Bakteri Gram positive

pembentuk spora
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 Spores
• Why do bacteria produce spores?
Survival
• Definition
a resting cell, highly resistant to dessication,
heat, and chemical agents; when returned to
favourable conditions bacteria re-activated,
the spores germinate to produce single
vegetative cells.
• The gram-positive spore-forming bacilli are
the Bacillus and Clostridium species. The
bacilli are ubiquitous, and because
they form spores, they can survive in the
environment for many years. Whereas the
Bacillus species are aerobes, the Clostridium
species are anaerobes.
 Gram-positive Bacilli
Spore-Forming Non-Spore Forming
Bacteria Bacteria
Bacillus Corynebacterium
Clostridium Actinomyces
Sporolactobacillus
 Bacillus
• Aerobic, G+ rods in chains, spores are located in
center of the non-motile bacilli
• Found in soil, water, air and vegetation
• Spores are viable for decades.
• B. cereus – produce enterotoxin and cause food
poisoning.
• B. anthracis – infection in human through injured
skin (cutaneous anthrax), mucous membranes (GI
anthrax), or inhalation of spores into lung.
 Typical Organisms

• The typical cells, measuring 1 × 3–4 µm, have

square ends and are arranged in long chains;
spores are located in the center of the
nonmotile bacilli.
 Culture

• Colonies of B anthracis are round and have a

“cut glass” appearance in transmitted light.
Hemolysis is uncommon with B anthracis but
common with B cereus and the saprophytic
Bacilli.
• Gelatin Is liquefi ed, and growth in gelatin
stabs resembles an Inverted fir tree.
 growth Characteristics

• The saprophytic bacilli use simple

sources of nitrogen and carbon
for energy and growth.
• Th e spores are resistant to
environmental changes, withstand
dry heat and certain chemical
disinfectants for moderate periods,
and persist for years in dry earth.
• Animal products contaminated with
anthrax spores (eg, hides, bristles,
hair, wool, bone) can be sterilized by
autoclaving
Bacillus anthracis
 Bacillus
• Spores germinate in the tissue of entry, and
growth of vegetative organisms result in
formation of a gelatinous oedema and
congestion.
• Spread via lymphatics to bloodstream and
multiply freely in blood and tissues.
• Capsulated, poly-D-glutamic acid capsule is
antiphagocytic
 Bacillus
• Anthrax toxin is made up of three
proteins:
Protective antigen (PA),
Oedema Factor (EF) and
Lethal Factor (LF).
• PA binds to specific cell receptors, and
after proteolytic activation, it forms a
membrane channel that mediates entry
of EF and LF into the cell.
• EF is an adenylate cyclase; with PA, it
forms a toxin known as edema toxin. LF
plus PA form lethal toxin, which is a
major virulence factor and cause of
death
 Cutaneous anthrax
• generally occurs on exposed surfaces of the arms or hands
followed in frequency by the face and neck.
• A pruritic papule develops 1–7 days after entry of the
organisms or spores through a scratch.
• The papule rapidly changes into a vesicle or small ring of
vesicles that coalesce, and a necrotic ulcer develops.

