Lesson 1: STAPHYLOCOCCUS Infections associated with in-dwelling
medical devices such as IV catheters,
GENUS STAPHYLOCOCCUS
Morphological Characteristics
• Perfectly spherical Gram (+) cocci
approx. 0.5 to 1u in diameter
• Grow in clusters like a bunch of
grapes (Greek word “staphylo”)
Two pigmented colony types
1. S. aureus – golden yellow
2. S. albus/S.epidermidis – white
Habitat
1. S. aureus – nasal passages, skin,
oral cavity and intestinal tract
2. S. epidermidis – inhabitant of the
skin
Pathogenesis
Virulence factors
1. Surface proteins – promote
colonization of host tissues such as
those that promote attachment to
host
a. Surface proteins promote
attachment to host proteins
such as laminin and
fibronectin that form part of
the extracellular matrix
• Fibronectin present on
epithelial and endothelial
surfaces as well as
component of blood clots
b. Fibrinogen/fibrin binding
(clumping factor) which
promotes attachment to
blood clots and traumatized
tissues
c. Collagen-binding factor
prosthetic joints, replacements heart valves)
• Biomaterial implanted in human
body becomes coated with complex
mixtures of host proteins and
platelets (fibrinogen is dominant
component) -> adherence of S.
aureus
• If material is in the body for long
periods, fibrinogen is degraded and
no longer promote bacterial
attachment
• Fibronectin becomes predominant
ligand promoting attachment
• Factors that inhibit phagocytosis -
capsule, immunoglobulin-binding
protein A
a. Capsular polysaccharide –
surface polysaccharide of
serotype 5 or 8
• Called microcapsule because it can
be visualized only by electron
microscopy after antibody labelling
• Polysaccharide rapidly lost upon
laboratory subculture
• Function not clear but may impede
phagocytosis
b. Protein A surface protein of
S. aureus which binds with
IgG the wrong way by non-
immune mechanism and
disrupts opsonization and
phagocytosis
c. Leukocidin – toxin that
specifically acts on
polymorphonuclear
leukocytes
• Protein toxins
• Responsible for symptoms
during infections
• Membrane damaging toxins

a. A – toxin
• Best characterized and most potent
membrane-damaging toxin; major
virulence factor
• Binds to membrane of susceptible
cells, its subunits oligomerize to form
hexameric rings, a central pore
through which cellular contents leak
• Human platelets and monocytes
have a high affinity to toxin
• A complex series of secondary
reactions cause release of
eicosanoids and cytokines which
trigger the production of
inflammatory mediators that produce
the symptoms of septic shock
b. B-toxin
• Sphingomyelinase C damages
membranes rich in this lipid
• Classical test is the lysis of sheep
RBC, at 4’C-15’C produces
incomplete hot-cold lysis
c. Delta- toxin
• Small peptide toxin produced by
most strains of S. aureus
• A phospholipase which role in
disease is not known
d. Gamma – toxin and
leucocidin
• Two-component toxin that damages
membrane of susceptible cells
• Produced separately but act
together to damage membranes
• Expresses 3 proteins: B and C form
leukotoxin with poor hemolytic
activity whereas A and B are
hemolytic and weakly leukotoxic
• Panton and Valentine (PV)
leucocidin distinct from leukotoxin by
gamma locus; has potent
leukotoxicity but non-hemolytic
• PV leucocidin is an important factor
in necrotizing skin lesions such as
dermonecrosis wnen injected SC in
rabbits
e. Superantigens – enterotoxins and
toxic shock syndrome toxin (TSST1)
1. Enterotoxins – 6 serotypes A, B, C,
D, E ad G
• Cause diarrhea and vomiting
(staphylococcal food poisoning)
• Can cause TSS
2. TSST1 – weakly related to
enterotoxin and does not have
emetic activity
• Responsible for 75% of TSS
including menstrual cases; tampon-
related TSS not true infection
• Superantigens stimulate T-cells
without normal antigenic recognition,
results to release of cytokines in
large amounts causing symptoms of
TSS
f. Epidermolytic (exfoliative) toxin – ET
• Causes scalded skin syndrome in
neonates with widespread blistering
and loss of epidermis
• 2 forms: ETA and ETB
• Protease activity
• Possibly the toxin targets a specific
protein involved in maintaining the
integrity of the epidermis
g. Other extracellular proteins
1. Coagulase
• Extracellular protein which binds
with prothrombin in host to form
staphylothrombin which causes the
activation of the protease activity of
thrombin resulting to conversion of
fibrinogen to fibrin and formation of
clot in plasma
• Traditional marker for identifying S.
