 1 to 5 days. produces no gas from glucose.

Sur. The best identification method is the poly- merase chain reaction. In the labora- 91. B. mallei has not been isolated recently in the U. putrefa- ciens will produce acid from sucrose. aeruginosa is the most common gram-negative bacillus that is not in 89. the mother as it passes through the birth canal. Occasionally tation and the production of H2S. Screening procedures for the recovery of the A. . is caused by phomonas maltophilia the second most common.form of conjunctivitis. forming 93. nonmotile. Neisseria meningitidis and N. P. Bordetella pertussis. The infection is transmitted to the newborn by 90. It is important to be able to quickly differentiate those organisms resem- bling Salmonella from other H2S-producing organisms such as Citrobacter freundii and Edwardsiella tarda. C.. some strains of S. B. encapsulated.species is easily accomplished by demonstrating itive lysine and a negative urease reaction. unlike Salmonella. a minute. recommended for the isolation of this agent. cepacia is the most common Burkolderia spp. the family Enterobacteriaceae and Stenotro- A. gonorrhoeae enteric pathogen Salmonella rely heavily on dif. face structures such as pili aid in attachment to mucosal epithelial cells and invasion of submu- cosa to produce infection. with the agar would resemble Salmonella in that the paired cells having adjacent walls that are flat- organism is not able to ferment lactose and does tened. yellow colonies on Hektoen enteric agar. produce a significant amount of H2S. gram-negative. Ophthalmia neonatorum. ney bean" cellular morphology. These gram-negative coccal organisms samples on a medium such as Hektoen enteric appear characteristically as diplococci. It is an impor.S. will exhibit this morphol- Shewanella putrefaciens recovered from stool ogy. or pertussis. Whooping cough. Differentiation of these two seen in Color Plate 27 •. pleomorphic bacil- lus. D. which phenyl-beta-galactopyranoside) positive test. However. which the fermentation of lactose or an ONPG (o-nitro- differentiates them from Proteus spp. is associated with Neisseria gonorrhoeae. are most commonly described as having a "kid- ferential media. Isolates of some Moraxella spp. 92. which indicate lactose fermen. B. Neisseria gonorrhoeae is a primary The use of an ophthalmic solution of erythromy- pathogen of the urogenital tract. they demonstrate a pos. Most serotypes of Salmonella produce Neisseria meningitidis. a form of conjunc- tivitis. D. an organism of signifi- hydrogen sulfide in triple sugar iron agar. As cant pathogenicity. in clinical specimens. also produce H2S. Neisseria are important human pathogens. ANSWERS & RATIONALES 717 88. Regan-Lowe medium is 94. tory this agent may be mistakenly identified as D. B. 95.cin is recommended for the prevention of this tant cause of sexually transmitted diseases. Neisseria lactamica is part of the normal nasopharyngeal flora of humans.

recovery of Brucella are blood and bone marrow. C. Acinetobacter spp. The incubation 104. Many strains of A. Acinetobacter spp. Hemor- rhagic colitis is characteristic of infection. These organisms are known as one of the animal B. attending daycare centers. but it is not asbated at 42°C under microaerophilic conditions serious an infection." This form of conjunctivitis is highly contagious and is frequently seen in children 101. A. The largest number of human cases occurs of nosocomial infections. baumannii can form pink to purple colonies on MaConkey agar that 103. It has been isolated from a variety of clinical specimens but is generally considered clinically insignif- 100.718 • CHAPTER 6: BACTERIOLOGY 96. icant. are all oxidase positive. They are oxidase neg- between May and November and seems to be ative and will grow on most laboratory media. The characteristics of this organism are positive motility and lysine. They are predomi- nantly animal pathogens. curved. highly associated with exposure to water or soil. Hafnia alvei is the only species in the genus. are fastidious. Hafnia alvei is a member of the family aches. 102. teristically self-limited and does not demonstrate significant pulmonary symptoms. Brucella spp. The usual specimens for . and KCN reactions. Aeromonas. undercooked ground beef or raw milk.Shiga toxin produced by Shigella dysenteriae. can be mistaken for lactose fermentation. The agent is an aerobic gram-negative bacillus that is nonmotile and A. motile gram-negative rods that are hippurate hydrolysis positive. E. are opportunistic pathogens for humans and are important causes pathogens that cause "red leg disease" in frogs. agar is used for isolation from stool and is incu- mophila. infection can also lead to hemolytic uremic syn- drome resulting from toxin-mediated kidney A. Chryseobacterium. are nonfermenters. ornithine. Campylobacter jejuni are small. are found in bodies of fresh water and salt water that can be flowing or stag- 98. Enterobacteriaceae and is oxidase negative. baumannii will oxidize glucose. but A. fever. is short. coccobacillary organisms. as is Legionnaires disease. and headache. Pontiac fever is caused by Legionella pneu. A. including MacConkey agar. coli O157:H7 produces a toxin similar to requires both hemin (X factor) and nicotine ade. but occasionally pro- duce disease in humans. muscle C. but 97. gram-negative. unlike that for Legionnaires disease. chills. with the latter considered the more sensitive. V factor) for growth. It is most commonly transmitted by ingestion of nine dinucleotide (NAD. bacteriaceae. trointestinal tract of a variety of animals. Aeromonas spp. They are found in the gas- 99. nant and contaminated with sewage. Hafnia is a member of the family Entero- damage. followed by symptoms of malaise. period. Haemophilus aegyptius is the causative agent of "pinkeye. This febrile illness is charac-(10% CO2. ONPG. 5% O2 with balance N2) for 72 hours. and Vibrio A. Campy B.

cellosis is suspected. often produce inflammation resulting in bloody diarrhea. coli and epithelium. The most commonly used method for serogrouping Shigella is based on the somatic oligosaccharide or O antigens. supporting the theory of a fecal-oral or oral-oral route of transmission. the cause of death in untreated patients. requiring increased osmotic V. 112. Proper therapy begins with intravenous fluids to A. The other bacteria antimicrobial agents limits the selection of ther- listed produce toxins or invasive diseases that apeutic agents. Because Shigella spp. The capsule (K) antigens are used to the age of 60 years are infected. ANSWERS & RATIONALES • 719 105. or salt water. Growth in the presence of 1 % NaCl but no growth in media without the added Na+ is the test for the . especially in agar but not sheep blood agar. Incubation under aerosol-generating procedures. halophilic Vibrio sp. N.and commonly colonizes hospitalized patients. dence of gastritis increasing with age. parahemolyticus is halophilic. D. invade the intestinal mucousa but is attached to Its resistance to many of the commonly used the surface of enterocytes. H antigens are used for E. Marine water is the normal habitat of most Vibrio spp. Severe dehydration is usually 111. Most laboratories send iso- lates to a reference laboratory for confirmation 106. the H or flagella antigens cannot be used. It is important CO2 is required for recovery. Vibrio parahemolyticus is found in brackish 108. gonorrhoeae will grow on chocolate C. Neisseria gonorrhoeae is the causative agent of gonorrhoea and is very sensitive to drying. 107. in the form of salt. Acinetobacter is widely distributed in nature restore the patient's water volume and elec. gastric mucosa colonizing the mucous layer of the antrum and fundus but does not invade the however. for growth. or definitive identification because they lack spe- cialized media and containment facilities. trolyte balance. A. temperature variations. The microorganism does not Infection occurs mainly in compromised hosts. This makes routine biochemical test media less than optimal because of their low NaCl content. pylori has been cultured from feces and dental plaque. differentiation of halophilic organisms. and fatty acids in clinical 109. D. The mode of transmission is the C. Vibrio parahaemolyticus is classified as a ingestion of contaminated water or seafood. are also used to serogroup Escherichia coli and B. C. are non- motile. and selective for the laboratory to be notified whenever bru- media like Thayer-Martin are recommended. Approximately 50% of adults over Salmonella. Helicobacter pylori is found in the human Salmonella. Brucella spp. H. with the inci- serogroup Klebsiella pneumoniae. material. pressure. are harzardous. Vibrio cholerae produces an exotoxin that causes infected individuals to lose massive amounts of fluids. The O antigens 110.

Salmonella. The Voges-Proskauer (VP) test is a broth test those that do not degrade lactose. the Vi antigen has been secretion of large volumes of fluids into the removed. Pathogenic mechanisms of V. A red color of E. 117. 119. A. colonies tive. In the U. would produce red colonies on XLD agar. on Hektoen enteric agar. dominant epidemic strains.S. Brucellosis presents as an A. Enterobacteriaceae that are VP positive are would be orange. The most common 24 hours on xylose-lysine-desoxycholate (XLD) clinical isolates from the genera Providencia. sonnei is ONPG positive but is a delayed lac. is characterized as producing outbreaks of cholera. extraintestinal symptoms. The Vi antigen is a strains colonize the small intestine where they heat-labile capsular antigen associated with elaborate choleragen. Ogawa and Inaba strains are considered the pre- B.720 • CHAPTER 6: BACTERIOLOGY 113. 118. and Serratia are posi. agar. tion of humans. motil- lent antiserum composed of antibodies against ity. After heating a Salmonella produced when the enterotoxin stimulates the Typhi suspension. Isolates metabolism of glucose in the medium. Non-lactose fermenters such as Shigella Escherichia. and Proteus are gener.S. enzymes such as protease and mucinase. the somatic grouping antisera. On ally VP negative. is mainly job related or S. disease characterized by its ability to ferment lactose. the strain O139 has also been associated with unlike C. Campylobacter fetus subsp. Classic epidemic strains of Vibrio cholerae are included in the antigenic O group 1. . decarboxylation.. and the commonly isolated strains. Salmonella Typhi demonstrates a positive agglutination reaction in D-grouping sera./eftw is occasion. Brucella spp. urease. Most strains of E. D. such as in tose fermenter. beta-hemolytic on sheep blood agar. jejuni. Disease is Salmonella Typhi. motility. for the differentiation of lactose fermenters from A. A. including antis. In the last few years ally implicated in human disease. In some media that detects the presence of acetoin from the H7S production may be demonstrated. coli are typically methyl red negative. are small. include adherence to enterocytes via pili. coli would produce yellow colonies at indicates a positive reaction. Organisms biochemically resembling C. involves food or animal associations. and the organism can now react with intestinal lumen. and lysine 120. lactose fermenters produce Klebsiella. Epidemic era against the Vi antigen. the production of an enterotoxin. Those persons most at risk of infection are those with preexisting disease who are in a debilitated condition. The genus Shigella is characterized biochemically by being negative for citrate. The 114. hunters or those who drink raw milk. pink colonies. This is the most commonly iso. D. Shigella sonnet is a group D Shigella and is undulant febrile illness. This organism. Vari- ous selective and differential agars are available 116. caused by Brucella sp. lated species of Shigella in the U. gram-negative intra- cellular parasites implicated in zoonotic infec- 115. whereas most members of the MacConkey agar. cholerae Salmonella are typically tested using a polyva. an enterotoxin. Escherichia coli is a lactose-fermenting member of the family Enterobacteriaceae. Enterobacter.

