Download - Gram Negative Bacillus
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8/18/2019 Gram Negative Bacillus
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GRAM NEGATIVE BACILLI
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8/18/2019 Gram Negative Bacillus
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Salmonella spp
Gr –ve bacilli
Non sporing , motile bacilli
All exept S.typhi are non – capsulate bacilli
Facultative anaerobes
haracteristics
Animal gut
Human is the only natural host for S. typhi & S
paratyphi
Habitat
Foodborne roote
Faecal-oral route
Transmission
Endotoxin=cause fever , leukemia , hypotension
&shock
Capsule=antiphagocytosis
Can survive within macrophages
Pathogenesis
Typhoid fever (Enteric fever)
1. bacteria rich small intestine and then enter
lymphatics& blood streem
2. cause fever , malaria , headache ,
constipation etc
3.
spleen &liver become enlarged
Enterocolitis (gastroenteritis or
food poisoning )
septicemia
Diseases
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
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
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Shigella
Gram –Ve bacillus
Faculitatie anaerobic
Most shigella pathogen is S.sonnei
Non lactose fermentive .
Non motile .
haracteristics
Endotoxin
Capsule
Antigenic phase variation .
Pathogenesis
Gastroenteritis (( shigellosis ))
S.dysenteriae cause bacterial dysenteria
Diseases
Oxidase – Ve
Isolation from specimens requires use of
selective media .
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
4/31
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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 stomachacids. 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
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8/18/2019 Gram Negative Bacillus
5/31
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Vibrio cholerae
Small , curved (comma – shapped )
Gr –ve bacilli
Non sporing , motile bacilli , non capsulate
bacilli
Facultative anaerobes
Were separated from Enterobacteriaceae on the
basis of positive oxidase reaction and the
presence of polar flagella.
haracteristics
Contaminated water (usually) or food
(sometimes )
Habitat
Foodborne route
Faecal – oral routeTransmission
Choleragen enterotoxin = hypersecretion of
electrolytes andwater.Pathogenesis
Cholera
1. bacteria colonize the intestinal tract in
very high numbers
2. bacteria attach to but do not invade the
intestinal mucosa
3. intese vomiting & diarrhea
4.
severe loss of water &electrolytes*gastroenteritis
Diseases
Oxidase +ve
Commercial kits available for full identification
Special selective media for vibrio thiusulfate
citrate bile salts sucrose(TCBS).
Do not require salts for growth. But can tolerate
it.
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
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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.
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8/18/2019 Gram Negative Bacillus
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8/18/2019 Gram Negative Bacillus
8/31
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-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 bemeasured 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
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8/18/2019 Gram Negative Bacillus
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granulomatis can not be cultured on traditional
media.
-the ability to ferment lactose has been used todifferentiate 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.
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8/18/2019 Gram Negative Bacillus
10/31
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-there are other species of Enterobacteriaceae :
1)Enterobacter
2)Citrobacter
3)Serratia4)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.
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8/18/2019 Gram Negative Bacillus
11/31
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Escherichia coli
Gr –ve bacilli
Non sporing , motile , non capsulated bacilli but
some strains are capsulate
Facultative anaerobes
haracteristics
Common normal flora of human intestinal tractabitat
Foodborne route
Faecal – oral route
Endogenous spread
Transmission
Endotoxin = cause fever , leucopenia
Capsule (in some strains ) = antiphagocytosis
Enterotoxin = cause diarrhea
Pathogenesis
Urinary tract infection (UTI)
Diarrhoeal disease
Neonatal meningitis
Septicemia
Gastroenteritis.
