Anaerobes :Characters Anaerobic bacteria will not grow in the presence of oxygen and are killed by oxygen( O2 (metabolite

They lack Catalase , Peroxidase and !peroxide oxide dism!tase( O"# en$yme The main so!rce of energy is fermentation CLOSTRIDIUM %Characters of Clostridium species (a-Anaerobic spore forming &m '(e large bacilli , grow mainly on blood agar ()-hemolysis b-They fo!nd mainly as commensals in the intestine of man and animals or as saprophytes in soil c-They prod!ce exotoxins that play a role in pathogenesis d-C!lt!re media !nder anaerobic coditons % *- +,agar 2--lood agar .-/obertson cooked meat :Classification of Clostridium accordin to patho enesis #istotoxic "eurotoxic !nterotoxin Prod!ce toxin Ca!se tiss!e Prod!ce ne!rotoxin( act Prod!ce exotoxin act mainly damage as myonecrosis (gas (mainly on ne!rons on &0T (gangrene 1x%Cl,tetani, Cl,bot!lin!m 1x% Cl,perfringens , Cl,difficle 1x% Cl,perfringens, Cl,septic!m

Differentiation bet$een different t%pes of Clostridium species that infect human lostrid!m 3otility caps!l +itrate spore ,&l! ,2ac ! "isease in man e red!ction ,c tani ' ' terminal Tetano!s erfringens ' & ' 1ccentric ' ' ' &as gangrenefood poisonongot!lin!m ' s!bterminal ' ' -ot!lism Cl, difficile ca!sing pse!domembrano!s colitis5 antibiotic associated diarrhea 4 Clostridium tetani Tetanus :(atho enesis contamination of wo!nds with street d!st )spores*6 germination 6 bacteria 6 release exotoxin (potent neurotoxin +tetanospasmin,*6reaches C+ directly by retrograde axonal system or indirectly from the blood 67 release of inhibitor% mediators l%cine and -A.A/ generali$ed )m!sc!lar spasm, hyperreflexia, sei$!res and spastic paral%sis )trismus ' loc01a$ The in8!ry may be a contaminated s!rgical wo!nd , or an infection of !mbilical st!mp leading to + 9tetanus neonatorum Dia nosis% On clinical s!spicion treatment with antitoxin sho!ld be started witho!t waiting for laboratory diagnosis, which is done for confirmation Sample: :o!nd ex!dates 2ilm: stained with &ram showing &ram '(e bacilli swollen at one end d!e to terminal spherical ,pro8ecting spores% +drum'stic0 appearance (Cultural characters a#n!trient agar b#blood agar(- hemolysis-tetanolysin
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,c# /obertson cooked meat medi!m Animal patho enicit%: :hen mice are in8ected 0,3 with c!lt!re of the organism 6 tail become ,stiff, the in8ected limb shows spastic paralysis which spreads to rest of body The administration of tetanus antitoxin protects the animal (re3ention and control ,Tetan!s is a totally pre(entable disease, Prophylactic meas!re is the only way to control tetan!s Acti3e immuni5ation: Al!m precipitated tetan!s toxoid is gi(en in combination with diphtheria '4 ,toxoid (accine9 "PT9 in three intram!sc!lar in8ections at the age of 2,; and < months A booster dose is gi(en to military personnel , pregnant women (assi3e immuni5ation: a-0,3, h!man tetan!s imm!ne glob!lin =T0& i,e, tetan!s antitoxin made ,in h!man to a(oid hypersensiti(e reactions that occ!r when antitoxin made in horses is !sed b- Antitoxin prophylaxis sho!ld be accompanied by acti(e imm!ni$ation with tetan!s toxoid at different sites i,e, passi3e 6 acti3e immuni5ation Treatment ,Antitoxin sho!ld be gi(en at once to ne!trali$e any toxin that did not fix to the C+ ,* Penicillin or metronida$ol to eliminate the organism from tiss!es -2 ,An ade>!ate airway and respiratory s!pport ?, -en$odia$epines to pre(ent spasm -. Clostridium perfrin ens'7 They present in large intestine of h!man and animals ,its presence in water indicate fecal pollution Diseases: a-gas gangrene b-food poisoning )-as -an rene)M%onecrosis Sacchrol%tic proteol%tic Mixed @erment carbohydrate with "igest protein release =2 prod!ction of acid and gas which blacken the wo!nd and prod!ce bad odo!r (Cl,welchii(perfringens Cl,sporogenes Cl,histolytic!m Cl,septic!mCl,no(yii (atho enesis: wo!nd are contaminated with d!st (spores#6 germination 6(egetati(e bacteria b-Sacchrol%tic m8o8 ferment s!rgars prod!cing large amo!nt of gas which interferes with their blood s!pply leading to their death, +ecroti$ing toxin, Collagenase and hyal!ronidase fa(o!r ,necrosis and spreading of infection (roteol%tic clostridia digest dead tiss!e leading to dark colo!r , fo!l odo!r of the wo!nd, The wo!nd is oedemato!s, with crepition in the ad1acent tissues8 The condition is accompanied by ,toxaemia, shock, m!ltiple organ fail!re and may be fatal :9irulence 2actors Alpha toxin is a lecithinase )phospholipase* which destroy cell membrane6 cell death,* , Theta toxin has a heamol%tic and necroti$ing effect,2 8D"ase: h%aluronidase and colla enase ,.

