Gram negative organisms

and their pathogenesis

Dr. Phoo Pwint Maw Khine

List
Characteristics

Genus

Representative
diseases

a. Cocci
b. Rods
(1)Facultative
(a) Straight
i. Respiratory
organisms

Neisseria

Gonorrhea, meningitis

Haemophilus
Bordetella
Legionella
Brucella
Yersinia
Escherichia
Enterobacter
Klebsiella
Salmonella
Shigella
Proteus
Campylobacte
r
Helicobacter

Meningitis
Whooping cough
Pneumonia
Brucellosis
Plague
Urinary tract
Urinary tract infection
Pneumonia, UTI
Enterocolitis, typhoid
fever
Enterocolitis
UTI
Enterocolitis
Gastritis, peptic ulcer

ii. Zoonotic
organisms
iii. Enteric and
related
organisms
infection,
diarrhea

Neisseriea
Neisseria gonorrhoeae
Neisseria meningitidis

Etiology
Gram negative cocci, in pairs,
kidney shaped
The flat or concave sides are
adjacent
Polysaccharide capsules
meningococci (+) gonococci (-)
Plasmids meningococci (rare)
gonococci (+)
- Best grow under aerobic
- Ferment carbohyadrates,
producing acid not gas
- Oxidase (+)
- Grow best on media containing
heated blood, hemin and animal
proteins
- Rapidly killing by drying, sunlight,
moist heat and many disinfectants

Neisseria gonorrhoeae

Pathogenesis and clinical

findings

- Transmitted sexually
- Attack mucous membrane
of the genitourinary tract,
eye, rectum and throat
producing suppuration
followed by chronic
inflammation and fibrosis
- In males, urethristis (yellow
creamy pus and painful
urination), epididymitis and
fibrosis
- In females, mucopurulent
discharge (cervix, urethra
and vagina)

Pathogenesis and clinical

findings
In prepubertal girl,
vulvovaginitis occurred
either in poor hygiene
and by sexual abuse
Gonococcal opthtalmia
neonatorum

Diagnostic laboratory tests

Specimen - Pus and secretion from the

urethara, cervix, rectum, conjunctiva, throat
and synovial fluid
Smear gram negative diplococci within
pus cells
Culture modified Thayer- Martin medium,
incubate for 48 hrs
Colony appearance smooth, grey,
translucent and slightly raised
Oxidase positive, ferment glucose
Serology - ELISA which detects gonococcal
antigens

- DNA probe assay which

detects gonococcal ribosomal
genes

Prevention and control

Avoiding multiple sexual partners
Early diagnosis and treatment
Mechanical prophylaxis (condom)

Neisseria meningitidis

Pathogenesis and clinical

findings
- Human
- Nasopharynx: portal of
entry (nasopharygitis)
- Reach bloodstream to
meninges or joints
- Fulminant
meningococcemia (high
fever with hemorrhagic
rash)
- Weterhouse Friderichsen
syndrome
- Meningoccemia
- Meningitis (sudden with
intense headache,
vomitting and stiff neck)

Diagnostic laboratory
tests
Specimen blood, CSF,
nasopharyngeal swab for carrier
Smear gram (-) typical neisseriae
within polymorphonuclear
leukocytes
Culture chocolate agar, modified
Thayer Martin medium
Oxidase positive
Ferment glucose and maltose
Serology - antibodies by latex
agglutination test,
hemagglutination test.

Prevention and control

By air borne droplets

Chemoprophylaxis for household and other close droplets
Vaccination in selected population(eg., the militrary; civilian
epidemics)
Reduction of personal contacts in a population with a high
carrier rate

Enterobacteriacae

Etiology

Large family of gram negative bacilli

Primarily in the colon of humans and
other animals
Normal flora
Motile by means of peritrichous
flagella or nor motile, capsule +/Aerobes or facultative anaerobes
Grow readily on ordinary culture
media (eg., peptone water)
On MacConkeys agar produce pink
colonies (lactose fermenters) pr pale
colonies (non lactose fermenters)
With production of acid and gas, or
some produce gas only
Oxidase negative

Commensal enterobacteria
Comprise of 1. Escherichia coli
2. Klebsiella or other
coliforms
3. Proteus sepcies
Pathogens (organisms related to enteric tract
infection)
1. Salmonella
2. Shigella
3. Campylobacter
4. Vibrio
5. Yersinia

Escherichia coli

Etiology

Normal intestinal flora

Usually non pathogenic in their habitat
Most frequent sites of clinically important infections are the
urinary tract, biliary tract and
Gram negative bacilli, motile by means of peritrichous flagella,
aerobe and facultative anaerobe
Culture on blood agar some strain produce haemolysis
- on MacConkeys agar pink colonies (LF)
Ferment sugar with the production of acid and gas
Indole test +, methyl red test +, Voges Proskauer reaction - ,
citrate
Urease test (-)

Clinical findings
1.

