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Avian mycobacteriosis (AM) is a chronic and contagious disease of pet birds, captive exotic, wild and domestic fowl, and mammals. Mycobacterium avium subsp. avium is the most common cause of AM in poultry. For the first time, we report a chronic outbreak of AM in an Iranian breeder flock of 250 45-week-old turkeys (Meleagris gallopavo) with a morbidity and mortality rate of 91.6% and 80%, respectively. A well-defined clinical feature of the outbreak included a progressive weight loss, decreased egg production, listlessness, and lameness. Tuberculous nodules were seen on liver, spleen, ovary, and ribs. Granulomatous inflammation and acid-fast bacilli were confirmed by using Ziehl-Neelsen method on hepatic lesions. M. avium subsp. avium was identified by polymerase chain reaction techniques based on the presence of 16S ribosomal RNA gene and insertion elements IS1245 and IS901. In this report, we not only describe the epidemiological, pathological, and molecular characteristics of the outbreak in detail, but we also discuss multiple factors influencing the introduction and development of AM critically. In this case, wild feral pigeons might have been the source of infection, but further molecular-epidemiology studies are needed to understand the role of wild birds in the persistence and transmission of Mycobacterium.RESEARCH HIGHLIGHTS First report of avian mycobacteriosis in an Iranian commercial turkey flock is described in detail.Risk factors intrinsic to the bird and mycobacteria, as well as extrinsic factors influencing the introduction and development of avian mycobacteriosis in birds, are critically discussed.

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BACKGROUND AND OBJECTIVES: Paratuberculosis (PTb) (John's disease) is an incurable chronic intestinal infection that mainly affects ruminants. PTb is caused by Mycobacterium avium subspecies paratuberculosis (MAP) with a global distribution. Despite evidences on MAP contribution in Crohn's disease its causal role is still a matter of controversy. In ruminant farming, vaccination is broadly accepted as an effective control measure of PTb. This article describes preparation and field trial of an inactivated PTb vaccine made from the MAP 316F strain. MATERIALS AND METHODS: Formulation of the vaccine was conducted based on the method traditionally used in the UK. Identity of the MAP strain was authenticated by PCR-IS900 and PCR-F57 tests. In the field, a group of 100 lambs (3-8 weeks old) were subcutaneously inoculated with the vaccine preparation under study. These animals, pre-vaccination, were all PTb ELISA negative. Serum level of antibody was determined by ELISA on days 0, 30, 60, 120 and 240, post-vaccination. RESULTS: In PCR-900 and PCR-F57, the MAP 316F strain produced two fragments of 560 and 704 bp length respectively, a confirmation of its identity as MAP bacterium. In the field trial and at the arranged time intervals, the achieved blood serum levels of antibody, attributable to the vaccine formulation, displayed considerably high values. CONCLUSION: Given that the PTb-caused economical losses in the Iranian environment are dramatically high and also the fact that future of state policy on control of PTb remains unknown, we belive vaccination of animals is the best recommendable practice.

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INTRODUTION: Mycobacterium avium ssp paratuberculosis (MAP) causes paratuberculosis (Johne's disease) in ruminants. As a species, M. avium comprises M. avium subsp. hominissuis and a number of clones that are known to have evolved from this subspecies, namely M. avium subsp. avium (MAA), M. avium subsp. silvaticum, and MAP. Despite the very high genomic similarity of MAP and MAA, the insertion sequence IS900, which is 1,451-bp long, is now understood to be exclusively present in 10-20 copies in the genome of MAP. In the present study, a multidiscipline polymerase chain reaction (PCR)-based algorithm targeting16SrRNA, IS6110, IS901, IS1245, and IS900 markers has been employed to differentiate between six laboratory strains of M. avium complex (including MAP 316F, III&V, and 2e plus MAA D4), Mycobacterium tuberculosis DT, and Mycobacterium bovis AN5 strains used at the Razi Institute (Tehran, Iran) for the preparation of paratuberculin, avian, human, and bovine tuberculin, respectively. MATERILS AND METHODS: Three laboratory strains of III&V, 2e, and 316F were subcultured on Herrold's egg yolk medium, whereas the MAA strain of D4 along with M. bovis AN5 and M. tuberculosis DT were subcultured on Lowenstein-Jensen slopes. All the inoculated culture tubes were incubated for 8weeks at 37°C. Eventually, their genomic DNA was extracted according to the method of van Soolingen. Five individual PCRs were conducted on these templates to amplify 16SrRNA (genus-specific marker shared by all mycobacteria), IS900 (MAP-specific marker), IS901 (MAA-specific marker), IS1245 (M. avium complex (MAC)-specific marker), and IS6110 (M. tuberculosis complex (MTC)-specific marker) loci. RESULTS: Consequently, a 543-bp amplicon was amplified by all the six strains in PCR against 16SrRNA, an indication of their identity as members of Mycobacterium genus. A 245-bp fragment was detected in only IS6110-PCR with M. bovis AN5 as well as M. tuberculosis DT. In the IS1245 assessment, the MAA strain of D4 produced a 427-bp amplicon, whereas none of the other studied strains produced this amplicon. A 1,108-bp amplicon fragment of the IS901 marker was successfully produced by MAA strain, whereas no PCR product was achieved in amplification of all the three MAP strains. In IS900-nested PCR, the three MAP strains produced the expected 400-bp and 298-bp fragments CONCLUTION: However, no amplification was observed with other strains. Two main achievements of this work are the development of an efficient means of differentiation between the six Razi laboratory mycobacterial strains and characterization of the genomic profile of these strains, a capability that is vital when cross contamination is potentially an important concern.

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In 1931 Carpantier reported bovine TB (BTB) in Iranian cattle. Some eighty years on with a national test-and-slaughter programme in place for over four decades, the efforts to vanquish Mycobacterium bovis (M. bovis) infection in cattle have been in vain as the vast majority of the 30 Iranian provinces still have reports of BTB in their cattle herds every year. This paper reviews the present epidemiology of BTB in Iran and in the region and evaluates the success of government policy in controlling this disease.

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Restriction fragment length polymorphism (RFLP) genotyping was employed to analyze the population genetics of Mycobacterium bovis in Iran. One hundred and twenty-three isolates collected from slaughtered tuberculosis-suspect cattle and one clinically asymptomatic buffalo were subjected to RFLP analysis with probes of the polymorphic GC-rich sequence (PGRS) and the direct repeat sequence (DR) using DNA digested with PvuII and AluI. All these methods detected a large homogeneous population in which only a few isolates had variant genotypes. Only AluI-based RFLPs of both the PGRS and DR sequences were able to clearly differentiate between BCG and field strains of M. bovis. As in previous reports, these findings seem to reflect a recent dispersal of one or a few strains in Iran following the substantial expansion of Holstein-Friesian cattle over the last few decades.

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