= Sp eet ‘odent to person by the —_——— ed from 1 y the fleas, The « 2. Bndemic typhus (Rickettsia yphi)-— Wis transite put the disease is milder ana Farah Clinica, epidemic tY ly fatal picture of endemic typhus is mostly same as epider heomapieh meme ee 291 .¢ transmitte tes, This 3. Serub typhus (Orientia tsutsugamushi)— It 18 IT re jg the esehar, the punched-out ulcer covers disease resembles epidemic typhus clinically. One ave mite bite. Generalized lymphadenopathy ang with a blackened scaty that indicates the locatiol’ nt may be severe etyral involven ‘ : lymphocytosis are common. Cardiae and cerebral i (Refi Jawets's-281n) Q. Short note:Rocky Mountain spotted fever (NTR) Ans. Rocky Mountain spotted fever R rickets Arthropod Vecto ‘Mammalian Reservoir: Dogs, rodents. Nomenclature: The name "Rocky Mountain spotted fever" is der which the disease was first found Organist ‘cks, (See the picture on right) ived from the region in Clinical features 1. Acute onset of nonspecific symptoms, eg, fever, severe headache, myalgias, and prostration. 2. Typical rash, which appears 2 to 6 days later, begins with macules that frequently progress to petechiae. The rash usually appears first on the hands and feet and then moves inward to the trunk, (See the picture) 4% CNS symptoms: Delirium, coma. & DIC (Disseminated intravascular coagulation) €& Edema (8 Circulatory collapse (Ref: Review-17th) Diagnosis: The diagnosis must be made on clinical diagnosis is delayed until arise in antibody titer can be coo Bromelltp besaupe tia ab atory Skin biopsies taken from Patients between the qn a immunofluorescence stain, hs days of illness ‘ * PCR: The sensitivity of PCR is 700% meavdnvSAl ickettsiag by grounds and therapy (Refi Review-1 7th; Jawetz 's-28th)Whey are true bacte Classificatio Systeme Bactertology Ee 289 gate Important Properties 1. Rickettsiae are 2, Barely visible in theiaht ‘Cell wall resembles that 6 4, They do not stain we Koccobacilliappearing either as{short 1048-9 e\ aitfonegatiy ell with Gi cope when stained ar the light microscope Wh e unde n stain but are readily visible © arene to «With Giemsa’ stain, Gimener’s Stain, acridine orange, or ther st, produ eae, ee Rickettsiae are eblignteintracelllar parasites, wecaute they cell culture, embryona Teolicate extracellularly, Therefore, rickettsiae mast be grown. | experimental animal & Rickettsiae divide by inary Tiggaa within the host cel ctor or host. Rickettsi 7. Most rickettsiae survive ee short times outside of the vec destroyed by heat, drying, and bact Antigenic structures: 1. Specific soluble antigens. 2. Species specific corpuscular antigen, 3. Alkali stable polysaccharide a are quickly wete's-28th) icidal chemicals Jaa (Ref: Review-17th: » because- 1. They divide by binary fission, 2 ‘They contain both DNA and RNA. Sensitive to antibiotics genera - 1. Rickettsia - causes typhus fever, spotted fevers, and scrub typhus. 2. Coxiella - causes Q fever. 3. Rochalimaea- causes trench fever eR cin) Transmission 1. The rickettsiae with one exception, lice, fleas, and mites. % The essen t9 airopod transmission is C. bum, which is transmitted by aerosol and inhaled into the lungs. fre transmitted to humans by the bite of the arthropods such as ticks, Q. Name some Rickettsial diseases with etiological agents, Q. Enumerate the diseases caused by Ricketsia, (SU-18M) Ans. Selected Rickettsial Diseases: ey Organism Spotted fevers iis Rocky Mountain spotted fever | R rick Rickettsia |e Typhus group eee Mammatian Reseryoir265 Q Whats the abbrebiation of MOTT? (RUIN) MOTT: MOTT refers to Mycobacteria Other Than Tuberculosis Ans. Morphology osis and atypical mycoba Systemié Bacteriology wcteria. (RU-O9N) yPteria other than tubercurosis, (RU-155u) Rod shaped! with beaded appearance. Acid fastness More acid fast Long filamentou Tess acid fast Alcohol fastness Alcohol fast Not Pigment production _ | Absent, Many of them produce pigments. Reservoir Found only in human, Widely distributed in the environment. Tuberculin test Strongly positive Weakly positive Culture Grow in 6-8 weeks (slow growth). Dry heaped buff color colony. Grow in 1-3 weeks (rapid growth). Smooth, soften & regular colony. tubercular drugs Pathogenic to guinea | Yes No pigs 9. Human tohuman | Occurs Not usual transmission 10. Sensitivity to antic [Sensitive Less sensitive / resistant, 11. Basic lesion Granulomatous lesion. (Granulomatous lesion may not be seen. 12, Niacin test Positive Negative except M. simiae Q. State the differences between human & bovine type of mycobacteri Q. Write down the differences between M. tuberculosis & M. Bovis. (DU-978) Pi Ans. Differences between human & bovine type of Mycobacteria: ee | Pee temvebateaion | mauogaapenacaa —— Sterns) yahoo - Morphology lender, long, straigl | oe 12, Straight or slightly Stout, shorter, straight 2. Staining Beaded 3. SonRee Infected person pair Route and portal ofinfeetion — [Primarily by inal ‘Contaminated cow's milk, 5. Culture in Lowenstein-Jensen | Euyenio pore Drinking OF infected milk, one luxarian growth, Dysgtnic, 6. Action of giveerol in Ld media [Gye \ ilyceral enka | 7. Action of pyruvate Cee Glycerol inhibits ea Growth snot enhanced Edina 9. f Sar 19.0; requirement See More than 3 Wk 10. Niacin test Bu z 1. Nitrate reduction test ne 12. Usual disease in human vette tony 7 ere cin aves, Skin ~ Tenastnes ‘yes. pass, contactue, (anaesthesia, is, contra iibues fess deetuston Face, hands, feet Lepromatous Borde Tuberculold BL aT Figure, Leprosy: mechanisms of damage and tissue affected. Mechanisms uncer the broken line are characteristic of disease near the lepromatous end of the spectrum, and those under the solid line are characteristic of the tuberculoid end. (BL = borderline lepromatous; BT = borderline tuberculoid) Ans. a. Bacterial agent: Mycobacterium leprae b. Laboratory test: Histopathology of the biopsy material ‘* In the tubereuloid leprosy: Typical granulomas is seen, ‘In lepromatous leprosy: Foam cells (Lipid-laden macrophages) are seen, Ans, 2 Probable diagnosis: Leprosy i ined pacar ral nation: Zichl-Neclsen staining with 5% HaSO, ri hich cannot in conventional culture media; 2. Rickettsiae 3. Treponema pallidum 4. Mycobacterium leprae : Ei “ological agents Mycobacterium leprae Stain needed f ae for staining: Ziehl-Neelsen ‘Staining with 59% 4, SO. bSOxBlueprint Microbiology : 262 6 NA based technique: The diagnosis can be confirmed by using the 7. Others: © Cultures are negative because the: * No serologic tests are useful polymerase chain reaction (PCR), > organism does not grow on artificial media, (Ref: Review:-17th; Davidson’s-21su344) Nice 10 kno M, leprae can be cultured in mouse foot pad. 9 months time 1. Detection of drug resistance 2, To see the potency of anti-leprosy drugs. 3. Detection of viability of the bacidli during treatment, (Lepr in test(DU-01J, RU-095) used to determine the type of leprosy in a patient. It involves injection ‘Mycobacterium leprae intradermally. 2 vis required, The purposes are ~ Q.Shos Zepromin test: The lepromin skin test of standardized antigen (Ag)of inactivat Procedure: , 1. Anextract sample of M. leprae is injected intradermally, usually in the forearm, so that a small Jump pushes the skin up. The lump indicates that the antigen has been injected at the correct depth. 