• The lesions typically are 1–3 cm in diameter and have a

characteristic central black eschar. Marked edema occurs.
• Lymphangitis and lymphadenopathy and systemic signs and
symptoms of fever, malaise, and headache may occur.
• After 7–10 days, the eschar is fully developed.
• Eventually, it dries, loosens, and separates; healing is by
granulation and
leaves a scar. It may take many weeks for the lesion to heal
and the edema to subside
 inhalation anthrax
• The incubation period 6 weeks
• hemorrhagic necrosis and edema of
the mediastinum. Substernal pain
• Hemorrhagic pleural effusions
• Cough
• Sepsis
• may be hematogenous spread to the
gastrointestinal tract, causing bowel
ulceration, or to the meninges,
causing hemorrhagic meningitis.
• The fatality rate in inhalation anthrax
is high
 gastrointestinal anthrax
• Animals acquire anthrax through ingestion of
spores and spread of the organisms from the
intestinal tract. This is rare in humans, and
gastrointestinal anthrax is extremely
uncommon.
• Abdominal pain, vomiting, and bloody
diarrhea
• Treatment: ciprofloxacin, penicillin G along
with gentamicin and streptomycin.
• Vaccine with live spores and a toxoid used to
protect livestock in endemic areas.
 Laboratory diagnostic
• Specimens to be examined are fluid or
pus from a local lesion, blood, pleural
fluid, and cerebrospinal fluid in
inhalational
• anthrax associated with sepsis and stool
or other intestinal contents in the case of
gastrointestinal anthrax.
• Stained smears from the local lesion or
of blood from dead animals often show
chains of large gram-positive rods.
Anthrax can be identified in dried smears
by immunofluorescence staining
techniques.
• blood agar plates, the organisms produce
nonhemolytic gray to white, tenacious colonies with a
rough texture and a ground-glass appearance. Comma-
shaped outgrowths (Medusa head, “curled hair”) may
project from the colony.
• Demonstration of capsule requires growth on
bicarbonate-containing medium in 5–7% carbon
dioxide. Gram stain shows large gram-positive rods.
• In semisolid medium, anthrax bacilli are always
nonmotile, but related organisms (eg, B cereus) exhibit
motility by “swarming.”
 Clostridium
• Anaerobic, G+, motile rods, typical tennis racquet
morphology is created by terminal spores that swell
the sporangium
• Clostridial diseases from wounds inc tetanus (NM
disease) and gas gangrene (soft tissue infection that
damages muscle)
• Found in soil, animal faeces.
• Spores is placed centrally, subterminally or
terminally; most species are motile with flagella.
 Clostridium
• Many form colonies with a zone of
haemolysis on blood agar. C perfringens
typically produce multiple zones of
haemolysis around colonies.
 Clostridium botulinum
• C botulinum causes botulism, from eating poorly
preserved food (canned).
• Spores are resistant to 100°C for many hours,
diminished at acid pH or high salt.
• C botulinum is distinguished by antigenic type of
toxin
• Toxin - 7 antigenic varieties (A →G). A, B, E (F) mainly
harmful to human.
• Botulinum toxin is absorbed from gut and binds to
receptors of presynaptic nervous system and cranial
nerves.
 Clostridium
• Toxin acts by blocking release of
acetylcholine at synapses and
neuromuscular junctions → flacid
paralysis.
• Symptoms such as visual disturbances,
inability to swallow, speech problem;
seldom with no apparent GI symptoms;
no fever.
• ‘floppy baby’ = infant botulism. C
botulinum spores ingested from babies’
food. Present with weak sucking
response, loss of tone and respiratory
complications.
 Clostridium
• Treatment – antitoxins raised in
horses.
• Trivalent (A, B, E) antitoxin must be
promptly administered intravenously
with precautions; plus adequate
ventilations.
• Treatment to control the microbe’s
growth and toxin production.
• Clostridium tetani cause tetanus or
lockjaw.
Clostridium Tetani

Pathogenesis of tetanus caused by C

tetani
General introduction

• C tetani is found worldwide.

Ubiquitous in soil, it is occasionally
found in intestinal flora of humans
and animals
• C.tetani is the cause of tetanus,or
lockjaw. When spores are
introduced into wounds by
contaminated soil or foreign objects
such as nails or glass splinters
 BIOCHEMICAL CHARACTERISTICS
• Morphology: long and
slender; peritrichous
flagella,no capsule, terminal
located round spore(drum-
stick apperance), its diameter
greater than vegetative cell.
• Culture:obligate anaerobic;
Gram(+); swarming occures
on blood agar, faint hemolysis.
• Biochemical activities:does
not ferment any carbohydrate
and proteins.
• Resistance: tolerate boiling
for 60 min.alive several ten
years in soil.
• Classification and Antigenic 2-5 x 0.3-
Types: C tetani is the only
species. There are no
0.5um
serotypes
 Pathogenicity
• No invasiveness; toxemia  retrograde
(exogenous infection） transport to (CNS)
• produces two exotoxins:
tetanolysin, and
 delitescence：a
tetanospasmin(a kind of few days to several
neurotoxin, toxicity weeks
strong)  The two animal
• The actions of species most
tetanospasmin are
susceptible to this
complex and involve
three components of the toxemia are horses
nervous system: central and humans.
motor control, autonomic
function, and the
neuromuscular junction.
Clostridium tetani -Tetanospasmin

