Colstridium
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Transcript of Colstridium
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Anaerobes of clinical Importance
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Oxygen
ClassDefinitionExamples
Obligate aerobe
Grow only in the presence of O2
Mycobacterium tuberculosis
Obligate anaerobe
Can not grow in the presence of O2
Clostridia
Facultative anaerobe
Can grow in the presence or absence of O2
Most bacteria of medical importance
Microaerophilic bacteria
Require low O2 tension
Campylobacter
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22222 OO2H2OO2H
In the presence of oxygen, two toxic substances to the bacteria are produced which are hydrogen peroxide and superoxide anion.
In obligate aerobes and facultative anaerobes:
Catalase and peroxidase enzymes degrade hydrogen peroxide.
Superoxide dismutase enzyme degrades superoxide anion.
BUT
In obligate anaerobes: These enzymes are not present. So, the presence of oxygen is toxic to them.
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CLASSIFICATION
Anaerobic spore forming bacilli (Clostridia)Gram negative bacilli non-sporing forming
(Bacteroides)Anaerobic streptococci (Peptostreptococcus)Anaerobic staphylococcus (Peptococcus)Gram negative diplococci (Veillonella)Gram positive bacilli (Actinomyces)
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HABITAT I:
These organism are normal flora in:• A. Oropharynx eg. . Fusobacteria and
Veillonella • B. Gastrointestinal tract
– Found mainly in the large colon in large numbers– Total number of anaerobes = 10 11
– While all aerobes (including E. coli) = 10 4
– examples are (1) B acteroides fragilis (2) Bifidobacterium species
• C. Female genital tract (mainly in the vagina)
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Features of anaerobic infections• Infections are always near to the site of
the body which are habitat.1.Infection from animal bites.2.Deep abscesses3.The infections are also polymicrobial 4.Gas formation, foul smell5.Detection of "Sulphur granules"' due to
actinomycosis 6.Failure to grow organism from pus if not
culture anaerobically.7.Failure to respond to usual antibiotics.
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Infections begin :• Disruption of barriers
–Trauma–Operations–Cancerous invasion of tissues
• Disruption of blood supply–Drops oxygen content of tissue–Tissue necrosis
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what are the infection caused by these anaerobic organisms
Post operative wound infectionBrain, dental, lung abscessIntra abdominal abscess, appendicitis,
diverculitis Infection of the female genital tract: Septic
abortion, endometritis , , puerperal infection and endometritis , pelvic abscess or breast abscess
Diabetic foot infections
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Laboratory diagnosis:
• When anaerobic infection is suspected;a) Specimens have to be collected from the site
containing necrotic tissue.b) Pus is better than swabs.c) Specimens has to be send to the laboratory
within 1/2 hour d) Fluid media like cooked meat broth are the
best culture media.e) Specimens have to incubated anaerobically
for 48 hours.
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Organism groupsGram negative rods
BacteroidesPrevotellaPorphyromonasFusobacterium ButyrivibrioSuccinomonas
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Bacteroides
Strict anaerobePleomorphicGram negative bacilli (cocco bacilli)Normal flora in
Oropharynx Gastrointestinal tractVagina
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Bacteroides
• B. Fragilis, B. Vulgaris, B. Uniformis–Account for 1/3 of all isolates–Resistant to 20% bile–Resistant to many antibiotics
• Penicillin, kanamycin, vancomycin, colistin – and many more
–No pigmentation of colonies or fluorescence
•
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Bacteroides other species
Bacteroides species other than b. Fragilis groupBile sensitiveResistant to kanamycin onlySome pigmented
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Other gram negative rods• Fusobacterium necrophorum• Gram negative bacilli• Peritonisillar intrnal jugular vein
thrombosisemboli to the lung• Peptococcus • Gram positive cocci in clusters• Peptostreptococcus• Gram positive cocci in chains• Brain abscess• Veillonella parvula• Gram negative cocci
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Clostridium SPP
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Clostridium spp. Anaerobic Gram-Positive Spore-Forming Bacilli Four broad types of pathogenesis:
1. Histotoxic group — tissue infections (C. perfringens type A, exogenously acquired more commonly than endogenously) (C. septicum; endogenously-acquired)
a. cellulilitis b. myonecrosis c. gas gangrene d. fasciitis
2. Enterotoxigenic group — gastrointestinal disease a. clostridial foodbome disease (8-24h after ingestion of large numbers of organisms on con-taminated