2. The injection site is labeled and examined at 48 hours and 21 days to see if there is a reaction. Types of reaction: 2 types of reaction: 1. Early or Fernandez reaction: If red area is > 10 mm in diameter at the end of 48 hours, the testis Positive. Early reaction is more pronounced with refined Ag, ie, leprosin (the ultrasonicates of tissue free bacilli), Late or Mitsuda reaction: If red area is > 5 mm in diameter at the end of 21 days, the test is positive. Late reaction is more pronounced with crude Ag, ie, lepromin (boiled bacilli rich lepromatous lesion whieh also includes tissue elements), Interpretation of test: 1. A positive reaction indicates allergic tissue response against invasion by leprosy bacilli, In other words, it indicates relative resistance of the host. ‘The test is used as a tool for classification of leprosy- Positive reaction in tuberculoid leprosy & negative in lepromatous leprosy. It is of prognostic value, A good prognosis is indicated when a ne} i it licated wi -gative reaction turns into positive reaction and a bad prognosis when itis the other way round, bi ‘The reaction has not much value as a diagnostic test as many rearton healthy individuals may give a positive Q. Write down the differences between tuberculoid & lepré ‘pre - c : loid & lepromatous leprosy, (DU-06M, SU-12Ju), ). Mentic important differences between tuberculoi i DU-19. Q. Mention two imps #¢s between tuberculoid & lepromatous leprosy. (DU-19Ju) Differences between tuberculoid & lepromatous leprosy: Feature A, Type of lesion eof oft ie ted response to M. leprae | Presont (5. Lepromin skin test eee ea Reduced or absent — Negative EY ‘One or few lesions with little tissue destruction (RTI Many lesions with marked tissue destruction (REF Review 170)ss es [eg Tuberculln skin ta 3-8 weeks "| Primary comptes, pase berlin sk | Meningeal, mitiary and pleural d ea : Gastrointestinal, bone and join, aM Re Post-primary diseas al tract disease reactivation or reinfection — (REF. Davidson imary tuberculosis? Q. Define primary tuberculosis, What are the sites of pri Q, What are the primary si (CU-163) AS poem defined as infection of an individual lacking previous contact or Primary tuberculosis: Primary tuberculosis is defined as inf initial infection (fist eX immune responsiveness 0 tubercle bail, Primary tuberculosis occurs after iit ‘posure) with Mycobacterium tuberculosis or Mycobacterium bovis Primary sites of tuberculosis BS Lungs- are the usual site of primary tuberculosis with M. tuberculosis, Ap\CON & The tonsil > Intestine. > Skin. Q. What are the sites of tuberculosis? (CU-16)) " Q. What are the common sites of extra-pulmonary TB? (CU-18M, RU-17N) Ans. s of tuberculosis: = Pulmonary TB: in lungs. aa + Extra-pulmonary TB: Occur almost in any organ including pleura, pericardium, lymph nodes, larynx, GIT, kidney, genitor-urinary tract, bone and joints, skin, meninges, CNS, tonsil, etc, Q. °M. tuberculosis does not produce any exotoxin, endotoxins or enzymes, still it produces damaging disease”- why and how? Ans. M. tuberculosis produces no exotoxins, enzymes and does not contain endotoxin in it antigens elicit delayed hypersensitivity reaction, Itis the de damage and other pathologic manifestations of tuberculosis. ts cell wall. Mycobacterial lelayed hypersensitivity reaction that results in tissue such as cascating granulomas and cavitation, Q. How do the tubercle bacilli spread within the body? a of the tubercle baelll within the body occurs by two mechanisms: nee ene an ne! solo oerpouansifoxpe eas Ne tani 6 2 Itean diseminate vi the bloodstream to many internal organs, Disse if cell-mediated immunity fails t0 contain the intial infection or ‘mination can occur at an early stage immunocompromised ata late stage if a person becomes (Ref: Review-17th) What isa ‘tubercle’ Tubercle: A tuberc I le is a granuloma surrounded Tubercles heal by fibrosis and calcification, It odnge coe undergone caseation necro lonary tuberculosis : (Ref: Review-17th)L be ino |Yellow-orange pigmented colony only tuberculosis especially in cae | Wher exposedto light immunocompromised ace _ © M marinwn Swimming pool granuloma (fish tanl AMNLPALOTA PIO MAZEL granuloma) Group-I. Scotochromogens: Produce the] * ML. soroflaceum | Scrofula (a granulomatous cervical adenitis, | pigment only in the dark usually in