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CHAPTER 18 Haemophilus and Other Fastidious Gram Bacilli
CHAPTER 18 Haemophilus and Other Fastidious Gram Bacilli
CHAPTER 18 Haemophilus and Other Fastidious Gram Bacilli
Kingella
Coccobacillary to short bacilli with
squared ends
Occurs in pairs/short chains
Resist decolorization in gram stain
Nonmotile
Nutritionally fastidious
Oxidase (+)
Other Gram (-) Bacilli
Catalase (-)
1.)Capnocytophaga
Fermenters of glucose and other
Req. CO2 for enhanced growth
sugars (no gas)
Fastidious
Facultative anaerobe
Gram(-) bacilli
Thin & fusiform (pointed ends),
spindle-shaped, coccoid & curved
filaments 3.)Brucella
Flagella is absent Small gram(-), aerobic, nonmotile,
Haze (Gliding motility) on solid unencapsulated bacteria w/c do not
surface form spores & may appear
Opaque, shiny; pale beige or yellow- coccobacilli or bacilli
orange Smooth, raised & translucent colony
-hemolytic except C. haemolytica Facultative intracellular pathogens
ferment sucrose, lactose, glucose & Oxidase(+), Catalase(+), Urease(+)
maltose Require CO2
indole(-) Using lead acetate=H2S(+)
reduce nitrate & hydrolyze esculin Biosafety level 3
C. ochracea most clinical isolate Category B select biological agents
Brucellosis/Undulant Fever
2.)Pasteurella Acquired through aerosol,
Gram(-) percutaneous & oral routes of
Cat scratch exposure
Animal or feline bite/animal licking Transmitted by unpasteurized
Nonmotile milk products
Facultative anaerobe coccobacilli 3 stages:
appearing ovoid, filamentous or Acute
bacilli Subchronic
Bipolar staining (safety pin chronic
appearance)
Catalase (+); Oxidase (+)
Weak glucose fermentation
Will grow on SBA & CHOC agar w/c
will producing grayish colonies.
P. multocida:
Most frequent isolate
-hemolytic (SBA), mucoid,
narrow green-brown halo
around the colony after 24-
48hours of incubation at 370C
P. bettyae
solated from placenta,
amniotic fluid, blood, rectal
sites, abscess & urogenital
specimens
fastidious, capnophilic
coccobacilli & bacilli w/c are
facultative anaerobe
ferment glucose & fructose
catalase(+), oxidase(V), indole
(+) nonmotile
Growth in
Serum
Natural Dyes H2S (lead
Agglutina Urease CO2
Host Thioni Fuch acetate)
tion
ne sin
B. Goat or
+ V - - - -
melitensis Sheep
Cattle or +<2hou
B. abortus + + - + +/-
Cow rs
Swine or +<0.5h
B. suis + - + + -
Pig our
+<0.5h
B. canis Dog - - - - -
our
Legionella
Legionella
4.)Francisella
ubiquitous gram (-) bacilli
Small, nonmotile, non-sporeforming,
gram(-) bacilli or coccoid bacteria & acquired through inhalation
strictly aerobic multiply in free-living amoebas &
Facultative intracellular pathogen coexist w/ them in biofilms
Fastidious & require supplementation Found on bodies of water, also in air-
w/ cysteine, cystine or thiosulfate for con
growth
3.1 Virulence factors
MAC/EMB (-)
- ability to enter, surive,and multiply
Gray-white, raised colonies w/
within hosts cells esp.
smooth appearance bronchoalveolar macrophages
Negative: oxidase, urease, satellite - ability to produce proteolytic enzymes
or X & V - survive in an extracellular
Weakly positive for catalase & - environment
lactamase activity
Virulence Factor: Endotoxin 3.2. Infections
Causative agent: rabbit 1. Legionnaires disease
Category A; biosafety level 3 febrile disease w/ pneumonia
Tularemia flu-like symptoms
Rabbit fever usually produces lobar,
Deerfly fever segmental or patchy
Lemming fever pulmonary infiltration
Water rat trappers disease incubation of 2-10 days
Focal ulcer (ulceroglandular) 3 major patterns:
at site of entry of organism & Sporadic cases
enlargement or regional lymph most common and
nodes usually occur in
community
2) Epidemic outbreaks
short duration and low
attack rates
3) Nosocomial clusters
occurring in
compromised patient
population
Serogroup 1 = L.
