ESCHERICHIA COLI

Gram ve, non-sporing

bacillli.most strains are
motile.

Grow on ordinary media,

Mac Conkeys agar.
Colonies are rose
pink(lactose fermenter)

KLEBSIELLA
K. Aerogenes
K. pneumoniae
K. rhinoscleromatis
K. ozanae
Gram ve, non-motile, non-sporing
bacilli. Freshly isolated strains
produced well defined polysaccharide
capsule.
Grow on ordinary media,
Optimum temp, 37C.
Colonies are large raised, moist, &
mucoid, depend on loose slime
characters produced by klebsiella.
Strains usually ferment lactose &
produce pink colonies on Mac
Conkeys agar.

Epidemiological typing of proteus.

Many gram-ve bacteria produce bacteriocins which are
bactericidal subs which are produced by certain strains of
bacteria are active against some other strains of same or
closely related bacteria.
Bacteriocin-producing strains are resistant to their own
bacteriocin, thus bacteriocin can be used for typing or
organisms.
Bacteriocin production & sensitivity has been used as
good tool for typing strains of proteus & this method
could be made more discriminating when used in
combination with O serotyping.

PROTEUS
Pr. Vulgaris
Pr. mirabilis

MORPHOLOGY
Gram ve bacilli, nonencapsulated, highly motile.

CULTURAL CHARACTERS
Fishy odour, swarming
appearance in non-inhibitory
solid media.(appear as thin
colorless, transparent film
extend from margins of
young colony & spread in
several waves demarcated
by raised margin.
Medium inhibit swarming:
-Blood agar, 2-3X usual
conc.(5-6%)
-Mac Conkey agar & Desoxy
cholate citrate agar (DCA)
-Electrolyte deficient media
like CLED medium
-Media contain inhibitory
chemicals
Dienes phenomenon
2 identical proteus cultures
inoculated at different point,
swarming w/out sign of
demarcation.

PSEUDOMONAS
Pseudomonas aeruginosa

ENTEROCOCCI
Enterococci faecalis

Gram ve motile bacillus

Gram +ve cocci in

pairs or in short
chains, non-motile &
non-capsulated

-obligate aerobe, grows well at 3742C.(helps to differentiate Ps

aeruginosa than others species)
-grows readily on many types of media
& most strains hemolyse blood on
blood agar.
-produces several colonial types, most
virulent is mucoid type.
-usually produces sweet grape like
odour.
-almost all strains characteristically
produce on agar diffusible pigments.
Pyoverdin:greenish
Pyocyanin: bluish
Pyorubin: red pigment
Pyomelanin: black pigment
-grow on special selective medium,
cetrimide agar. Cetrimide is chemical
disinfectant that is well tolerated by Ps,
aeruginosa but not by others.

-grows readily on
ordinary culture media
& on Mac Conkeys
agar as minute lactose
fermenting colonies,
on blood agar,
colonies form zones of
B-haemolysis,
sometimes ahaemolysis, nonhaemolysis.
-grow on media with a
high salt content 6.5%
NaCl.
-E. faecalis grows
within a wide
temperature range 1045C.

Distinguished from other

enterobacteria & defined as
species(e.g: fermentation of sugars,
indole production (+ve), citrate
utilization (-ve), methy red
reaction(+ve))
Distinguished from other coliforms
by its ability to form acid & gas
from lactose incubated at
44C(Eijkman test), use for
detection of E.coli in studies of
faexal pollution of water.

Most species are:

-weak urease producers
-indole negative
-citrate positive

In two different strains, they

fail to coalesce and remain
separated by narrow easily
visible area.
BIOCHEMICAL REACTIONS
Ability to hydrolyse urea
rapidly(within 24 hr) by its
production of urease enzyme
Ability to deaminate phenyl
alanine to phenyl pyruvate
acid by its production of
enzyme phenyl alanine
deaminase.

-does not ferment sugars but may

oxidise glucose.
-oxidase positive
-generally a biochemically inactive
organisms
Identification:
-presence of characteristics
pigment(pyocyanin & pyoverdin)
-oxidase positive
-growth at 42C

ANTIGENIC STRUCTURES
3 kinds of surface Ag myb
Identification:
Classified into several O
Strains of Ps. Aeroginosa have been
demonstrated & observed for
Differentiated into several
groups by their O antigen &
classified into several O serotypes by
serotyping E.coli: O(osmotic),
capsular serotypes by
gps are further subdivided
their O antigens.
K(capsular),
identification of their
according to different H
-possess many flagellar H antigen for
H(flagellar) Ag.
capsular Ag.
antigens into many
further serotyping.
Best done by microscopical
serotypes.
-epidemiological purpose strains are
demo of capsular swelling in Determination of serotype of also phage & bacteriocin typed.
wet films with capsular anti- isolate is useful in
serum.
epidemiological studies.
PATHOGENESIS AND DISEASES PRODUCED
Intestinal commensal
K. pneumoniae is present in resp tract
Produce infections in human -Ps aeruginosa produced infection of
strains of E.coli cause
& faeces of about 5% of normal
when bacteria leave the
wounds & burns
opportunistics infections
individual.
intestinal tract:
-meningitis
in other parts of body
Causes 3% of bacterial pneumonias.
Pr. vulgaris is important
-UTI(catheter)
when there is
Ocassionally produced UTI and
nosocomial pathogen
-resp tract infection(esp through

Ferments mannitol
with gas production.

abnormality or
impairment of defense.
They are commonest
cause of UTI. Only limited
no of serogroups of E.
Coli are uropathogenic.
UPEC possess certain
virulence factors that
allow colonization &
invasion of urinary tract.
Most important are type
1 fimbrae that bind to
uroepithelial cell & a and
b haemolysins that lyse
host erythrocytes.
Commonly present in:
Appendicular abscess
Peritonitis
Cholecystitis
Septic wound
Bed sores
May infect lower resp
passage, cause
bacteraemia &
nosocomial infections of
various body site.
Ocassionally cause
meningitis in neonates
(K1 strains)

nosocomial.
Associated with inflammatory
conditions of upper resp. tract:
K. ozanae: has been isolated from
nasal mucosa in ozanae a foetid
progressive atrophy of mucous
membranes
K. rhinoscleromatis: has been isolated
from rhinoscleroma a destructive
granuloma of nose & pharynx.
-Specimen: urine, blood, pus, sputum
-Smear: presence of large capsules by
special staining is suggestive of
Klebsiella, direct capsule swelling test
can be performed on Klebsiella visible
in fresh specimens to save time.
-Culture: specimens are plated on both
blood agar and differential media.
Characteristics mucoid lactose
fermenting colonies are observed after
24 hrs incubation.
-Serotyping: colonies can be further
identified biochemically & by capsular
swelling reaction using appropriate
antisera.

May produce bacteremia,

pneumonia or mengitis
May lead to focal
lesion(debilitated pts + IV)
Common cause of UTI

contaminated resp resulting in

necrotizing pneumonia, & certain
mucoid type produce chronic infective
condition-cystic fibrosis of lung)
-external otitis in swimmers
-infection in eye after surgery or
surgical operation
-fatal septicemia

LABRATORY DIAGNOSES
-Specimen: urine, blood, pus, -Specimen: should be obtained as
sputum
indicated by type of infection
-Smear: gram ve bacilli
-Smears: gram stained smears often
-Culture: plated on blood
reveal gram ve rods
agar & Mac Conkeys agar,
-Culture: specimen are plated on blood
incubated at 37C for 24 hr.
agar, Mac Conkeys agar & cetrimide
Reveal: feature of swarming
agar if selection is required.
on blood agar & inhibition on Ps aeruginosa does not ferment lactose
Mac Conkeys agar where
& can be easily recognised by its
Proteus grow as non-lactose pigmentation on nutrient agar & by
fermenting colonies + fishy
being oxidase positive & its growth at
odour.
42C.
Urease test is performed to
demonstrate strong urase
production.
-Serotyping: slide
agglutination technique
using O antisera.