aureus
• Coagulase and clumping factor
separate entities
• Coagulase extracellular protein
though small fraction is tightly bound
on bacterial cell surface and reacts
with prothrombin
• Clumping factor is fibrinogen-binding
determinant on S. aureus cell
surface, sometimes referred to as
bound coagulase
2. Staphylokinase
• Plasminogen activator
• Complex formed between
staphylokinase and plasminogen
• Activates plasmin-like proteolytic
activity which dissolves fibrin clot
3. Enzymes
• Proteases, lipases,
deoxyribonuclease (Dnase) and fatty
acid modifying enzyme (FAME)
• FAME important in abscesses where
it could modify antibacterial lipids
and prolong bacterial surviva
• First three provide nutrients for
bacteria
4. Hyaluronidase – hydrolyzes
hyaluronic acid (cementing
substance)
• Spreading factor
S. epidermidis
• Most important coagulase-negative
staphylococci (CNS) which is a
common common of the skin
• Major cause of infections associated
with prosthetic devices and catheter
• Production of characteristic slime
(biofilm)
Cultural and Biochemical Characteristics
• S. aureus coagulase and catalase
(+), S. epidermidis coagulase (-) but
catalase (+)
• Salt tolerant, S. aureus ferments
mannitol, S. epidermidis does not
Epidemiology
• Epidemiological tracing traditionally
phage typing but has limitations
Diseases
S. aureus
1. Botryomycosis in horses
• Stump of spermatic cord after
castration become infected,
enlarged and sclerotic with pockets
of pus containing granules similar to
actinomycosis
2. Mastitis – suppurative lesions in
cattle, organisms colonize tips of
teats
3. Tick pyemia in lambs
• Acute septicemia or bactermia along
with toxemia kills lamb
• Ixodes ricinus
4. Facial or periorbital eczema in sheep
5. Purulent synovitis in poultry
• Bumble foot in turkey
6. Cutaneous staphylococcosis
 • Exudative dermatitis in rabbits
and subcutaneous abscesses in
older animals
7. Porcine necrotizing staphylococcal
endometritis
S. epidermidis
• Opportunistic invader
S. intermedius
• prevalent in dogs and carnivores
S. hyicus subsp. hyicus
• Exudative epidermitis of swine
• Thru breaks on the skin
• Moist, greasy exudate on entire
body of animal
• Biotin requirement
Diagnosis
1. Smears
2. Culture on BAP
3. Mannitol salt agar (MSA)
4. Coagulase
5. Deoxyribonuclease
Immunity
• No effective vaccine
Antimicrobial Resistance
1. Beta lactamase
2. Methicillin resistant S. aureus
(MRSA) or multiple-drug resistant
Lesson 2: STREPTOCOCCUS
GENUS STREPTOCOCCUS
Morphological Characteristics
• Gram (+), non-motile, non-spore
forming coccus in pairs or chains,
capsulated
• Individual cells are round to ovoid,
0.6-1.0 um in diameter
• Catalase (-) facultative anaerobe
• Habitat: saprophytes in milk,
parasites of mucous membranes
and intestines
Classification:
1. Sherman
a. Pyogenic – pathogenic species
b. Viridans – a/green hemolysis
c. Lactic – associated with milk
d. Enterococcus – intestinal
inhabitant
2. Lancefield
• Serologically active carbohydrate (C
substance) which is antigenically
different from one species to another
• Six groups – A,B,C,D,E, and N
• Add 14 – F, G, H, K, L, M, O, P, Q,
R, S, T, U, V
• CHO or polysaccharide Ag found in
cell wall or between the cell wall and
cell membrane teichoic)
Cultural Characteristics
• More fastidious organisms require
enrichment with blood or serum
• Small, delicate, translucent colonies
about 1 mm in diameter
• Grow well in milk producing lactic
acid
S. agalactiae
• Long chains in secretions from
infected udder
