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  1. zutalaba.com Family MycobacteriaceaeGeneral characters of mycobacteria1-Special staining(Zhiel Neelsen) acid fast M.O. +strong carbol fuchsin wash by acid alchol sol. + methylene blue Red (acid fast)2-Slow growth Long incubation period (21-45 days)3-growth medium is poured in screw capped bottles4-Sample preparation before culture(sample processing)5-First diagnosis by allergic reaction (tuberculin test)6-specific lesion (Granuloma formation)7-No treatment in animals ( only test and slaughter) but only in human

  2. 21 Mycobacterium tuberculosis Smear of concentrated sputum smear stained with Ziehl-Neelsen showing acid –fast bacilli zutalaba.com Acid-fast bacilli Pink bacilli surrounded against blue background Zagazing University Fac. Of Vet. Med. Dep. Of Microbiologyv

  3. zutalaba.com Species Tuberculosis group (typical) 1-Mycobacterium tuberculosis(MTB)2-M.bovis , 3-M.avium, Jhons disease M.paratuberculosisLeprosy disease M.lepraeOppurtunistic diseases Atybical mycobacteria Specimens 1- sputum (pulmonary infection) urine ,peritoneal or pelural aspirate 2-lymph node (after slaughter) , milk tracheo-bronchial aspirate 3-bone marrow Specimen processing:- tracheobronchial exudate + NaoH4%(0.5h.) lymph nodes + ,, milk Culture centrifugation sediment Z.N.

  4. Pathogenesis (lesion)Route of infection inhalation , ingestion ,skinbacillus entry odema,eosinophil, macrophages surronded by fibrin (Exudative lesion) multinucleated giant cells + epitheliod cells + fibroblasts + lymphocytes +fibrous capsule + calcium (productive lesion) Heal or or Continue to productive zutalaba.com Still as Tubercle or or as Miliary tuberculosis

  5. CultureLowenstein Jensen medium egg medium malachit green (selective) cycloheximide (inhibit fungus) lincomycin (antibiotic) glycerol (inhance growth of MTB) sod. Pyruvate ( , of M.bovis) incubate up to 8 weeksDiff. on culture and biochemical tests zutalaba.com

  6. 22 zutalaba.com Mycobacterium tuberculosis Culture on Lowenstein-Jensen medium Niacin production test Yellow colonies Yellow colour Zagazing University Fac. Of Vet. Med. Dep. Of Microbiologyv Negative Positive

  7. zutalaba.com Diagnosis of pathogenic mycobacteriaField diagnosis tuberculin test (65% true positive) tuberculin (old tuberculin or PPD) PPD MTB , M.bovis , M.avium inoculate in glcerol brothincubate(37C,7week) centrifugationsupernatant + trichloroacetic acid purification0.1 ml. ID.(skin)I.D. Single or double comparative using mamlian and bovine PPDLaboratory diagnosisconfim by culture, by DNA probeVaccinationBCG(bacillus Calmette and Gurin) attenuated strain of M.bovis(since 100 year) Same skin Thickness (-Ve) Thickness >4 mm over (+Ve)

  8. zutalaba.com Diff. acc. To pathogenicityMycobacterium paratuberculosisemaciation ,diarrhoea, (sheep,goat,cattle)Specimen life animal(15 gm. Faecal sample) dead , (mesentric L.N.) histopathology or specimen processing for microscopy , then cultureCultureegg yolk medium + mycobactin ( incubate for 10 weeks)Field diagnosisJhonin test(I.D.) give false +Ve up to 75%

  9. Atypical MycobacteriaRunyon classification1-acc.to pigment prod.(colonial morphology)-Schotochromogens (yellow pigment in absence or presence of light) -Photochromogens(pigment In light only-Non chromogens as (M.avium)2-acc. To growth rate-Rapid grower(less than 7 days)M.fortutum -Slow grower(over 7 days) + Scoto. AsM.scrofolacium+photo. AsM.marinum zutalaba.com

  10. Mycobacterium scrofulaceum Scotochromogen zutalaba.com 24 Yellow pigment was produced even when incubated in the darkness Zagazing University Fac. Of Vet. Med. Dep. Of Microbiologyv

  11. 23 zutalaba.com MycobacteriummarinumPhotochromogen Both tubes were incubated in the dark at 37 C for 10 days While this tube was incubated in the dark for another 5 days Then this tube was incubated for 5 days in sun light So producedYellow pigment So no pigment was produced Zagazing University Fac. Of Vet. Med. Dep. Of Microbiologyv

  12. Members of mycobacteria are difficult to be stained due to :a-lack of cell wall. b-has waxy coat. c-unstainable. d-a and b.Pathogenic Mycobacteria are mainly.a-acid and alchol fast. b-alchol fast. c-non acid, non alchol fast.Atypical mycobacteria differ from typical one in : a-pigmentedb-rapid grower c-acid and alcohol fast d-a and bCulture for pathogenic mycobacteria on L.Jensen media still in incubator maximally for .a-one month at 37c b-24hr at 37 c c-2 weeks. d-non of themMycobacteria are mainly scanty in sputum samples, it must be treated with. a-bile b-4 % Na hydroxide c-H2so4Tuberculin material contains . a-tuberculo proteins b-antituberculous Ab c-T.B cell wall d-non of them

  13. Farm of milking cows ,examined by tuberculin test , only 10 cows were positive, then turned to slaughtering.1-what are samples should be turned to diagnostic laboratory2- laboratory precaution of sample handling3- one confirmatory laboratory test for the principle bacterial agent4-Differential confirmation of the causative agent5- write the laboratory report that should be sent to the farm.Two lymph nodes(L.N.) from sheep and 5 L.N. from 5 cows were liquified and granulomatous ,were sent from slaughter house for bacteriological exam.1-enumerate suspected chronic bacterial causative agents2- bacteriological media required for first isolation3-confirmatory identification of each pathogen in each L.N.

  14. Mamary lymph node from slaughtered cow was turned to bact. Lab.1-Describe the node in case of tuberculosis2-write on isolation media required for the suspected bacteria3- sample preparation for staining and stains used4-How do proof that the isolate is typical mycobacteriumBuffalo lung with cavitation and casiated noduls was turned bact. Lab. 1.Enumerate isolation media required for suspected isolates2.Colony ,shape and characters on each medium3.Biochemical confirmatory tests for one of themFive milk samples from tuberculin positive lactating cows were turned to bact. Lab.1.wite on materials and readu ing results of or tuberculin test2.specimen processing for staining and record shape of +ve Z.N. slides3.Confirmation of typical mycobacterial infection of these nodes

  15. 1- Photochromogens produce yellow pigment only in the presence of light.  2- Photochromogens can not produce pigment in the darkness.  3- The left tube is not considered photochromogen.  4- The medium used here is Lowenstein Jensen medium  Zagazing University Fac. Of Vet. Med. Dep. Of Microbiology

  16. 1- In the left part , the medium used is Dorset egg medium.  2- In the left part , the M.O produce yellow pigment.  3- In the left part , the right tube is considered +ve.  4- Yellow colour in the right tube is due to pigment production.  www.zutalaba.com

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