meat products, spores germinate, enterotoxin produced (C. perfringens type A) b. necrotizing enteritis (beta toxin-producing C.perfringens type C)
(C. difficile endogenously-acquired or exogenously-acquired person-to-person in hospital) c. antibiotic-associated diarrhea d. antibiotic-associated pseudomembrane colitis
3. Tetanus (exogenously acquired) — C. tetani neurotoxin
a. generalized (most common) b. cephalic(primary infection in head, comnnonly ear) c. localized e. neonatal (contaminated umbilical stump)
4. Botulism (exogenously acquired) — C. botulinum neurotoxin
a. foodborne (intoxication,1-2days incubation period) b. infant (ingestion of spores in honey) c. wound (symptoms similar to foodborne, but 4 or more days incubation)
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Spores Clostridium form endospores under
unfavourable environmental conditions
Spores are a survival mechanism
Spores are characterized on the basis of position, size and shape
Most Clostridium spp., including C. perfringens and C. botulinum, have ovoid subterminal near an end) ( (OST) spores
C. tetani have round terminal (RT) spores
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Clostridium Associated Human Disease
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Clostridium perfringens
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Clostridium perfringens — histotoxic or enterotoxigenic infections
Morphology and Physiology • large, rectangular bacilli (rod) staining gram-positive • spores rarely seen in vitro or in clinical specimens (ovoid, subterminal) • non-motile, but rapid spreading growth on blood agar mimics growth of motile organisms • aerotolerant, especially on media supplemented with blood • grow at temperature of 20-50°C (optimum 45°C) and pH of 5.5-8.0
Pathogenicity Determinants (note that toxins include both cytolytic enzymes and bipartite exotoxins) • four major lethal toxins (alpha (), beta (), epsilon (), and iota () toxins) and an enterotoxin • six minor toxins (delta(), theta(), kappa(), lambda(), mu(), nu()toxins) & neuraminadase • C. perfringens subdivided into five types (A-E) on basis of production of major lethal toxins • C. perfringens Type A (only major lethal toxin is alpha toxin) responsible for histotoxic and
enterotoxigenic infections in humans; Type C causes necrotizing enteritis (not in U.S.)
Lab Identification • direct smear and Gram stain, capsules upon direct examination of wound smears • culture takes advantage of rapid growth in chopped meat media at 45° C to enrich and then
isolate onto blood agar streak plate after four to six hours • gas from glucose fermentation • in vivo toxicity testing and identification of the specific toxin types involved • double zone of hemolysis on blood agar (p-hemolytic theta(e) toxin, a-hemolytic alpha(oc) toxin) • Nagler rxn; precipitation in serum or egg yolk media; oc -toxin (phospholipase C) is a lecithinase • "stormy" fermentation (coagulaltion) of milk due to large amounts of acid and gas from lactose
Diagnosis/Treatment of systemic infection — Early diagnosis and aggressive treatment essential • removal of necrotic tissue (surgical debridement) • Penicillin G in high doses if more serious infection
Of poorly defined clinical value are: • administration of antitoxin • hyperbaric oxygen (dive chamber) adjunct therapy (??inhibit growth of anaerobe??)
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Summary of C. perfringens
Infections
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Micro & Macroscopic C. perfringens
NOTE: Large rectangular gram-positive bacilli
Inner beta-hemolysis = θ toxin Outer alpha-hemolysis = α
toxin
NOTE: Double zone of hemolysis
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Clostridium perfringens • Pathogenesis: Traumatic open wounds or
compound fractures lead to muscle damages and contamination with dirt etc,
• Mainly in war wounds, old age, low blood supply and amputation of thigh (required prophylaxis with penicillin
• Prevention and Treatment• Remove dead tissue , debris and foreign
bodies .Penicillin and hyperbaric oxygen in some cases
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Clostridium perfringens • Can leads to the following diseases • 1) Wound Contamination• 2) Wound infection• 3) Gas Gangrene - most important disease• 4) Gas Gangrene of the uterus in criminal
abortion• 5) Food Poisoning : Spores are swallowed
Germinate in gut after 18 hours(Toxin production) abdominal pain and diarrhoea
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Clostridial Cellulitis
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C. perfringens Virulence FactorsM
ajor
Min
or
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Exotoxins Associated with C. perfringens Types A-E
Major
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C. perfringens Nagler Reaction
NOTE: Lecithinase (α-toxin; phospholipase) hydrolyzes phospholipids in egg-yolk agar around streak on right. Antibody against α-toxin inhibits activity
around left streak.