• Brick reddish growth on solid
medium containing starch
• CAMP Test (Christie, Atkins, Munch-
Peterson)
• CAMP test: synergistic hemolysis
produced by sequential action of
staphylococcal sphingomyelinase
(B-toxin) and ceramide-binding
protein (N-acyl sphingosine) of
Streptococcus agalactiae
Pathogenesis
• In mammary glands of cows, sheep
and goats
• Spread thru milker’s hand,
contaminated milking machine,
mouth of calves
• Enters thru teats and colonize
mammary glands -> inflammation
and fibrosis of adjoining area -> milk
becomes alkaline and WBC counts
exceeds 500,000/ml -> milk
reduced, thin and watery
Prevention
• Vaccination not an effective means
of controlling
Diagnosis
1. Direct smears
2. Colony characteristics and
biochemical test
Streptococcus dysagalactiae
• Acute, severe mastitis
• Lancefield group C
• Often produces a distinct greenish
discoloration
Pathogenesis
• Infection much less frequent than
Streptococcus agalactiae
• Produces hyaluronidase
S. zooepidemicus
• Normal commensal of the skin and
upper respiratory tract, tonsils and
lymphoid tissues of horses
• Closely related to Streptococcus
equi and S.equisimilis
Pathogenesis
• Most common cause of wound
infections in horse
• Secondary invader of viral
infections of upper respiratory tract
of foals and young horse
• Mastitis in cows
• Fibrinous pleuritis, pericarditis and
pneumonia in lambs
S. uberis
• Bovine mastitis
• Does not react in Lancefield
grouping system
• Acute but mild
S. equisimilis
• Closely related to S. zooepidemicus
Pathogenesis
• Most frequent cause of suppurative
arthritis in pigs -> lameness, swelling
of joints and necrosis of joint
surfaces
• Tonsillar tissues and draining lymph
nodes
Prevention
• Vaccine shown to stimulate
protective immunity
S. equi
• Causes strangles, a severe purulent
infection of the upper respiratory
tract and draining lymph nodes in
horses
• Long chains in exudates and fluid
cultures
• Sometimes with definite capsular
material
• Outer surface of organism with
peach-fuzz like coating of protein
under electron microscope
Cultural and Biochemical Features
• Colonies matt or mucoid
• Matt colonies with irregular surface
folding and look dried -> due to
phage-controlled hyaluronidase
action on the hyaluronic acid
capsule
• O2 sensitive Streptolysin O-like
hemolysin produces a wide zone of
B-hemolysis
 • carries R antigen (acid, heat and
trypsin resistant but pepsin sensitive,
cross-reacts with S. zooepidemicus)
and M antigen (provides
antiphagocytic action)
Immunity
Pathogenesis
• Obligate parasite of family Equidae
• Thru nasal and oral route, ingestion
most common portal of
entry;inhalation of infective droplets
• Incubation period -6 days to 3 Diagnosis
weeks or longer
• Epithelial cells of oropharynx ->
lesion interiorized -> lymph drainage
to submandibular and
retropharyngeal
Prevention
Lymph nodes -> abscess formation
• Complement-derived chemotactic
factors C3a and C5a) release and
attract PMN cells -> powerful
cytotoxin produced by
microorganism damages phagocytes
-> extracellular multiplication of
microorganisms Other Streptococci
• Other virulence factors: M protein
and hyaluronic acid capsule -> high
rate of intracellular survival
• Symptoms: high fever, serous nasal
discharge -> mucopurulent ->
nasopharyngeal mucosa inflamed
and small abscesses develop in
lymphoid follicles of soft palate ->
submandibular and retropharyngeal
lymph nodes
• Abscesses rupture in 1-2 weeks
and animals recover quickly and
completely after drainage of purulent