children) Group-Iil. Nonchromogens: Produce little| “Mf avium Lung disease clinically resembling or no yellow pigment, irrespective off ¥7M. intracellulare tuberculosis [presence or absence of light Group-IV. Rapid grower: Grow rapidly, * M. fortuitum ‘Skin infections and skin abscess in producing colonies in fewer than 7 days | * M. chelonei immunocompromised patients (at the site of Classes ‘Species Diseases $3 heen OM. tuberculosis: sa dogs OOM. bovis TB * M. africanum = MM leprac Group-l. Photochromogens: Produce a] = M. kansasil Lung disease clinically resembling, : inctured et = M. smegmatis See Non-pathogen (Ref: Review-17ih) Q. What is Mf, tuberculosis complex? (CU-15D) Q. Enumerate 4 organisms under M. euberculosis complex? (DU-173) Q. Enumerate the members of M. tuberculosis complex. (DU-195,17N) Ans, M. tuberculosis complex: Mycobacterium tuberculosis comp! Mycobacterium species that can cause tuberculosis © Mycobacterium tuberculosis © Mycobacterium africanum © Mycobacterium bovis © Mycobacterium microti ete, lex refers to a genetical humans or other organisms. I pce esas isms. Tt includes: Q. Write some special features of mycobacteria. (DU-07Ju) Q. Name important properties of mycobacteria, (RU-I9N) Ans: . a (Ref: Wikipedia) te231 Systemic Bacteriology Q. Short note: Vibrio cholerae 0139 Bengal. (DU-0SM) Ans. Vibrio cholerae O139 Bengal: In December 1992, « large epidemic of cholera took place in Bangladesh, and large numbers of people have been affected, The organism has been characterized as ‘Vibrio cholerae O139 Bengal’. tis derived genetically from the Fi-Tor strain but it has changed its antigenic structure such that there is no existing immunity and all ages, even in endemic areas, are susceptible, ‘The epidemic has continued to spread & Vibrio cholerae 0139 Bengal’ has affected at least 11 countries in southern Asia. (Ref: textbookofbacteriology.net) Q. Write down the lab diagnosis of cholera, (CU-19N,181,17N, RUAOD) Q, State microscopic findings of a cholera stool, (DU-161) Q. How Vibrio cholerae can be isolated and identified from stool sample? (DU-18M) Q. What is the appearance of Vibrio cholerae in Gram stain? (CU-21N) Ans. m Lab diagnosis of cholera: The approach to laboratory diagnosis depends on the situation, During an epidemic, a Glinieal judgment is made and thee i litle need forthe laboratory. nan area where the dsense is endemic oF for the detection of carriers, itis based on demonstration of Vibrio cholerae by microscopic examination. Isolation and identification is done by bacteriological techniques, Biochemical and Serological tests ar also helpful Steps! A. Specimen collection: = Stool, = Vomit = Rectal swab First day B. Microscopic examination: . = Gram staining: Gram-negative comma shaped ba 5 fe ZF Hanging drop preparation: Typical motility (a fish in steam appearance). . Dark ground inition (DGI): Shows the rapidly motile vibrios like shooting star in dark sky. “ . Isolation and culture: yaa , “& Monsur's media (Tellurite taurocholate gelatin agar media): Small black colonies surrounded by ‘characteristic hollows. Thiosulphite-Citrate-Bile-Sucrose agar (TCBS agar): Large yellow colored colony. On TSI agar, an acid slant and an acid butt without gas or HS are seen because the organism ferments sucrose. “ ‘MacConkey agar. Colorless colonies, because lactose is fermented slowly. al tests: The organism is oxidase-positive, which distinguishes it from members of the iaceae. polyvalent O1 or non-O1 antiserum, ‘retrospective diagnosis by detecting a sein antibody iter in acte- and convalescent-phas lescent-phase sera. (Ref Review-17th; Jawetz “s-28th) diagnosis: Cholera, due to Vibrio cholerae, ‘and laboratory diagnosis;ee } 229 Systemic BacteriolOKy Vibrio Cholerae Properties of Vibrio choterag: Vibrios are curved, comma ‘ u shaped Gram. Inia means onan ae Vibrios are orgie partion, they have ssh: tr team appearance Nee aes Which differentiates them from enteric gram-negative bacteris “Antigenic structures. © & pe ie, maxi ef, Reviews17h; Jawete 5:28) Q. What is the appearance of Vib ‘Ans. On Gram stain, Vibrios are rio cholerae om Gram stain? (CU-21N,12)) curved, comma-shaped Gram-negative rods. . What are the cultural and biochemical characteristics of Vibrio cholerae? (SU-1S1011))) Culture and growth characteristics of Vibrio cholerae: 1. ¥. cholerae produces convex, smooth, round colonies that are opaé 2. Grow well at 37°C in aerobic condition. 