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Dia nosis is made clinically, laboratory is !sed for confirmation Sample: 1x!date aspirated from deep sites of the wo!nd 2ilm: stained by gram, &m'(e bacilli non motile , caps!lated , spore is o(al , s!bterminal Culture % !nder anaerobic condition on +,agar, -lood agar (do!ble $one of hemolysis#, robertson cooked meat .iochemical test: a'They ferment gl!cose,lactose ,maltose and s!crose with acid and gas b'Storm% clot reaction: The organism ca!se rapid fermentation of lactose in litm!s milk and the gas prod!ced splits the clot c-"e lar;s reaction: All types of Cl,perfringens prod!ce opalescence in egg yolk media (?A egg ,yolk agar# d!e to prod!ction of lecthinase which ca!se a (isible precipitate aro!nd the colonies Such opalescence can be specificall% inhibited b% specific Cl,perfringens antitoxin Animal inoculation: I8M8 in8ection of * ml of /obertson cooked meat broth c!lt!re into a g!inea , pig ca!ses death in * to 2 days,The in8ected limb shows oedema and crepitation The in1ection of Cl8perfrin ens antitoxin before in1ectin the culture fluid protects the animals Treatment , !rgical debridement of de(itali$ed tiss!es, Amp!tation may be needed ,* Penicillin in big doses in combination with metronida$ole ,2 ,=yperbaric oxygen therapy may be of (al!e, ;, Poly(alent antitoxin are of do!btf!l (al!e ,. :(re3ention ,a- "epends !pon ade>!ate cleaning of contaminated wo!nd and s!rgical debridement b- administration of antibiotic 8"8.: "o Antitoxic serum for passi3e proph%laxis8 "o Toxoids a3ailable for acti3e immuni5ation Cl8 (erfri ens food poisonin (atho ensis: ingestion of large n!mbers of clostridia that ha(e grown in reheated meat dishes, The enterotoxin forms when organisms spor!late in the g!tB its mode of action is the same as that of the ,enterotoxin of taph, a!re!s i,e, 0t acts as a s!perantigen (Clinical features: watery diarrhea no (omiting nor fe(er)inc!bation period is <-*C ho!rs Treatment: @l!id replacement therapy, The condition is self limited in *-2 days Clostridium botulinum'< .otulism different t%pes of botulism are identified < .otulism food poisonin >ound botulism Infant botulism= flopp% child 9classical t%pe+ ;s%ndrome ingestion of canned food spore germination and toxin' pore germination and toxin containing the preformed prod!ction by m,o, present prod!ction by m,o,present in &0T ne!rotoxin prod!ced by Cl, in the wo!nd of the infant bot!lin!m associated with dr!g ab!se, - "!e to presence of the spore in specially skin- popping with ,honey ,black tar heroin They de(elop poor feeding, 2?

