clinically important gram positive bacilli spore forming 1. bacillus 2. clostridium non spore...
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Clinically important Gram positive bacilliSpore forming1. Bacillus2. Clostridium
Non spore forming1.Corynebacterium2.Listeria3.Lactobacillus
Bacilli w/ branching filaments
1.Actinomyces2.Nocardia
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1.BACILLUSBacillus anthracis
Human pathogenIsolation also considered to be clinically significantZoonosis
Bacillus cereusEnvironmental organismContaminates foodCommon cause of food poisoning
Bacillus stearothermophilusTolerates very high temperaturesUsed for quality control of autoclaves
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a.Bacillus anthracisLarge bacilli of 1-3
mHistorical
importanceSingle or paired in
clinical isolatesIn vitro –
prominent capsuleHighly resistant
spores
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Anthrax Pathogenesis and clinical presentations
Virulence factorsCapsule
(antiphagocytic)Toxin
(oedema & death)
Cutaneous anthraxAbout 20% mortality
Cutaneous anthraxAbout 20% mortality
Gastrointestinal anthraxHigh mortality
Gastrointestinal anthraxHigh mortality
Inhalation anthraxHigh mortality
Inhalation anthraxHigh mortality
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Anthrax - Epidemiology
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Anthrax - DiagnosisSpecimen
Aspirate or swab from cutaneous lesionBlood cultureSputum
Laboratory investigationGram stainCultureIdentification of isolate
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Anthrax – treatment and prevention
Penicillin(Tetracycline /chloramphenicol)
Erythromycine,Clindamicine
PreventionVaccination of animal herdsProper disposal of carcasses
Active immunisation with live attenuated bacilli
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b.Bacillus cereusLarge, motile, saprophytic bacillusHeat resistant sporesPre formed heat and acid stable toxin
(Emetic syndrome)Heat labile enterotoxin (Diarrhoeal disease)Lab diagnosis – Demonstation of large
number of bacilli in food
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Gastroenteritis
Gastroenteritis
Bacillus cereus clinical presentation
Incubation period < 6 hoursSevere vomitingLasts 8-10 hours
Incubation period > 6 hoursDiarrhoea
Lasts 20-36 hours
EMETIC FORM DIARRHOEAL FORM
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CLOSTRIDIUM(ANAROBES)AnaerobicSporingGram positiveDiameter of the spore is larger than the
cell resemble a spindleClostridium is derived from Kloster
meaning spindle
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Spores Pleomrhic (elongated, spindle)
Most are obligate anaerobes produce neurohisto toxins
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Saprophytes - Most
Some are opportunists - tetanus/gas gangrene/food poisoning
Cl. perfringens - commensal of the intestine
Cl. sporogenes - -do- Can invade the intestine after the death
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CLASSIFICATION BASED ON THE TYPE OF
DISEASE PRODUCED A . Tetanus Cl. tetani - Present in soilB. Gas gangrene
Established Cl. perfringens ‘gut’ organism
Cl. septicum Cl. novyi
- Less pathogenic Cl. histolyticum Cl. fallax
- Doubtful Cl. bifermentans Cl. sporogenes
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C. Food poisoning 1. Gastroenterritis - Cl perfringens Type
A
2. Botulism - Cl. botulinum/ Soil 3. Pig-bel Cl. perfringens type C
D. Acute colitis - Cl. difficile / gut’ organism
(pseudomembranous colitis)
Commonest cause of ‘nosocomial’ diarrhoea
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Gas gangrene C. perfringens type A (Principal),
Capsulated, non-motile
Lecithinase C - toxaemia
Nagler reaction
Colonies with haloes
Colonies withouthaloes
Incorporated withAntitoxins
GAS GANGRENE
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Dead tissue, blood clots, foreign matter aerobic organisms
In an injury DEVELOP ANAEROBIC CONDITION
(Exogenous infection) Germination of spores
Gas gangrene
oedema, necrosis, gas production,
toxaemia, myositis
Crepitus
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C PerfringensC histolyticum
C septicumC novyii
C Perfringens Alpha toxin(lecithinase)
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TETANUS Cause tetanus in both man and animals
disease which effect the nervous system
of the host.