Diseases
Oxidase –ve , lactose fermentive
Grow well on macConkey agar , DCA , XLD
agar
Commercial kits available for full identification
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
12/31
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Proteus spp
Gr –ve bacilli
Non sporing highly motile , non capsulate bacilli
Facultative anaerobes
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.
haracteristics
Common normal flora of the human intestinal tractabitat
Foodborne route
Faecal – oral route
Endogenous spread
Transmission
Endotoxin = cause fever , leucopenia
Pathogenesis
UTI
SepticemiaDiseases
Oxidase –ve , non lactose fermentive
Grow well on macConkey agar , DCA , XLD
agar
Swarming colonies on solid media
Commercial kits available for full identification
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
13/31
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Yersinia pestis
Gr –ve bacilli (cocobacilli ) (facultative
anaerobes)
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.
haracteristics
Animal reservoirs are rodents such as ratsHabitat
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 )
Transmission
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
Pathogenesis
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8/18/2019 Gram Negative Bacillus
14/31
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PLAGUE :
1. is normally azoonotic disease of rodents
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
Diseases
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8/18/2019 Gram Negative Bacillus
15/31
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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
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 .
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
16/31
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Pasteurella
Garm –Ve bacillus
Non spore forming
Non motile
Faculitative anaerobic
Fermentative coccobacilli .
Most common P.multocida and P.canis
haracteristics
abitat
Transmission
Pathogenesis
Bite wound infection .
Chronic pulmonary disesase
Bacteremia
Meningitis
Diseases
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
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
17/31
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Klebsiella pneum oniae
Gr –ve bacilli
Non sporing , non motile , prominent capsulate
that responsible for the mucoid.
Facultative anaerobes
Formerly called (Donovaina granulomatis)
haracteristics
Common normal flora of the human intestinal
tract & oropharynxHabitat
Foodborne route
Faecal – oral route
Endogenous spread
Transmission
Endotoxin = cause fever , leucopenia
Capsule = antiphagocytosisPathogenesis
UTI
Neonatal meningitis
Septicemia
Pneumonia (Rare )
Diseases
Oxidase –ve , lactose fermentive
Grow well on macConkey agar , DCA , XLD
agar
Large mucoid colonies
Commercial kits available for full identification
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
18/31
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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 4)Brucella canis
haracteristics
Chronic infection in domestic animalsHabitat
Contaminated milk or other unpasteurized dairy
products
Direct contact
Transmission
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
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
Pathogenesis
Undulant fever (malta fever or brucellosis )
1.
patients frequently present with PUO
2. chronic , debilitating febrile illness usually
without any localizing signs
3.
intracellularly , bacteria localize and cause
Diseases
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8/18/2019 Gram Negative Bacillus
19/31
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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 )
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
Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
20/31
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Francisella tularensis
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.
haracteristics
Widely found in animal reservoirsHabitat
biting arthropods
direct contact with infected animal tissue
ingestion of contamination food or water
inhalation of aerosols
Transmission
F.tularensis is highly infection ; penrtration of the
skin or mucous membranes or inhalation of 50
organism can result in infection
F.tularensis is an intracellular pathogen of
phagocytosis cells
Capsule= phagocytosis
Pathogenesis
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8/18/2019 Gram Negative Bacillus
21/31
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tularemia (sometimes called rabbit fever or
glandular fever or tick fever or deer fly fever ):
1-clinical presentation dependes on the route of
infection2- a plague – like disease of rodents
Diseases
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.
Laboratory
Identification
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emophilus
-the family pasteurllaceae spp are1)Haemophilus
2)Actinobacillus3)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 growthneeds, requiring enriched media for growth.
the mostare Haemophilusmembers of the genus-
commonly and significant human pathogen.
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Haemophilus influenzae
Gr –ve bacilli (coccobacilli)
Non sporing , non moitile ,
The surface of many(but not all) strains of
H.influenzae is covered with polysaccharide
capsule.
Facultative anaerobes
Haemophillic bacteria
Pyogenic (pus –producing ) bacteria
Plemorphic.
Is the most species of Haemophilus that cause
disease.
The cell wall structure is typical of other gran-
negative rods(lipopolysaccharide with endotoxin
activity is present in thecell wall.
Is fermentive.
haracteristics
Human upper respiratory tract as anormal flora
but many also cause respiratory disease , usually
as secondary invader
Habitat
Airborne routeTransmission
Endotoxin = causing damage to the epithelial cell
of the upper respiratory tract
Capsule = antiphagocytosis
IgAase = degrades IgA antibodies that is key host
defense mechanism in mucous membrane
H.influenzae type b is most virulent withRPR(polyribitol phosphate) in capsule
Pili=adhere to host cells.
