MT 111

CLINICAL BACTERIOLOGY 1 - LECTURE

MISCELLANEOUS GRAM-NEGATIVE BACILLI

Capnocytophaga - The ability to multiply over the temperature
range of 20° to 43° C and survive for varying
● Normal microbiota of the oral cavity of humans periods at 40° to 60° C
● Causes septicemia in patients with neutropenia - capacity to adhere to pipes, rubber, plastics,
● Common sites of clinical isolation and sediment and persist in piped water
- Blood cultures from patients who have systems, even when flushed
neutropenia with oral ulcers (source of the ● Infections:
Capnocytophaga) 1. Legionnaire’s Disease
- Soft tissue infections - febrile disease with pneumonia
- Peritonitis - Pneumonia- predominant
- endocarditis manifestation
1. Capnocytophaga orchracea- most common clinical - Community acquired bacterial
isolate pneumonia
2. Capnocytophaga canimorsus and Capnocytophaga - Incubation period: 2-10 days
cynodegmi- normal inhabitants of the oral cavity of - nonproductive cough, fever,
dogs and cats headache, and myalgia
- Infection from dog or cat bites - sputum may be bloody or purulent
● fastidous , facultative anaerobe - may lead to extrapulmonary
● Thin and often fusiform (pointed ends) resembling infection
Fusobacterium spp. 2. Pontiac Fever
● No flagella but can produce gliding motility on solid - influenza-like febrile disease
surfaces - nonpneumonic form of legionellosis
● Ferment sucrose, glucose, maltose, and lactose - incubation period of 2 days
● (-) indole - inhalation of bacterial toxins or an
acute allergic reaction to the
bacteria
Legionella spp. - previously healthy individuals who
complain of flu like symptoms of
fever,headache, and myalgia that
● non–spore-forming, faintly staining, thin, last 2 to 5 days and then subside
gram-negative bacilli without medical intervention
● first recognized to cause human disease during an 3. Asymptomatic infection
epidemic of pneumonia that occurred among ● Laboratory Diagnosis
members of the Pennsylvania American Legion who ➔ Legionnaire’s disease: combination of culture
had gathered in Philadelphia to celebrate the 1976 and urine antigen detection
bicentennial ➔ Pontiac fever: serology
● Caused by Legionella pneumophila ➔ Specimen for culture and direct examination:
● Found in the environment especially water - sputum, bronchoalveolar lavage,
● Tolerate chlorine concentration of 3 mg/L and bronchial washings
● Transmission: - Other tissues or fluids, such as
- Exposure to contaminated water (faucets, pleural fluid, are generally
shower heads, public fountains, aircon acceptable when suspicion is high
water) - Freeze specimens at −70° C if
- Hot water systems processing will be delayed for
- Cooling towers several days
- Evaporation condensers are major reservoirs ➔ Microscopic examination:
● Virulence Factors - pleomorphic, weakly staining,
- Ability to exist as intracellular pathogens gram-negative bacilli found outside
(amebae and mammalian cells) of and within macrophages and
- Survive inside phagosomes, prevent the segmented neutrophils
formation of phagolysosome - other stains that can be used:
Diff-Quik and Giemsa

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➔ Isolation methods:
- fastidious, aerobic bacteria that do
not grow on SBA and require
L-cysteine for growth
- Acid treatment of specimen before
inoculation
- aliquot of the specimen is first
diluted 1:10 with 0.2 N potassium
chloride–hydrochloric acid
- allowed to stand for no more than
4 minutes
- medium is then inoculated with a
portion of the acid-treated
specimen Bordetella
- specimens from normally sterile
sites should be diluted 1:10 in
● Bordetella pertussis and Bordetella parapertussis
tryptic soy broth or distilled water
● Causes whooping cough or pertussis
- BCYE (buffered charcoal yeast
● Transmission:
extract) Agar with L-cysteine: best
- occurs person to person through inhalation
for Legionella isolation
of respiratory droplets
● colonies appear as grayish
- highly contagious, acute infection of the
white or blue-green,
upper respiratory tract
convex, and glistening,
- caused by Bordetella pertussis
measuring approximately
1. Catarrhal stage
2 to 4 mm in diameter
● symptoms are the same as for a mild cold
● central portion of young
with a runny nose and mild cough
colonies has a
● lasts several weeks
“ground-glass”
2. Paroxysmal stage
appearance, light gray and
● severe and violent coughing
granular
● 15 to 25 paroxysmal coughing episodes can
● the periphery of the colony
occur in 24 hours
has pink or light blue or
● Vomiting and “whooping” (the result of air
bottle green bands with a
rapidly inspired into the lungs past the
furrowed appearance
swollen glottis)
- Selective BCYE agar contains
● Lasts 1-4 weeks
polymyxin B, anisomysin, and either
3. Convalescent Phase
vancomycin or cefamandole (inhibit
● Begins within 4 weeks of onset with a
the growth of other bacteria)
decrease in frequency and severity of the
- Incubate at 35° C in air; increased
coughing spells
CO2 can enhance the growth of
● Cause mild illness and asymptomatic infection,
some of the more fastidious species
primarily in household contacts and in a number of
unvaccinated and previously vaccinated children
● Vaccine is available

