Pasteurellaceae:

Haemophilus, Pasteurella
Fastidious Gram-Negative Rods
 Pasteurellaceae: Genera
• Small, gram(-) bacilli; pleomorphic (cocci,
filamentous rod)
• Medically important genera:
– Haemophilus - human oropharynx NF; infection
of unimmunized children
– Pasteurella - animal oropharynx NF; bite wound
infection, RTI, bacteremia, meningitis
– Aggregatibacter (former Actinobacillus) -
human oral NF; periodontitis, endocarditis, bite
wound
Haemophilus: Characteristics
• Aerobic, facultative anaerobe
• Optimal growth 35-370 C
• Atmosphere of 5-10% CO2
• Catalase(+)
• Oxidase(+)
• Nonmotile
• Most fastidious, require enriched media
• Susceptible to drying and chilling; autolytic
• Clinical specimens should not be
refrigerated, processed immediately
Haemophilus: Characteristics
• Obligate parasites
• Mucous membranes of human, animals
• “blood-lover”
• Most require growth factors
– X (hemin) factor - synthesis of iron containing
respiratory enzymes (cytochrome, cytochrome
oxidase, catalase, peroxidase)
– V (NAD) factor - coenzyme for oxidation-
reduction
 Haemophilus: CBA
• Grow 1-4 days
• Some β-hemolytic
• Most grow poorly, or not
at all on CBA
• Satellitism - grow as
small colonies on CBA
around Staphylococcus
aureus (hemolysis
secrete NAD into
medium, hemin diffuses
from RBC)
Haemophilus: Chocolate Agar
• Grow best on highly enriched CHOC
• Heat RBC 80°C, 15 min. - destroys
NADase, releases NAD into medium
• Can make selective - add bacitracin,
vancomycin, clindamycin to eliminate NF
• Colonies grayish:
– Capsulated strains = smooth
– Non-encapsulated strains = rough
ID Haemophilus: X & V Factor
Requirement
• Only V (NAD) = H. parainfluenzae
• Both X (Hemin) and V (NAD) = H.
influenzae
Haemophilus influenzae: Lab ID
• Six serotypes (a-f)
– by capsular polysaccharide
– Type b most virulent (>95% of all invasive
infections)
– Hib ~20,000 pediatric infections/year prior to
vaccine
– Capsule contains polyribitol phosphate (PRP);
used for vaccine, elicits host protective
antibody response
• Eight biotypes (I-VIII)
– Encapsulated type I – pathogens, more invasive
disease
– Nonencapsulated type II, III - opportunists
Haemophilus influenzae:
Virulence Factors
• PRP capsule – protects against
phagocytosis
• LPS – endotoxin; induces inflammation
• Protease – IgA specific; facilitates
colonization of mucosal surfaces
• Pili and outer membrane proteins –
adhesion, attachment
 Nonencapsulated
Haemophilus influenzae
• Colonize URT within first few months of
newborn
• Opportunistic pathogen - spread locally
and cause disease:
– Otitis media
– Sinusitis
– Bronchitis, pneumonia – elderly, patients with
chronic pulmonary disease
• Disseminated disease uncommon
 Encapsulated
Haemophilus influenzae
• Infrequently found in URT as NF
• Infect, able to penetrate nasopharynx
(NP) submucosal, into bloodstream
• Common infection of infants and young
children
Encapsulated H. influenzae:
Infection and Disease
• Meningitis:
– Follow bacteremic spread from NP
– Usually infants 3-18 months age
– Generally preceded by 1-3 days URT disease
– High mortality
• Epiglottitis:
– Occurs in 2-4 year olds, mostly boys
– Cellulitis, tissue swelling, life-threatening
emergency
– Pharyngitis, fever, breathing difficulties
– Progress to complete obstruction of airway
leading to death
Encapsulated H. influenzae
Infection and Disease
• Cellulitis:
– In very young children
– Fever, reddish-blue color on cheek, periorbital
area
• Arthritis:
– Usually children <2 years age
– Infection of large joints, secondary to
bacteremia
• Conjunctivitis:
– H. aegyptius
– Acute, contagious conjunctivitis
– Commonly called “pink eye”
 Haemophilus ducreyi
• DNA studies – not Haemophilus; but
related to Pasteurellaceae
• STD – genital ulcers, soft chancre or
chancroid
• Common in hot, tropical countries
(Africa, Asia); less common in Europe,
North America
• Diagnosed in men, usually AS in women
• Following 5-7 days exposure, tender
papule, progress to painful ulcer,
inguinal lymphadenopathy
• Does not spread further
• Autoinoculable, resulting in multiple
lesions
Haemophilus: Treatment and
Prevention
• Prompt treatment or causes high mortality
• Antibiotic resistance:
– ~30% penicillin (β-lactamase)
– <1% chloroamphenicol (chloramphenicol
acetyltransferase)
• Drug of choice – ceftriaxone (β-lactamase-
resistant cephalosporin), good CSF
penetration
• For chancroid – treat with erythromycin
• Purified HIB vaccine:
– 3 doses purified PRP-conjugate
– Given 2-6 months of age
– In USA, dramatically reduced disease in infants
 Pasturella multocida:
Characteristics
• Bipolar staining
• Grows well on CBA, Chocolate agar
• Poor growth on Mac and other media for
Gram(-) rods
• Fermentative (glucose, sucrose), little or
no gas
• TSI confusing because of weak acid
production = “sick” appearance
P. multocida: Infection and
Disease
• Three forms of disease
• Animal bite wound infection - localized
cellulitis and lymphadenitis
• Exacerbation of chronic respiratory
disease of patients with underlying
pulmonary dysfunction
• Systemic infection in immunocompromised
patient (bacteremia, meningitis)
• Exquisitely sensitive to penicillin
– 2 units of penicillin
– may be used for presumptive ID)
• Tetracycline, chloramphenicol also used
 Class Assignment
• Textbook Reading: Chapter 18 Haemophilus
and Other Fastdious Gram-Negative Rods -
Haemophilus, Pasteurella (Omit HACEK Group,
Capnocytophaga)
• Key Terms
• Learning Assessment Questions
 Case Study – H. influenzae
• A 78-year-old man confined to a nursing
home awoke with a severe headache and
stiff neck.
• Because he had a high fever and signs of
meningitis, the nursing home staff took
him to a local emergency department.
• The CSF specimen was cloudy.
 Case Study – H. influenzae
• Analysis revealed 400 white blood cells per
mm3 (95% polymorphonuclear neutrophils),
a protein concentration of 75 mg/dl, and a
glucose concentration of 20 mg/dl.
• Small gram-negative rods were seen on
Gram stain of the CSF, and cultures of
CSF and blood were positive for
Haemophilus influenzae.
 Case Study - Questions
• 1. Discuss the epidemiology of H.
influenzae meningitis,and compare it with
the epidemiology of meningitis caused by
Streptococcus pneumoniae and by
Neiserria meningitidis.
• 2. Compare the biology of H. influenzae
strain that is likely to be the cause of this
patient’s disease with the strains that
historically caused pediatric diseases
(prior to vaccination).
 Case Study - Questions
• 3. What other diseases does this organism
cause? What other Haemophilus species
cause disease, and what are the diseases?
• 4. What diseases are caused by Pasteurella
multocida? What is the source of this
organism?