• disseminates systemically
• binds to ganglioside receptors
– inhibitory neurones in CNS
• glycine
– neurotransmitter
• stops nerve impulse to muscles
• spastic paralysis
• severe muscle contractions and
spasms
• can be fatal
 Clinical Manifestations
• The initial symptom is cramping and
twitching of muscles around a wound. The
patient usually has no fever but sweats
profusely and begins to experience pain,
especially in the area of the wound and
around the neck and jaw muscles (trismus).
• Portions of the body may become extremely
rigid, and opisthotonos (a spasm in which
the head and heels are bent backward and
the body bowed forward) is common.
• Complications include fractures, bowel
impaction, intramuscular hematoma, muscle
ruptures, and pulmonary, renal, and cardiac
problems
 Clinical Manifestations
DISEASE CLINCAL MANIFESTATIONSA

Generalized Involvement of bulbar and paraspinal

muscles(trismus or lockjaw, risus sardonicus,
difficulty swallowing, irritability,
opisthotonos);involvement of autonomic
nervous system(sweating, hyper thermia,
cardiac arrhythmias, fluctuations in blood
pressure)
Cephalic Primary infection in head,particularly
ear;isolated or combined involvement of
cranial nerves, particularly seventh cranial
nerve; very poor prognosis

Localized Involvement of muscles in area of primary

injury; infection may precede generalized
disease; favorable prognosis

Neonatal Generalized disease in neonates; infection

typically originates from umbilical
stump;very poor prognosis in infants whose
mothers are nonimmune
 Epidemiology
• 1 million cases of tetanus occur annually in the
world,with a mortality rate ranging from20% to
50%. But rare in most developed countries.
• In some developing countries, tetanus is still
one of the ten leading causes of death, and
neonatal tetanus accounts for approximately
one-half of the cases worldwide.
• In less developed countries, approximate
mortality rates remain 85% for neonatal
tetanus and 50% for nonneonatal tetanus.
• In the United States, intravenous drug abusers
have become another population with an
increasing incidence of clinical tetanus
• In untreated tetanus, the fatality rate is 90%
for the newborn and 40% for adults.
 Immunity

• Humoral immunity(antitoxin)
• There is little, if any, inate immunity
and the disease does not produce
immunity in the patient.
• Active immunity follows vaccination
with tetanus toxoid
 Diagnosis
• Diagnosis is primarily by the clinical
symptoms (above). The wound may not
be obvious.
• C tetani can be recovered from the
wound in only about one-third of the
cases.
• It is important for the clinician to be
aware that toxigenic strains of C tetani
can grow actively in the wound of an
immunized person.
• Numerous syndromes, including rabies
and meningitis, have symptoms similar
to those of tetanus and must be
considered in the differential diagnosis.
 Vaccination