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Clostridium tetani
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Clostridium tetani — agent of tetanus
Morphology and Physiology- • long thin gram-positive organism that stains gram negative in old cultures • round terminal spore gives drumstick appearance • motile by peritrichous flagella • grow on blood agar or cooked meat medium with swarming • beta-hemolysis exhibited by isolated colonies • spores resist boiling for 20 minutes
Antigenic Structure- flagella (H), somatic (0), and spore antigens. Single antigenic toxin characterizes all strains.
Pathogenicity Determinants" • play a role in local infection only in conjunction with other bacteria that create suitable
environment for their invasion • systemic-acting, plasmid-mediated A-B neurotoxin (tetanospasmin) produced intracellularly
Mode of Action — one of most poisonous substances • binds gangliosides in synaptic membranes (synapses of neuronal cells) and blocks
release of inhibitory neurotransmitters; continuous stimulation by excitatory transmitters • muscle spasms (spastic paralysis) (trismus (lockjaw), risus sardonicus, opisthotonos),
cardiac arrhythmias, fluctuations in blood pressure
Lab Identification" • use characteristics of resistance to heat, motility, and toxin production to help identify
Diagnosis/Treatment/Prevention • empirical diagnosis on basis of clinical manifestations • treat to prevent elaboration and absorption of toxin
clean wound (debridement), control spasms metronidazole administered to eliminate vegetative bacteria that produce neurotoxin passive immunity (human tetanus immunoglobulin); vaccination (active) as preventative antitoxin administered to bind free tetanospasmin
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C. tetani Infections
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Summary of Clostridium tetani Infections (cont.)
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Clostridium tetani Gram Stain
NOTE: Round terminal spores give cells a “drumstick” or “tennis racket”
appearance.
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Clinical Forms of Tetanus
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Opisthotonos in Tetanus Patient
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Risus Sardonicus in Tetanus Patient
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Clostridium botulinum
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CLOSTRIDIUM BOTULINUIM
• Found in soil ponds and lakes• Toxin is exotoxin (protein) heat labile at 100
OC and resist gastrointestinal enzymes• It is the most powerful toxin known Lethal
dose 1 µg human and 3 kg kill all population of the world .It dictated for by lysogenic phage
• Botulism• From canned food., sea food e_g. salmon
when it is not well cooked (Spores resist heat at 100 oC ) then multiply and produce toxin
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CLOSTRIDIUM BOTULINUIM
• Symptoms• Abnormal eye movement as if cranial nerve
affected when bulbar area of the brain affected. Finally the patient might develop respiratory and circulatory collapse
• Enfantile Botulism• Ingestion of Spores germination in the
gutBotulism .• Botulism Patogenesis• Attacks neuromuscular junctions and prevents
release of acetylcholine that can leads to paralysis
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CLOSTRIDIUM BOTULINUIM
• Laboratory diagnosis• Suspected food and the patient faeces culture
or serum toxin detection by mice inoculation after weeks paralysis and death
• Treatment • Maily supportive and horse antitoxin in sever
cases• Prevention• Adequate pressure cooking autoclaving and
heating of food for 10 minutes at 100 OC
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Clostridium dificile
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Clostridium Difficile
• Normal flora in gastroentestinal tract after exposure to antibiotics and killing of other normal flora, this organism will multiply witch then produce toxin that has two components– A–Subunit enterotoxin (cause diarrhea) – B-Subunit Cytotoxic ( kill the cells ie necrosis)– Hyaluronidase and spore forming so bacteria can
survive for a month in hospital environment • PSEUDOMEMBRANE COLITIS is the clinical
manifestation of this disease which composed of bacteria , fibrin , WBCs and dead tissue cells
• Sever dehydration , intestinal obstruction and perforation are some of complication of this syndrome
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Clostridium Difficile
Laboratory diagnosis: this organism hard to grow in the laboratory required special media and growth of the organism in solid media required cell line culture to demonstrate cytotoxicity of the organism.
The simplest method for diagnosis by detection of the toxin in the stool by immunological testing (ELISA)
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Clostridium Difficile
• Treatment : Metronidazole or and oral vancomycin in sever cases
• Prevention: This organism form spores and hard to control in the hospital because they are resistant to alcohol decontamination ( use Na hypochloride instead).
• Patient need to be isolated and contact need to be screened to find out if they carrying the toxic strain of the bacteria.