SPECIAL BACTERIOLOGY

GRAM POSITIVE RODS

Dr.Ghulam Shah Nizamani
Associate Professor of Pathology
 GRAM POSITIVE BACILLI
Sporing
Non-sporing

Non- Filamentous Anaerobic Aerobic

filamentous

•Actinomycetes
Clostridium Bacillus
•C. diphtheriae
• C. tetni •B. anthracis
• L. monocytogens •Nocardia
•C. botulinum •B. cerus
•Lactobacillus
•C. perfringes
•C. difficle
Corynebacterium
 Corynebacterium
 Bacilli show round, solid ends like clubbing,
therefore also “club-shaped bacilli”.
 Have beaded appearance because of the
presence of “metachromatic granules,
granules appear brown or red, rest of the
body appear violet
 The bacilli show typical pattern of
arrangement (“V” or “Y” configurations) or in
clumps resembling “Chinese letters”
 C. diphtheriae
 Also called “Klebs Loeffler’s bacillus’’

 Habitate
Not found in our body but organism
resembling it are present on skin,
tongue, nasopharynx etc, which
resemble morphologically with C.
diphtheriae but are non-pathogenic,
called “diphtheriod”
 C. diphtheriae
 Found in patients and in carriers

 Mode of infection: Droplet infection

 Resistant to dryness and light

 Can contaminate fomites and can

spread through fomites
 C. diphtheriae
PATHOGENESIS

 Invasion
 Exotoxin which inhibits protein
synthesis by ADP-ribosylation of
elongation factor 2 (EF-2).
 PATHOGENESIS
INVASION
 Invade the mucus membrane, form a
pseudomembrane in the throat
(composed of fibrin, leukocytes,
necrotic epithelial cells and C.
diphtheriae) which bleed on touch, it
can extend down to larynx and may
cause death due to asphyxia.
Pseudomembrane Throat
 PATHOGENESIS
EXOTOXIN (DISEASES)
 Airway obstruction
 Acute myocarditis
 Acute heart failure
 Damage both cranial and spinal nerves
 Loss of accomodation, Diplopia (squint)
 Dysphagia
 Nasal rgurgitation of food
 Faliure of diaphragm, Dysponea
 Peripheral neuropathy
 C. diphtheriae

Diphtheria is a serious disease, if

not properly treated, pt may die
because of formation of
psuedomembrane leading to
asphyxia and heart failure.
 Lab Diagnosis
Throat Swab

Smear Culture Screening

 Methylen Loeffler’s media • Schick
blue • Earlier growth test
 Important • 6-12 hr
stain for Tellurite media
volutin
granules • Growth in 24
hrs
 Typical
Chinese • Differentiate
letter between diff
colonies biotypes of
C.diphtheriae
 Schick Test
 Not a diagnostic test but a screening test
 Done to detect in population, which persons
are immune and which persons susceptible
 0.mL I/D inj. Of purified standardized toxin. If
patient has no antitoxin, the toxin will cause
inflammation within 4-7 days and viceversa.
 Positive reaction shows person is susceptible
to diphtheria and needs active immunization
 Negative reaction shows person is immune to
diphtheria and no need for immunization
ACTINOMYCETES
 ACTINOMYCETES
 Filamentous bacteria acting like fungi
 Found in the oral cavity and gut as normal
flora
 Causes opportunistic infection
 Granulomatous inflammation
 Important species is Actnomycetes israelii
 Diseases
 Chronic suppurative granulomatous
inflammation
 Causes tissue destruction, deformities, scarring
and discharging sinuses
 Discharge contain yellow granules
(sulfur granules) composed of colonies of
actinomycetes
 Diseases
 Initial lesion appears as hard non tender
swelling that drain through sinus tract. It may
appear on
 Face and neck (Cervicofacial Actinomycosis)
 Chest (Thoracic Actinomycosis)
 Abdomen (Abdominal Actinomycosis)
 Pelvis (Pelvic Actinomycosis)
Cervicofacial Actinomycosis with
discharging sinus
NOCARDIA
 NOCARDIA
 Filamentous, non-sporing bacillus
 Not found in the body but in external
environment
 Causes opportunistic infection
 Granulomatous inflammation
 Important species is Nocardia asteroids
(produces star shaped colonies)
 Diseases
 Skin infections (Madura foot)
Usually in the feet of farmers and who work in
the fields and have cuts in their feet, develop
multiple discharging sinuses
 Pulmonary Nocardiasis
Immunodeficient people when inhale Nocardia,
develop firm adherent swelling in the chest
wall “Mycetoma” with pneumonia like
symptoms
Madura Foot
CLOSTRIDIUM
 CLOSTRIDIUM
 Gram +ve spore forming, non filamentous
bacilli
 Obligate anaerobe
 Most of them are saprophyte
 Found in external environment
 CLOSTRIDIUM
Important species are
 C. tetani
 C. botulinum
 C. perfringes (welchi)
 C. difficile
 1. Clostridium tetani
 Disease:
Tetanus (lock jaw)
 Habitate:
Bacilli are found in gut of human and animal
Spores are found in soil, air, feces excreted by
animal
 Portal of entry: Wound, skin popping,
contaminated umbilicus or circumcision wound
 Clostridium tetani
Virulent factor
 Produced a powerful exotoxin Tetanospasmin”
 It is actually a neurotoxin
 Produced by bacilli at the site of wound and
travel in a retrograde fashion within nerves
 Bacilli are non-invasive and will never enter the
blood
 PATHOGENESIS
Tetanus toxin (tetanospasmin) neurotoxin produced by
bacilli at site of wound