2.
3.
4.
5.

Urinary tract infection (most

common, 90% of first UTI in
young women)
Meningitis and brain abscess
Gastroenteritis
Haemorrhagic colitis, HUS
Subacute bacterial
endocarditis
Urinary frequency, dysuria,
haematuria and pyuria
High incidence in pregnant
women
In men, prostatic
enlargement is the most
common predisposing factor
Catheterization and
cystoscopy risky

Diagnostic laboratory
tests

a.
b.
c.
d.
e.

Specimen mid stream urine collected, urinalysis

Gram stain
Culture
E.coli associated diarrhoeal disease
Enteropathogenic E. coli (EPEC): diarrhea in infants
Enterotoxigenic E. coli (ETEC): travelers diarrhea
Enterohaemorrhagic E. coli (EHEC): haemorrhagic
colitis, HUS
Enteroinvasive E. coli (EIEC): similar to shigellosis
Enteroaggregative E. coli (EAEC): acute and chronic
diarrhea in developing countries

Prevention of E. coli
enteritis

The early correction of fluid and electrolyte imbalance is the

most important single factor in preventing the death of the
patient
Proper food hygiene
Pasteurization of milk
Safe water supply
Spread in hospitals and nurseries is mainly from patient to
patient

Klebsiella
Klebsiella pneumoniae
Klebsiella aerogenes
Klebsiella ozaenae
Klebsiella rhinoscleromatis

Etiology

Tends to shorter and thicker

Gram negative rods, 1-2m long
x 0.5- 0.8m wide, capsulated,
non motile abundant loose
extracellular microcapsule,
facultative anaerobes
Growth on MacConkeys agar
pink colonies (lactose fermenter)
produce large mucoid colonies
Grow rapidly on ordinary culture
media
Grow between 12 43C, killed
by moist heat at 55C for 30 mins

Pathogenesis

Fairly common cause of UTI

Occasionally bronchopneumonia,
multiple lung abscess formation in the
lungs
Mortality rate is high
K. pneumonia
Friedlanders
pneumonia
K. aerogenes
urinary tract infection
K. ozaenae ozaena (progressive atrophy
of nasal mucosa)
K. rhinoscleromatis
rhinoscleroma
(chronic upper respiratory disease,
prolonged exposure to crowded and
unhygienic conditions, lesions occur in
the nose, larynx and throat)

Proteus
Proteus vulgaris
Proteus mirabilis
Proteus rettgeri
Proteus morganii

Etiology
Gram negative rods, highly
motile by means of peritrichous
flagella and producing
swarming overgrowth on blood
agar,
Faculatative anarobes
On MacConkeys agar
colorless colonies
Gowth emit fishy odor
Spreads progressively from the
edge of the colony and
eventually covers surface of the
medium
Expanding rings-waves

Pathogenesis

Urinary tract infection

Both community and
hospital acquired
Saprophytes in the
environment
Nosocomial infection
Chronic otitis media
Pneumonia
Infection type of food
poisoning
Wound infection, infection of
burns and bedsores

Shigella
Shigella dysenteriae
Shigella flexneri
Shigella boydii
Shigella sonnei

Etiology
Slender gram (-) non motile
bacilli, coccobacillary forms
Non lactose fermenting
facultative anaerobes bur grow
best aerobically
Convex, circular, transparent
colonies with intact edges
Enrichment media selenite F
borth, tetrathionate broth
Selective media MacConkeys
agar, Salmonella- Shigella
agar, Deoxycholate citrate
agar
Ferment glucose except
Shigella sonnei
No gas, no H2S formation

Pathogenesis and clinical

findings

IP 1- 4 days
Limited to gastrointestinal tract
Bloody diarrhea (dysentery) by
invading the cells of mucosa of
distal ileum and colon
Lead to mucous membrane,
superficial ulceration, bleeding
and formation of
pseudomembrane
Sudden onset of fever and
abdominal cramp followed by
diarrhea (watery at first and
later contains blood and mucus)
Accompanied by straining and
tenesmus (rectal spasms) with
lower abdominal pain
Recurrent bouts of disease

Diagnostic laboratory
tests

Specimen fresh stool, mucus flecks, rectal swab

Culture
Biochemical test sugar fermentation test
Serology slide agglutination test using species
specific sera
Detection of antibody

Prevention and control

Transmitted by food,
fingers, faeces and flies
from person to person
Eliminated by
- Sanitary control of water,
food and milk; sewage
disposal and fly control
- Isolation of patients and
disinfection of excreta
- Detection of subclinical
cases and carriers,
particularly food handlers

