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o Temp…35 – 37°C
o Minimum temp for growth is 20- 25°C
o H influenzae killed at 55 for 30min
o Grows better in aerobic than in anaerobic
o conditions
o Raised CO2 enhance growth of many spp.
This is responsible for the phenomenon of “Satellitism” in which Hemophilus
influenzae forms colonies in the vicinity of Staphylococcal colonies.
X Factor V Factor
1 H influenzae
+ + -
2 H parainfluenzae
- + -
3 H ducreyi
+ - -
4 H hemolyticus
+ + +
5 H parahemolyticus
- + +
6 H aphrophilus
- - -
X = Hematin, V = NAD
Virulence factors of Haemophilus Spp
• Capsule
• Fimbrial adhesin
• HMW1 and HMW2 adhesin
• Hap adhesin
• Hia and Hsf adhesins
• Haemocin
– More than 90% of H influenzae type b
– Inhibits same or related spp.
– Not produced by encapsulated non type b or non typeable H influenzae
strains
Virulence factors cont…
• IgA protease
• Lipooligosaccharide
• Meningitis
• Epiglottis
• Acute pharyngitis
• Pneumonia
Pathogenesis
• H influenzae acts opportunistically as a secondary invader in a variety of
Respiratory Tract Infections usually preceded by viral infections.
• Acute Epiglottitis
– A life-threatening infection involving cellulitis of the epiglottis.
Typically occurs in older children
– Can lead to upper airway obstruction if attempt to take a sample is
made.
– Sore throat and fever rapidly progress to dysphagia, drooling and
airway obstruction
Clinical Manifestations, Hib
• Cellulitis
– Occurs in young children
– Face and/or neck area are commonly involved
– Bacteremia is usually present and 10% patients have additional
focus of infection elsewhere
• Pneumonia
– Also occurs in young children (<2years)
– Clinically indistinguishable from other bacterial pneumonias
• Otitis media
– One of the three most common causes of childhood OM
– Infants present with fever and irritability, older children
complain of pain.
– Viral URTI often precedes OM
– Tympanocentesis can be diagnostic but rarely done
Clinical Manifestations, Non-typable Strains
• BRAZILIAN PURPURIC FEVER (BPF):
– H influenzae biovar aegyptius causes a fulminant pediatric
disease manifested by fever, hemorrhagic skin lesions,
septicemia, vascular collapse, hypotensive shock and death
usually within 48hours.
– Similar cases have been reported from Australia also but the
organism is genetically different from BPF strains.
Diagnosis
• Gram stain of the clinical material may be suggestive.
• CSF
• Positive Gram stain for Gram Negative cocco-bacillary organisms
and subsequent culture of Gram Negative pleomorphic organism
is diagnostic.
• Gram stain is positive in ~80% of untreated cases
• Latex test for PRP is positive in ~95% cases
• PRP can be positive in other sterile body fluids
Diagnosis
• With the exception of H ducreyi and H aphrophilus, all
Haemophilus spp require V factor and grow as satellite colonies
around Staphylococcal streak.
• A heavy suspension is made from agar plate and incubated for 4hours at
37°C. The mixture is exposed to UV light in a dark room and observed for
red fluorescence.
Haemophilus - + + - + +
haemolyticus
Haemophilus + - + - + V
parahaemolyticus
Haemophilus - + - V - -
ducreyi
Haemophilus + - + V - V
parainfluenzae
Haemophilus + - - + - -
aphrophilus
Haemophilus + - - + - -
paraprophilus
*
On horse and rabbit blood.
Biotyping
• Three tests are used for biotyping of H influenzae
biotyping.
– Indole Test
– Urease Test
– Ornithine De-Carboxylase (ODC) Test
BIOTYPE I + + +
BIOTYPE II + + -
BIOTYPE III* - + -
BIOTYPE IV - + +
BIOTYPE V + - +
BIOTYPE VI - - +
BIOTYPE VII + - -
BIOTYPE VIII - - -
*H. influenzae and H. aegyptius can be differentiated by analysis of OMP profile
Haemophilus influenzae, Management
• Meningitis due to H. influenzae is treated with one of the newer
cephalosporins such as Ceftriaxone, Cefotaxime, or Cefuroxime.
• Steroids Administration
– Studies have provided evidence that administration of steroids in meningitis
due to H. influenzae is beneficial with regard to reducing the frequency of
subsequent sequelae.
Haemophilus hemolyticus:
• It is the most markedly hemolytic of Haemophilus spp.
Haemophilus parainfluenzae:
• It is part of normal flora and resemble H influenzae. It
requires V factor only.
Chinese Proverb