2.
Genus Klebsiella
By Mr.Hamse Barud
(a) Klebsiella pneumoniae
Pathogenicity
Klebsiella pneumoniae causes chest
infections and occasionally severe
bronchopneumonia with lung abscesses.
Infections are often opportunistic,
occurring in those with existing chest
disease or diabetes mellitus, or in
malnourished persons.
Klebsiella aerogenes is associated with
hospital acquired infections of wounds and
of the urinary tract.
It is also found in the respiratory tract
where it may cause infection, particularly in
LABORATORY DIAGNOSIS
Specimens:
Depending on the site of infection,
specimens include urine, pus, sputum
and infected tissue.
Morphology
Klebsiellae are Gram negative, non-
motile, usually capsulated rods.
Culture
Klebsiellae are aerobes and facultative
anaerobes.
Blood agar:
Klebsiellae produce large grey white
MacConkey agar and CLED medium:
Most klebsiellae are lactose-
fermenting,producing mucoid pink colonies
on MacConkey agar and yellow mucoid
colonies on CLED medium
Biochemical tests
Indole negative
Ornithine decarboxylase negative
Do not produce H2S
Antimicrobial susceptibility
Klebsiellae often produce beta-lactamases
and are resistant to ampicillin.
Cephalosporins and aminoglycosides are used
to treat Klebsiella infections.Some Klebsiella
3.Genus Salmonellae
By Mr.Hamse Barud
Species of medical importance are:
Salmonella typhi
Salmonella paratyphi
Salmonella enteritidis
All pathogenic salmonellae belong to a
single species, Salmonella enterica which
is subdivided into 7 subspecies
S. enterica subsp. enterica has over 2000
serovars which can cause disease in
humans.
Pathogenicity
The following diseases are caused by
Salmonella:
It is caused by S.typhi and S.paratyphi, and
transmitted by fecal-oral route via
contaminated food and drinks
Incubation period: 10-14 days
Both manifest with persistent fever,
headache, malaise, chills, enlargement of
liver and spleen, and skin rashes.
Paratyphoid fever is milder than typhoid
fever
2.Bacteraemia caused by S. Typhi (most
serious form) and S. Paratyphi A, B, C.
These salmonellae are usually found only in
humans, being excreted in the faeces and
Infection is by ingesting the organisms in
contaminated food or water or from
contaminated hands
(S. Typhi is mainly water-borne, S. Paratyphi is
mainly food-borne).
3. Gastroenteritis. It is caused by S. enteritidis
and S. typhimurium
It manifests with initial watery diarrhea, and
later bloody mucoid diarrhea associated with
crampy abdominal pain and tenesmus.
Bacteremia is rare (2-3 % of cases). It usually
resolves in 2-3 days
S. Typhimurium and S. Enteritidis causes
Salmonella enterocolitis infection is by
ingesting salmonellae, in food that has become
contaminated from animal or human intestinal
sources, directly or indirectly.
LABORATORY DIAGNOSIS
Specimens:
1.Blood, Bone marrow, stool, urine and serum for
enteric fever(typhoid)
Blood – 80% positive in the first week.
Stool- 70-80% positive in the second and third
week.
Urine- 20% positive in the third and fourth
week.
2. Stool for gastroenteritis.
Morphology
Salmonellae are Gram negative rods.
With the exception of S. Pullorum-
gallinarum, all salmonellae are actively
motile.
They are non-sporing and with the exception
of S. typhi, non-capsulate.
Culture
Salmonellae are aerobes and facultative
anaerobes.
They grow between 15–45 ºC with an
optimum temperature of 37 ºC.
Columbia agar-broth diphasic medium
Is recommended because this can be used for
all salmonellae and other pathogens that cause
bacteraemia.
Blood cultures are subcultured on to blood
agar.
Blood agar (subculture):
Salmonellae produce grey-white 2–3 mm
diameter nonhaemolytic colonies, similar in
appearance to those of many other
enterobacteria.
Some strains appear mucoid.
XLD agar:
Hydrogen sulphide (H2S)-producing
Salmonellae that do not produce H2S, e.g. most
strains of S. Paratyphi A, form pink-red colonies
without black centres, similar in appearance to
shigellae on DCA and MacConkey agar:
Salmonellae produce non-lactose fermenting
pale coloured colonies which on DCA, have
black centres(H2S-producing salmonellae)
Biochemical tests
Salmonellae like shigellae can be presumptively
identified biochemically using KIA (Kligler iron
agar) medium
KIA medium is used to help identify salmonellae
following isolation on a primary selective
medium.
Salmonellae produce pink-red (alkaline) slope
and yellow (acid) butt,indicating fermentation
of glucose but not lactose.
Blackening in the medium due to H2S unless
serotype does not produce H2S, e.g. S.
Paratyphi
Only a small amount of blackening is seen
with S. Typhi.
Urease and indole negative
Lactose negative
Gas produced from glucose fermentation (S.
Typhi does not produce gas)
Citrate positive (S. Typhi and S. Paratyphi A
are citrate negative)
Lysine decarboxylase (LDC) positive (S.
Paratyphi A is LDC negative)
Beta-galactosidase (ONPG) negative
S. Typhi can be biochemically differentiated
from other salmonellae by being citrate
negative, not producing gas and forming only
small amounts of H2S
Serotyping
Based on their O and H antigen
composition,typhoid fever can be diagnosed
serologically.
Detection of a specific antibody response with
typical clinical symptoms is suggestive of
To make a definitive diagnosis, culture is
required.
Serological tests currently in use to assist in
the diagnosis of enteric fever(typhoid) include:
Widal test
Ig M antibody immunoassays
(a) Widal test
The diagnostic value of the Widal test remains
controversial.
Most agree that the test is not sufficiently
sensitive or specific to be clinically useful when
only a single acute-phase serum sample is
tested
The Widal test measures agglutinating
In acute typhoid fever, 0 agglutinins can
usually be detected 6–8 days after the onset of
fever and H agglutinins after 10–12 days.
High or rising titer to O antigen (≥ 1:160)
suggests active infection.
High or rising titer to H antigen (≥ 1:160)
suggests past infection or immunization.
In endemic areas the Widal test produces
many false positive and false negative test
results.
False positive results occur because S. Typhi
shares O and H antigens with other
Salmonella serovars and crossreactions also
occur with other enterobacteriacae.
Causes of raised O and H agglutinins other
than typhoid fever include;
Liver disease associated with raised globulin
levels
Previous Salmonella infections
Chronic salmonellosis associated with
schistosomal infection
Vaccination with typhoid vaccine.
Acute falciparum malaria(particularly in children)
Chronic diseases such as rheumatoid arthritis
Myelomatosis
Nephrotic syndrome.
False negative widal tests may be due to
antibody responses being blocked by early
antimicrobial treatment or following a typhoid
(b)IgM antibody assays to diagnose
typhoid fever
These assays detect IgM antibodies to S.
Typhi which develop early in acute typhoid.
They suggest current infection, are more
sensitive and specific than the Widal test,
and can be performed more rapidly.
In the absence of culture facilities, IgM
antibody tests are more useful in helping to
diagnose typhoid in endemic areas
particularly after 7 days following the onset
Antimicrobial susceptibility
Antimicrobials with activity against S.
Typhi include chloramphenicol, co-
trimoxazole, and ampicillin.
Chloramphenicol resistant strains,
however, have been reported from
developing countries
S. Typhimurium multi-drug resistance is
causing a major public health problem
in several developing countries and other
parts of the world where the incidence of
salmonellosis (transmitted from animals
to humans) has increased greatly.