Organ- is perhaps the most accurate means of detecting isms such as Citrobacter freundii. Collection and processing of speci- B. is the etiologic agent because this organism is sensitive to drying and of the sexually transmitted disease granuloma low temperatures. and erythromycin is the drug of the culture media in order to grow. is nonmotile at 20-25°C. plague. which is grossly observed for the deter. When streaked on DNase test medium. although it does not Gram stain well. Pneumonic legionellosis and the nonpneu. The hanging-drop method sole source of carbon as an energy source. vation is 35°C. mens must be done under optimal conditions. aerogenes. flooded with a 0. inguinale. D. medium such as modified Thayer-Martin or GC- Lect should be used for primary isolation. are characteristically nonmotile. formerly Calymma. encapsulated bacillus. bacteriaceae that are Voges-Proskauer positive are C. Direct fluorescent antibody is often used ism requiring the addition of serum or blood to diagnostically.1% solution of HC1. also citrate positive. unlike other Yersinia for DNase activity. Enterobacter motility of nonfermentative microorganisms. Of the Entero- bacteriaceae. espe- cially from sites that may be contaminated with 124. The optimal temperature for culti. after which the plates are for tLS and urease. these organisms are extremely difficult to recover. This marcescens will demonstrate a positive reaction oxidase-negative organism. First seen as inclusions (Donovan bod- ies) in mononuclear cells from genital ulcers stained with the Giemsa or Wright stain. It is also negative bated 18 to 24 hours. tobacterium granulomatis. . Klebsiella granulomatis. B. Citrate utilization is a common test used for Motility of the Enterobacteriaceae can normally the differentiation of members of the family be detected by the use of a semisolid motility Enterobacteriaceae. Inoculated plates are incu- spp. gram-negative. A. the colonies.. and cold enrichment is not appro. B. Many Entero- 123. and Serratia marcescens are able to grow on citrate agar and produce a color change from green to blue in the medium. Motility can be important in the identi- fication of microorganisms. It is a pleomorphic. the genera Klebsiella and Shigella 126. A selective choice for therapy. 125. DNase- producing organisms are differentiated by the development of a clear zone in the agar around 122. 127. Neisseria gonorrhoeae is a fastidious organ- priate. ANSWERS & RATIONALES • 721 121. Shigella boydii are incapable of using citrate as the mination of motility. Yersinia pestis is the causative agent of D. monic illness known as Pontiac fever are the two clinical forms of disease caused by Legionella pneumophila. normal flora. Both Escherichia coli and medium. The organism is endemic in rodents and colonies of Staphylococcus aureus and Serratia is transmitted to humans by the rat flea. as is Tatumella.

Moraxella catarrhalis possesses the enzyme observed. Campylobacter jejuni is an important human pathogen most commonly associated with cases nia. also an important cause of meningitis. colony morphology.134. and blue-green tive. Organisms such as E. Special handling of cultures suspected of the Enterobacteriaceae. which possess the enzyme tryptophanase. Indole represents the "I" in the IMViC reac- of bloody diarrhea. A. Most P. Pyocyanin is the nonfluorescent. Neisseria meningitidis. Wet reaction on TSI agar that indicates that glucose mounts demonstrate the typical "darting" motil- and either lactose or sucrose or both have been ity of this isolate. which rapidly dark- A. would be . oxidase positive. C. because they are generally not able to indophenol oxidase. and both are uncommon isolates. to contain this organism is required for optimal recovery. would for differentiating lactose-positive Escherichia likely grow on both SBA and chocolate agar. but growth is enhanced by the addi- 131. a battery of tests used for the identification humans. All of the bacteria listed are fastidi- ous. are able to 132. and weakly pigment. E. gram-negative bacillus that is nonmotile.722 • CHAPTER 6: BACTERIOLOGY 128. hominis will grow on sheep blood agar. Shigella can be ruled out when such a reaction is C. coli. catalase negative. The enteric pathogens Salmonella and 133. tetramethyl-p-phenylenediamine (oxidase reagent) is applied to colonies of these organisms. the 130. in a microaerophilic atmosphere and examined at 24 and 48 hours for spreading nonhemolytic A. coli produces an acid over acid (A/A) colonies. which may be slightly pigmented. Haemophilus influenzae would be expected to grow on chocolate agar but not sheep blood agar (SBA). duce this pigment. diffusable pigment produced by Pseudomonas aeruginosa. colonies turn a purple color. however. When a 1% solution of use either lactose or sucrose within 18 hours. aeruginosa strains can be identi. ens. pyruvic acid.that is recovered predominantly from cases of fied presumptively by their characteristic grape. coli from other members of the family Entero- bacteriaceae. indole positive. fever. blue-green. Bacteria that ferment lactose or sucrose produce large amounts of acid in the medium. Cultures should be incubated at 42°C 129. expected to grow on SBA incubated in CO2. C. It is characterized as a fermenta- like odor. Cardiobacterium hominis is a rare pathogen green. tion of yeast extract to media. fermented. and abdominal pain in tions.endocarditis. and ammo. metabolize the amino acid tryptophan with the production of indole. It is the only bacterium able to pro. The indole reaction is a widely used method Bordetella parapertussis and Brucella sp. The other species listed are oxidase negative. Color Plate 28 • is a Gram stain of a cere- brospinal fluid specimen revealing many white blood cells. Pyocyanin mixes with the yellow pigment fluorescein to turn culture media C. D.

tum.positive cultres for Legionella pneumophila. The bacteria are seldom recovered from blood specimens. The isolate is associated with two distinct clinical conditions: primary 139. blood agar. therefore. Unlike 5% sheep most cases with preexisting hepatic disease. usually in association with an environmental source of A.cedures and are not commonly performed. Legionella pneumophila requires the use of belongs to the family Enterobacteriaceae and is special laboratory media for cultivation and does not stain well by the conventional Gram stain. are biochemi- cally inert. and Morganella morganii. it provides both hemin (X factor) Septicemia due to V. produces a fulminant disease with a high mortal- H. pigment. D. Providencia stuar- tii. are obligate par- asites of animals and are found primarily in the 136. which includes Pseudomonas aeruginosa. Cationic salts such as magnesium sul. 140. and P. putida. are appropriate for the isolation of L. Chocolate agar is the preferred culture with this organism appears to be correlated in medium for Haemophilus. upper respiratory tract and oral cavity. MacConkey agar. cein (pyoverdin) is produced by members of the Pseudomonas fluorescent group. This includes Proteus vulgaris. Wound infection with this organism is Mueller-Hinton-based chocolate agar. tract (lung biopsy) have the greatest yield of ultraviolet light source. rescent pigments is dependent upon nutritional However. Septicemia B. of transmission is by the airborne route. supple- usually associated with trauma and contact with mented with 1% IsoVitaleX and 3 |ag/mL of a marine environment. The water-diffusible yellow pigment fluores- bacteria. and is oxidase-positive. such as cetrim. ity rate. K. The production of pyoverdin can be detected when a culture of the C. also PDA positive. and autofluoresce. P. Tissue samples from the lower respiratory organism is exposed to a short-wavelength. denitrificans can be isolated from the human upper respiratory tract. C. will grow on modified Thayer-Martin agar.such as bronchial wash and expectorated spu- fate intensify luminescence. ANSWERS & RATIONALES 723 135. ide. fluo. Most Legionella spp. should be used when trying to detect the Cultures of lower respiratory tract specimens. Kingella spp. The production of fluo. Vibrio vulnificus is a halophilic lactose- fermenting organism. are gram-negative bacilli or coccobacilli that may appear in short chains. vulnificus characteristically and NAD (V factor) required for growth. special media. ducreyi grows best in a special medium.141. vancomycin. are motile. pneu- mophila. Members of the tribe Proteae are characteristi- cally positive for phenylalanine deaminase (PDA). these specimens require invasive pro- factors. Haemophilus spp. Tatuella ptyseos A. C. and growth is poor on triple sugar iron agar. rescens. 138. The growth of this organism is inhibited by . septicemia and wound infection.The primary mode 137.

and characteristically resistant to 150. Modified C. This species will oxidize but not ferment ornithine decarboxylase negative. It is commonly associated with polymicrobial . animals. Proteus mirabilis is commonly associated are based on this species. mens. wound infection. Cephalexin is often added to make the medium selective. Penicillin is an effective drug Some strains of E. The bac- terium requires both X and V factors and is 145. nonmotile. suppurative pigment that aids in its presumptive identification. similar to Neisseria gonorrhoeae both on Gram D. infections following bite or clenched-fist wounds. This modified Thayer-Martin medium and its posi- organism. The ability of flora of humans. glucose. and phenylalanine deami. Acinetobacter spp. influenzae biotype III in D. D. and they are oxidase negative. multocida is the Skirrow's medium is a primary plating media for species commonly recovered in clinical speci- Campylobacter spp. cowanii are also pigmented. in other parts of the body. It resembles H. Enterobacter sakazakii produces a yellow duces a rapidly progressing. Proteus are characteristically rapidly urea posi- tive. It is morphologically 146. Regan-Lowe. stain and colonies on culture media. It is a motile organism that characteristically swarms across the surface of sheep blood agar plates. therefore negative for delta-aminolevulinic acid (ALA). D. for the treatment of Pasteurella infections. B. Humans most often become infected from a bite or scratch of a cat or dog. pneumophila will also autofluoresce. pneumophila is not with urinary tract infections as well as infections the only one associated with human disease. sakazakii is an occasional clinical isolate that has been linked to respiratory tract infections and wounds. Members of the genus 149. denitrificans to reduce nitrates is a key test for its differentiation from N. Legionella pneumophila is able to hydrolyze 144. Confusion is further compounded by its ability to grow on bic gram-negative bacillus that requires hemin in the culture medium to grow aerobically. The diagnosis of pertussis. This gram-negative coccobacillus is a normal inhabitant of the oral cavity of domestic 143. gonorrhoeae. and nitrate. Acinetobacter baumannii is not able to reduce that it is indole negative. K. ease known as Brazilian purpuric fever. lactose negative. hippurate. is seldom found in pure culture. Although most of the studies done on legionellosis C. E. which produces pitting of the agar cough. penicillin. Kingella denitrificans is most often associ- ated with endocarditis. provides the best results. which pro- C. Eikenella corrodens is a facultatively anaero. which is a part of the normal indigenous tive oxidase and glucose reaction. L. is confirmed by culture. 148. or whooping this organism. Isolation of the etiologic agent is best 147. L. are able to grow on MacConkey agar. painful. P. Haemophilus influenzae biogroup aegyptius causes a conjunctivitis followed by invasive dis- nase positive. a on which it is isolated. In the genus Pasteurella.724 • CHAPTER 6: BACTERIOLOGY 142. charcoal-based medium. urease positive. done within the first week of the illness. Infections of the face and neck may also involve C.

Infection in humans teria produce buboes in the groin and can cause a has manifested mainly as diarrheal diseases. canis. D. 155. and gelati. and septicemia. The production of DNase. P. Posterior nasopharyngeal cultures are recom- 152. . These small. a serious sexually transmitted dis- genie studies. 157.. Wampole Laboratories) is infections such as septicemia and meningitis and more sensitive than a biphasic culture bottle.ease. Serratia can produce severe tion method (Isolator®. P.recommended specimens for the isolation of nase differentiates the genus Serratia from other Brucella spp. nature of Pseudomonas aeruginosa in the envi. Cephalexin is added In addition. range of temperatures (4-42°C). B. Serratia spp. B. Ingestion of contaminated unpasteurized probably not involved in the pathogenesis of B V. The lysis-centrifuga- comial infections. sumptive diagnosis. "nonspecific vaginitis. B. vaginalis is B. Brucellosis is found worldwide. and B. 154. The disease is more prevalent in the tropics ily Enterobacteriaceae.senstitive recovery method. mended for the recovery of Bordetella pertussis C. especially for a continuous monitoring system is the most S. Inoculation of a blood culture bottle Enterobacteriaceae. Swabs of the nasopharynx are inoculated on the ronment and its resistance to many disinfectants. melitensis. The bac- only species in this genus. 158. selective agar Regan-Lowe. Based on nucleic acid and anti. Cultures of blood and bone marrow are the D. Haemophilus ducreyi is the causative agent Vibrionaceae. a waterborne mode of transmission is often the source. aeruginosa is an opportunistic pathogen commonly associated with hospital-acquired infections. (raw) milk or cheese is one of the primary routes its presence in high numbers is considered a pre- of infection. B. gram-negative and symptoms vary from asymptomatic to a bacilli are frequently seen in great numbers on debilitating systemic infection. 156.of chancroid. B. suis. ANSWERS & RATIONALES 725 151. __ C. are frequently difficult to eradicate because of the characteristic antimicrobial-resistant strains found in the hospital environment. shigelloides is the than in temperate parts of the world." Although G. abortus. The recalls described illustrate the ubiquitous in suspected cases of pertussis (whooping cough). it was recently moved to the fam. Gardnerella vaginalis is associated with cases of bacterial vaginosis (B V) formerly called 153. the bacterium has minimal nutritional to the culture medium to inhibit the growth of requirements and the ability to tolerate a wide contaminating indigenous flora. lipase. have a close association with noso. Only four of the the surface of epithelial cells ("clue cells") taken six species are typically pathogenic for humans: from the vagina. Plesiomonas was previously in the family A. marcescens.