material
• Bastard strangles – formation of
abscesses in thorax or abdomen ->
rupture results to death
Complications
1. Laryngeal hemiplegia
2. Guttural pouch empyema
3. Purpura hemorrhagica – side effect
of vaccination
• Immunity after recovery from first
vaccination
• Animals can contract the disease
again
1. RIA
2. ELISA
3. Mouse protection test
4. Gel diffusion precipitin test
• Heat inactivated bacterin or M-
protein rich extracts -> not useful
• Avirulent, genetically modified strain
stimulates local nasopharyngeal
antibodies
1. S. porcinus – cervical lymphadenitis
• Group E strepiococci
• Swine strangles
• Abscessation of mandibular,
retropharyngeal and parotid nodes -
> jowl abscessation
2. S. suis
• Meningitis and septicemia in young
pigs
• Lancefield group D
• Zoonotic
3. S. canis
• Mucous membranes of dogs and
cats
• Lancefield Group G
• Metritis and vaginitis in bitches
4. S. equinus
• Abundant in feces of horses
• Not known to be pathogenic
5. S. lactis
 • No pathogenic properties but
omnipresent in milk
• Common milk-souring organism ;
short chains
6. S. bovis
• Always present in mouth and
intestinal tract of cattle
Lesson 3: CLOSTRIDIUM
Toxin-forming, non-invasive group
1. Clostridium tetani
• Causative agent of tetanus
• Best known of all anerobic spore-
forming bacilli
Morphology
• Straight slender rod, 0.4-0.6 um x 2-
5 um
• Occurs singly in tissues and culture
• Spores formed after 24-48 hours at
end of rods 2-3x width of the rod ->
badminton-racket or drumstick-like
• Gram variable, peritrichous flagella
Cultural Characteristics
• Colonies in deep agar fluffy, cottony
spheres
• Brush-like effect in gelatin
• Does not ferment CHO
Resistance
• Highly resistant to sporicidal agents
and technics
• Boiling kills spores of most strains in
15 mins
• Antigen and Serotypes
• glycopeptide antigen, heat labile
flagellar or somatic antigen
• 9 serotypes
Pathogenesis
• Found in the soil, common in horse
manure
• Contamination of wounds and
umbilicus especially deep
penetrating wound which become
necrotic with reduced O2 and
lowered Eh (measure of redox,
measurement of electrical potential
expressed in volts)
• Produces 3 toxins
▪ Tetanospasmin
▪ Tetanolysin
▪ Peripherally active
non – spasmogenic
toxin
• Tetanospasmin responsible for
characteristic feature of tetanus
• Tetanolysin causes local tissue
necrosis which is favorable for
multiplication; hemolytic for RBC and
lethal in lab animals
Tetanospasmin
• Two subunits, heavy and light chains
which are non-toxic when separated
• Destroyed by gastric juices, heat
resistant, poorly absorbed across
mucous membranes
• Extremely potent, LD50 for mice is
equivalent to 2 x 10-8 mg
• Binds gangliosides which are bound
to cerebrosides, 2 gangliosides to 1
molecule toxin
• Reaction difficult to reverse and Ab
ineffective when toxin is bound
• Travels along nerve trunk to spinal
cord
• Action: prevents the release of
glycine, transmitter substance
responsible for inhibitory nerve
network of spinal cord -> prevents
contraction of muscle when its
opposite counterpart contracts ->
continuous stimulation and tetanic
spasms of groups of muscles
• Paralytic action on PNS and
inactivation of inhibitory nerve
network -> inhibition of protein
synthesis in brain
• Neuromuscular activity favors
migration of toxin along both motor
and sensory nerves
 • Toxin travels up regional motor
nerves where tetanus develops first -
> spreads upward in opposite limbs
and subsequently in muscles of the
trunk (ascending tetanus)