Pathogenesis
Cause severe infection in children ( acute
meningitis )
H.influenzae serotype was responsible for more
than 95% of all Haemophilus infection.
Serotypes C and F and noncapsulated
Diseases
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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
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.
Laboratory
Identification
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Pseudomonas aeruginosa
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[raddish-
brown]
P.aeruginosa is the most common type.
haracteristics
Widley distributed in water , soil & moist areas
Carriage as a part of normal gut flora in higher
proporation of hospital inpatients
Habitat
Direct contact
Endogenous spreadTransmission
Exdotoxin = cause fever , leucopenia
Exdotoxin A = inhibit protein synthesis
Exdotoxin S = inhibit protein synthesis
Protease & Elastase = destruction of tissue
Capsule=inhibits antibiotics+suppresses
neutrophil and lymphocyte activity.
Pili=adhesin
Pathogenesis
UTI
Septicemia
Pneumonia
Burn wounds & wound infection
Ear and eye infection
Diseases
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8/18/2019 Gram Negative Bacillus
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Oxidase +ve , lactose fermentive
Identified by colonial characteristics(e.g.
hemolytic, green pigment, grape like odor)
Grow well on macConkey agar , DCA , XLDagar
Commercial kits available for full identification
Laboratory
Identification
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cinetobacter spp
Gr –ve bacilli (plump coccobacilli)
Non sporing , non moitile , capsulated
coccobacilli
Strictly aerobic
The genus can be subdivided into two groups:
1)glucose oxidizing species(A.baumannii)
2)glucose nonoxidizing species(A.lwoffii)
haracteristics
Survive in moist surfaces. Including respiratory therapy
equipment .And survive on dry surface including human skin(the
latter feature is unusual for gram-negative rods)
Habitat
Direct contact
Indirect contact via invasive devicesTransmission
It has ability to colonize on internal surfaces of
the invasive devicesPathogenesis
*pulmonary infection.
*opportunistic pathogens that cause:
Respiratory tract unfection and urinary tract infection and
wound and septicemia.
Diseases
Oxidase –ve
Grow well on simple mediaLaboratory
Identification
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Bacteriodes
Gram –Ve bacillus .
Obligate anaerobic .
Most common species is b.fragilis .
Polymorphic in size and shape .
haracteristics
Intraabdominal infection .
Gynecologic .
Skin and soft tissues infection .
Bacteremia .
Diseases
Grow rapidly (( detected in 2 days )) .
Stimulated growth in 20% bile .Laboratory
Identification
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8/18/2019 Gram Negative Bacillus
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Bordetella pertussis
Gr –Ve extremely smaal bacilli
Non sporing , non motile bacilli
Capsulate bacilli
Obligate aerobes
haracteristics
Human respairatory tract
Habitat
airborne route
direct contactTransmission
tracheal cytotoxin = inhibits DNA synthesis in
ciliated cells
fimbrial antigen (pili ) = permit the adherence of
bacteria to the ciliated epithelium of the
respiratory tract
endoxin = causing damage to the epithelial cells
of the upper respiratory tract
pertussis toxin
Pathogenesis
whooping cough (pertussis ) by Bordetella
pertussis.
Mild pertussis by Bordetella para pertissis.
Respirtory disease in dogs by Bordetella
bronchiseptica.
Diseases
slow growing & fastidious in its growth
requirement
bordet – gengous agar or blood – charcoal agar
requires media supplemented with charcoal,
starch, blood, or albumin.
Laboratory
Identification
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Differential characteristics of Bordetell species
B.bronchisepticaB.parapertussisB.pertussischaracteristics
+ - + oxidase
+ + - urease
+ - - motility
+ + - On sheep agar
+ /+- - On macConkeyagar
requires 3-5 day incubation in moist atmosphere
it dose not require X and V factors on culure
oxidase amino acids.
Not ferment carbohydrates.
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