Pneumophila
2. Pontiac fever
febrile disease w/o
pulmonary involvement
doesnt develop & infection Specimens: sputum,
does not spread beyond bronchoalveolar lavage, and
lungs bronchial washings.
incubation of 2 days Respiratory secretions (acid
caused by L. pnemuophila treatment) and body fluids (lysis
centrifugation) are submitted in
3. Pneumonia sterile, leak proof containers.
predominant manifestation of Saline and buffer should not be used
legionellosis. because of the inhibitory effects of
a. S. pneumonia most common sodium.
cause of bacterial pneumonia. When transport of specimens is
prolonged, specimens should be
4. Atypical Pneumonia refrigerated if more than 2 hours
disease caused by Mycoplasma pass between collection and
pneumoniae, Chlamydophila processing.
pneumoniae and Legionella spp If processing will be delayed for
several days, 700C
For blood cultures, the Isolator
3.4 Epidemiology system w/c uses the lysis
naturally occurring in aquatic sources centrifugation method is preferred.
(lakes,rivers,hot springs, and mud) Urine is an impt specimen to be
can tolerate chlorine conc. Up to collected for antigen detection.
3mg/L Specimens are collected in sterile,
resist water treatment leak proof containers and assayed
Hot water systems, cooling within 24 hours of collection
towers and evaporative If testing is delayed, specimens should
condensers major reservoirs be stored at 2 degrees to 8 degrees or
frozen at 200 C
Factors that contribute to 4.2 Microscopic Examination
colonizing ability to these Legionella spp. are pleomorphic,
sources: weakly staining, gram(-) bacilli
ability to multiply over the tempt L. micdadei is weakly acid fast in
range of 20 degrees to 43 tissue and stains best with the
degrees and survive for varying modified Kinyoun procedure.
periods at 40 degrees to 60 Giemsa, Diertle
degrees
capacity to adhere to pipes, 4.3 Isolation and Identificaiton
rubber, plastics, and sediment 4.3.1 Isolation Methods
and persist in piped water i. Acid treatment of specimens
systems even when flushed Aliquot of specimen is first
ability to survive and multiply diluted 1:10 w/ 0.2N KCL
w/in free living protozoa and in HCL and allowed to stand
the presence of commensal for 5 mins.
bacteria and algae ii. Inoculated medium is
incubated at 35 to 37 degrees
LABORATORY DIAGNOSIS in air for at least 7 days.(3 to
1. Combination of Urine Antigen 5 days), Legionella spp colonies
Detection and Culture best are visible
approached in diagnosis of
Legionnaires disease iii. Legionella spp are fastidious,
2. Serology diagnosis of Pontiac fever aerobic bacteria that will not
grow on sheep blood agar and
4.1 Specimen Collection and Handling require L cysteine for growth
iv. buffered charcoal yeast avian commensal
extract agar w/ L cysteine is B. holmesii and B. Trematum
best for Legionella isolation respective agents of immune
charcoal: acts as compromised bacteremia and
detoxifying agent wound or ear infection
B. avium, B. bronchisepica, B.
Henzii
oxidase positive
BORDTELLA
Bordetella pertusis and Bordetella
parapertussis are primary human
pathogen of respiratory tract
causing whooping cough or
pertussis.
Other Species
B. bronchiseptica
non fastidious, RT pathogens of
wild, domestic birds, and
mammals. (Opportunistic
human pathogen causing RT
and wound infection.)
B. avium Virulence factors
non fastidious, RT pathogens of Filamentous hemaglutinin (FHA)
wild, domestic birds, and and peractin (69 kDa ouermembrane
mammals proein)
B. hinzii
o Provides adherence to ciliated begins within 4 weeks of
epithelial cells onset with decrease of
frequency and severity of
Pertussis toxin (PT)
coughing spells.
o A Protein exotoxin
Colony Morphology
Charcoal-horse blood & Regan-
Lowe media
young colonies are smooth,
glistening, and silver (like
mercury droplets)
Colonies turn whistish gray as
they age.
Bordet Gengou agar
colonies of B. pertussis & B.
parapertussis are hemolytic
Identification Methods
On Gram stain:
appear as tiny gram negative
coccobacilli
maybe elongated if recovered from
media containing cephalexin
Necessary to increase the safranin
counterstaining time to 2 minutes
to see typical morphology.
Trimethroprim sulfamethoxazole
Zoonotic Diseases
Infected by tick
bite, direct contact
w/ infected animal
Tularemia (Rabbit fever, tissue, inhalation
Gram (-) coccobacilli
Francisella Lemming Fever,Water of aerosols or
Pinpoint colonies
tularensis rat trappers disease, ingestion of
(mercury droplet)
Deerfly fever) contaminated food
or water
High risk to lab.
Personnel (BSL-3)
Most common organism
Pasteurell
Gram (-) bacilli in human wounds
a Mousy odor
Bipolar staining inflicted by Dog & Cat
multocida bites
Characterized by
Streptobac Long filamentous gram(-) recovering fever,
bacilli w/ swellings arthritis-like
illus
(pleomorphic); Rat bite fever (Haverhill symptoms
monolifor Puffball or string of pearls Fever) (inflammation
ms or colonies on Thioglycolate pain, stiffness) &
Spirillum broth infection of the
lymphatic vessel