Treatment
1. Antitoxin asap in single IV or
intracisternal dose of 100,000 to
200, 000 units
2. Large doses of penicillin

• Tetanus develops first in susceptible Diagnosis

motor nerves of the head and neck
then descends to voluntary muscles
of forelimbs, upper trunk and
hindlimbs -> descending tetanus,
usually in humans and horses,
affecting nictitating membrane, facial
and jaw muscles causing lockjaw or
risus sardonicus
• Shorter incubation period, worse
prognosis
• Non-spasmogenic toxin
Portals of entry
1. Nails wounds in horses
2. Castration and docking in lambs
3. Calving, dehorning, castration and
nose ringing in cattle
4. Autointoxication
5. Castration in swine
6. Wound infection in dogs and cats
7. Umbilical cord in newborn
1. Characteristic symptoms
2. Drumstick-like spores
3. Injection to mice
Clostridium botulinum
• Source of potent neurotoxin which
causes botulism, a disease
characterized by flaccid paralysis
and eventual death due to
respiratory failure
• First discovered in Belgium from
imperfectly smoked ham
Forms
1. Intoxication
2. Wound infection
3. Infant botulism – intestinal
colonization
Morphology
• Spores oval, centric and excentric

Immunity Cultural and Biochemical Characteristics

1. Birds – naturally resistant, no Abs, • Acetic acid major metabolic product
brainhas no affinity of all strains
2. Sheep and goats neutralizing Abs • Non-proteolytic type acidify but do
3. Horse, dogs, pigs and human no not coagulate milk
antitoxin, power of binding with toxin • Proteolytic type slowly curdle milk
4. Hyperimmuned serum from horse, and partially digest and darken curd
1,500 units protective for 2-3 weeks • All strict anaerobes but not fastidious
5. Vaccines – 0.4% formalin -> • 6 serotypes, 7 toxin types
precipitated with aluminum
potassium sulfate suspended in Strain Hosts Disease
saline
o 3 doses at 3-week intervals A Humans, Cattle, Botulism,
o Horses: toxoids at 6 to 8- Horses, Chicken, Forage
week intervals followed by Mink poisoning,
booster 6-12 months later, Limberneck,
then annual booster Botulism.

B Humans, Cattle, Botulism

Horse, Chicken,
Mink, Foal
C Ducks, Wildbirds, Shaker foal functional change, vesicles containing
Humans syndrome, acetylcholine no longer able to release AC
Limberneck,
Westernduck • Toxin only affects cholinergic nerves
disease, of the PNSlood-brain barrier protect
Botulism cholinergic nerves
D Cattle Lamsiekte • Effect is flaccid paralysis which
progresses to involve the respiratory
Botulinum toxin muscles resulting to death
• Complex consisting of toxin • Disturbance in vision, locomotion
molecules and hemagglutiin moiety difficult, paralysis of the tongue,
• Toxins of non-proteolytic strains pharyngeal paralysis, respiratory
require proteolytic enzymes such as paealysis
trypsin to fully express its toxic • In poultry, nictitating membrane
activity • Swine and carnivores resistant to
• Toxin released during lysis of cells toxin

Pathogenesis Immunity
• From ingestion of food contaminated • Homologous antitoxin protective but
with preformed toxin not universally practiced in birds
• From decaying carcass, ingested or • Polyvalent toxoid
contaminating other food stuffs • Polyvalent antitoxin no longer
o Fly larvae carry toxins to effective once toxin is bound to
birds that feed on them nerve membranes
o Decaying vegetation on
edges of ponds and lakes -> Diagnosis
ducks can develop botulism • Demo of toxin in serum, intestinal
o Offal and fish meals contents and suspected foodstuffs