Salmonella
Salmonella typhi
Salmonella paratyphi A
Salmonella paratyphi B
Salmonella choleraesuis
Salmonella enteritidis

Etiology
Gram (-) bacilli, vary in length
Motile with peritrichous
flagella
Grow readily on simple media
Never ferment lactose or
sucrose
Form acid and sometimes gas
from glucose and mannose
Produce H2S
Pale and colorless colonies on
MacConkeys culture
Survive freezing in water for
long periods
Resistant to tetrathionate,
sodium deoxycholate

Pathogenesis and clinical

findings
Enter via oral route usually contaminated food or drinks
Host factors resistance gastric acidity, normal intestinal
flora, local immunity
Enterocolitis (food poisoning), typhoid and septicaemia
IP 10 14 days
Enter the lymphatics and blooodstream, reach small
intestine
Multiply in intestinal lymphoid tissue and excreted in stools
Clinical features
- Fever, malaise, headache, constipation, bradycardia and
myalgia
- Spleen and liver enlarge
- Rose spots

Clinical features of Enteric

fever
-

Fever, malaise, headache,

constipation, bradycardia and
myalgia
Spleen and liver enlarge
Rose spots on the skin of
abdomen and chest
Complication
Intestinal haemorrhage and
perforation
Multiply in MNL of Peyers
patches and cause necrosis
Spread to lover, gall bladder and
spleen
Bacteremia
Disseminate to periosteum, lungs

Diagnosis laboratory tests

During the 1st wk
i. Blood culture up to 10
days
ii. Widal test to get a base
line titre of Abs
During the 2nd wk
i. Widal test
ii. Stool culture
iii. Urine culture
Widal test
- Measuring the titre of
specific antibodies in the
patients serum by tube
agglutination test

During the 3rd wk

i. Widal test
ii. Stool culture
iii. Urine culture

During relapse
i. Widal test
ii. Stool culture
iii. Urine culture
iv. Bile or duodenal drainage
culture

Clinical features of
enterocolitis

Nausea, headache, vomiting and profuse

diarrhea
Low grade fever
Laboratory diagnosis
Specimen vomitus, faeces, any suspected
food stuff
Prevention and control
Sanitary measure must be taken to prevent
contamination of food and water by
rodents or other animals that excrete
salmoenlla
Provision of chlorinated water supply
Infected poultry, meats and eggs must be
thoroughly cooked
Pasteurization of milk
Strict hygienic precautions

Vibrios

Vibrio cholera
Vibro parahemolyticus

Etiology
Most common bacteria in
surface water worldwide
Comma shaped, curved rod
2-4m long gram (-) bacilli
Actively motile by means of
a polar flagellum
Alkaline nutrient agar,
thiosulfate-citrate-bile
(TCBS) agar yellow
colonies
Positive oixdase test
Halotolerant
Ferment sucrose and
mannose

Pathogenesis and clinical

findings
Colonize in small intestine,
not an invasive infection
Adhere to cells of brush
border of the gut, multiple
and secretes choleragen
IP 1- 4 days
Sudden onset of nausea and
vomiting and profuse
diarrhea with abdominal
cramps
Rice water stool
Rapid loss of fluid and
electropytes

Diagnostic laboratory
tests

Specimen faeces, rectal swab

Hanging drop preparation darting
motility
TCBS yellow colonies (V. cholerae)
TCBS green colonies (V.
parahaemolyticus)
BSA small translucent and raised
colonies
MacConkeys agar NLF colonies
Oxidase positive
Ferment glucose and sucrose with
production of acid only
Serology slide agglutination test with
antibodies

Prevention and control

Health education
Sanitation, particularly food and feces
Chemoprophylaxis killed vaccine
- live oral cholera vaccine

Campylobacters
Campylobacter jejuni
Campyloacter coli

Etiology

Gram (-) bacilli with comma,

S or gull wing shapes
Motile, with a single polar
flagellum, microaerophilic
Skirrows medium colorless
or gray, watery and
spreading round and convex
Oxidase and catalase
postive
Do not ferment
carbohydrates

Clinical findings

Acute onset of crampy abdominal pain, profuse diarrhea

(grossly bloody)

Diagnostic laboratory tests

Specimen stool
Smear gull wing shaped, Dark field or phase contrast
microscopy typical darting motility

Helicobacter pylori

Etiology
Spiral shaped gram (-)
rod, multiple flagella at
one pole, actively motile
Skirrows medium
translucent 1-2 mm in
diameter
Oxidase, catalase (+),
urease (+)