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● Laboratory Diagnosis:
➔ Specimen: Nasopharyngeal aspirates or a
nasopharyngeal swab (calcium-alginate or
Dacron on a wire handle)
➔ Specimen obtained from the throat, sputum,
➔ Transport time is critical
➔ Transport media that can be used:
1. Half-strength Regan-Lowe agar:
enhances recovery when used as a
transport and enrichment medium
2. Cold casein hydrolysate medium
3. Casamino acid broth
➔ Culture media:
1. Bordet-Gengou: Potato infusion agar
with glycerol and sheep blood with
methicillin or cephalexin (short
shelf-life)
2. Modified Jones-Kendrick charcoal:
Charcoal agar with yeast extract, starch,
and 40 μg cephalexin (2- to 3-month
● Erythromycin: drug of choice for pertussis
shelf-life but inferior to Regan-Lowe
agar)
3. Regan-Lowe: best for the recovery of B.
pertussis from nasopharyngeal swabs Pasteurella
- Charcoal agar with 10% horse
blood and cephalexin (4- to 8-week
● Zoonosis- a disease that humans acquire from
shelf-life)
exposure to infected animals
4. Stainer-Scholte: Synthetic agar lacking
● Normal flora of oral cavity in birds and mammals
blood products
● Causes septicemia, arthritis, endocarditis,
➔ Plates are incubated at 35°C in a humidified
osteomyelitis, meningitis
atmosphere without elevated carbon dioxide
● Soft tissue (cutaneous) infection from animal bites
for up to 12 days
1. Pasteurella multocida
➔ Young colonies of B. pertussis and B.
parapertussis:” mercury drops”
➔ colonies become whitish gray with age

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 - fresh cheese for several months
● a cause of devastating economic loss among
domestic livestock
● considered category B select biological agents by the
CDC
● occurs worldwide, especially in Mediterranean and
Persian Gulf countries, India, and parts of Mexico and
Central and South America
● Human can be infected by:
1. Ingestion of infected unpasteurized animal
milk products (most common means of
transmission)
2. Inhalation of infected aerosolized particles
(laboratory-acquired infection is the most
important source of transmission)
3. Direct contact with infected animal parts
through ruptures of skin and mucous
membranes
4. Accidental inoculation of mucous
membranes by aerosolization
● systemic, deep-seated disease resulting in various
2. Pasteurella canis- dogs long-term sequelae

● gram-negative, nonmotile, facultative anaerobic

coccobacilli
● bipolar staining: “safety pin” appearance
● Laboratory Diagnosis:
➔ grow on SBA and CHOC agar
➔ grayish colonies
➔ nonhemolytic colonies on SBA that may
appear mucoid after 24 hours of incubation
at 37° C
➔ production of a narrow green-to-brown halo
around the colony after 48 hours
● Laboratory Diagnosis:
➔ Specimen: blood, blood marrow,
cerebrospinal fluid (CSF), pleural and
synovial fluids, urine, abscesses, or other
tissues
➔ Culture media
1. Grow on Blood agar and Choc agar
2. Brucella agar or infusion base:
recommended for specimen types other
than blood
3. addition of 5% heated horse or rabbit
serum enhances growth on all media
4. incubated in 5% to 10% CO2 in a
humidified atmosphere
Brucella 5. colonies appear small, convex, smooth,
translucent, nonhemolytic, and slightly
yellow and opalescent after at least 48
● Brucellosis is a zoonotic disease
hours of incubation
● small, facultative, intracellular, nonmotile, aerobic,
gram-negative coccobacilli or short rods
● capable of survival for extended periods
- soil- 10 weeks
- aborted fetuses- 11 weeks
- bovine stool- 17 weeks
- milk and ice cream- 3 weeks

