Haemophilus Spp And Other Fastidious Gram Negative Rods

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General characteristics: Family Pasteurellaceae

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Gram neg coccoid to rod shape

non-motile

aerobic to facutlatively anaerobic

nitrate to nitrite (reduction)

oxdiase and catalase postive

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Haemophilus

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gram negative pleomorphic coccobaccilli or rods

non motile

oxidase and catalase positive

nitrate reduction

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Where do Haemophilus spp live

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obligate parasites of the mucous membrane

H. influenzae, H. aegyptius, H. ducreyi (STD)

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Haemophilus means

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blood loving

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Requirements for Haemophilus

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X factor in hemin, hamtin

V factor is (NAD)

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H. influenzae require both:

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X and V factors

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H. parainfluenzea requirements

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produces it's own X

requires V

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Aggregatibacter species

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formerly Haemophilus and Actinobacillus

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Haemophilus hemolysis

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H. hemolyticus and sometime H. ducreyi

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What does SBA contain

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only contains X factor

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Satellitism

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growth of a fastidious organism around other bacteria that release the necessary growth factors or breakdown toxic products

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Who does not do stallitism

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H. aphrophilus and H. ducreyi

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Growth patterns

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10% of normal flora of the upper respiratory tract in adults is Haemophilus

(2-6% in children; higher amount in daycare)-As they mature they convert from encapsulated to nonencapsulated strain

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When was H. influenzae first isolated

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1889-90 from a pandemic influenza virus victims but was not the cause

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Virulence factors of H. influenzae

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Capsule

IgA protease

LPS

Adherence

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H. influenzae capsule

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sixserotypes (a-f)

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H. influenzae IgA protease

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cleaves IgA on mucous membrane

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LPS H. influenzae

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role in virulence not well defined

antibody to the proteins is somewhat protective

Paralyzes the cilia

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Adherence H. influenzae

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pili and other structures

non-encapsulated strains

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Capsule serotpyes

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contains a unique polysaccharide: ribose, ribitol, and phosphate

vaccine against serotype B- in children causes meningitis and or bacteremia

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Nontypable strains (no capsule)

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invade the respiratory tract and tissues located around the same area

localized infection- pneumonia, otitis media, sinusitis

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Disease of Hib Strains

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Meningitis- invasion of respiratory tract, bacteremia spread to meninges and CSF

infants highly susceptible

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Epiglottitis

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acute inflammation and swelling causing airway obstruction

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Bacterial tracheitis

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life-threatening disease in young children

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Cellulitis

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cheek swelling in young children

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Other infections H. influenzae capsule

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acute pharyngitis and pneumoniaa

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Disease from non-capsulated

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otitis media, bronchitis, sinusitis, pneumonia in elderly

can cause invasive disease in older patients and immunocompromised patients

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H. aegyptius

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resembles H. influenzae

assoc with highly communicable form of pinkeye

Brazilian purpuric fever

purplish discolorization of the skin and mucous mem

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H. aphrophilus

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endocarditis and pneumonia

can be present as normal flora

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H. parainfluenzae

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normal inhabitant of human respiratory tract

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H. haemolyticus

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hemolytic

occurs in normal

nasopharynx and in assoc with rare upper resp. infections

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Vaccine selection

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Hib decrease of 99%

but invasive disease rises in adults

Countries with no vaccine: leading cause of bacterial pneumonia and meningitis in children

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H. aegyptius and H. influenzae biogroup aegypticus can cause

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Pinkeye

Acute contagious in children

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H. influenzae biogroup aegypticus fever caused:

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Brazilian purpuric fever a more virulent disease

high mortality rate, as high as 70%

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Sexually Transmitted infection

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H. ducreyi

strict pathogen

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Incubation of H. ducreyi

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4-14 days painful lesions

genital and perianal areas

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buboes

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enlarged and draining lymph nodes

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H. parahaemolyticus

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pharyngitis

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H. aphrophilus

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bite wound infections

endocarditis

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Haemophilus species processing

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die rapidly

H. ducreyi- clean site with steril saline and gauzze

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H. influenzae processing

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CA

CA with bacitracin- inhibits other organism

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H. aegypticus plates

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CA supplemented with 1% isovitaleX

hold for 4 days

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H. ducreyi plate

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Nairobi plate

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Nairobi plate

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GC agar with 2% BOVINE hgb

MH with 5% horse blood lyzed

both sides have vancomycin

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Air levels

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Capnophilic and moist

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Other plates

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BAP for H. aphrophilus

MacConkey agar (no growth)

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Colony morph of Haemophilus

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translucent, moist, smooth convex colonies

mousy or bleach like odor

encapsulated strain appear more mucoid

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H. influenzae gram

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coccobacili or small rods

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H. ducreyi gram

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coccobacili arranged in groups "school of fish"

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Taking a colony of Haemophilus

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don't touch growth media

place colonies in saline or nutrient broth

place on nutrient agar or MH agar plate: add v and x

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Normal growth temp and times on agars

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35-37 deg 5-10% co2 18-24 hours

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Porphyrin test that don't require X

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porphyrin + organism

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What does porphyrin test test?

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the ability of organism to convert ALA to porphyrins or protoporphyrins

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Kovacs reagent

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red color production in lower part of the tube when porphobilinogen is present

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Wood UV lamp

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fluoresce reddish orange under uv light

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Pro and cons of Porphyrin test

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Pro: X not required, eliminates carryover contam

Con: id of H. influ based on neg test

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Haemophilus quad plate

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perferred over Porphyrin since it's better for H. influenzae

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H. influenzae factors

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Requires X, V

Hemolysis negative

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H. paprinfluenzae factors

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Require V

Hemolysis negative

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H. hemolyticus

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require v and x

hemolysis positive

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H. parahaemolyticus

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requires V

hemolysis positive

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H. ducreyi

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require x

hem neg

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H. aegypticus

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requires v and x

hem neg

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H. aphrophilus

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has v and x

hem neg

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Treatment

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Cefotaxime or ceftriaxone: recommended

Alternative treatments:

Sulfamethoxazole, imipenem, ciprofloxacin, and chloramphenicol

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Drug resistance

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beta-lactams and altered PBP's

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H. ducreyi treatment

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Erythormyocin

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Chancroid

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gential ulcer disease: "soft chancre"

open sores faciltate the transmission of other STDs