• Aphosphorosis causes depraved
appetite for decaying carcasses ->
occurrence of botulism
• Lameness associated with II. Tissue-invading, enterotoxigenic
phosphorus deficiency -> lamsiekte group
or lame sickness
• Toxicoinfectious botulism, toxin in Clostridium Disease
necrotic areas of the body (foal) -> Perfringens
shaker foal syndrome -> A Yellow lamb
characterized by dysphagia, disease
tremors, weakness, recumbency B Lamb dysentery,
followed by death hemorrhagic
enteritis of sheep
Mechanism of Infection and goats
C Necrotic enteritis,
Ingestion -> intestinal wall (protected by Struck in sheep
hemagglutinins from digestive processes) -> D Enterotoxemia –
ruminal bacteria inactivate substantial overeating ds,
quantity of ingested toxin -> bloodstream -> pulpy kidney ds
peripheral nervous system -> binds to E Enterotoxemia in
gangliosides at neuromuscular junction -> lambs and
inside surface of cell membrane -> calves
 • Alpha toxin in small intestine is
Clostridium Red water disease absorbed into circulation causing
hemolyticum Hemorrhagic massive intravascular hemolysis and
disease, Inf. capillary damage
icterohemoglobinuria
C. novyi Big head in rams C . perfringens Type B
Black diseases • Lamb dysentery
(necrotic hepatitis in • Beta toxin produces hemorrhagic
sheep) zones and ulcerations
C. chauvoei Blackleg, black • of the small intestines
quarter, Quarter evil,
symptomatic anthrax C. perfringens Type C
C. septicum Malignant edema • Enterotoxemia
Braxy or bradsot • Beta toxin, labile and denatured by
C. colinum Ulcerative enteritis enzymes in the intestines
or quail ds • Acute hemorrhagic enteritis in pigs
C. spiroforme Iota enterotoxemia • Struck or Romney Marsh disease in
in rabbits sheep
C. difficile Enterotoxemia in • Beta toxin produces necrosis of
hamsters mucosa in the abomasum and small
C. villosum Subcutaneous intestine
abscesses in cats
C. perfringens Type D
Clostridium Perfringens • Pulpy kidney disease, overeating
• Synonym. C. welchii, gas bacillus, disease
Welch bacillus • Epsilon toxin which requires trypsin
• Five types, A to E, four major lethal or chemotrypsin activation and has a
toxic permease effect on the intestinal
• Spores oval, not much swelling mucosa enhancing absorption ->
• Old cultures pleomorphic, clubbed results to foci of liquefactive
types, ballooned cells and filaments necrosis, perivascular edema and
• Capsules in tissues, non-flagellated hemorrhages especially in the
meninges -> receptor site on
Cultural and Biochemical Features vascular endothelium of the brain ->
• Inner zone of complete and outer breakdown of intercellular junctions
zone of incomplete hemolysis and escape of fluids
• Produces lecithinase (phospholipase • Activated epsilon has pressure
C) activity which increases blood
• Stormy fermentation of litmus milk pressure
• 4 toxins: alpha (phospholipase),
beta, epsilon, iota C. perfringens Type E
• Hemorrhagic, necrotic enteritis in
Pathogenesis calves
• Ubiquitous in nature, part of normal
intestinal flora Immunity
• Strain A in soil, B-E in intestines • Alum precipitated type-specific
• Most common cause of necrotizing toxoid
myositis in horses
• C. perfringens Type A Diagnosis
• Toxin in intestinal contents
Clostridium hemolyticum
• Closely related to C. novyi (type D)
• Spores are oval and subterminal,
causes blging
• Motile
Cultural and Biochemical Features
• Exacting in cultural requirements
good anaerobic conditions, medium
must contain tryptophan
Antigens and Toxins
• C. hemolyticum, beta toxin only; C.