Pathogenesis and clinical

findings

Invade the epithelial cell

surface and damage the
mucosal cells of gastric
Antral gastritis, duodenal
(peptic) ulcer, gastric ulcer
Major risk factor for gastric
cancer

Diagnostic laboratory test

Specimen gastric biopsy
Smear Giemsa or special silver stain curved or spiraled
organisms
Serology assay to detect serum antiboides
Urease test
Urea breath test

Pseudomonas
Pseudomonas aeruginosa

Etiology
Present in moist
environments in hospitals
Gram (-) bacilli, motile with
multiple flagllea as single
bacteria, in pairs and short
chains
Oligate aerobes
Culture sweet or grape
like or corn taco-like odor
Nonfluorescent bluish
pigment Pyocyanin
Fluorescent bluish pigment
Pyoverdin(greenish) , dark
red pigment Pyorubin, black
pigment Pyomelanin
Do not ferment sugar

Pathogenesis and clinical

findings
Is pathogenic only when
introduced into areas of devoid
of normal defences
Colonize the mucous membrane
and skin invade locally and
produce systemic disease
Fever, shock, oliguria, DIC,
ARDS
Produce infection of wounds
and burns, giving rise to bluegreen pus
Haemorrhagic necrosis of skin
in sepsis ecthyma
gangrenosum (surrounded by
erythema no pus)

Diagnostic laboratory
tests
Specimens from skin leisions, pus, urine, blood and
spinal fluid sputum
Smear
Culture characteristic pigment

Control
Is primarily a nosocomial pathogen
Control of infection in hospitals sinks, water baths,
showers, hot tubs and other wet areas

Bordetella
Bordetella pertussis

Etiology
Small coccobacillary
encapsulated gram (-),
non motile
Toludine blue stain:
bipolar metachromatic
granules (+) capsule (+)
Culture Bordet-Gengou
medium
- charcoal
containing medium
Strict aerobes
Highly contagious disease
Portal of entry: air borne
droplet, respiratory route

Pathogenesis and clinical

findings
Attach to and multiply rapidly
on the epithelial surface of
the trachea and bronchi,
interferes with cilia activity
Not invade to blood
Causing coughing (whooping
cough)
Explosive character, series of
hacking coughs by
production of copious mucus
IP 2 wks
Acute
laryngotracheobronchitis
(ALTB)

Diagnostic laboratory
tests
Specimens saline nasal wash, nasopharyngeal wash,
cough droplets
Staining
Direct fluorescent antibody test

Prevention and control

Pertussive vaccine

Hemophilus
Hemophilus influenzae
Haemophilus ducreyi

Etiology
Very small, gram (-)
coccibacilli with
polysaccharide capsule, in
pairs or short chains
Facultative anaerobes,
very pleomorphic forms
Chocolate agar satellite
phenomenon
Blood agar small
transparent droplike
colonies

Pathogenesis and clinical

findings
Only human, habitat upper
respiratory tract
Mode of infection inhalation
Local extension to sinus and
middle ear, may reach blood
stream and to the meninges
Meningitis (most common cause
in children), pneumonia
Chronic bronchitis, otitis media,
sinusitis
6 months to 6 yrs

Diagnostic laboratory
tests
Specimen nasopharyngeal swabs, pus, blood and
spinal fluid
Immunofluorescence bright organisms against dark
background
Serology to detect capsular antigen

Prevention and contol

Hib Conjugate vaccine

Haemophilus ducreyi

Causes chancroid (soft chancre)

Haemophilus aphrophilus

Normal oral and respiratory tract flora

Legionella pneumonia
Fastidious areobic gram (-)
BACILLI
Legionnairess disease,
pontiac fever
Nonproductive cough, sore
throat

Brucella
Zoonotic infection
Small gram (-) coccbacilli
In soil, in dried foetal
material, in carcass meat,
pork, ham
Brucellosis
Undulating fever, malaise,
weakness

Yersinia pestis

Etiology
Gram (-) rod, safty pin
appearance, non motile
Wayson or Giemsa stain
bipolar staining
Facultative anaerobe
Blood agar, MacConkeys
agar

Pathogenesis and clinical

findings
Sylvatic cycle transmission
among wild rodents by fleas
Urban cycle
Organisms phagocytosed by
PML, and monocytes and
spread to regional lymph
nodes which become swollen
and tender are called buboes
(Bubonic plague)
DIC (black death)
Bubonic plague
Septicaemia plague
Pneumonic plague
IP 2 7 days

Diagnostic laboratory tests

Specimens blood, pus. Sputum

Smears bipolar appearance
Culture
Serology

Control

Spread of rats in urban area

Avoiding flea bites
Notified

Thank you!!!