3. Grow ata very high pH (8.5 to 9.5) and are rapidly killed by acid. Ba ret ca con which it produces ‘4 V. cholerae grows well on thiosulfate-citrate-bilesucrose (TCBS) agar. on hie P yellow colonies that are readily visible agaist the darkegreen background Of The ABT yg b. On Monsur’s media (Tellurite gelatin agar media): Dark colonies with ot around them. shar On MacConkey agar: Ferments lactose slowly (colorless / pale colonies) ‘4. On iutrient agar: Circular, smooth afd translucent colonies que and granular in transmitted tight. ‘Biochemical characteristics of Vibrio cholerae anes il ic from enteric gram-negative bacteria. “ie. Vibrios are oxidase-positive, which differentiates them from enteric gr ey Q.How can you differentiate ¥, cholerae from Enterobacterinceae? i -holerue and Enterobacteriaceae: Enterobacteriaceae 1, Motile with [2>-Straight bacilli |a-"Oxidase ‘Oxidase positive om ‘Acrobe Q. Name the virulence factors of V: cholera. (DU-210) Be 2 Virulence factors of ¥ cholerae: @ Enterotoxin (cholerage): & Pili, ‘8° Mucinase. (Ref: Review-17th) —EEE asSystemic Bacteriology 3 System Viva Q. How can you “fferentiate san =~ : = te Difference between salmon, egative rods that can be distingnuis Wa anting Gramieneaains a oe Salmonelta pa eet Shigeltae are non-lactose ferment"® Shigella criteria - 2 Thep do nynet Prottice gas on fear 2 Y do not produce Fg m fermentation of glucose. 3. They are non-motil (Ref: Review-17th) i 11d e eee pres ‘liagnosis of shigeosis, (DU-22N,18N/J,17).1 110. CUT!" sU-1L) Q. How do you diagnose Patllary dysentery, (DU-06M,05), RU-16)) #€ 4 case of shigella dysentery in the laboratory’ Lab- diagnosis of bactllary dysentery: yn, Isolation and ic examinatio! see on demonst ive agent. by. microscopi helpful. identification is done by bacteriological t Teshigue,Biseheneal ad Seo} cal tests are also help! Steps: i. Specimen collection: © Fresh stool 2 Rectal swab ii, Microscopie examination of stool: Findings are- © Plenty of pus cells. “© Macrophage “© Some RBC ii, Isolation and identification from culture: Culture is done in- ee = MacConkey agar media or + DCA media or : + Salmonella Shgelnagsmedi ¥ q ~Sansy (At 37°C temperature for 24~48 hours) «Oe teraper tata They eause allaling slant agian acid. but with no,ens and no Has: inegioregneiaston:. 4. ee al Odoriess, mucus & biotin soo. 2 Chemical- Alkaline stool Serological mation test to detect the endotoxin, 2 oe test for confirmation of shigell; i + + Slide agglutination fshiglls and for determination ofits group. (Ref: Review-17th) psis: Bacillary dysentery (Shivetiog, @. Probable diagnosis: Bae ty (Shigeli >. Probable pathogen: Shigella ei © Impartane findings in stool under micros d. : ope: Laboratory tests: rte sromabove, Pe PHeMty of pus cota Enterocolitisy Macrophage, ee Some RBCSystemic Bacteriology Important properti 8 of shigelt 3 \egative rods, Non-lactose fermenter, Form colourless colonies on MacConkey agar media. Fimbriae present (incase of Sh fletnorh. Somspore forming and non-capsulated Shigella Aston ate almost always imited to the c * Shigellac are facultative anaerobes but grow s ae colonies with intact edges reach a diameter of about 2mm in 24HOUTS- (2. jyyers's-28th; Review 17¢h) wane uite rare. sion is iO circular, transparent IT; bloodstre=™ ly. CONVEX, ‘pest aerobically: ae Q. Classify Shigella, Q. Name the species of Shigella. (DU-18N) pan aj! walls, and these antigens Cassifieation of Shigelia: ll shigetie have O antigens (polysaccharide) in their cell Wa are used to divide the genus into four groups: A, B, C, and D. | Leonie eae ~ Lactose (Ornithine fermentation | Decarboxylase Species ee [1 Shigella dysenteriae | 2._ Shigella flexneri 3._Shigella boydit (4. Shigella sonnei a5 (Ref: Jawetz’s- 28th) lo|a}el>| Q. What are the virulence factors of shigella? Q. What are the virulence factors of the causative agent of dysentery? (RU-16J) Q. What are the exotoxins of Shigella dysenteriae? (DU-19Ju) Ans, Determinants of Pathogenecity / Vi tors of shigell 1. Invasiveness— 2 Exotoxin: Shigella toxin. 2 types of action- ‘a. Produces watery diarrhea, 4B Produces colonic ulcer by inhibiting protein synthesis and by eytotoxicit 3. Endotoxin. (Ref: Review-17th) Q. Enumerate the disease caused by Shigella species, Disease caused by Shigella species: Shigella specieg Sshigeta sp aon Besar yet cars een pail passage or ete (Shigeloss Bases "he leading cause of infant Seg oe mucus. Bacil ing countries like u-2y The disease varies from mild to severe depending on 1. Species of Shigella: Shigella dysenteriae tWo major factors: (Ref: Revi i at, with, Lride causes the Pa * Review-17th) 2. Age of the patient, with young children a ae ae See dase, “ing the most Severely affected,a Mlueprint Microbiotayy aie + Mplonomey * Cough © Abdominal + Bienes ad death Uf wureatedy sell And of And week |" « Ltirtum, complications, then com. am Th Doar iwnaton Complications of (yphold fever + Howel © Perforuion = + Hemorrhage Septicemie fuel © Hone and joie infeetion + Meningis *_Cholecystidy Myorardits Nephritis Persistent galtbladder carriage Toxle phenomena Chronke carriage (Hef, Davidson ‘e278 Ans, Principle: Diagnosis is based on isolation of aalmonelta *Pesies by blood culture and antigen detection (recently introduced), crea collection: (Important far Viva) ® Ilood for blood oulture (1M & 20" @ Serum for Widal test (20 ‘woek) Stoo! for stool culture (2 sand 30 week) vw Urine for uring culture (4! week) Bone marrow for culture week) By Mleroscopl examiontion: iy not help Fositive ln 1" week of infection, Can be done in 3 pr - (sing liquid media on biphasic media) eer *(afler processing in Itc solution in wolid Automated (very rapid. within @ hour) a * Glotl culture: Positive from 2m! or 3 Rance Fel eck, Mis done ine ” Deoxycholite citrate ar (DCA, Shigell-salmonclis agar media Urine cut ac ‘Traditional re Positive in 26, gar and 4th Week. Culture is dong ine Peoxyeholate citrate ugar (DEA)Blueprint Microbiology s 196 2. Clostridium histolyticum 3. Clostridium Septicum 4. Clostridium novyi 5. Clostridium sordelli. (Ref: Revie Predisposing factors: ee 1. Reduced blood supply 2. Presence of aerobic organ 3. Reduced O-R potentials. Toxins and enzymes responsible for gas gangrene: 1. Alpha toxin (Lecithinase) /phospholipase C: It degrades lecithin (an important constituent of cejj membranes), 2. Theta toxin: It has hemolytic and necrotizing effects. 3. Hyaluronidase 4. Collagenase, DNAase & other proteinases. Pathogenesis of gas gangrene / myonecrosis: Spores of Clostridium are introduced in traumatized tissue (especially muscle). 4 Aerobic organisms multiply, utilize 3; and produce anaerobic environment: L In anaerobic environment, C. perfringens spores germinate into vegetative forms. t They start to multiply and liberate the toxin and enzymes which have lethal, necrotizing and hemolytic properties. Lecithinase damages cell membrane including those of RBC causing hemolysis. Degradative enzymes produce gas in tissues. L Wound rapidly expands in size. + Toxins and organisms enter into blood. L Severe toxemia J Circulatory failure and shock 4 Death (Ref. M.R, Choudheiry-5th/273,274: Review-17th) Any & Clinical diagnosis: Gas gangrene, D. Etiological agemts: See above, Pathogenesis: See above,Q. How ean How can You diagnoses qacor—s ———_098 ; “ase of food poisoning? (DU-97M) AX Diagnosi Pris on dem Shstration of toxin by neutralization test with specific antitoxin Specimen collection: + Leftoy. 