- Agricultural workers and gardeners and
are more prone because the spores are
present in the soil.- At birth under unhygienic conditions baby’s
can get – tetanus neonatorum.
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Soil/Intestine/Vagina
Drum stick appearance
Motile with peritrichous flagella
Obligatory anaerobes
Grow on Robertson’s cooked medium
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All types produce the same toxin
C. tetani – 10 types based on the H antigens
(CP – 5 types based on the type of toxins, alpha, beta, epsilon, iota).
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Susceptibility -
Some strains can withstand boiling for 3hrs/dry heat 1600C for 1hr. but all will destroy at 1210C/15 min.
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COMMON FEATURES FOR BOTH CT AND CP
All CP’s produce alpha toxinAll CT’s produce same exotoxin – plasmid mediatedHowever, CP’s got enterotoxins.
Exotoxin of CT has got two components .Tetanolysin – both heat and O2 labile – may act as a leucocidin
.Tetanospasmin – heat labile, but O2 stable (Therefore, can you give an edvantage ? will not get destroyed in the blood).
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Spores germinate -------toxin-----motor nerve endings--------along the motor neurones of the peripheral nerve to the anterior horn cells------local tetanus (in the proximity of the wound).
Ascending tetanus – when toxins spreads upwards along the spinal cord towards C.N.S. Gives generalized spasms.
Descending tetanus – when toxin is given IV , spasms will appear in the muscles of the head, neck and spreads downwards.
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Clinical symptoms
Early symptom is trismus (lock jaw) – spasms of the masseter muscle- difficulty in opening of the mouth and masticating - rigidity spreads to muscles of the face, neck and truck- risus sardonicus – contraction of the frontails and muscles at the angle of the mouth- back is usually slightly curved (Opisthonotus ?) - Insevere cases violent spasms will last for few seconds to 3-4 mins.- If convulsions appear soon after the initial symptoms, it is very serious.- The spasms gradually intensify and patient may die of
.exhaustion, b. asphyxia or aspiration peumonia - If local tetanus after a wound at the neck, you might think of tuberculous meningitis (irritation and
paralysis is common).
What happens
Toxin acts at the synaptic junction – prevent the synthesis of acetylcholine. Thus, prevents synaptic transmission.
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ToxinsTetanolysin - heat and oxygen labile/lyse
RBC/Tetanospasmin - heat and oxygen
stable/highly lethal (for mice 0.0000001 mg) dies within 1 - 2 days
get easily neutralize with antitoxin
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GABA GLYCINE
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Tetanus
Treatment - 10, 000 units of human tetanus immunoglobulin(HTIG)
Prevention and control
a. Immunization - HTIG 250 - 500 units (to immune patientsonly)
To non-immune adsorbed toxoid followed by
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Clostridial food poisonngC. perfringensCarriers for food poisoning strainsSurvival of heat resistant spores in
bulk mealsSporulation in gut - Short IP and
watery diarrhoea for 24-48 hoursBeta toxin production in C.
prerfringens type C – Necrotizing enteritis(Pig bell)
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BOTULISMSausage
Food borne botulism(IP 1-2 days)
Infant botulismWound botulism
(IP > 4 days)
8 toxins (A-G)
DiagnosisIsolation of organism in food/faecesDetection of toxin in faeces / serum
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Produces BotulismWorld wide distributionFound in soil and occasionally in animal fecesSporese are highly heat resistant ,withstand 100C for 3-5 hrs.Heat resistance is reduced by acid pH or high salt concentrations
ToxinReleased during growth and autolysis of bacteria.It is found in 7 antigenic varieties.A-GThe principle cause for human disease A,B,E/F
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A,B - Variety of foodsE - Fish productsC - Limberneck in birdsD - botulism in mammalsToxin is neurotoxic proteinDestroyed by heating at 100C for 20 mins.Action :Block release of Acetylecholine at synapsesand NMJ causing flaccid paralysis.PathogenecityIllness is not an infection.Botulism is an intoxication resulting from the ingestion of food in which C.botulinum has produced toxin.