726 • CHAPTER 6: BACTERIOLOGY 159. 166. appearance. but cultures are not recom.A. C. Gardnerella vaginalis is associated with bac. of the water. 161. The organism most commonly dicitis. vaginal epithelial cells with fuchsin or basic fuchsin be substituted for the gram-variable bacilli attached to their surface. in which the adjacent sides are flattened. can be recovered from kidney bean on Gram stain because of its charac- respiratory tract secretions. after which further identification logically similar. Mesenteric lymphadenitis is one of the com. Cefsulodin-irgasan-novobiocin (CIN) agar is 164. and it is recommended that carbol- detecting "clue" cells. like Francisella tularensis. the motile with a typical darting pattern on wet isolation of the organism may not be clinically mounts. Y. The cul. Fresh isolates of Campylobacter jejuni on terial vaginosis. Contaminated food products and supresses the growth of normal fecal flora and dif. coli can pro- dark red center surrounded by a translucent bor. These gram-negative bacilli are vaginalis as normal vaginal flora. 165. Neisseria gonorrhoeae is said to resemble a of Legionnaires disease. They stain poorly using the Gram stain significant. be interpreted with caution. B. water in foreign countries seem to be the major ferentiates colonies of Y.Gram stain characteristically reveal a "gull-wing" mended for diagnosis. This gram-negative coccobacillus grows associated with this condition is Pseudomonas well on routine culture media and has an optimal aeruginosa. Symptoms produced by pletely dry their ear canals when they get out this agent closely resemble those of acute appen. ined. Smears from the female genital tract must which is incubated in a moist chamber at 35°C. enterocolitic produces toxin-producing invasive or enteropathogenic "bull's-eye" colonies at 48 hours. enterocolitica. D. common to persons who swim and fail to com- tuberculosis infections. The bacterium is teristic gram-negative diplococcal morphology fastidious and. The disease can be diagnosed by method. This is a selective and differential agar that one is heat labile. . opportunistic pathogen and one that favors a watery environment. colonies show a Escherichia coli. Traveler's diarrhea is caused by strains of Yersinia and Aeromonas. Enterotoxigenic E. counterstain safranin. recommended for the primary isolation of B. B. It is an organism known to be an growth temperature of 25-30°C. however. because Growth on this medium may not be visible for other normal flora microorganisms are morpho- 3 to 4 days. 162. the causative agent C. therefore. duce one or two exotoxins: one is heat stable and der. 160. 163.cally these organisms are found intracellularly ture medium most commonly recommended is when direct smears of clinical material are exam- buffered charcoal yeast extract (BCYE) agar. procedures may be carried out. Many women carry G. D. vehicle for human infection with these agents. Typi- requires cystiene or cystine for growth. Legionella pneumophila. Swimmer's ear is a form of external otitis mon manifestations of human Yersinia pseudo.

A rapid positive urease is C. Bartonella bacilliformis is the causative B. and capsule resulting in mucoid colonies. Neisseria gonorrhoeae is identified in the after addition of the reagent is a positive result. Klebsiella spp. Proteus vulgaris is able to deaminate pheny- a key test for the identification of//. is able to liquefy gelatin. Nucleic acid is also nonmotile. This bacterium is not thought to be a primary pathogen and is only . adding a 10% solution of ferric chloride. The The disease is rare and occurs primarily in South mode of transmission generally involves trau- matic inoculation of the organism through the America. Pasteurella multocida is the species in the agent of Oroya fever and verruga peruana. identification procedures necessary. Salmonella is urease negative and H2S posi- ating this isolate from Serratia marcescens. It is a pleomorphic. 174.genus most often encountered in the clinical lab- cellular parasite of red blood cells and can be cul. all are oxidase positive. D. menin- positive and generally attacks carbohydrates gitidis ferments both glucose and maltose. nine and. are all nonmotile. amplification tests are also used frequently to diagnose gonorrhea. produce a logically. gram-negative rod that is an intra. Shigella. made only by the isolation and identification of W. 170. A. 171. Neisseria meningitidis is a human pathogen that can be pathogenic for humans. P.oratory. tive. clinical laboratory by its ability to ferment only glucose. Chromobacterium violaceum is a motile. pylori. are identified either by fermentation tests or sero- Chromobacterium is catalase and oxidase logic methods that use specific antisera. skin. multocida grows on sheep blood agar but not on MacConkey agar. humans. A dark green color on the slant A. The diagnosis of the sexually transmitted disease caused by this agent can be definitively 173. fermentatively. and it is an opportunistic pathogen. 175. The test is performed by inoculating the isolate on a slant of medium containing phenylala- 168. lalanine. Serratia liquefaciens. Morpho- in their identification. Klebsiella spp. after incubation. 169. gram-negative bacillus found in soil and water C. Yersinia enterocolitica and Y. A. ANSWERS & RATIONALES • 727 167. all members of the genus are alike. rarely isolated from clinical specimens. gram-negative diplococci by these organisms aids in their identification. as its name implies. The pro- most commonly associated with meningitis. The pos- itive arabinose reaction is one way of differenti- A. which aids gonorrhoeae in the clinical laboratory. It is normal oral flora in animals. pseudotuberculosis 172. which makes definitive another genus in the family Enterobacteriaceae. B. are both urease positive. N. The negative urease reaction would help to differentiate it from an H2S-positive Proteus spp. not tured from blood in the acute stage of the disease. duction of a non-water-soluble violet pigment These oxidase-positive.

Colonies of some organisms. Pseudomonas aeruginosa has not only a that the organism possesses the enzymes to con- vert aminolevulinic acid (ALA) into porphyrins and. resemble Salmonella in many ways. influenzae to grow. this rapid test will show green color. colonies are agar. such as agent can be easily recognized by its fermenta- the Enterobacteriaceae. 179. A. such as laboratory. Isolates are ONPG and catalase posi- positive indole test. bronchiseptica may marcescens is the most clinically significant of cause problems for researchers because it can the genus and is frequently involved in nosoco- cause outbreaks of bronchopneumonia in exper- mial infection. duces either a respiratory illness or wound infections. B. On sheep blood agar medium. 182. The red pigment produced is not imental animals. On eosin methylene blue tive. Edwardsiella tarda is a motile member of the zae on sheep blood agar medium around family Enterobacteriaceae and as such is char- colonies of organisms that provide an essential acteristically peritrichously flagellated. This other fermentative gram-negative bacilli. would not require hemin. 180. negative citrate reaction. 183. C. S. B. Escherichia coli produces characteristic beta-hemolytic. "Satellitism" is the name given to the appearance of colonies of Haemophilus influen- A. H. Aeromonas can be differentiated from many agent of community-acquired cystitis. produce NAD. Escherichia coli is frequently the etiologic C. ferment lactose. It also causes kennel cough in water soluble and is demonstrated more readily cannines. A. Bordetella bronchiseptica in humans pro- 177. Biochemically they may initially Staphylococcus and Neisseria. C. and water and has been isolated from a variety of human infections. rabbits and guinea pigs. hydrophila is found in soil dark colonies with a metallic sheen. motile organisms red fluorescence under UV light after a 4-hour are oxidase positive and are able to grow at incubation period. such as which diffuses into the surrounding agar and hydrogen sulfide production and the inability to enables H. B. In humans these opportunistic organisms cause many types of infections. therefore. The organism is a part of the normal A. influenzae requires both hemin organisms are infrequently isolated in the clinical and NAD. These growth factor. 728 • CHAPTER 6: BACTERIOLOGY -. 178. Serratia marcescens is a chromogenic respiratory flora of laboratory animals such as member of the family Enterobacteriaceae. A positive test result indicates . The porphyrin test is commonly used to test for the X factor (hemin) requirement of Haemophilus spp. 176. but they are pri- marily associated with burn wound infections. If por- characteristic grapelike odor but also a blue- phyrins are produced. These oxidative. by incubation at room temperature than at 35°C. in that they are oxidase tion of lactose. 42°C. and positive. 181.

C. C. accounted for as much as 10% of nosocomial infections. Hektoen enteric agar was developed to chronic carriers. 185. Pseudomonas aeruginosa is a major cause of pertussis. A. respectively.GC-Lect. Carriers are usually asymptomatic.include Martin-Lewis. . isms turn the medium yellow. Both diseases are spread by droplet infec. These oppor- Gengou or Regan-Lowe agars. Salmonella Typhi is commonly spread by A. A variety of media has been developed to aid known as epidemic cerebrospinal meningitis. On Bordet. The medium also gallbladder. Pontiac fever is caused by Legionella pneumoniae. H2S- 187. 186. this enteric improve the isolation of Shigella and Salmonella bacillus can be carried throughout a person's from stool specimens. B. The microorganisms are present in isolation of N. or whooping cough. Ferric ammonium citrate and sodium thiosulfate are included in the medium to detect H2S production. sucrose. Without treatment. greatest numbers in the upper respiratory tract. ANSWERS & RATIONALES • 729 184.of hospital-acquired infections. Cefsulodin-irgasan-novobiocin (CIN) is a selec- and specimens for isolation and identification tive and differential medium for the isolation of should be collected on nasopharyngeal swabs. 188. mallei. and the presence of the organism can be con. Milk is able to sup- port the growth of many clinically significant microorganisms. which may often be ingested in unpasteurized dairy products. The most tion or fomites contaminated with respiratory commonly used nonselective medium for the secretions. None of these is transmitted by milk. Bordetella pertussis is the etiologic agent B. modified Thayer-Martin. Fermentative organ- clinical laboratory. and New York City media. A nasopharyngeal swab is recom. in the isolation of Neisseria gonorrhoeae from Infection with Bordetella pertussis produces the specimens containing mixed flora. gonorrhoeae is chocolate agar. of one form of inflammation of the meninges. contains three carbohydrates—lactose. Brucella infects cattle and may be transmit.transferred to immunocompromised patients.and salicin—along with a pH indicator to detect firmed only by isolation and identification in the carbohydrate fermentation.environments for prolonged periods and may be cury droplets. the organism tunistic organisms are able to survive in moist forms small. The selective nature of lifetime and is sequestered most often in the this agar is due to bile salts. round colonies that resemble mer. Neisseria meningitidis is the etiologic agent 189. producing organisms appear as black-centered A. ted to humans by the ingestion of contaminated milk or other dairy products. Yersinia enterocolitica and Aeromonas.colonies. mended as the optimal specimen for the recovery Pseudomonas infections in recent years have of this agent. tussis. Meliodosis and glanders are caused by Burkholderia pseudoma- llei and B. Examples highly contagious upper respiratory infection per. 190.

