[Genomic structure of Burkholderia mallei Razi 325, the strain used for industrial production of Mallein in Iran]

Background and Objective: Iran remains a major stronghold for glanders in the Middle East. In Iran, the non-indigenous Burkholderia mallei Razi 325 strain is used in manufacturing of the mallein, required for malleination of animals. Multi Locus Variable number tandem repeat analysis is currently the standard globally accepted genotyping system for Burkholderia mallei. This study was done to survey the genomic structure of Burkholderia mallei Razi 325, the strain used for industrial production of Mallein

Methods: In this descriptive study, a MLVA genotyping system with 4 previously-characterized loci VNTR140, VNTR1367, VNTR2065, VNTR2971 along with two new loci of VNTR24, VNTR41 was used

Results: Optimization of PCRs resulted in a single protocol that enabled simultaneous amplification of all the six loci. Sequencing of PCR products revealed there were 2, 3, 12, 6, 1 and 2 copies of the unit repeat hold in the genome of the Burkholderia mallei Razi 325 strain. This observation was extended to include the already-whole genome sequenced Chinese Burkholderia mallei ATCC 23344 and Burkholderia mallei BMQ and also Burkholderia mallei SAVP1 strains

Conclusion: The Burkholderia mallei Razi 325 strain is distinguishable from the other three strains through MLVA genotyping method

Mycobacterium tuberculosis genotyping by MIRU-VNTR method

st and ardized genotyping systems in molecular epidemiology of tuberculosis in the world. This sudy was done to determine the Mycobacterium tuberculosis genotyping by MIRU-VNTR method. Methods: This descriptive study was done on sputum, gastric lavage clinical specimens of 53 tuberculosis suspected patients. Fifty-three isolates were identified by 16S rRNAandRv-typing followed by RD typing. They were then subjected to a 12-locus [ETRA, ETRB, ETRC, ETRD, ETRE and ETRF, MIRU-10, MIRU-26, MIRU-39, MIRU-30 plus QUB-11b] MIRU-VNTR typing system. Results: In MIRU-VNTR typing, forty-four types were identified with 13 isolates classified in 4 clustered and the remaining 40 isolates representing 40 orphan patterns. In comparative analysis of MIRU-VNTR loci, MIRU-26 with 7 alleles displayed the highest diversity level [Simpson's diversity index = 0.767. Out of the 53 isolates, only one was identified as Mycobacterium bovis. All the remaining isolates were characterized as Mycobacterium tuberculosis. None of the samples was affected to Mycobacterium complex strain. No evidence of either double or co-infection of the patients with more than one species/strain was detected. Conclusion: While the genomic diversity observed by MIRU-VNTR typing sounds extensive, the population genomic structure on the whole however, seems to be homogenous. Recent transmission between studied patients does not appear to be a frequent event as only 13 isolates representing 4 MIRU-VNTR types, were assumingly epidemic

Identification of mycobacterium tuberculosis complex, using molecular methods

A high level of homogeneity observed within all bacteria in the Mycobacterium tuberculosis complex makes a property that seriously challenges traditional biochemical-based identification methods of these pathogens in the laboratory. The work presented here was conducted to characterize Mycobacterium tuberculosis complex isolates in Golestan, Northern Iran. Between 2008 and 2010, 42 mycobacterial isolates were collected from clinical tuberculosis-suspected patients in Golestan province. The isolates were sub-cultured on fresh Mycobacterium-specific culture media including glycerinated and pyruvated Lowenstein-Jensen slopes. The isolates were subsequently subjected to a PCR-based identification scheme coined Huard-Warren method. This strategy consisted of three individual algorithms namely, 16SrRNA; RV typing [Rv0577, Rv3877.8, Rvl970, Rv3120, Rvl510 and IS 1561] and RD typing [RD1, RD 4, RD9 and RD12]. All isolates were proved to be M. tuberculosis. Furthermore, none of the patients were being infected with any other member of the M. tuberculosis complex or simultaneously co-infected with two mycobacteria. This fundamental observation was independently obtained by specific culture media, RV typing and also RD typing. Considering the fact that cattle and sheep farming play an important role in the economy of the region, absence of Mycobacterium bovis in the studied isolates can be unexpected to some extent. Huard-Warren which is a simple and cost-effective identification method can be used in both reference and regional laboratory for differential diagnosis of tuberculosis

[Detection of somatic antigens in Mycobacterium avium subspecies paratuberculosis]