• infant
• DPT (diptheria, pertussis,
tetanus)
• tetanus toxoid
– antigenic
– no exotoxic activity
 Control
• The offending organism must be removed by
local debridement
• toxoid
• TAT (tetanus anti toxin) ; Metronidazole (For
more serious wounds)
• AIDS patients may not respond to
prophylactic injections of tetanus toxoid
 C. perfringens
• soil, fecal
contamination
• gas gangrene
– swelling of
tissues
– gas release
* fermentation
products
• wound
contamination
 Toxins
toxin Biological Feature Types of Toxins
A B C D E
 lecithinase; increase + + + + +
the vascular
permeability;
hemolytic; produces
necrotizing activity
 Necrotizing activity, － + + － －
induces
hypertension by
causing release of
catecholamines.
 increase the － － － + －
permeability of
gastrointestinal wall
 Necrotizing activity; － － － － +
increase the
vascular
permeability
 Toxins
• Many of these toxins have lethal,
necrotizing, and hemolytic properties;
• The alpha toxin produced by all types
of C. perfringens, is a lecithinase that
lyses erythrocytes, platelets,
leukocytes, and endothelial cells. And
its lethal action is proportionate to the
rate at which it splits lecithin to
phosphorylcholine and diglyceride.
• The theta toxin has similar hemolytic
and necrotizing effects. DNAase,
hyaluronidase, a collagenase are also
produced
 Enterotoxin
• Many strains of type A produce
enterotoxin, which is a heat-labile
protein and destroyed immediately at
100 ℃.
• Trypsin treatment enhances the toxin
activity threefold.
• The toxin is produced primarily by type
A strains but also by a few type C and D
strains.
• It disrupts ion transport in the
ileum(primarily) and jejunum by
inserting into the cell membrane and
altering membrane permeability.
• As superantigen.
 Pathogenesis

•Tissue degrading enzymes

– lecithinase [ toxin]
– proteolytic enzymes
– saccharolytic enzymes
• Destruction of blood vessels
• Tissue necrosis
• Anaerobic environment
created
• Organism spreads
Without treatment death
occurs within 2 days
 effective antibiotic therapy
 debridement
 anti-toxin
 amputation & death is rare
 Gas gangrene
• Gas gangrene is a life-threatening disease with
a poor prognosis and often fatal outcome.
• Initial trauma to host tissue damages muscle and
impairs blood supply----lack of oxygenation
• Initial symptoms : fever and pain in the infected
tissue.; more local tissue necrosis and systemic
toxemia. Infected muscle is discolored (purple
mottling) and edematous and produces a foul-
smelling exudate; gas bubbles form from the
products of anaerobic fermentation.
 Gas gangrene
• As capillary permeability increases, the
accumulation of fluid increases, and venous
return eventually is curtailed.
• As more tissue becomes involved, the
clostridia multiply within the increasing area
of dead tissue, releasing more toxins into the
local tissue and the systemic circulation.
 Food poisoning

• Enterotoxin producing strains.

• These bacteria are found in mammalian
faeces and soil.
• Small numbers of the bacteria may also be
found in foods and they may propagate
rapidly to dangerous concentrations if the
food is improperly stored and handled.
 Food poisoning

• more than 108 vegetative cells are

ingested and sporulate in the gut,
the toxins can act rapidly in the body,
causing severe diarrhea in 6-18
hours, dysentery, gangrene, muscle
infections
• The action of C. perfringens
enterotoxin involves marked
hypersecretion in the jejunum and
ileum, with loss of fluids and
electrolytes in diarrhea.
 Cellulitis, Fasciitis

• Cellulitis, Fasciitis
• Fasciitis : a rapidly progressive, destructive process in
which the organisms spread through fascial plan es.
• Fasciitis causes suppuration and the formation of gas
• Absense of muscle involvement
• Necrotizing Enteritis
• Rare, acute necrotizing process in the
jejunum
• Abdominal pain, bloody diarrhea, shock,
and peritonitis
• Mortality: 50%
• Beta-toxin-producing C. perfringens type
C
• Septicemia
 Who is at risk?
• Surgical patients; patient after trauma with
soil contamination

• People who ingest contaminated meat

products (without proper refrigeration or
reheating to inactivate endotoxin)
 Epidemiology
• C. perfringens type A: the intestinal tract of
humans and animals, soil and water
contaminated with feces. forms spores under
adverse environmental conditions and can
survive for prolonged periods.
• Type B to E strains colonize the intestinal tract
of animals and occasionally humans.
 Epidemiology
• Type A: gas gangrene, soft tissue infections
and food poisoning
• Type C: enteritis; necroticans
 Laboratory identification