V cholerae and V. produce the enzyme cytochrome oxidase. Exotic pets such as iguanas. A. influenzae. Helicobacter pylori is implicated as an etio- logic agent of gastritis and peptic ulcer disease. B. parahemolyticus is unable to Salmonella have been implicated in cases of sal- ferment sucrose and exhibits colonies with blue to monellosis. Strains of Haemophilus able to synthesize positive urease test result. rhea.or tetra-methyl-p-phenylene- loss of vision. most common agent of bacterial keratitis associ- and the test organism is then rubbed into the ated with lens-cleaning solution. a naturally occurring substance containing dren who do not practice good handwashing bile salts and sodium cholate. or food poisoning. Natural medicinal products made nolyticus ferment sucrose and appear as large from snakes or other animals known to carry yellow colonies. Pseudomonads impregnated area. whereas 198. horse serum-based medium that is selective by the B. 193. and tur- medium to inhibit gram-positive organisms is tles are known to carry Salmonella. the quality of the antibody. Species such as H. H. tericin B. vancomycin. Pseudomonas aeruginosa is the diamine dihydrochloride is applied to filter paper. The oxidase test detects those organisms that characterized by inflammation of the cornea. pylori produces a strong B. This organism can be demonstrated in gastric biopsy specimens. loop or wooden applicator stick should be used to pick the colony. V. The selective agent in this C. inoculated is a positive oxidase test. New York City (NYC) medium was developed by the New York City Public Health Laboratory for the isolation of Neisseria gonorrhoeae. Material collected from the . Direct fluorescent antibody (DFA) test addition of colistin. another com- sumptive. 196. A red color is indicative of a positive reaction in this test. B. may lead to solution of dimethyl. green centers. The rapid development of a dark purple color in the area where the organism was 197. 192. snakes. a platinum or plastic associated with contaminated fluids. Because Nichrome wire may are opportunistic pathogens that are commonly cause a false-positive result. Sucrose is the carbo- after touching family pets are particularly at risk hydrate in the medium. which require heme. Both positive and negative test results monly used selective medium for N. gonorrhoeae. heme are identified by the porphyrin test. D. The quality of the is chocolate based. Thiosulfate-citrate-bile salt-sucrose (TCBS) agar is recommended for use in the selective isola- tion of Vibrio spp.730 • CHAPTER 6: BACTERIOLOGY 191. Modified Thayer-Martin. 195. diar. and the microscope. Young chil- oxgall. parainfluenzae would be positive. It is a 194. and ampho- results for Bordetella pertussis are rapid but pre. A 1% which. associated with cholera. would give a negative test result. algi- for infection. test result depends greatly on the experience of the microscopist. Keratitis is a serious clinical condition that is A. if not appropriately treated. __ H. nasopharnyx with calcium alginate swabs for DFA and culture is recommended. must be confirmed by culture.

aeruginosa is by far the most impor. ANSWERS & RATIONALES • 731 199. these bac. and acids. teria do not stain well with the Gram stain. Thus. Both of these A MacConkey agar plate is inoculated with a bacteria are oxidase positive and will grow on 7-day broth culture of the test organism. This slowly Mycobacteria growing Mycobacterium is a cause of cervical 201. (NALC-NaOH) method is recommended because the addition of NALC allows the concentration 202. 203. Plates duces a green discoloration of the medium it is are checked for growth at 5 days. in that crystal violet is omitted. aeruginosa typically pro. scotochromogen because of its characteristic of producing pigmentation in the dark. Eikenella corrodens can be normal flora of in 5 days. they are checked daily until day 11. The MacConkey agar used for mycobacteria tant cause of lower respiratory tract infections in identification is a different formulation than that patients with cystic fibrosis. tent acts to protect these organisms from dehy- dration and the lethal effects of alkali. . Although P. The optimal treatment reduces the numbers and replication as an environmental contaminant of indigenous microorganisms present in the in hot water systems. various germicides. chelonei are the only 200. M. P. and the NaOH acts as a decontami- terium xenopi is 42°C. The TV-acetyl-L-cysteine-sodium hydroxide an acid-fast staining technique must be used. The optimal growth temperature ofMycobac. those not known to cause cutaneous infections. significant Mycobacterium spp. significant cause of morbidity. Like Pseudomonas aeruginosa. E. Growth on MacConkey agar is a test used for cepacia is a ubiquitous opportunistic organism. the sample. fortuitum and M. corrodens will grow on 204. the oral cavity of humans. and if no grown on. Human infections caused sample without significantly reducing the number by M. B. B. differentiation of rapidly growing mycobacteria. adenitis and other types of infections predomi- nantly in children. Therapy may require suscep- A. Some strains D. high lipid content.205. growth is detected.. alcohol. at which time they are discarded as nega- tive. which enables its survival nant. The NALC is a mucolytic agent that frees trapped organisms in D. cepacia is also a used for enterics. The MacConkey agar. Mycobacteria characteristically possess a tibility studies that include the secondary drugs. Mycobacterium scrofulaceum is defined as a will produce pitting of the agar. Burkholderia B. xenopi are rare. inoculated plate is then incubated at 37°C. have an optimal growth temperature of 37°C. The high lipid con. of NaOH to be reduced to 2%. unlike gram-positive cocci because the organism is known in some cases to and gram-negative bacteria. mycobacteria able to grow on MacConkey agar A. sheep blood and chocolate agars. and resistant to isoniazid and streptomycin. It is a weak pathogen that is associated with polymicrobial abscesses of the oral cavity. The majority of clinically of tubercle bacilli.

NaOH. 208. are treated generally with a combination of at C. Skin or skin infections. Up to nine acid-fast bacilli per field should be 212. Their predilection for surface biopsy specimens taken from within the margin areas of the body is related to their optimal of a lesion will demonstrate the causative agent. Hansen disease (leprosy) is caused by C. and pyrazinamide. if at all. they published in 1981. B. these organisms other mycobacteria. Mycobacterium ulcerans and Mycobac- Mycobacterium leprae. growth temperature range of 30-32°C. unlike temperature (37°C) or higher. reported as a positive. induration following an intradermal injection of because of the high lipid content of their cell PPD. NALC (W-acetyl-L-cysteine) is a mucolytic agent used in decontamination and digestion procedures for the recovery of mycobacteria. such as isoniazid. between 2 and 4%. example of a hypersensitivity reaction. wall. exhibit acid fastness when stained. tion has adopted the diagnostic standards recom.rifampin. 211. Because mended by the American Thoracic Society as of the slowly growing nature of the bacteria. NALC liquefies mucus. A positive tuberculin skin test reaction is an stain does not need to be heated. Most species pathogenic for humans are slowly growing. Acid-fast bacilli can be demonstrated in teria. acid and are known as purified protein derivative C. the number of acid-fast bacilli observed in fuchsin-stained smears of clinical material. This is a method of reporting are innately resistant to a number of agents. is frequently stained smears of clinical material using the used as a bactericidal agent to prevent the over- Ziehl-Neelsen or Kinyoun acid-fast stains. Cultivation can be accomplished by injecting bacilli into the foot pads of mice or systemically into armadillos. Patients 207. Therefore. At body Cultures of this agent on artificial media. A positive test demonstrates an area of characterized as obligate aerobic bacilli that.least two of the primary drugs. The growth of normal flora in clinical specimens. the Kinyoun D. unlike the Ziehl-Neelsen stain. Mycobacterium kansasii is a slowly growing photochromogen. Rapid development of drug resistance is a concern in the treatment of tuberculosis. .732 • CHAPTER 6: BACTERIOLOGY I 206. Members of the genus Mycobacterium are (PPD). 210. B. at 3+. . Chronic skin lesions and terium marinum have both been implicated in sensory loss characterize this disease. The Centers for Disease Control and Preven. have not been successful. grow poorly. 213. which are precipitated with trichloroacetic 209. Tuber- culin preparations are prepared from culture fil- trates. releasing trapped bac- C. C. ethambutol. Kinyoun carbol-fuchsin method uses a higher concentration of phenol in the primary stain to accelerate the staining process.

and mortality in these patients. and it must be differ- close to the organism's optimal growth tempera- entiated from M. should be incubated at 30°C. tuberculosis. Growth of this well-known human pathogen appears in 2 to 3 weeks when 222. haemophilium. behind M. These bacteria cause disseminated infections in patients with acquired immunodeficiency syn- drome and are important causes of morbidity 220. Mycobacterium marinum is the causative these slow growers is 30°C. M. Auromine-rhodamine is a fluorescent stain used to visualize the mycobacteria. Mycobacterium gordonae has been recov. M. kansasii is the most commonly isolated photochromogen in the U. marinum. and 218." These organ. These niacin-positive mycobac- teria form dry heaping colonies that are buff B. M. which is why it has been called a fuschin-based stain (e. B. because the skin temperature is culosis in humans as well. faucets. Lesions generally occur on cattle. isms are not considered to be pathogenic for humans. tuberculosis when recovered ture of 25-32°C. from clinical material. 217. ug/mL of thiophene-2-carboxylic acid hydrazide A. they are slow growing and form yellow-orange colonies B. this stain is more sensitive than water in nature. agent of "swimming pool granuloma. The bacteria 216. and bodies of to see the bacilli. M.S. This agent is an etiologic agent of tuber- the extremities. The the "tap water scotochromogen. Members of Runyon group II. tiated from other mycobacteria by its suscepti- bility to T2H. avium complex. bovis is negative for niacin production and nitrate reduction. Mycobacterium bovis causes tuberculosis in matous skin lesions. Ziehl-Neelsen). avium complex is sometimes (T2H). Growth occurs only at 35°C and is differen- uncommon in immunocompetent individuals. The optimal temperature for C. The colored. mycobacteria associated with these type of infections include M. retain the stain and will appear bright yellow B. The human tubercle bacillus is Mycobac. terium tuberculosis." Typically. Skin cultures for the recovery of Mycobac- terium spp.221.S.resembling classic tuberculosis.. their identification is recom. patients with abraded skin come in contact with water containing this agent and develop granulo- A. kansasii produces chronic lung disease C. . This Mycobacterium is associated with cattle and is rarely isolated from humans in the referred to as Mycobacterium avium-intracellu- lare complex. 219. but because they may be recovered as contaminants. mended. A. It is the second that do not depend on exposure to light. ulcerans. incubated at 35°C.against a black background. ANSWERS & RATIONALES 733 214. Because it is easier ered from water stills. Unlike M. These slowly growing bacilli are U. M. most commonly isolated nontuberculosis Myco- bacterium sp. Mycobacterium bovis is susceptible to 5 215. The M.g.calcofluor white stain is a fluorescent stain used to visualize fungi.


223. under anaerobic conditions stimulate toxin
formation. The patient becomes ill following
D. Lowenstein-Jensen, Lowenstein-Jensen-Gruft,
the ingestion of food that contains nanograms of
and Middlebrook media are commonly used for
preformed toxin.
the isolation of the mycobacteria. Chocolate
agar will support the growth of Mycobacterium
haemophilium; however, chocolate agar is not 227.
routinely used for cultures of mycobacteria. This
B. Zoonotic diseases are diseases of animals that
species requires ferric ammonium citrate or
are transmissible to humans. Leptospirosis is pri-
hemin for growth and also has an optimal tem-
marily a disease of small animals such as rabbits.
perature of 30°C.
It is contracted by humans through contact with
infected carcasses or contaminated water. Bacillus
224. anthracis is found in the environment. Anthrax is
transmitted to humans by exposure to contami-
A. The mycobacteria are only slightly more
nated animal products such as cattle hides, goat
resistant to the decontamination procedures than
hair, or wool. Brucellosis is associated with a vari-
other bacteria. Therefore, it is only appropriate to
ety of animals.
decontaminate specimens for mycobacteria that
are contaminated with normal flora. Because
sputum passes through the oral cavity, it contains 228.
a large amount of normal oral flora. The other
A. Isolates of the anaerobic, spore-forming
specimens listed are typically sterile and lack
bacillus Clostridium perfringens characteristi-
normal flora.
cally produce a pattern of double zone hemoly-
sis on sheep blood agar plates. A Gram stain of
225. such colonies should demonstrate a medium-
sized gram-positive bacillus that does not con-
C. Mycobacterium leprae is the causative agent
tain spores. For further identification the isolate
of Hansen disease (leprosy). This bacterium
should be inoculated on an egg yolk agar plate to
cannot be grown on artifical media and requires
detect lecithinase production.
laboratory animals for cultivation. The optimal
temperature for M. leprae is lower than the core
body temperature of 37°C; therefore, infections 229.
generally occur in the skin in the extremeties.
D. Prevotella melaninogenica was isolated from
The bacteria are likely spread from nasal secre-
this cervical abscess. This anaerobic organism is
tions and not the lesions; they are not highly
part of the indigenous microflora of the respira-
contagious, as most people believe.
tory, gastrointestinal, and genitourinary tracts and
Anaerobic Bacteria is considered a significant human pathogen. The
black pigment appeal's after several days when
226. growing on laked blood agar plates. Prior to
B. Improperly home-canned foods, especially pigmentation, this isolate can be presumptively
low-acid-content vegetables, cause the majority identified by its brick-red fluorescence under
of the cases of food-borne botulism. The ubiq- UV light. Pigmented Porphyromonas spp. are
uitous nature of Clostridium botulinum enables asaccharolytic.
the spores to contaminate a variety of foods.
Contamination and subsequent germination