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➔ Gram stain: small coccobacilli that resemble fine
grains of sand
➔ (+) oxidase, catalase, urease

● Brucella spp. is differentiated by the rapidity with

● Laboratory Diagnosis:
which an organism hydrolyzes urea, its relative ability
➔ F. tularensis is a biosafety level 2 pathogen
to produce Hydrogen sulfide, its requirements for
➔ Organism is designated Biosafety Level 3
Carbon dioxide, and its susceptibility to the aniline
when the laboratorian is working with
dyes: Thionine and Fuchsin
cultures
➔ Performed in a Biosafety Cabinet wearing
PPEs
Francisella ➔ Specimen:
- Scrapings from infected ulcers,
lymph node biopsies, and sputum
● are facultative, intracellular pathogens that require
- Whole blood is acceptable but
cysteine, cystine, or another sulfhydryl and a source
false-negative results may occur
of iron for enhanced growth
during early stages of disease
● (-) oxidase
- Serum: early stage of the disease,
● (-) urease
convalescence
● (+) catalase
➔ Amie’s transport medium containing
● Nonmotile, non-spore-forming, strict aerobes
charcoal- swab specimen
● Infection is also called rabbit fever, deerfly fever,
➔ Cultavation:
lemming fever, and water rat trappers’ disease
- Isolation is difficult
● Francisella tularensis
- enhanced by enriched media
- Causes human and animal tularemia
containing sulfhydryl compounds
- wild rodents, rabbits, beavers, and muskrats
(cysteine, cystine, thiosulfate or
in North America
IsoVitaleX) for primary isolation
- Humans become infected by handling the
- glucose cystine agar
carcasses or skin of infected animals
- cystine-heart agar
- inhaling infective aerosols or ingesting
- chocolate agar supplemented with
contaminated water
IsoVitaleX
- through insect vectors (primarily deerflies
- nonselective buffered
and ticks in the United States)
charcoal-yeast extract agar (BCYE)
- being bitten by carnivores that have
- modified Mueller- Hinton broth and
themselves eaten infected animals
tryptic soy broth supplemented
- tularemia is one of the most common
with 1% to 2% IsoVitaleX
laboratory-acquired infections

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 ● rodent bites, ingestion of
contaminated food, or traumatic
injury
- Local lymphangitis and lymphadenitis,
migratory polyarthritis
- fever, chills, malaise, and, later, a general
morbilliform maculopapular or petechial
rash
● Laboratory Diagnosis:
➔ Specimen: blood, joint fluid, and abscess
material
➔ Incubated at 35° C in a humid environment
containing an atmosphere of 5% to 10%
CO2

Gardnerella

● Gardnerella vaginalis
● Thin, gram-variable rod or coccobacillus
● part of the anorectal flora of healthy adults of both
sexes, as well as of children
● part of the endogenous vaginal flora of women of
reproductive age
● Causes bacterial vaginosis
● has been called Haemophilus vaginalis and
Corynebacterium vaginale
● Nonmotile
● (-) catalase
● Growth is best observed after 48 hours of incubation
in a 5% CO2-enriched atmosphere
● Colonies are small and exhibit β-hemolysis on media
containing rabbit or human blood
● Laboratory Diagnosis:
➔ direct examination of vaginal secretions for
the presence of clue cells, small
gram-negative rods and coccobacilli, the
absence of lactobacilli
➔ Clue cells: epithelial cells covered with
bacteria on the cell margins
➔ pH greater than 4.5
➔ fishy amine odor after addition of 10%
potassium hydroxide (KOH) to the secretions

Streptobacillus

● facultatively anaerobic, fermentative,

nonencapsulated, and nonmotile pleomorphic
gram-negative rod
● string-of-beads appearance
● Streptobacillus moniliformis
- rat-bite fever or Haverhill disease
- indigenous flora in the upper respiratory
tract of wild and laboratory rodents
- infection in human is caused by: