Gram Negative Bacillus

GRAM NEGATIVE BACILLI
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Characteristics
Habitat
Salmonella spp
Gr ve bacilli
Non sporing , motile bacilli
All exept S.typhi are non capsulate bacilli
Facultative anaerobes
Animal gut
Human is the only natural host for S. typhi & S

paratyphi
Transmission
Foodborne roote
Faecal-oral route
Endotoxin=cause fever , leukemia , hypotension

&shock
Capsule=antiphagocytosis
Can survive within macrophages
Typhoid fever (Enteric fever)
Pathogenesis
1. bacteria rich small intestine and then enter
Diseases
lymphatics& blood streem

2. cause fever , malaria , headache ,
constipation etc
3. spleen &liver become enlarged
Enterocolitis (gastroenteritis or
food poisoning )
Laboratory
Identification
septicemia
Oxidase ve , non lactose fermentation
Produce acid &gas from glucose
Grow well on macConnkey agar , DCA , .S.S

agar
Commercial kits available for full identification
Shigella
Characteristics
Pathogenesis
Gram Ve bacillus
Faculitatie anaerobic
Most shigella pathogen is S.sonnei
Non lactose fermentive .
Non motile .
Endotoxin
Capsule
Antigenic phase variation .
Gastroenteritis (( shigellosis ))
S.dysenteriae cause bacterial dysenteria
Diseases
Laboratory
Identification
Oxidase Ve
Isolation from specimens requires use of

selective media .
Vibrio
-the second major group of gram-negative ,facultative anaerobic, fermentative rods.
- Were separated from Enterobacteriaceae on the basis of positive oxidase reaction
and the presence of polar flagella.
-the most important members of Vibrio spp are:

1)Vibrio cholera.
2)Vibrio parahaemolyticus.
3)Vibrio vulnificus.
-Vibrio species can grow on a variety of simple media within temp rate (14-40C).
V.cholerae can grow in the absence of salts(most other species of vibrio that are
pathogenic in human require salt [halophilic species]).
-Vibrio tolerate a wide range of PH (e.g.PH of 6.5-9.0) but are susceptible to stomach
acids. If gastric acid production is reduced or neutralized , patients are more
susceptible to Vibrio infection.
-most vibrio have asingle polar flagella (in contrast with peritrichous flagella in the
family Enterobacteriaceae.
-vibrio have also various pili that are important for virulence.
-epidemic strains of V.cholerae the etiologic agent of cholera
Characteristics
Vibrio cholerae
Small , curved (comma shapped )
Gr ve bacilli
Non sporing , motile bacilli , non capsulate

bacilli
Were separated from Enterobacteriaceae on the

basis of positive oxidase reaction and the
presence of polar flagella.
Habitat
Contaminated water (usually) or food
(sometimes )
Transmission
Pathogenesis
Foodborne route
Faecal oral route
Choleragen enterotoxin = hypersecretion of

electrolytes andwater.
Cholera
1. bacteria colonize the intestinal tract in
very high numbers
Diseases
2. bacteria attach to but do not invade the

intestinal mucosa
3. intese vomiting & diarrhea
4. severe loss of water &electrolytes
*gastroenteritis
Laboratory
Identification
Oxidase +ve
Special selective media for vibrio thiusulfate

citrate bile salts sucrose(TCBS).
Do not require salts for growth. But can tolerate

it.
Vibrio cholerae are rarely seen in Gram-stained

stool or wound specimens.
By using darkfield microscope may able to detect

the motile bacteria in stool specimen.
Enterobacteriaceae SPP.
-the family Enterobacteriaceae is the largest, most
heterogeneous collection of medically important
gram-negative rods.
-fewer than 20 species are responsible for more
than 95% of the infection.
-Enterobacteriaceae are ubiquitous organisms,
found worldwide in soil,water,vegetation.
-Enterobacteriaceae part of the normal intestinal
flora of most human.
-these bacteria cause a variety of human diseases
including 30-35% of all septicemia, more than 70%
of urinary tract infection , and many intestinal
infections.
-some organisms(e.g.salmonella typhi, shiglla,
yersinia pestis) are always associated with disease.
-while other organisms(e.g. E.coli, klebsiella
pneumoniae, protus mirabilis) are members of the
normal commensal flora that can cause
opportunistic infections.

-members of the Enterobacteriaceae family are

moderately in size, gram-negative rods .
-they share a co
mmon antigen (enterobacterial common antigen)
-are either nonmotile or motile with peritrichous
flagella
-do not form spore.
-all members can grow rabidly, facultative
anaerobic) on variety of nonselctive
media(e.g.blood agar) and selective
(e.g.macConkey agar).
-the Enterobacteriaceae have simple nutritional
requirements, ferment glucose.
-catalase +ve, oxidase ve.
-the absence of cytochrome oxidase activity is
important characteristic , because it can be
measured rabidly with simple test and is used to
distinguish the Enterobacteriaceae from many other
fermentative and nonfermentative gram-negative
rods.
-a few exceptions to these rules exits (e.g.
plesiomonas shigelloides is oxidase +ve, klebsiella

granulomatis can not be cultured on traditional

media.
-the ability to ferment lactose has been used to
differentiate lactose-fermenting strains(e.g.
Echerichia, klebsiella, enterobacter, citrobacter ,
serratia spp) from strains that do not ferment
lactose or do so slowly(e.g. proteus, salmonella,
shiglla, yersinia spp).
-resistance to bile salts in some selective media has
been used to separate enteric pathogen(e.g.
shigella, salmonella) from commensal organisms
that are inhibited by bile salts(e.g. gram+ve and
some gram-ve bacteria present in the
gastrointestinal tract)
-some Enterobacteriaaceae have prominent
capsules (e.g. most klebsiella, some Enterobacter,
Escherichia strains),
-the heat-stable lipopolysaccharide(LPS) is the
major cell wall antigen.
-most the Enterobaceriaceae are motile with the
exception of the common isolates Klebsiella,
shigella, yersinia.
-the motile strains posses peritrichous flagella.

-there are other species of Enterobacteriaceae :

1)Enterobacter
2)Citrobacter
3)Serratia
4)Morganella
-These species subdivide as lactose fermenter
Enterobacteriaceae.
-these species caused infections rarely in
immunocompetent patients.
-they are more common causes of
immunocompromised patients.
-e.g. citrobacter koseri cause meningitis and brain
abscesses in neonates.
Escherichia coli
Gr ve bacilli
Non sporing , motile , non capsulated bacilli but

some strains are capsulate
Characteristics
Habitat
Transmission
Pathogenesis
Diseases
Laboratory
Identification
Common normal flora of human intestinal tract
Foodborne route
Faecal oral route
Endogenous spread
Endotoxin = cause fever , leucopenia
Capsule (in some strains ) = antiphagocytosis
Enterotoxin = cause diarrhea
Urinary tract infection (UTI)
Diarrhoeal disease
Neonatal meningitis
Septicemia
Gastroenteritis.
Oxidase ve , lactose fermentive
Grow well on macConkey agar , DCA , XLD

agar
Characteristics
Proteus spp
Gr ve bacilli
Non sporing highly motile , non capsulate bacilli
P.mirabilis is the most common species.
Produce large quantitiesof urease.(this process

raises the urine PH and facilitates the formation
of renal stones , the increased alkalinity of the
urine is also toxic.
Habitat
Transmission
Common normal flora of the human intestinal tract
Foodborne route
Faecal oral route
Endogenous spread
Pathogenesis
Diseases
UTI
Septicemia
Laboratory
Identification
Oxidase ve , non lactose fermentive

agar
Swarming colonies on solid media
Yersinia pestis
Gr ve bacilli (cocobacilli ) (facultative

anaerobes)
Characteristics
Have a tendency for bipolar staining (in

which the endof the bacilli stain darker
than the central part)
Nonsporing , non motile bacilli . capsulate
bacilli
Some species (e.g.Y.enterocolitica) can
grow at cold temp.
Habitat
Transmission
Animal reservoirs are rodents such as rats
Bacteria spreads between anmals by fleas

Major outbreaks in humans result from
exposure to infected rats
The rat flea carries infection from rat to rat
& from rat to human
Bubonic plague is not transmitted from
person to person
Pneumonic plague spread from person
to person by droplets (airborne route )
Pathogenesis
Intracellular murine toxin = causing

irreversible shock and death
Capsule (f1) = antiphagocytosis
The V/M antigen = specific protein act as
antiphagocytosis & promote intracellular
growth of the bacteria
PLAGUE :
1. is normally azoonotic disease of rodents
Diseases
that exists in 2 kinds of epidemic centers

2. permanent but relatively resistant rat
population (wild rat)
3. temporary but susceptible rat population
(domestic rats )
4. epidemics of plague usually occur in
crowded areas poor sanitation
bubonic plague :
1. bacteria multiply at the entery
site in the skin
2. bacteria
spread
via
the
lymphatics to lymph nodes

3. 2 to 6 days after the flea bite
occurs , lymph nodes become
very tender , and enlarge to form
buboes
with
haemorrhagic
inflammation
4. death rate is about 75% in
untread bubonic plague
pneumonic plague (black
death) :
1. bacteria
disseminates from
lymph nodes by
way of the
bloodstream to
the spleen , liver ,l
lungs
2. pneumonic plague
can be transmitted
by
respri
atory
route but Iis
3. death tate is nearly

100%in
untreaed
pneumonic plague
stained smears should show bipolar

Laboratory
Identification
staining
organism can be grow from aspirtes of
enlarged buboes , or from sputum in case
of pneumonic plague
colonies are grayish , mucoid & rough
colonies may be noted
the organism are not fastidious ; it can be
rrown on routine media
oxidase ve .
Pasteurella
Characteristics
Garm Ve bacillus
Non spore forming
Non motile
Faculitative anaerobic
Fermentative coccobacilli .
Most common P.multocida and P.canis
Habitat
Transmission
Pathogenesis
Diseases
Bite wound infection .
Chronic pulmonary disesase
Bacteremia
Meningitis
Laboratory
Identification
Grow well in blood and choclate agar . and

poorly in macConkey agar .
After over night incubation in blood agar

large buttery colonies with musty odor cuase
by product indole
Klebsiella pneumoniae
Gr ve bacilli
Non sporing , non motile , prominent capsulate
Characteristics
that responsible for the mucoid.
Formerly called (Donovaina granulomatis)
Common normal flora of the human intestinal
Habitat
Transmission
Pathogenesis
Diseases
tract & oropharynx
Foodborne route
Faecal oral route
Endogenous spread
Capsule = antiphagocytosis
UTI
Neonatal meningitis
Septicemia
Pneumonia (Rare )
Laboratory
Identification
Oxidase ve , lactose fermentive

agar
Large mucoid colonies
Characteristics
Brucella spp
Gr ve small coccibacilli
Non sporing ,non motile , non capsulated bacilli
Intracellular pathogen
Slow growth requirements
Fastidious.
Is member of the alpha proteobacteria group
Strictly aerobic
Does not ferment carbohydrates.
Four species:
1)Brucella abortus 2)Brucella melitensis.

3)Brucella suis
Habitat
Transmission
4)Brucella canis
Chronic infection in domestic animals
Contaminated milk or other unpasteurized dairy

products
Direct contact
Invasion of the body by brucella results in

lymphatic dissemination of the bacteria
As regional lymph nodes become infected , the

brucella are phagocytized , but they are capable
of survive & intracellular multiplication inside
Pathogenesis
phagocytic cells
As phagocytic cells die , brucella , are released

into the blood stream and intracellularly ,
establish localized infection in bone marrow ,
liver &spleen , and thus hide from host defence
Undulant fever (malta fever or brucellosis )

1. patients frequently present with PUO
Diseases
2. chronic , debilitating febrile illness usually

without any localizing signs
3. intracellularly , bacteria localize and cause

granulomas lesions in the spleen , liver
bone marrow & lymph nodes
4. patients presents with intermittent fever &
profound weakness , chills , sweats ,
headache , backache
5. two types of infection occur ; (acute
infection ) or (chronic infection lasting
12 months )
Laboratory
Identification
requiring complex growth media like :glucose

serum or liver infusion broth or agar
usuakky require 3-5 days incubation
blood provides the best material for culture
grow occasionally on MacConkey agar.
Oxidase +ve
Urease+ve
Francisella tularensis
Characteristics
Gr Ve small bacilli (coccobacilli)
Often showing bipolar staining
Non motile , non sporing , capsulate , coccobacilli
Strick aerobic
Fastidious.
Two species of Francisella spp:
1)F.tularensis
2)F.philomiragia
*require cystein for growth.
Widely found in animal reservoirs
Habitat
Transmission
biting arthropods
direct contact with infected animal tissue
ingestion of contamination food or water
inhalation of aerosols
F.tularensis is highly infection ; penrtration of the

skin or mucous membranes or inhalation of 50
organism can result in infection
Pathogenesis
F.tularensis is an intracellular pathogen of

phagocytosis cells
Capsule= phagocytosis
tularemia (sometimes called rabbit fever or

glandular fever or tick fever or deer fly fever ):
Diseases
1-clinical presentation dependes on the route of

infection
2- a plague like disease of rodents
Laboratory
Identification
blood is taken for serological tests
growth dose not occur in most ordinary

bacteriological media but small colonies appear in
1-3 day on glucose , blood agar incubated at 37 C
under aerobic conditions
culture on cystein-supplemented
media(e.g.choclate agar, BCYE agar ) is specific.
Culture on cystin blood agar.
Culture on cystin glucose agar
BCYT= buffered charcoal yeast extract.
Haemophilus
-the family pasteurllaceae spp are
1)Haemophilus
2)Actinobacillus
3)Pasteurella
-the members of this family are:
*Small gram-negative
*Non-spore
*Nonmotile
*Aerobic or facultative anerobic rods
-most members of this family have fastidious growth
needs, requiring enriched media for growth.
-members of the genus Haemophilus are the most
commonly and significant human pathogen.
Haemophilus influenzae
Characteristics
Gr ve bacilli (coccobacilli)
Non sporing , non moitile ,
The surface of many(but not all) strains of

H.influenzae is covered with polysaccharide
capsule.
Haemophillic bacteria
Pyogenic (pus producing ) bacteria
Plemorphic.
Is the most species of Haemophilus that cause

disease.
The cell wall structure is typical of other grannegative rods(lipopolysaccharide with endotoxin
activity is present in thecell wall.
Is fermentive.
Human upper respiratory tract as anormal flora
Habitat
but many also cause respiratory disease , usually

as secondary invader
Transmission
Airborne route
Endotoxin = causing damage to the epithelial cell

of the upper respiratory tract
Pathogenesis
Capsule = antiphagocytosis
IgAase = degrades IgA antibodies that is key host

defense mechanism in mucous membrane
H.influenzae type b is most virulent with

RPR(polyribitol phosphate) in capsule
Pili=adhere to host cells.
Cause severe infection in children ( acute

meningitis )
Diseases
H.influenzae serotype was responsible for more

than 95% of all Haemophilus infection.
Serotypes C and F and noncapsulated

H.influenzae diseases.
Cause chronic , less severe infection beyond age 6

(,pneumonia,Otitis media , sinusitis , bronchitis )
in older children and adult
This bacteria is the most common cause of

bacterial meningitis in children from 5 months to
5 years of age
Mortality rate of untreated H. influenzaw

meningitis may be up to 90%
Act synergistically with viral infection ,

predisposing to severe haemophilus infection , the
incidwnce increase during influenza outbreaks
Laboratory
Identification
Require enriched media such as blood or

chocolate agar or(Levinthals agar)
Need both od tow growth factor (factor V-X)

1. heat stable X factor = hemin or some
other iron containing prophrin
2. heat stable V factor = nicotinamide
adenine nucleotide (NAD)
grow poorly in the absence of oxygen & growth

is enhancedin an atmosphere with add CO2
on chocolate or blood agar : a streak of staph .

aureus across the plate produce V factor and
enlarges the size of adjacent colonies of H .
inflluenzae = satellitism phenomena
colonies are small , translucent , non hemolytic
microscopy is sensitive test for detecting

H.influenzae in(cerebrospinal fluidCSF .synovial
fluid, and lower respirtory specimens.
Pseudomonas aeruginosa
Characteristics
Gr ve bacilli arranged in pairs.
Non sporing , motile , capsulate (polysaccharide)
Obligae aerobes
Nonfermentative.
The presence of cytochrome oxidase(detected in

rabid,5minute test) in pseudomonas species is
used to differentiate them from the
Enterobacteriaceae
Some pseudomonas produce diffusible pigments

(e.g.pyocyanin
[blue],fluorescein[yellow],pyorubin[raddishbrown]
Habitat
P.aeruginosa is the most common type.
Widley distributed in water , soil & moist areas
Carriage as a part of normal gut flora in higher

proporation of hospital inpatients
Transmission
Direct contact
Endogenous spread
Exdotoxin = cause fever , leucopenia
Exdotoxin A = inhibit protein synthesis
Exdotoxin S = inhibit protein synthesis
Protease & Elastase = destruction of tissue
Capsule=inhibits antibiotics+suppresses
Pathogenesis
neutrophil and lymphocyte activity.
Diseases
Pili=adhesin
UTI
Septicemia
Pneumonia
Burn wounds & wound infection
Ear and eye infection
Laboratory
Identification
Oxidase +ve , lactose fermentive

Identified by colonial characteristics(e.g.
hemolytic, green pigment, grape like odor)

agar
Acinetobacter spp
Gr ve bacilli (plump coccobacilli)
Non sporing , non moitile , capsulated

coccobacilli
Characteristics
Strictly aerobic
The genus can be subdivided into two groups:
1)glucose oxidizing species(A.baumannii)

2)glucose nonoxidizing species(A.lwoffii)
Survive in moist surfaces. Including respiratory therapy
Habitat
equipment .
And survive on dry surface including human skin(the
latter feature is unusual for gram-negative rods)
Transmission
Direct contact
Indirect contact via invasive devices
It has ability to colonize on internal surfaces of

the invasive devices
Pathogenesis
*pulmonary infection.
Diseases
*opportunistic pathogens that cause:

Respiratory tract unfection and urinary tract infection and
wound and septicemia.
Laboratory
Identification
Oxidase ve
Grow well on simple media
Bacteriodes
Characteristics
Gram Ve bacillus .
Obligate anaerobic .
Most common species is b.fragilis .
Polymorphic in size and shape .
Diseases
Intraabdominal infection .
Gynecologic .
Skin and soft tissues infection .
Bacteremia .
Laboratory
Identification
Grow rapidly (( detected in 2 days )) .
Stimulated growth in 20% bile .
Bordetella pertussis
Characteristics
Gr Ve extremely smaal bacilli
Non sporing , non motile bacilli
Capsulate bacilli
Obligate aerobes
Human respairatory tract
Habitat
Transmission
airborne route
direct contact
tracheal cytotoxin = inhibits DNA synthesis in

ciliated cells
fimbrial antigen (pili ) = permit the adherence of

bacteria to the ciliated epithelium of the
Pathogenesis
respiratory tract
endoxin = causing damage to the epithelial cells

of the upper respiratory tract
pertussis toxin
whooping cough (pertussis ) by Bordetella

pertussis.
Diseases
Mild pertussis by Bordetella para pertissis.
Respirtory disease in dogs by Bordetella

bronchiseptica.
Laboratory
Identification
slow growing & fastidious in its growth

requirement
bordet gengous agar or blood charcoal agar
requires media supplemented with charcoal,

starch, blood, or albumin.
requires 3-5 day incubation in moist atmosphere
it dose not require X and V factors on culure
oxidase amino acids.
Not ferment carbohydrates.
Differential characteristics of Bordetell species

characteristics
B.pertussis
B.parapertussis
oxidase
+
urease
+
motility
On sheep agar
+
On macConkey
-/+
agar
B.bronchiseptica
+
+
+
+
+
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Gram Negative Bacillus

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GRAM NEGATIVE BACILLI

Non sporing , motile bacilli

All exept S.typhi are non capsulate bacilli

Human is the only natural host for S. typhi & S

Endotoxin=cause fever , leukemia , hypotension

Can survive within macrophages

Typhoid fever (Enteric fever)

1. bacteria rich small intestine and then enter

lymphatics& blood streem

Oxidase ve , non lactose fermentation

Produce acid &gas from glucose

Grow well on macConnkey agar , DCA , .S.S

Commercial kits available for full identification

Most shigella pathogen is S.sonnei

Non lactose fermentive .

Antigenic phase variation .

S.dysenteriae cause bacterial dysenteria

Isolation from specimens requires use of

-the most important members of Vibrio spp are:

Small , curved (comma shapped )

Non sporing , motile bacilli , non capsulate

Were separated from Enterobacteriaceae on the

Contaminated water (usually) or food

Faecal oral route

Choleragen enterotoxin = hypersecretion of

2. bacteria attach to but do not invade the

Commercial kits available for full identification

Special selective media for vibrio thiusulfate

Do not require salts for growth. But can tolerate

Vibrio cholerae are rarely seen in Gram-stained

By using darkfield microscope may able to detect

-members of the Enterobacteriaceae family are

granulomatis can not be cultured on traditional

-there are other species of Enterobacteriaceae :

Non sporing , motile , non capsulated bacilli but

Common normal flora of human intestinal tract

Faecal oral route

Endotoxin = cause fever , leucopenia

Capsule (in some strains ) = antiphagocytosis

Enterotoxin = cause diarrhea

Urinary tract infection (UTI)

Oxidase ve , lactose fermentive

Grow well on macConkey agar , DCA , XLD

Commercial kits available for full identification

Non sporing highly motile , non capsulate bacilli

P.mirabilis is the most common species.

Produce large quantitiesof urease.(this process

Common normal flora of the human intestinal tract

Faecal oral route

Endotoxin = cause fever , leucopenia

Oxidase ve , non lactose fermentive

Swarming colonies on solid media

Commercial kits available for full identification

Gr ve bacilli (cocobacilli ) (facultative

Have a tendency for bipolar staining (in

Animal reservoirs are rodents such as rats

Bacteria spreads between anmals by fleas

Intracellular murine toxin = causing

that exists in 2 kinds of epidemic centers

lymphatics to lymph nodes

route but Iis

3. death tate is nearly

stained smears should show bipolar

Non spore forming