component of selection media for the
Bacteroides group, it is important to note that,
A. Clostridium tetani is an obligate anaerobe.
in general, gram-negative, non-spore-forming,
Spores are widespread in nature and cause disease
anaerobic bacilli are susceptible to penicillin.
by contaminating puncture wounds. The exo-
The B. fragilis group is an exception in that it is
toxin, tetanospasmin, produced by this organism
known to be resistant to penicillin.
is one of the most powerful bacterial toxins
B. The second most commonly encountered
group of anaerobes in human infections is the
B. The gram-positive, non-spore-forming,
anaerobic, gram-positive cocci. They may
anaerobic bacillus Actinomyces israelii is a
account for one-fourth of all anaerobes isolated in
slowly growing organism that is considered to
clinical laboratories. Estimating their clinical sig-
be an opportunistic pathogen. Colonies may not
nificance, however, is often difficult. Important
be visible before 5 to 7 days or longer. When
isolates include Fingoldia magna (formerly Pept-
colonies are seen, they appear white, opaque,
ostreptococcus magnus) and Peptostreptococcus
lobate, irregular, and shiny and are described as
resembling a molar tooth. A. israelii is part of
the indigenous flora of the human mouth, and a
few Actinomyces spp. have been found to inhabit 235.
the vagina. Pathogenesis generally involves
A. The closed chest abscess described is char-
trauma to tissues of a mucous membrane and the
acteristic of human actinomycosis, which is
introduction of this endogenous organism.
caused by Actinomyces israelii, an anaerobic,
gram-positive, non-spore-forming bacillus. The
232. organism is not acid-fast, which helps to differ-
entiate it from Nocardia spp. Actinomycotic pus
A. The predominant indigenous flora of the
characteristically shows "sulfur granules" or
human intestinal tract is anaerobic, gram-negative,
solid yellow particles made up of masses of the
non-spore-forming bacilli. The Bacteroides frag-
filamentous bacilli seen on the Gram stain in
ilis group, in particular, predominates in the fecal
Color Plate 29 •.
flora. Trauma involving the intestinal area or
bowel surgery predisposes patients to an endoge-
nous anaerobic infection. Although these organ- 236.
isms are present in large numbers, their routine
A. Kanamycin-vancomycin laked blood (KVLB)
identification in fecal cultures is of no diagnostic
agar is selective for the Prevotella and Bacteroides
spp. Presumptive identification of B. fragilis group
can be accomplished utilizing its antimicrobial
233. resistance pattern. Bacteroides spp. are resistant to
vancomycin and kanamycin, unlike Fusobac-
A. Members of the Bacteroides fragilis group,
terium spp., which are resistant to vancomycin but
the most commonly isolated anaerobes and a
predominant part of the indigenous fecal flora susceptible to kanamycin. A KVLB agar plate
should be part of the primary plating media for
in humans, are not inhibited by the presence
anaerobic cultures.
of bile. Bile-esculin agar plates are used for
the selection and presumptive identification of
the B. fragilis group. Although not used as a

k 738 • C

M\?C. C. difficile is an important cause of a hospital-
common contaminant of blood cultures because
of its presence on the skin. Care in the prepara-
tion of the skin before venipuncture helps to
acquired infection commonly called pseudomem-
eliminate confusion caused by the recovery of
branous colitis. Hospitalized patients treated with
this anaerobic isolate.
broad-spectrum antimicrobial agents become
colonized when their normal intestinal flora is
diminished. The most rapid and accurate diagnos- 241.
tic method is detecting toxins A and/or B in stool D. The spore of Clostridium tetani is located
specimens. Cycloserine-cefoxitin-fructose agar terminally and is larger than the sporangium.
(CCFA) is the recommended selective medium Characteristically, when seen on Gram stain, the
for C. difficile, although few laboratories attempt cells of Clostridium tetani resemble a drumstick
isolation. or tennis racket. Spores can be readily seen in
late growth phase cultures incubated at 37°C.
D. Although it has a gram-positive-like cell 242.
wall, Mobiluncus stains gram-variable to gram- C. Fusobacterium nucleatum, a gram-negative,
negative. This curved and motile bacillus seems anaerobic bacillus, is part of the indigenous
to contribute to the pathology of bacterial vagi- microbial flora of the respiratory, gastrointesti-
nosis (BV). A Gram stain of the discharge that nal, and genitourinary tracts. It is frequently
is produced in this condition can be used for the implicated as the causative agent in metastatic
detection of these distinctively curved organ- suppurative infections such as brain abscesses.
isms. The presence of "clue cells," gram-variable These pale-staining bacilli characteristically
pleomorphic bacilli on vaginal epithelial cells, is appear as long, thin bacilli with pointed ends.
diagnositic of BV.

239. C. Propionibacterium spp. are non-spore-form-
B. Infant botulism or "floppy infant" syndrome ing, anaerobic, gram-positive bacilli. Clostridium
is seen in children up to 6 months of age. This spp. typically form spores, although it is diffi-
infectious process begins with the ingestion of cult to induce some species to form spores in
food contaminated with spores of Clostridium vitro. Veillonella is a gram-negative coccus, and
botulinum. Following ingestion, viable spores Fusobacterium is a gram-negative bacillus.
are carried to the lower bowel, where they ger-
minate and elaborate the powerful neurotoxin
that produces the characteristic flaccid paralysis. 244.
C. Clostridium botulinum is the causative agent
of botulism, a disease produced by an exotoxin
240. that acts on the central nervous system. Types A,
D. Propionibacterium acnes is the most fre- B, E, and F are causes of human botulism; types C
quently isolated of all the gram-positive, non- and D and less commonly types A and B are asso-
spore-forming, anaerobic bacilli. It is a part of ciated with disease in animals and birds. Type G
the normal human bacterial flora and predomi- has not been associated with disease in humans or
nates on the surface of the body, but may also be animals. This anaerobic organism produces oval,
recovered from the upper respiratory tract, intes- central, or subterminal, spores that germinate in
tines, and urogenital tract. This organism is a food products or less commonly in wounds.


245. C. septicum, C. novyi, C. sordellii, and C. his-
D. Resazurin is an Eh indicator used in anaerobic
culture media. When the oxygen concentration is
reduced, the resazurin indicator is colorless. A 250.
pink color in the medium indicates aeration and
an unsuitable environment for the preservation of D. Eubacterium, Bifidobacterium, and Propi-
onibacterium are all anaerobic, gram-positive,
obligate anaerobic organisms.
non-spore-forming bacilli. This group of anaero-
bic microorganisms is difficult to identify in the
246. clinical laboratory and often requires the use of
gas chromatography. These organisms are rarely
B. Fusobacterium nucleatum is the most fre-
quent clinical isolate within the genus isolated. Suttonella wadsworthensis is an anaer-
obic, gram-negative bacillus.
Fusobacterium. These anaerobes are part of the
indigenous flora of human mucous membranes,
oral cavity, intestine, and urogenital tract. F. 251.
necrophorum is, however, much more virulent. D. Clostridium tetani is an obligate anaerobe.
Spores are widespread in the soil. When intro-
duced into a puncture wound, the spores require
the reduced oxygen environment produced by
B. Clostridium septicum is isolated in the clini- the necrotic tissue and poor blood supply in the
cal laboratory in cases of serious or often fatal wound. Cleaning and debridement of the wound
infections. Bacteremia is seen in association is important, as is the administration of a tetanus
with an underlying maglignancy. The most com- toxoid booster. Many clostridia require anaero-
mon types of cancer are colon or cecum, breast, bic conditions for spore formation.
and leukemia or lymphoma.

248. B. Most Clostridium spp. are gram-positve, and
A. Botulinal toxin is the most potent exotoxin they generally form spores. Because they are
known. When absorbed, this exotoxin produces obligate anaerobes, they will not grow on sheep
the paralyzing disease botulism. Toxin acts in the blood or chocolate agars incubated aerobically.
body by blocking the release of acetylcholine in Bacillus spp. also form spores, but they are fac-
the neuromuscular junction of the peripheral ner- ultative anaerobes and would therefore grow on
vous system, causing muscle paralysis. media incubated aerobically. Lactobacillus is a
non-spore-forming, gram-positive bacillus, and
Prevotella is a gram-negative bacillus.
B. Clostridium perfringens is the species
most commonly associated with clostridial 253.
myonecrosis or gas gangrene. These soil and C. Lactobacillus spp. are normal flora of the
water saprophytes most frequently gain entrance vagina and digestive tract and are rarely patho-
to the human body through traumatic wounds. genic. They are aerotolerant anaerobes and will
Once they have been introduced into injured tis- produce alpha-hemolysis on sheep blood agar
sue, the characteristic syndrome of myonecrosis plates incubated aerobically. These organisms
due to the elaboration of exotoxins may occur. can also produce a green discoloration on choco-
Other species involved with myonecrosis are late agar.

ered clinically significant. I 738 •CHAPTER CH 6: BACTERIOLOGY H\. Alpha-toxin replaced the use of cell culture monolayers. D. The clinical manifesta. single straight streak of Streptococcus agalac- with the rare exception of glucose. A reverse CAMP test aids in the identifica- motile and produces terminal round spores.. D. This organism is known rarely seen in foods or on laboratory cultures. pected C. A are not inhibited by kanamycin. or lecithinase is produced by all strains of which were examined after incubation for evi- C. perfringens isolates are inoculated at olytic and H2S negative. Its growth is inhibited by a anaerobius is made easier by the use of the 1-ug kanamycin disk and the presence of bile. tion of Clostridium perfringens. perfringens. C. 255. to cause pseudomembranous colitis associated This organism is divided into five types. These tests have generally of myonecrosis and food poisoning. 258. appears on Gram stain as a gram-negative rod A. The identification of Peptostreptococcus with pointed ends. they may lose their curved light enables a more rapid differentiation because appearance and appear as thin rods. nuclea- of the appearance of a brick red fluorescence tum is found in human specimens and is consid- before the pigment is demonstrated. Bio. laked blood with the use of an ultraviolet light C. Clostridium perfringens produces spores C. zone of inhibition of 12-18 mm around the disk is considered sensitive and a presumptive identi- fication of this organism. cytopathic effect). In this test. Fusobacterium nucleatum is a thin gram- source. The The Bacteroides fragilis group and the pig. dence of cytotoxicity (i. The symptoms of Clostridium difficile infec- that are oval and central in location but that are tion are toxin mediated. Type A is responsible for human cases immunoassays. After tions of tetanus are the result of the release of a anaerobic incubation. C. . Fusobacterium nucleatum characteristically 260. Sus- gelatinase and indole positive but is nonprote. 256. F. perfringens will exhibit neurotoxic exotoxin. 257. This important anaerobic pathogen can negative rod with pointed ends and a slightly be differentiated after 5 to 7 days' incubation by curved appearance in fresh isolates. enhanced hemolysis at the intersection where the two species meet. sodium polyethanol sulfonate (SPS) disk. with the use of antimicrobial therapy. 259. As the bac- its black pigmentation. The use of ultraviolet teria are subcultured.e. Clostridium tetani is a strict anaerobe that is B. C. tetani is tiae is made down the center of the plate. C. Cytotox- based on the quantities and types of exotoxins ins can be directly detected in stools by enzyme produced. a chemically it does not utilize carbohydrates. A to E. Prevotella melaninogenica can be rapidly presumptively identified on media containing 261. agalactiae inoculum. right angles to the S. test is performed by growing the organism in mented species Prevotella and Porphyromonas the presence of a disk impregnated with SPS.