novyi, type B, alpha and beta toxins
Pathogenesis
• Predilection for alkaline water
• Site of toxin production is liver
• Tissue destruction caused by
migration of liver flukes which
provides suitable microenvironment
for germination of spores
• Beta toxin (phospholipase C) causes
massive intravascular hemolysis and
capillary damage -> hemorrhage into
lumen of intestine and Hb in urine ->
dark red or port-colored, clear but
foamy -> no intact RBC
• At time of hemoglobinuria, 40 to
50% of hemoglobin destroyed
• Death caused by anoxemia due to
massive destruction of
RBC
• Most characteristic lesion is large
infarct in liver as a result of
occluding thrombosis
Immunity
• Alum-precipitated, formalinized
whole culture
Diagnosis
1. Culture from liver lesions
2. Demonstration of phospholipase C
in liver lesions
Clostridium novyi
• Novy’s bacillus edematis maligni II
• Three types: A, B, C
Morphology
• Spores oval and subterminal
• Motile
Cultural and Biochemical Features
• More strictly anaerobic and requires
• Cysteine in reduced form
Pathogenesis
• In soil and intestinal tract of
herbivores
• Multiply in wounds contaminated by
soil and cause gas gangrene
• Big head in rams as result of
infection of wounds sustained
In fighting
• Alpha toxin damages capillary
endothelium at site of invasion,
brain, muscle, liver, heart ->
elevation of intracellular enzymes
lsctic dehydrogenase and glutamic
oxaloacetic transaminase
Black disease – necrotic hepatitis in
sheep
• Liver damage from migration of
immature Fasciola hepatica
• Alpha and lesser beta toxin
• Necrotic areas on surface of liver
• Extensive blood-stained edema
under skin -> black disease
• Immunity
• Alum precipitated, formalinized
whole culture
• Vaccination before heavy fluke
activity
Diagnosis
1. Demo of alpha toxin in tissues and
exudates
• FAT – fluorescein antibody
technique
Clostridium chauvoei (Synonym
Clostridium feseri)
• Blackleg in ruminant
Morphology
 • Spores oval and excentrical,
swelling rods into lemon-shaped
structures
Cultural and Biochemical Features
• Strictly anaerobic, blood or serum
favors growth in ordinary media,
high requirement for cysteine
Antigens
• Flagellar, somatic and spore
antigens
• Toxins include alpha, hyaluronidase
and deoxyribonuclease
Pathogenesis
• In soil, unknown if it lives in the soil
and multiplies in the intestine
• Entry via oral route during grazing ->
multiplies in the intestine ->
lymphatic and blood circulation ->
muscle and liver -> dormant until
muscle mass becomes altered of
damaged
• May enter thru alveoli of the teeth
• necrotizing, leukocidic and
spreading factor of alpha toxin and
hyaluronidase promote development
of myonecrosis
• Area reddish brown to black,
crepitant with spongy texture due to
entrapped gas, dry on cut surface
• Affected area first swollen, painful
and crepitant -> sensation lost and
skin becomes tighter
Immunity
• Formalinized whole culture or
anacultures with alum
Diagnosis
1. Organism in heart, liver and
peritoneal fluid
2. FAT
Clostridum septicum
• Ghon-Sachs bacillus, Malignant
edema bacillus
• Malignant edema
Morphology
• Spore oval, excentrical and swell
cells
Cultural and Biochemical Feature
• Grows readily on all ordinary media
with good anaerobic condition
Antigens
• Somatic and flagellar agglutinogens
Patogenesis
• Common in soil and intestinal tract
• In lambs thru wounds, umbilicus and
abomasal lining (braxy or bradsot)
• Wound infection -> malignant edema
-> sewllings are soft and pit on
pressure -> large amounts of
gelatinous exudate, muscular tissue
dark red but contains little or no gas
• Parablackleg bacillus
• 4 toxins: alpha, beta, gamma, delta
Lesson 4: BACILLUS
Genus Bacillus
• Almost all are non-pathogenic and
saprophytic
• Most pathogenic is B. anthracis
Bacillus anthracis
• Causes anthrax, milzbrand
(German), charbon (French)
• Herbivores highly susceptible,
carnivores and birds resistant
• Fatal in humans
Morphology