Johne's disease or Paratuberculosis as a chronic granulomatosis enteritis in ruminants will be caused by Mycobacerium avium subsp . Paratuberculosis . Detecting whole bacterial cell wall antigens would be helpful in potential applications for diagnosis, vaccine production, and elucidation of pathogenesis . To determine secreted somatic cell antigens of Mycobacterium avium subspecies Paratuberculosis . Standard strain [III-V] of Mycobacterium avium subspecies Paratuberculosis DNA was extracted from the cultured and gene analysis was done using PCR to confirm bacterial purity . On the other hand, protein concentrations in both media and cell entracts were determined . Furthermore, all proteins pattern were shown by SDS -PAGE . Electrophoretic findings showed some somatic antigens in the range of 19 - 100 KDa. These purified somatic antigens can be used for further study and potential application in vaccine production

Outbreak of avian mycobacteriosis in flocks of domestic pigeons: an epidemiological approach

Pigeons are extensively kept for homing and racing purposes in Iran. The main objective of this study was to investigate dissemination of M. avium subsp. avium [MAA] in pigeon aviaries in Tabriz, North-western Iran. Postmortem pathologic specimens from thirty-nine out of 140 birds collected from private flocks [n = 3], were subjected to bacterial culture out of which 3-4 mycobacterial isolates were recovered. Applying a five-PCR diagnostic algorithm targeting short but definitive stretches of 16S rRNA and RV0577 genes, IS6110, IS901 and IS1245 genomic loci, proved all the isolates were MAA. They were either IS901+/IS1245+ [n = 22] or IS901+/IS1245- [n = 12]. When four healthy cattle sensitized against Mycobacterium bovis AN5 and Mycobacterium avium D4 were tuberculinated, the results confirmed the observed skin reactions against bovine tuberculin in animals sensitized with M. avium were large enough to complicate test interpretation. We believe the extent of such epidemiological impact deserves further investigation if progress in control of bovine tuberculosis is intended

[Hepatitis B "e" antigen [HBeAg] status and its relation to transaminase level in patients chronically infected with hepatitis B surface antigen [HB5Ag] in southwestern Iran [1999-2001]]

Background: There is a wide variation in the frequency of HBeAg among HBsAg infected patients in the world with possible prognostic significance

Patients and Methods: In this prospective study, three-hundred and seven patients who were found to be HBsAg positive either during a blood donation or incidentally in a routine check-up, were evaluated by testing HBeAg, HBeAb, serum alanine transaminase [ALT] and serum alfa-fetoprotein. Liver ultrasound was performed in all patients. Patients were then divided into two groups: Group A [HBeAg positive] and Group B [HBeAg negative]. The correlation of this viral marker [HBeAg] with a necroinflammatory index [ALT] was evaluated

Results: HBeAg was positive in 50 cases [16.3%] and its antibody [HBeAb] was detected in the remaining 257 patients [83.7%]. Serum ALT was abnormal [>2.5 times the normal level] in 16 [32%] of HBeAg positive and 39 [15] of HBeAg negative cases. Among 55 patients with an increase in ALT, 16 [29%] cases were positive for HBeAg while 39 [71%] patients were HBeAg negative. The rate of HBeAg in patients with a normal enzyme level was 13.5% [34 out of 252]

Conclusion: The rate of HBeAg negativity among chronic HBsAg infected patients in our area [southwestern Iran] is in the range reported from other parts of the world: Additionally, HBeAg has a positive correlation with the level of ALT

Ascorbic acid status in 6-60 months old children in Sirjan villages

Six- to sixty months old children were studied in Sirjan villages, divided into villages with or without a health house [HH]. All the 893 children were medically examined and their food intake was assessed, using the 24-hours dietary recall method. Ascorbic acid was determined in blood samples of 199 children. The results obtained are as follows: 1; the average vitamin C intake for all the children were more than the respective RDA, being 32.3+32 mg. In the 13-24 months group, however, the intake was 25.1+17.8 mg. 2; As compared with the standard, a smaller percentage of 6-12 months old subgroup suffered from vitamin C deficiency relative to other subgroups, which might be because of breast feeding. 3; Altogether 30% of the children had a low intake. 4; the mean total blood ascorbic acid in all the subgroups was significantly higher than the standard. 5; There was no significant difference between the two regions with regard to blood ascorbic acid, there was no difference in two sexes either. 6; only 1.5% of all the children had a low blood vitamin C level. 7; A low dietary intake of vitamin C was a nutritional problem among the children, but biochemical and clinical findings did not confirm this