• lecithinase production

Double Hemolysis Circles

C. botulinum
 Biological Features

• Anaerobic
• Gram-positive
• rod-shaped
• sporeformer
• produces a protein neurotoxic.
• soil, sediments of lakes, ponds,
decaying vegetation.
• intestinal tracts of birds, mammals
and fish.
 Division

---A, B, C1, D, E, F,
and G.
---type A. 62%
---Not all produce
toxin.
---C and D not
 Transmission
---spores heat resistant.
canning.
anaerobic environment
---Botulism
eating uncooked foods
spores
---GI, duodenum, blood stream,
neuromuscular synapses.
 Virulence factors

---bacterial
protease
---light
chain,A,50 kDa;
heavy
chain,100kDa.
---disulfide bond.
---A potent toxin
• binds peripheral nerve receptors
– acetylcholine neurotransmitter
• inhibits nerve impulses
• flaccid paralysis
• death
– respiratory Botulinum toxin
– cardiac failure
 Botulinum toxin
• Bioterrorism
– not an infection
– resembles a chemical attack
– 10 ng can kill a normal adult
 Clinical syndromes

---18-36 hours:
---weakness, dizziness,dryness of
the mouth.
---Nausea,vomiting.
---Neurologic features: blurred
vision, inability to swallow,
difficulty in speech, descending
weakness of skeletal muscles,
respiratory paralysis.
 Botulism

• food poisoning
– rare
– fatal

• germination of spore
• inadequately sterilized canned
food
– home
• not an infection
Infection with C. botulinum

• Neonatal botulism
– uncommon
– the predominant form of
botulism
– colonization occurs
• no normal flora to compete
• unlike adult
 Wounds

– extremely rare
– an infection
 Immunity

---specifically neutralized, antitoxin.

---toxoided, make good antigens.
---does not develop, amount toxic.
---Repeated occurrence.
---Once bound, unaffected by antitoxin.
---circulating toxin ,neutralized ,
injection of antitoxin.
---treated immediately with antiserum.
---multivalent
toxoid,unjustified,infrequency.
experimental vaccine.
 Diagnosis

---by clinical symptoms alone

---differentiation difficult.
--- most direct and effective:
serum or feces.
---most sensitive and widely
used: mouse neutralization
test. 48h.
Culturing of specimens 5-7d.
 Treatment

• Individuals known to have ingested

food with botulism should be
treated immediately with antiserum.
• antibiotic therapy (if infection)
– Vaccination will not protect hosts
from botulism, however passive
immunisation with antibody is
the treatment of choice for cases
of botulism.
 Prevention

---proper food handling and

preparation.
--- spores survive boiling (100 degrees
at 1 atm) 1h.
---toxin heat-labile, boiling or intense
heating, inactivate the toxin.
---bulge, gas, spoiled.
 C. difficile

• After antibiotic use

• Intestinal normal flora --greatly
decreased
• Colonization occurs
• Enterotoxin secreted
 Pseudomembranous Colitis

• Pseudomembranous colitis (PC) results

predominantly as a consequence of the
elimination of normal intestinal flora
through antibiotic therapy.
• Symptoms include abdominal pain with
a watery diarrhea and leukocytosis.
"Pseudomembranes" consisting of
fibrin, mucus and leukocytes can be
observed by colonoscopy.
• Untreated pseudomembranous colitis
can be fatal in about 27-44%.
 Therapy

• Discontinuation of initial antibiotic

(e.g. ampicillin)

• Specific antibiotic therapy (e.g.

vancomycin)
CLOSTRIDIA
Non-Spore Forming Bacteria
• Generally, members of normal flora of
skin and mucous membranes of
humans.
Aerobic G+ with Aerobic G+ with
High G+C content, lower G+C content,
irregularly shaped regularly shaped
Corynebacteriu Listeria
m Lactobacillus
Propionibacteriu Erysipelothrix
m
Actinomyces
Rhodococcus