Absorbed intra-axonally (retrograde) to CNS

anterior horn of spinal cord

Block the release of inhibitory neurotransmitter (e.g.,

Glycine & GABA)

Persistent spasm (spastic paralysis)

 CLINICAL FEATURES
 Strong muscle spasms (spastic paralysis)
 Lock jaw (trismus)
 Risus sardonicus (typical facial expression)
 Exaggerated reflexes
 Opisthotonos (arching of back due to spasm of
muscle)
 Death may occur due to paralysis of respiratory
muscle (respiratory failure)
Spastic paralysis and Lock jaw
Opisthotonos (arching of back due
to spasm of muscle)
 Clostridium tetani
Diagnosis
 No microbiologic or serological
diagnosis
 Organisms are rarely isolated from
wound site
 It produces a terminal spore which
gives it “tennis racket” appearance
 2. Clostridium botulinum
 Disease:
Botulism
 Habitate:
Spores are found in soil, contaminate vegetable
and meat, when canned or vacuum-packed,
spores germinate, exotoxin is produced within
canned food and ingested preformed
 Most important target is peripheral nervous
system, doesn’t affect CNS
 Portal of entry: contaminated vegetables, meat,
fish. Toxins are killed by sufficient cooking
 PATHOGENESIS
Botulinum toxin as an exotoxin, taking preformed in
food

Absorbed through blood from gut

Carried to peripheral nerve

(Neuromuscular junction)

Blocks release of acetylcholine

Flaccid paralysis
 DISEASES
 Descending weakness and paralysis
 Paralysis of eye muscle
Drooping of eyelid (ptosis)
Blurred vision
Diplopia (double vision)
 Dysphagia
 Paralysis of respiratory muscle
Dysponea , may lead to death
 DISEASES
1. Wound botulism
2. Infantile botulism
 Botulism in less than 6 months
 Usually due to honey contaminated with
spores
 Weakness and flaccid paralysis called
“floppy baby syndrome”
 May cause death due to respiratory failure
Floppy baby syndrome
 3. Clostridium perfringens
 Disease:
Gas gangrene and food poisoning
 Habitate:
Bacilli form normal flora of colon and vagina
Spores are found in soil
 Portal of entry: usually a wound
contaminated with soil, specially war wound,
road accident and septic abortion, ingestion of
contaminated food.
 PATHOGENESIS & CLINICAL
FEATURES
Gas gangrene
Spore contaminate the wound and
produces a toxin called “alpha toxin
(lecithinase)” → hemolysis
 Degradative enzymes → gas in tissues
 Pain, edema, cellulitis in wound area
and crepitation
 Hemolysis → jaundice
 Shock and death may occur
Gas Gangrene
 PATHOGENESIS & CLINICAL FEATURES
Food Poisoning
 Member of normal flora of colon and vagina
but not small intestine
 Food specially meat, chicken, fish
contaminated with spores → enterotoxin,
which are heat resistant so not killed by
cooking
 Causes cramps, vomiting, watery diarrhea, etc
 4. Clostridium difficile
 Disease:
Antibiotic associated
pseudomembranous enterocolitis.
Clindamycin was the first than
cephalosporins, ampicillin,
fluoroquinolones and anticancer drugs.
 Habitate:
In approximately 3% of general
population in GIT, 30% of hospitalized
patients
 PATHOGENESIS
Heavy and prolonged antibiotic therapy suppress
normal flora

overgrowth of C. difficile

Powerful exotoxin

Damage to intestinal mucosal cells

Formation of a pseudomembrane over intestinal

wall
 CLINICAL FEATURES
 Watery diarrhea with
pseudomembranes (yellow-white
plagues) on the colonic mucosa
 Fever
 Abdominal cramps
 Toxic megacolon can occur
 Bacillus anthracis
 Zoonotoic Disease: Anthrax

 Pulmonary anthrax (woolsorter’s

disease)

 Cutaneous anthrax

 Intestinal anthrax
 Bacillus cereus
 Endospores are present in the environment
 Contaminate food like cereals specially reheated
fried rice
 Produce enterotoxin which cause ADP-
ribocylation of G protein which stimulates
adenylate cyclase and leads to increase cAMP
within the enterocyte.
Disease
 Food poisoning (Chinese rice poisoning)