weakness or paralysis and may need respiratory s!pport b!t reco(er spontaneo!sly :).otulism food poisonin )classical t%pe 0ngestion of preformed toxin in canned food6 absorbed from g!t 6 blood 6peripheral ner(o!s system 5cranial ner(es67 release of acetylcholine at ne!rom!sc!lar 8!nction 6flaccid paralysis Clinical si ns% 0tDs characteri$ed by symmetrical descending flaccid paralysis ("ilated !n reacti(e p!pil (oc!lar paralysis# 5 respiratory and cardiac fail!re ("eath )There is no fe3er nor -IT s%mptoms)"o 3omitin or nausea ((roperities of Cl8botulinum exotoxin:) .otulinum toxin a- (ery potent ne!rotoxin, 0t is a polypeptide encoded by a lysogenic phage ,b- There are C antigenic type (A-=#type A,-,1, and occasionally @ ca!se bot!lism in man ,c- The toxin is heat labile can be destroyed by boiling for 2E min!tes d- -ot!lin!m toxin is considered to be a ma8or agent for bioterrorism and biologic warfar Dia nosis : is clinical one but ma% need lab8 confirmation Specimens% @ood reminants :Test for toxin detection'4 , a*Animal patho enicit%: food extract in8ected in mice, the animal will die with flaccid paralysis b#120 A% detect bot!lin!m toxin , less sensiti(e than mo!se bioassay c#PC/ to detect gene :Isolation and identification of C8botulinum'7 @ood remnants are inoc!lated on /obertson cooked meat media and blood agar anaerobically colonies is identified as &m stained film ,-iochemical test and animal pathogenicity ,Treatment : Tri(alent antitoxin (types A,- and 1 is gi(en together with respiratory s!pport ,The antitoxin is made in horses and ser!m sickness occ!r *?A of recipients , (re3ention is by caref!l sterili$ation of food before canning, wollen cans sho!ld be discarded ,=oney sho!ld not be gi(en to infants Clostridium difficile Source: The organism is carried in the &0T in abo!t .A of the general pop!lation :Disease and transmission 0t ca!ses pse!do-membrano!s colitis and antibiotic associated diarrhea and is the commonest of , ,hospital ac>!ired diarrhea it is transmitted by the faecal F oral Causes of disease: a complication of antibiotic therapy% clindamycin ,The antibiotics s!ppress dr!g sensiti(e normal flora allowing Cl, difficile to m!ltiply and prod!ce an exotoxin A, which is an enterotoxin that ca!ses O!tpo!ring of fl!ids i,e, watery diarrhea, and an exotoxin -, which is a ,cytotoxin that ca!ses damage to the colonic m!cosa, leading to the pse!domembrane formation :Dia noses can be done b% "etection of the toxin in stool s!pernates by a latex aggl!tination, 120 A or by cytotoxic effects G
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,on h!man embryo fibroblast cell c!lt!res ,0solation of the organism on special mediaG ,Treatment : a- :ithdrow the ca!sati(e antibiotic b- Treat with metronida$ole or (ancomycin and replacement fl!id(3etronida$ole is preferred (beca!se !sing (ancomycin my select for (ancomycin resistant enterococci,(H/1 ? Choose the most correct ans$er Cl8perfrin ens infections are commenl% associated $ith '4 a- Contamination of deep lacerated wo!nd b- Antibiotic treatment c-Cons!mption of water contaminated with sewage d-0mm!no-s!ppression : 2ood poisonin due to Clostridium botulinum is most often caused b% '7 a- Contaminated po!ltry prod!cts b-Canned food c-Poorly cooked seafood d-"airy prod!cts >hich of the follo$in tests used to detect lecithinase en5%me of Cl8perfrin ens '< a- +eglarDs reaction b-1lekDs test c- AscoliDs test d- +iacin test >hat;s the t%pe of toxin produced b% Cl8tetani in tetanus '@ a- Cytotoxin b- =istotoxic c- 1nterotoxin d- +e!rotoxin >hat=s the t%pe of Clostridium that cause disease associated $ith antibiotic use 'A a-Cl,tetani b-Cl,perfringens c-Cl,tetani d-Cl,difficle >hat=s the t%pe of Clostridium that has drum stic0 appearance of spore 'B a- Cl,tetani b-Cl,perfringens c-Cl,tetani d-Cl,difficle Cloastridium tetani causes loc01a$ paral%sis because 'C a-m,o enters m!scle cells 5 blocks transmission b- m,o enters m!scle cells 5 stim!late transmission c-0ts ne!rotoxin blocks release of inhibitory mediator leads to generali$ed m!scle spasm d- 0ts ne!rotoxin stim!lates release of inhibitory mediator leads to locali$ed m!scle spasm : >hich of the follo$in not a reEuirement for Clostridium culture media 'D a- Presence of anaerobic condition b-Thioglycolate is media !sed for growth of m,o c-Prod!ce )-heamolysis on blood agar d-They can grow on "orset egg media II* (ut true or false for the follo$in statements Clostridi!m perfringens are acti(ely motile by peritricho!s flagella -* Tetanolysin 5 =yl!rindase are the most prominent toxin 5en$ymes prod!ced by Cl,tetani -2 -ot!lism food poisoning is characteri$ed by na!sea , (omiting and diarrhea , b!t no fe(er -. Tetan!s can be pre(ented by administration of tetan!s toxoid and antitoxin -; o!rce of Cl,tetani spore is soil or d!st contaminated with h!man or animal feces -?

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