2 Sitever food and patients serum * Vomitus 2. Microscopic exami: ination: Gram-staining of spe com the colony. ie 18 of specimen fr * Isolation from leftover f food * Blood agar media + MacConkey agar media * Robertson's cooked meat media, ; ifie antitoxin. | 4. Toxin identification: By animal pathogenecity test in mice and by neutralization with specific antito Important properties of Clostridium perfringens: Morphology: a. Gram-positive bacilli b. Non-motile. ¢. Capsulated, 4. Oval, sub-terminal / central spore. 2. Saccharolytic. . ¥ 3, It grows overnight under anaerobic conditions, producing hemolytic colonies on blood agar. ; 4, In broth containing fermentable carbohydrate, growth of C perfringens is accompanied by the production of large amounts of hydrogen and carbon dioxide gas, which can also be produced in necrotic tissues; hence the term gas gangrene 5, Transmission: Spores arc located in the soil; vegetative cells are members of the normal flora of the colon and vagina. Gas gangrene is associated with war wounds, automobile and motorcycle accidents, ‘and septic abortions (endometritis). a. (Ref: M.R. Choudhury-Sth/270; Sherris-8th) ise i ae 1, Gas Gangrene 2. Anaerobic cellulitis 3. Food poisoning. in (Ref: Lippincott's-2"4/150) Ans. J Myonecros Causative organisms / histolytic or histoto conser ric species of Clostridi 4. Clostridium perfringens (most important, 90%)Blueprint Microbiology Q. Why botulism is not food poisoning? (RU-17)) a & Q. Why botulism is not considered as a classical example of food poisoning? a botulinum differs from other causes of food poisoning? (CU-03)) s 194 Botulism is not considered as a classical example of food poisoning, Lseerell i 1. Tthas no action on GIT but normally food poisoning: occurs due to intestinal upset. 2. Flaccid type of muscle paralysis occurs but does not produce vorniting or diarrhea. Q. Write down the laboratory diagnosis of botulism. ” Q. Two people shared a canned fish during a pienic. Both developed dysphasia, diplopia and Subse, developed flaccid paralysis. State your probable diagnosis. Name the recommended samples, collected and the tests you like to perform to diagnose the condition. (DU-10J) Ans Probable diagnosis is Botulism and left over canned fish should be collected if available. of botulism: r : a x Principle: Diagnosis of botulism is based on demonstration of toxin by neutralization test with specific antitoy Botulinum toxin is demonstrable in uneaten food and the patient's serum by mouse protection tests Specimen collection: Leftover food and patient's serum. In infant botulism, Cl. botulinum and toxin ca, be demonstrated in bowel contents but not in serum. 2 Culture: Culture is not usually done. 3. Toxin identification: a. ByELISA b. By passive hemagglutination or radioimmunoassay. c. By mouse protection test- Mice are inoculated with a sample of the clinical specimen and will de unless protected by antitoxin. (Ref: Review-17th; M.R. Choudhury-Sth'27?, Q. Define food poisoning. Q. Name the bacteria causing food poisoning, (DU-09J, CU-03J, RU-17: 'J,15Ju, SU-19M,08)w/ ). Short note: Food poisoning. (CU-17J,15J) u Ans. Eood poisoning: It is an acute gastroenteritis caused by ingestion of food or drink rith either livi tas oh ox or organ chee sence ene eck cemented with ier Bacteria A. Toxin mediated: 1. Staphylococcus aureus. 2. Clostridium botulinum, 3. Clostridium perfringens. | 4. Escherichia coli Verotoxin producing) | B. Non-toxin mediated: Salmonella typhi murium Salmonella enteritidis Clostridium botulinum Campylobacter jejuni, Bacillus cereus. Listeria monocytogenes, __ ae