• Large, Gram (+) rods, 1um x 3-6 um
• In cultures, forms long chains which
appear as solid filaments because of
square ends that fit closely together
• In tissues, short chains with ends of
cells rounded
• Single capsule enclose organisms in
chain
 • Spores formed at 15-40’C in the
presence of air -> sporulation
inhibited by high CO2 tension such
as carcasses
• Spores central to subterminal and
not swollen
Cultural and Biochemical Features
• Grows well on most lab media
• Spores germinate at 65’C for 15
mins
• Ground glass-like appearance on
surface colonies on agar with
irregular margins -> under LPO
resemble locks of wavy hair known
as Medusa-head colonies
• Deep colonies small, ragged and
stringy
• Biochemically, much less active than
other bacilli
• Non-motile
• Capsule made up of polymers of D-
glutamic acid and some
polysaccharide
• Spores resistant to boiling but killed
by autoclaving
• Resistant to disinfectants and
remain viable for more than 50
• years
• Vegetative cells killed at 60’C for 30
mins and quickly destroyed by
enzymes and putrefactive bacteria’
Pathogenesis
• Major anthrax enzootic zones in
tropics and subtropics such as
India, Pakistan, Africa, South
America
1. Alkaline soil with high nitrogen level
caused by decaying vegetation
2. Alternating period of rain and
drought
3. Temperatures beyond 5.5 ‘C
Source of infection
1. Soil
2. Bone meal and vegetable protein
3. Wool and hair
4. Tannery effluents
5. Bloodsucking flies and carrion
eaters
• Infection via the skin and respiratory
tract usually by ingestion of spores -
> germination of spores in the
mucosa of throat and intestinal tract
-> multiplication at site of primary
invasion -> lymphatic channel ->
lymph nodes -> multiplication ->
bloodstream -> spleen
• If splenic clearance exceeded,
animal dies -> 80% of organisms in
blood, 20% in spleen
• Death due to extracellular toxin
Toxin complex or holotoxin
1. Edema factor (EF)
2. Protective antigen (PA)
3. Lethal factor (LF)
• Edema factor – Adenylate cyclase ->
increases intracellular cyclic AMP
• Protective antigen – plasma-
encoded, necessary for biological
activity of EF and LF; probably a
receptor binding molecule
Net effect of holotoxin
1. Damage and kill phagocytes
2. Increased capillary cell permeability
3. Damage clotting mechanism
 4. Inhibit bactericidal activity of serum
• Capillary thrombosis -> fluid leaks
thru damaged endothelium -> blood
pressure falls -> shock
• Blocks opsonizing activity of C3
factor of complement -> reduced
phagocytosis
3. Ground glass-like non-hemolytic
colonies on BAP -> Medusa-head or
judge’s wig-type colnies
4. String of pearl test – in presence of
penicillin, organisms grow with cell
wall impairment -> organisms
resemble string of pearls

Symptoms: 5. Bacteriophage (gamma phage) ->

only B. anthracis lysed
1. Edema
6. Animal test – pathogenic in guinea
2. Shock pigs and mice, death within 24 hrs
3. Hemorrhaging before death Immunity
• Peracute form 1-2 hours, acute form • Recovered animals have permanent
less than 24 hours immunity
Forms of anthrax • Immunization with Sterne’s live,
noncapsulated, avirulent spore
1. Localized anthrax – thru wound on vaccine or culture filtrate of avirulent,
skin; cutaneous form; malignant noncapsulated strain
carbuncle or pustule
Prevention and Control
• More often in humans than in
animals • Reportable disease
• Often becomes generalized • Should be treated, uninfected
• In swine and dogs, localized form animals immunized
with pharyngeal involvement and
gastroenteritis -> inflammatory
edema of head and neck ->
suffocation
2. Generalized anthrax – airborne
spores; pulmonary or inhalation
anthrax; woolsorter’s disease
Diagnosis

• By methods that release minimum

number of anthrax organisms in the
environment; blood samples from
ears veins
1. Mc Fadyean’s stain – organisms
blue, capsules pink
2. Capsular antigen detected by agar
gel precipitation test /AGPT known
as Ascoli test