the methylene . The capsule is a contributing factor to the lecithin in the medium. Resazurin. Bacteroides. usually in polymicrobial infec. This is the only species in the lactamase production is responsible for their genus. 267. Mobilun- cus is not associated with aspiration pneumonia. Bacteroides fragilis stimulates abscess forma.lecithinase.S. 264. The Bacteroides fragilis group is among the ment that begins olive-green and gradually most antimicrobial-resistant anaerobes. B. Peptostreptococcus niger produces a pig- D. Methylene blue strips are the most commonly used oxidation-reduction (Eh) indicators. to make a presumptive identification of the pres- ence of anaerobic flora in this clinical case. is used in anaerobic transport systems human disease. producing an insoluble pathology produced by this anaerobe. when in an the intestinal tract. The bacterial spores can survive cooking pneumonia are oral anaerobes. 266. When the bacteria are ingested. important causes of food-borne diseases in the B. another Eh I Peptostreptococcus spp. ANSWERS & RATIONALES • 739 262. white. C. ogy of Fusobacterium would enable a physician the color fades to white. when reduced. and upon pigmented Prevotella and Porphyromonas. enable them to produce disease. When anaerobic conditions are achieved. B. These endogenous organisms. An area of precipitate in the agar around the 263. they sporulate in cocci.and anaerobic culture media such as the prere- tions. such as the black. 268. Lecithinase (alpha-toxin) cleaves tion. 269. The common agents in cases of aspiration U. The characteristic Gram stain morphol. Beta. Anaerobes are a major cause of brain abscess. indicating reduction. indicating anaerobiosis. A. Often these are polymicrobic infections mixing anaerobes with aerobic or facultative organisms such as Entero- bactericeae or Staphylococcus aureus. B. (typically found in meats and gravies). Clostridium perfringens is positive for lecithinase. fusobacteria. is the most common anaerobic gram-negative bacillus isolated in the clinical laboratory.becomes black. and cooling they germinate into vegetative cells. are associated with indicator. material. and can be seen on a Gram stain of clinical duced anaerobically sterilized (PRAS) system. It is a weak pathogen sometimes found in resistance to the penicillins. possess virulence factors that coat protein made in excess and released by the bacteria. also resistant to first-generation cephalosporins and aminoglycosides. The enterotoxin is a spore abnormal site. fragilis product. These anaerobes are polymicrobial infections.Resazurin when oxidized is pink. blue indicator will turn from blue (oxidized) to D. Clostridium perfringens is one of the most 265. colonies indicates that the organism produced C. and anaerobic strepto.

They can be tions can also be acquired by ingestion of unpas- cultivated in cell cultures. the vaginal pH will rise toward assays. the latter blindness worldwide.M. pleomorphic organ- atypical pneumonia. The Chlamydia and Chlamydiophila are vive for long periods in the environment. 277. C. Penicillin is Streptococcus pneumoniae. the organism pro. This pro- cess causes the upper eyelid to evert (entropion).740 • CHAPTER 6: BACTERIOLOGY 270. hominis. Mycoplasma pneumoniae causes primary A. bacterial vaginosis. Chlamydia. psittaci (psittacosis) occur after exposure to B.plemented with peptone. trachoma. The organism is an 272. This favors the growth of other bac. Unlike rickettsial diseases.festations of the disease include severe headache. yeast extract. Human infections with Chlamydiophila III. SP4 agar or broth can The bacteria produce acids from the metabolism be used for the growth of M. Trachoma is a primary cause of this organism may be mild or fulminant. C. Species of the genus Mycoplasma are well-known human pathogens that cause a vari- ety of disease processes. these bacteria by inhalation and contact with fomites. 274. Infec- cannot be grown on artificial media. resulting in an acid environ. A true zoono- agent of inclusion conjunctivitis. pneumoniae and of carbohydrates. 273. no rash occurs in Coxiella burnetti infections.of which has a high mortality rate. and isolation requires media sup- bilateral infiltrates. C. such as Mobiluncus. although physical examina. serum. The disease produced by loma venereum. and most infections are diagnosed by serologic bacilli decreases. .tory tract pathogen of humans. and Mycoplasma 275. pneumoniae is an important respira- neutrality. They require causes a zoonosis and is transmitted to humans ATP from their host cell. Lactobacillus spp. duces hypertrophy of the lymphoid follicles on weakness. that can result in humans and is typically spread person to person. C. The pneumonia is atypical isms that lack a cell wall and are best visualized in that it is milder than the pneumonia caused by by darkfield or phase microscopy. are found as normal flora D. ment in the vagina. Chlamydia trachomatis is the causative infected birds and their droppings. As such. teurized milk. which ultimately leads to blindness. but when it is not treated. the inner surface of the upper eyelid. If the population of lacto. Clinical mani- ble. M. including lymphogranu. It obligate intracellular parasites. and mild pulmonary symptoms. pneumoniae is a slow-grower. Chest X-rays of not an effective treatment because of their lack patients with atypical pneumonia may show of a cell wall. M. obligate intracellular parasite that is able to sur- A. Rickettsia. 276. It is found only in teria. Mycoplasma hominis. psittacosis is a disease of birds that may be genital tract infections. Provided that arginine is added for in the gastrointestinal and female genital tract. Mycoplasmas are small. The disease is preventa.contracted by humans. and tion reveals few chest findings.

directly using fluorescent antibody techniques or can be cultured on McCoy cells. Transovarian passage from generation to gen- cell culture and nearly 100% specific for the eration in ticks perpetuates Rickettsia rickettsii detection of Chlamydia trachomatis. A. which has romycin are effective treatments. this organism is able to resist heat and drying for long periods and does not rely C. These species can also tious fomites such as dust from contaminated cattle hides and fluids released during birth are be isolated from asymptomatic sexually active considered the primary modes of infection. 281. nary heart disease. Rodents and small is needed. C. Polymerase chain reaction DNA amplifica- tion has been shown to be more sensitive than D. When grown on culture media.nique calls for flooding a thin smear. ettsiae that cause this form of spotted fever. Clinical material can be examined more dense and is slightly raised. The organism is a human pathogen stain. 282. Unlike 279. Machiavello. malachite green is added for 6-9 seconds before the final washing with tap water. M. adults. 280. Chlamydiophlia pneumoniae is an impor. Mycoplasma hominis. Suitable for several generations outside an animal host. M. trachomatis ried by the arthropod vector. 285. When confirmation of C. The recom- ratory tract disease characterized as atypical mended procedure is the nonspecific Gimenez pneumonia. colonies. hominis. other rickettsiae. Most infections are against a green background. pneumoniae is primarily a respiratory tract pathogen. which colors the organisms a brilliant red spread person to person. The organism been air dried. A. C. can be detected in corneal scrapings of appearance because the central portion of the suspected cases of trachoma and inclusion con- colony has grown into the agar and thus appears junctivitis. or Gimenez. genitalium. pneumoniae. in particular. Chlamydia trachomatis. which has unique characteristics. Direct microscopic examination for Rick- B. After washing with tap water. M. Mycoplasmas are implicated in a variety of human infections. Infec- Ureaplasma urealyticum have been linked to human genital infections. with a solution of carbol-fuchsin has been associated epidemiologically to coro. that results in corneal involvement. Tetracycline and eryth. and on an arthropod vector for transmission. Trachoma is a chronic inflammatory process of the conjunctiva 283. is . Q fever is caused by infection with Coxiella burnetii. most A. a clinically important respiratory tract pathogen. The staining tech- diagnosed serologically. ANSWERS & RATIONALES • 741 278. 284. tissue culture remains the method of mammals are the natural reservoirs for the rick- choice. are said to have a "fried egg" blindness.ettsia organisms is possible using such stains as tant cause of sporadic and epidemic lower respi. a leading cause of notably M. A specimens for detection are cervical secretions blood meal serves to reactivate the rickettsiae car- and urine.Giemsa.for 1-2 minutes.

it is primarily a disease of bated in the dark at 30°C or room temperature for rodents and is sometimes transmitted to humans. a small zone of beta-hemolysis will be seen premature birth or fetal death. which causes lysis of red blood subsp. perinatal death is not an 290. agent Rickettsia prowazekii. Prevalent in added to 5 mL of culture medium. because of the activity of urease. chetes cross the placenta and infect the fetus. Humans who have had the classic form of serum and the Venereal Disease Research Labo- typhus may remain infected with the causative ratory (VDRL) for cerebrospinal fluid. or the pregnancy around the colonies. are asso. After the first week of disease and Control of disease outbreaks is related to rodent lasting for several months. taining guinea pig red blood cells.during the first week of illness by the direct cul- demic typhus. specimen of choice for isolation of the organism. Humans Spirochetes contract the disease through association with infected squirrels. Murine typhus is transmitted to humans by serum. 293. Congenital syphilis occurs when cells. 289. may go to term.. The in the U. when squirrels seeking D. of infection is known as Brill-Zinsser disease and A. Cardiolipin is a tissue lipid produced as a shelter enter dwellings. pneumoniae growing on SP4 a pregnant woman has a septicemia. C. C. The bacterium . are most reliably detected may serve as an interepidemic reservoir for epi. uncommon consequence of infection. After incuba. The symptoms of murine or endemic typhus are Direct microscopic examination is not reliable for similar to those of the classic epidemic form seen detection because of the low numbers of organ- isms normally present in body fluids. The latent form 292. up to 6 weeks.S. The disease is more common 291. The squirrel louse transmits the organ. The most commonly 287. which is incu- the southern U. the infant is most often born with lesions characteris- tic of secondary syphilis. The media of choice are Fletcher semisolid and Stuart liquid medium. Nontrepone- mal tests for syphilis take advantage of antibod- ies made to cardiolipin. the urine becomes the (rat) control and the related rat flea population. both of which are supplemented with rabbit D. D. Following in utero infection. and the spiro- agar can be overlayed with a molten agar con. Flying squirrels. Suspected M. byproduct of treponemal infection. pallidum. Mycoplasma pneumoniae produces hydro. The bacteria ciated with cases of the sylvatic form of typhus are slow growers and form tiny colonies. Glaucomys volans. 288. in the winter months. One or two drops of the patient's blood are fleas infected with Rickettsia typhi. Leptospira spp. in Europe. Syphilis is caused by Treponema pallidum gen peroxide.S. used tests are the rapid plasma reagin (RPR) for A. turing of a blood sample. Infection can affect fetal development and cause tion. 10 B broth is used with genital specimens to isolate Ureaplasma urealyticum. Relapses or recrudes- cence of disease may occur in these persons years or decades after the initial attack. ism among the squirrel population.742 • CHAPTER 6: BACTERIOLOGY 286. requires urea and produces a strong alkaline pH D. broth will typically not appear turbid.

centration methods are used. body that is not directed against T. Silver impregnation 295. cytoplasm. such as the fluorescent binds to cardiolipin-lecithin-coated cholesterol treponemal antibody-absorbed (FTA-ABS) assay. Confirmation with a protein.5 McFarland standard is used when adjusting Clinically the disease peaks in the summer and the turbidity of the suspension of test organism. Lyme disease was first described in 1975 fol. Pathogenic microscopy is recommended to view these deli- species not only have specific vectors but also cate. A to humans by the tick vector Ixodes dammini. malaise. The VDRL test detects an anti- 296. 299. direct observation using darkfield or phase is the vector for Borrelia recurrentis. Relapsing fever is caused by Borrelia. and detection is difficult even when con. indicating a positive test result. The eti. and lymphadenitis. pallidum C. such as cerebrospinal fluid and urine. Epi- demic relapsing fever is found in Ethiopia. extending along the organisms in suspected cases of leptospirosis. headache. coiled cells in body fluids or tissue sections. The B. particles. Borrelia burgdorferi. Infections other than syphilis can cause a pos- itive VDRL result. In this flocculation test. causing the particles to flocculate and is required. Pediculus humanus. The are examined by direct darkfield microscopy and fibrils are attached to the cytoplasmic membrane stained preparations for the presence of these close to the ends of the cell.A. but most do not stain with the Gram stain. The body under the outer membrane. B. antigens. tent with approximately 1. is produced by infected patients. The axial fibrils number of organisms present in clinical samples most closely resemble bacterial flagella and are is low.5 McFarland standard has a turbidity consis- characterized by skin lesions. C. 298. well-defined geographical distributions. Blood and membrane. Spirochetes are gram-negative. . cytoplasmic membrane-peptidogly- other fluids. an antibody-like but it is not highly specific. and Q fever is caused by Coxiella burnetti. Rat-bite of broth or saline. erythema. Connecticut. and axial fibrils. ANSWERS & RATIONALES • 743 294. Reagin specific treponemal test. The human body louse. Antimicrobial Agents and Antimicrobial Susceptibility Testing 297. Standardization of the susceptibility testing lowing an outbreak in Lyme. is transmitted tibility of an organism to antimicrobial agents. C. Sudan. reagin. associated with motility of the organism. #0. The antigen in the VDRL test is cardiolipin. can complex. can be used to visualize them in smears.procedure is essential for determining the suscep- ologic agent. The basic structure of spirochetes is an outer members of the genus Leptospira. A produces an epidemic inflammatory condition #0. fever is caused by Spirillum minus. and parts of South America. 301. Cultural and sero- logic tests are available for the diagnosis of disease produced by these organisms. A.5 X 108organisms/mL myalgia. 300. It is a good screening test for syphilis. The test must be read microscopically. The description given is characteristic of B.