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PSEUDOMEMBRANOUS COLITISVirulence factors
Enterotoxin(Toxin A)
Cytotoxin(Toxin B)
Diagnosis
Clinical suspicionCulture of faecesDetection of toxin
Management
Discontinue antibioticsAmpi/Tetra/ClindaOral metronidazoleOral vancomycin
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CORYNEBACTERIA(AEROBES)
- Causes localized inflammation (pseudomembrane, greyish white exudate ) and generalized toxaemia
- Prevalent in baby’s after 3-6 months (that’s why DPT is given at 3, 5, 7 months, boosters at 18 months and at school entry), very high in young children
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MorphologyGram/+ve/palisade/Chineseletter arrangement
Irregular swellings at one end -club shaped.
Corynebacteria tend to pleomorphism in microscopic
and colonial morphology.
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On blood agar Small granular & gray with irregular edges and may have small zones of hemolysis.
Grow aerobically on ordinary media
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a. Corynebacterium diphtheriae
Normal flora of nasopharynx in about 10%
Diphtheria caused when infected by lysogenic bacteriophage
b. DiptheroidsNormal flora of skinUsual contaminants of samplesCan cause disease in ‘compromised’ host
C. ulcerans C. haemolyticum
C. jeikeium
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Rare in developed countries/ third world countries
Nose, Nasopharynx, skin aerobic, facultatively anaerobic
Nasal carriers are very dangerous
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Loeffler's serum slope Blood telurite agar (black colonies)
Morphological differences
Three biotypes
Gravis (severe)
Inter-medius (intermediate)
Mitis (mild)
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Epidemiology
It is rare in developing countries, a disease of the third world countries. Still highly prevalent in the former Soviet Union.
Spread through droplets.
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Types of DiphtheriaFaucialLaryngealNasalConjunctivalVulvovaginalOtiticCutaneous around the mouth and the nose
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Effect of toxins1. Local2. General
Toxaemia and acts on the myocardium and on motor nerves and adrenals
Complicationsa, pseudomembrane may extend to larynx and
cause obstructionb.myocarditis /PolyneuropathyDegenerative changes in the liver adrenals, kidney's
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Pathology
•Toxin is absorbed in the mucus membrane and causes destruction of epethelium and causes a superficial inflammatory respons.
•Necrotic epethelium becomes embeded in exuding fibrin and red and white cells, with bacteria-
•Grayish pseudomembrane is formed over the tonsilasand pharynx and larynx.
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•Removal of pseudomembrane - capillary damage and bleeding..
•Regional lymphadynopathy with marked edema of the neck within the membrane bacilli produce toxin.
•This results in distant toxic damage paranchymatous degeneration fatty infiltration & necrosis in heart muscle liver kidney & adrenals.
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How to identify the immune persons
Shick test – suitably diluted stabilized toxin intradermally, localized erythema (1-3cm) in 2-4 days, means no or little antibodies
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DiagnosisDirect smear - Albert's stainCulture - Loffler's serum slope/blood agar/blood
telurite agar
Check the toxigenicityAnimal inoculation
Death within 96 hrs
Guinea pigs/rabbits
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Elek's plate test
Filter paper with antitoxin Precipitation
Strain
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Management –
1. Patients - isolation of the patient / bed rest/antibiotic treatment/antitoxins (horse serum)
Penicillin/erythromycin/teracycline/rifampicin/clindamycin
2. Contacts – immunize if not (toxoid) – adults should be shick tested or given low dose as immunization of immune adults can result in severe reaction.
- prophylactic antibiotic – erythromycin- swab nose and throats of contacts
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3. Community – immunization
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DIPHTHERIA
DIAGNOSIS
Clinical suspicionSwab for cultureToxin production
TREATMENT
PenicillinAnti-diphtheretic serum
Maintaining airwaySupportive
PREVENTION
Immunization(toxoid)
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4.Listeria monocytogenes