Certain drug when tested independently. Beta-lactamase production by strains of 305. D.resistance. the mechanisms of resis- of bacteria to use p-aminobenzoic acid. 303. influenzae done to evaluate a drug's activity. This test. B. organisms. The drug sul.tic combinations of antimicrobials are used teria for which susceptibility tests are usually primarily in the treatment of tuberculosis. espe- cially those caused by Escherichia coli. Therefore. The testing antimicrobial agent. . testing is obtained by the most effective drug in the com- not usually performed even when these organ. The therapeutic effect of antimicrobial ther- apy is often increased by the use of a combina- tion of drugs. which denotes a positive result due to the production of penicilloic acid. It is recommended that rapid beta-lactamase test- Bacteria unable to produce peptidoglycan for ing be performed on isolates in life-threatening their cell walls are subject to the effects of vary. tors: X factor (hemin) and V factor (NAD). Streptococcus agalac. penicillin act to inhibit cell wall development. The requirement of 99. A combination of antimicrobials is 304.decrease in the number of viable cells from that ety of antimicrobial agents. which 308.tests all rely on this enzyme's ability to act on a ponent of the cell wall protects the bacterium beta-lactamase ring and in turn produce a color from lysis. 307. A tenfold organisms are predictably susceptible to a vari. 306. which is tance are similar. Even though clindamycin and erythromycin D. In vitro growth of H. requires supplemented tional quantitative assessment of the killing effect media to support the growth of these fastidious of a drug on a specific patient isolate.bacillus infections. fisoxazole is a member of this group and is used in the treatment of urinary tract infections. and Neisseria meningitidis. change. 309. C.requested in cases of life-threatening infections. Hemophilias Test Medium is recommended C. The D-zone inhibition. Antimicrobial susceptibility testing is not effects is greater than that derived from either routinely performed on all bacteria. said to be synergistic when the sum of their D. Bac.744 • CHAPTER 6: BACTERIOLOGY 302. ente- not performed include Streptococcus pyogenes rococcal endocarditis. and certain gram-negative (group A Streptococcus).9% killing defines the for use in the disk-agar diffusion susceptibility minimum bactericidal concentration (MBC) of an testing procedure of Haemophilus. must synthesize folic acid for growth. The MBC test is an addi- of Haemophilus spp. by competitive can induce clindamycin resistance. The presence of erythromycin a part of the folic acid molecule. Sulfonamides act to interfere with the ability are in different classes. Synergis- isms are the etiologic agents of infection. is sometimes requires the presence of accessory growth fac. The peptidoglycan com. D.clinical infections such as meningitis. The rapid ing osmotic pressures. Haemophilus influenzae renders them resistant to C. tiae (group B Streptococcus). Inhibitors of peptidoglycan synthesis such as the antibacterial effect of penicillin and ampicillin. These chemotherapeutic agents are test is used to detect the presence of this inducible bacteriostatic and not bactericidal.bination is the definition of synergism.

pneumoniae. ANSWERS & RATIONALES • 745 310. The Kirby-Bauer or disk-agar diffusion sus. ESBLs cleave the antibiotic. ceptibility test requires that the pH of the agar be C. with the exception of spectinomycin. and is the only agent consistently bacte. Most fastidious bacteria do not grow satis- tested at room temperature to ensure an optimal factorily in standard in vitro susceptibility test range of 1.g. sis. ESBLs have only been found in gram-negative bacteria. Metronidazole. Haemophilus influenzae should be tested for treating anaerobic infections has gained empha.4 before use in the procedure. vating it. The test can be per- sis in light of resistance patterns of many of the formed directly. whereas bacteriostatic drugs only arrest . The use of this drug for B. All aminogly- cosides. The extended spectrum beta-lactamases slight batch-to-batch variability. of those listed. is a synthetic compound that acts by inhibit- ing DNA synthesis. Tobramycin. Streptococcus pneumo- trode is acceptable for direct testing. trum cephalosporins such as ceftriaxone and Streptococcus pneumoniae). Low in tetracycline and sulfonamide inhibitors. tested by macerating it in distilled water and testing with a pH meter electrode. an aminoglycoside. or a color change can be observed when the beta-lac- 313. in general. A pH indicator may be used to ricidal against susceptible isolates. is able to cross the blood-brain rely on a color change to detect the presence of barrier.315. For certain species. and other Streptococcus species. inacti- blood may be added. B. antibiotic.. cillin resistance. 5% lysed sheep cefotaxime. 5% lysed horse blood. a drug recommended for the treatment of amebic dysentery and trichomonia. current obtain a reading. Metronidazole reliable for the detection of penicillin and ampi- is consistently active against all gram-negative.316. C. is the only hydrolyzed by the enzyme. modifica- acceptable method is to allow the agar to solidify tions have been made to the standard Clinical around the electrode of a pH meter and then and Laboratory Standards Institute (CLSI) methods. 311.2-1. Neisseria gonorrheae. in both the disk diffusion and tube dilution sus- ceptibility test procedures is Mueller-Hinton.this enzyme. that is bactericidal. detect the penicilloic acid produced when the beta-lactam ring of penicillin is cleaved. B. and the methods are rapid and commonly recovered anaerobes. So far. 314. Another niae. In the case of S. A sample of the Mueller-Hinton medium can be systems that use unsupplemented media. tam ring of a chromogenic cephalosporin is D. anaerobic bacilli. For the suscep- (ESBLs) confer resistance to the extended spec- tibility testing of fastidious organisms (e.beta-lactamase production. a surface elec. this medium has been found to show only B. Bactericidal antibiotics actually destroy the bac- teria. Rapid test methods. CLSI broth dilution test conditions include cation-supplemented Mueller-Hinton broth with 312. such as Haemophilus influenzae. the growth of the microorganism. The recommended plating medium for use are bactericidal in their activity.

are the agar proportion method and the bone marrow toxicity is a rare but usually fatal BACTEC 460TB radiometric method. Clavulanic acid is a beta-lactamase inhibitor. gram-positive anaerobic bacilli and gram-positive D. The second form of U. The rise in antimicrobial-resistant isolates of wide variety of gram-negative bacilli. production of cytotoxic intermediates. Sulbactam and tazobactam are also Guidelines developed for the quality assurance beta-lactamase inhibitors. Tolerance is described as the ability of cer- anaerobic cocci are resistant to this agent. Testing should not be done olism. Klebsiella. Two such methods that have thrombocytopenia occurs as a direct result of the been described and are in common use in the agent on hematopoiesis. Vancomycin. Staphylococcus D. B. The accepted methods for determining the in vitro antimicrobial sus. The main activity of this drug is to inhibit peptidoglycan synthesis. and of a single drug. This mechanism of bacte- rial resistance is attributed to a deficiency of cell 322. Bone marrow toxicity is the major complica- growth of the microorganisms on solid or in liq- tion of chloramphenicol. and Haemophilus the Clinical and Laboratory Standards Institute. Escherichia coli (ATCC synthesis of susceptible bacteria. The mechanism of this response is not known. from stored cultures but rather from freshly grown 18. is produced by 25922). andEnterococcusfecalis (ATCC 29212). A.S. 323. organisms be used for this purpose: Pseudomonas aemginosa (ATCC 27853). Only some of the non-spore-forming. Mycobacteria tuberculosis has been an impor- tant public health crisis.lyophilized to maintain their antimicrobial sus- ceptibility pattern. leukopenia. which acts to inhibit cell wall aureus (ATCC 25923). 321. a nitroimidazole derivative. ceptibility of mycobacteria are based on the B.748 • CHAPTER 6: BACTERIOLOGY 317. an actinomycete. anaerobes. C. 324. These drugs act on the 318. Gentamicin is particularly effective against a C. 319. Reversible bone mar- uid media containing a specified concentration row suppression with anemia. wall autolysins. The growth of these organisms is only inhibited by these cidal drugs. Metronidazole. but it also Cultures of these organisms should be frozen or has an effect on other aspects of bacterial metab. Gentamicin is a member of the aminoglyco- side group of antibiotics. aplastic 24-hour cultures. of the disk-diffusion antimicrobial susceptibility test procedure recommended that the following 320. is active against most of the clinically significant . Standard quality control strains maintained cillin and is effective in treating infections caused by the American Type Culture Collection (ATCC) by beta-lactamase-producing bacteria such as should be tested routinely as recommended by staphylococci. It can be administered with amoxicillin or ticar- A. This tain strains of organisms to resist lethal concen- drug acts to disrupt bacterial DNA through the trations of antimicrobial agents like penicillin. influenzae. Vancomycin is a bactericidal antibiotic. 308 ribosomal subunit to inhibit protein synthe- sis.

Within the clot. Normal flora can offer the host protection optimal temperature ranges. This polyan- The bacteria are grown under standardized con. The most effective anticoag- ulant for use in the microbiology laboratory is sodium polyanetholsulfonate (SPS) in a concen- 328. Fluids known to clot on C. C. The portable Fyrite carbon diox- ide gas analyzer may be used for the daily moni- 327.05%. This yellow compound polyanetholsulfonate (SPS). pathogenic bacteria. Most commercially available blood culture most sensitive test for detecting the production of media contain the anticoagulant sodium beta-lactamase enzymes. toring of capnophilic incubators. 331.vive well within clotted blood. against infections by providing competition to and proper atmospheric conditions for growth. Anticoagulation is will become red if the organism produces the important because certain bacteria do not sur- enzyme that breaks the beta-lactam ring. Candle not contaminated by upper respiratory tract flora. procedures bon dioxide. The chromogenic cephalosporin method is the D. Gas liquid chromatography is used for other bacteria and yeasts. Capnophilic environments may be such as bronchoscopy or percutaneous transtra. jars produce only about a 3% concentration of carbon dioxide. Earlier and later in life. The MIDI system is based on the analysis of standing should be transported to the laboratory fatty acids in the cell wall of microorganisms. and the fatty acids are extracted. cefin. Microbiologic examination of body fluids is childbearing years. B. ionic anticoagulent is also anticomplementary ditions. phagocytosis can occur. Incubation of inoculated bacteriologic cul- 326. Procedures and Biochemical Identification of Bacteria 330. tration of 0. Results are compared to a computerized database. obtained by using incubators equipped with a cheal aspitate are used to obtain a specimen that is tank of carbon dioxide and a regulator.025-0. adequate moisture. The flora of the female genital tract changes with age and the associated effects of pH and estrogen concentration in the mucosa. has a high neutrophils and macrophages remain active and affinity for most bacterial beta-lactamases. When diagnosing nificant isolates is one that contains 5-10% car- lower respiratory track infections. in a sterile tube containing SPS. Nitro. are the predominant flora during B. 329. less effective when bacteria become trapped in staphylococci and corynebacteria predominate. ture media requires that attention be given to C. Mycobacterium fatty acids are analyzed by high- performance liquid chromatography. B. The and antiphagocytic. Lacto. ANSWERS & RATIONALES • 747 325. The lungs are not typically The optimal atmosphere for many clinically sig- colonized with bacterial flora. the commonly used compound. . clotted specimens. bacillus spp.

The yellow color indicates acid produc- bition of most bacteria other than staphylococci. color within 30 seconds. sugar iron agar is not a sensitive indicator. lase broth is inoculated with a test organism. which a positive reaction. are added to the medium. of an organism to use citrate as its sole carbon B. tion. It is used for the isolation and identifi. and a black (lead sulfide) color develops. A. Formation of a green nitrate to nitrogen gas. and nitrites are produced in the medium. The decarboxylase activities of members of the family Enterobacteriaceae are important 335. present. 338. organism first ferments the glucose present. sulfide-producing organisms. as indicated by a change activity will then be able to attack the ammo acid in the phenol red indicator. An organism that possesses decarboxylase Mannitol fermentation. The presence of A. The presence of liberated. the color change will deaminase enzyme is accomplished by adding a not occur. The 7. Detection of the activity of this gone on to nitrogen gas. One of the metabolites in the tricarboxylic acid cycle. Infectious aerosols put laboratory profes. duce infectious aerosols. or decanting may pro. color in the liquid on the agar slant indicates the presence of phenylpyruvic acid. The lowered ation of staphylococcal species because most pH (acid) activates the decarboxylase enzyme. B. the ferential. The addition of zinc dust with no 10% solution of ferric chloride to the growth on resulting color change confirms the reduction of an overnight agar culture. Bacteria frequently are D. The strip is inserted above the medium in isolate on the streak line or both are indicative of the tube and is secured under the closure. centrifugation. After incubation. oc-naphthylamine and sulfanilic acid 334. 337. The assessment of the ability 336. solved sulfide gas reacts with the lead on the strip. citrate can serve as an energy source for some bacteria. A blue color and/or growth of the acetate. When inoculated on a sulfur-containing source aids in the identification of the family medium. Escherichia coli is citrate seals the tube. yellow. aureus isolates ferment mannitol (changing The amines in turn raise the pH. When a decarboxy- C. . aids in the differenti. S. organisms that produce hydrogen sul- Enterobacteriaceae. nitrites is indicated by the production of a red lalanine results in the formation of phenylpyru. coagulase-negative staphylococci are unable to ferment mannitol. The deamination of the amino acid pheny. cation of staphylococcal species. and a color the color of the medium to yellow) and most change from yellow to dark purple results. An organism with this ability reduces nitrates. Mannitol salt agar is highly selective and dif. than in the liquid medium. producing alkaline amines. sionals at risk for acquiring many diseases. because the triple A. If the reduction has vic acid. dis- negative.748 • CHAPTER 6: BACTERIOLOGY 332. The handling of clinical specimens that require pipetting.5% which produces a color change from purple to concentration of sodium chloride results in inhi. Nitrate reduction is a general characteristic present in greater numbers in aerosol droplets of members of the family Enterobacteriaceae. tests for their identification. Klebsiella pneumoniae is fide will demonstrate a partial blackening of a able to use citrate with the production of alkaline strip impregnated with a 5% solution of lead by-products. This test is useful for the detection of weak hydrogen 333.

and test helps differentiate Staphylococcus aureus organisms may be present in low numbers. red color complex if acetoin is oxidized. Blood cultures should be held in the laboratory at least 1 week before being reported as negative.A. nique is recommended for the routine screening D. The prior addition of a-naphthol.B. the formation head space pressure. Bromthymol blue is the numbers of microorganisms in fluid and exudate pH indicator. Glucose metabolism by certain organisms of blood cultures and cerebrospinal fluid sedi- produces acetyl methyl carbinol (acetoin) as the ment smears because of its superior sensitivity as chief end product. tion. Demonstration is further facilitated by an increase in the concentration of 343. Knowledge of the various clinical condi. hydrogen peroxide is bers of the genera Klebsiella. Bacterial metabolism of carbohydrates in the first devised by Hugh and Leifson in an attempt to culture media produces the by-product carbon detect weak acid production from nonfermenta.345. Mem. Oxidative-fermentative (OF) medium was D. which terial metabolism in these systems within only a may neutralize the weak acids produced by the few hours of inoculation. which acts as a catalyst. Haf. Manometric systems measure the (0. compared to the Gram stain. Pyrrolidonyl-a-naphthylamide (PYR) is the tions that produce bacteremia is essential for substrate. The application of this fluorescent dye enables the microbiologist to screen samples at low-power microscopic magnification.dioxide. 342. is reduced. Between 10 and 20 mL of blood is recommended. carbohydrate (1. . In the calalase test. possible. The acridine orange stain is used to detect low semisolid consistency. cultures should be drawn before antimicrobial therapy is initiated and optimally before a fever spike. It is possible to detect bac- of oxidative products from amino acids. test. which oxi- dizes acetoin to diacetyl.reduced to water and oxygen. which is captured as head gas in sealed tive bacilli. B. Not all enzyme L-pyrrolidonyl arylamidase. produces a 344. The catalase test is used to differentiate the staphylococci (positive) from the streptococci (negative). If (positive) from S. 40% potassium hydroxide is added. Blood cultures are one of the most important specimen types sent for bacteriologic examina. and the metabolic reaction can be demonstrated. organism. There is a period of 1-2 hours from the time of the release of bacteria into the bloodstream and the subsequent physio- logic chill response. Enterobacter. lugdunensis (negative). ANSWERS & RATIONALES • 74S 339. The formation of nia. The test detects the presence of the optimal recovery of the causative agent.2%) used in conventional media. and Serratia produce a positive reaction in oxygen produces the bubbles seen in a positive this test. D. The PYR conditions produce continuous bacteremia. 341.0%) in the medium. To demonstrate this reaction. along with a A. This tech- 340. By decreasing the amount of peptone culture vials. samples.

C. 347. E. hematopoietic disease. C. The anatomy of the female urethra allows causing soft tissue infections in a wide variety of bacteria from the perirectal region to reach the farm animals. Disease is transmitted by selective medium for the isolation of Neisseria respiratory droplets among people in prolonged gonorrhoeae in addition to nonselective media. but most cases occur in a rural envi- pathogen in uncomplicated community-acquired ronment and include a history of abrasion or urinary tract infections.352. Neisseria meningitidis is a leading cause of from these patients should be inoculated to a bacterial meningitis. and the reticuloen. however. Chemo- Thayer-Martin agar has a chocolate agar base prophylaxis with rifampin is appropriate for those formulated to support the growth of fastidious in close contact with the patient: household mem- species of Neisseria while suppressing the bers. Vibrio vulnificus is responsible for sep- ticemia after consumption of contaminated raw oysters.D. The presence of organ. Only about 50% of patients with gonococcal arthritis will 351. Mortality in patients . Arcanobacterium pyogenes has been reclas- sified several times. A. growth of normal or indigenous flora by the addition of antimicrobial agents. 349. Other organisms are more undetected wounds with animal exposure.750 CHAPTER 6: BACTERIOLOGY Case Studies with septicemia can be as high as 50% unless antimicrobial therapy is started rapidly. Listeria monocytogenes is also a gram-postive bacillus that is CAMP positive. or chronic renal failure and those receiving immunosuppressive drugs. prevalent in nosocomial or recurrent infections. especially in the 350. major joints of the body. such as in daycare centers. A. daycare staff. The bacteremic phase is seen of tampons. is commonly associated with inva. Staphylococcus aureus has been isolated isms is the result of an extravascular site of from a majority of the reported cases of the clin- infection. Disseminated gonococcal infection pro- duces symptoms of arthritis. have positive synovial fluid cultures. 348. Infections are most severe in patients with hepatic disease. coli is the most common is unknown. C. Symptoms are associated with the before the organism can be recovered in stool. Salmonella Typhi. Mode of transmission to humans bladder easily. close contact.disease was linked to the use of a specific brand dothelial system. The extravascular sites in the case of ical syndrome described—toxic shock syn- typhoid fever are the small intestine. production of a pyrogenic exotoxin (toxic shock syndrome toxin-1. TSST-1) by the coagulase- positive Staphylococcus aureus. sion of the bloodstream. pyogenes is a well-known animal pathogen A. First reported in the late 1970s. and classmates. Samples of joint fluid D. the causative agent of typhoid fever. it is catalase positive. the lymph nodes of the intestine. 346. the regional drome. It was formerly a member of the genera Corynebacterium and Actinomyces.

usually pro- causing Lyme disease have not been demon. The etiologic agent in this case is Yersinia the organism is able to reproduce in red blood cells enterocolitica. A key finding for Y. The organism is by more serious complications. Most infections resolve in about a week. myocarditis. Escherichia coli 0157:H7 is associated with often used for serologic confirmation. infection characteristically follows trauma to the body in or around water. frequently mimics the symptoms of appendicitis. and humans become infected by eating another name for leptospirosis. animals like rats and mice or from water con- taminated with urine of these animals. Listeria monocytogenes has been associated U. predominantly in the northeast and mid-Atlantic B. These strains pro- duce verotoxin and are associated with out- breaks of diarrheal disease following ingestion 355. with human disease following the ingestion of toms of this disease may be followed months later unpasteurized daily products. and jaundice. and mesenteric lymphadenitis. go on for much longer and can cause fatal kid. Disease caused by this organism without causing lysis or a color change. The Gram stain is not helpful in making the diagnosis because of the poor staining quality of this 356. Human infections caused by Leptospim rants and contact with calves at petting zoos. pneumophila could pro- infections. Infections result products made from their meat or contaminated from contact with the urine or tissue of infected with their excretions. The spirochetes food. Contamination of f J stored blood units is not visually detected because D. refrigerator temperatures (4°C). although it has been implicated in a variety of clinical illnesses such as bacteremia. unless in large on the skin or the extremities of humans. The clinical presentation suggests the etio- ney and liver damage. characteristically produce the clinical symptoms Cattle infected with this strain serve as the reser- of fever. An indirect immunofluorescence test and an ELISA test are available for the detection of specific antibody in 359. enterocolit- A. Human is a positive urease. Yersinia enterocolitica causes a variety of fluorescent antibody to L. in uncompromised patients. and arthritis of the and is commonly found in low numbers in animal large joints. The initial symp. logic agent is Legionella pneumophila. enterocolitica ism found in salt or brackish water. Examination of the sputum using D. The western blot assay is A. Listeriosis associated with contaminated disease is Borrelia burgdorferi.C. Lyme disease is an inflammatory disease seen 358. This organism is able to grow at vide a rapid positive identification. hemolytic uremic syndrome. nonspecific febrile illness. A. cholecysti. This numbers or pure culture.S. the patient's serum. may be overlooked in species of Mycobacterium is a free-living organ- the laboratory. but they can 360. ANSWERS & RATIONALES 751 A I 353. microorganism. . 357. Y. strated in peripheral blood smears. anemia. of undercooked hamburger at fast-food restau- D. Weil disease is voir. during the summer months. 354. duces a self-limiting. The etiologic agent of this tick-borne products. capable of replicating at refrigerator temperatures goencephalitis. Mycobacterium marinum produces lesions ica grows slowly at 35°C and. such as menin. tis.

Rosenthal. Philadelphia: Elsevier Mosby. S.. R. Lehman. (2007).I 752 • CHAPTER 6: BACTERIOLOGY REFERENCES Forbes. R. (2005). Manual of Clinical Microbiology. Baron. 12th ed. E. A. C. M. and Pfaller. M. Washington. K. DC: American Society for Microbiology Press. Murray. J.. L. Landry. 5th ed. Louis: Saunders Elsevier.. St. Philadelphia: Mosby. P.. G.. Marion. D. C. Medical Microbiology. Jorgensen. A. D. Bailey and Scott's Diagnostic Microbiology. H. P... and Pfaller. J. (2008). R. 3rd ed. Textbook of Diagnostic Microbiology. and Manuselis. and Weissfeld. B.. A. E. (2007). Murray. . M. S. Sahm. A. 9th ed.