Molecular characteristics of Mycobacterium tuberculosis in the "closed" Russian town with limited population migration.

The aim of this study was to perform a molecular characterization of Mycobacterium tuberculosis strains circulating in one of the "closed" Russian cities under conditions of the limited population migration and high HIV coinfection rate. We analyzed 109 M. tuberculosis isolates recovered from TB patients in the Novouralsk municipality in the Ural area of Russia; 38.5% were from HIV coinfected TB patients and 19.3% patients were former prison inmates. The Beijing genotype was predominant (78.9%) while 57.8% and 17.4% of isolates belonged to the Beijing B0/W148 and Beijing 94-32-clusters, respectively. An atypical allele of the QUB26 VNTR locus (2 repeat units) was detected in 11 of 63 Beijing B0/W148 isolates. The non-Beijing isolates were subdivided into nine spoligotypes of the four genetic families (Ural, LAM, Haarlem, T), SIT35/Ural being the largest group (n = 9; 8.3%). Multidrug resistance (MDR) was detected in 63.6% and 83.7% of isolates from newly diagnosed and previously treated patients, respectively. Almost all isolates of the B0/W148-cluster were MDR (92.1%) compared to the Beijing 94-32-cluster (47.4%). No association was found between HIV status of patients and MDR-TB or particular genetic cluster. A combined contact and molecular investigation confirmed three family foci; in two of them, Ural SIT35 and Beijing B0/W148 strains were transmitted from HIV-infected sons to their fathers. To conclude, M. tuberculosis population in Novouralsk features an exceptionally high prevalence of the strongly MDR Beijing B0/W148-cluster and emergence of the B0/W148 substrain with unusual QUB26 allele. This situation was likely synergistically shaped by the limited population migration, high prevalence of the HIV coinfection and high proportion of the former prisoners. The existing organizational approaches to prevent TB transmission are insufficient and require a serious revision.

Distribution and diversity of Nosema bombi (Microsporidia: Nosematidae) in the natural populations of bumblebees (Bombus spp.) from West Siberia.

Nosema bombi is an obligate intracellular parasite of bumblebees (Hymenoptera, Bombus spp.), which has significant negative effect on individual bumblebees, colony fitness, and development. Recently, several new genetic variants of N. bombi without a defined taxonomic status were identified in natural bumblebee populations from Russia, China, and several European countries, as well as N. ceranae, originally isolated from honey bees, was described in bumblebee species. Thus, it is required to investigate more Nosema variability in bumblebee populations for identifying new genetic Nosema variants. In our study, we used several methods such as total DNA isolation, polymerase chain reaction (PCR) amplification, cloning, sequencing, and comparative and phylogenetic analysis to investigate a prevalence of N. bombi and its diversity in the natural populations of bumblebees across West Siberia. DNA was extracted from intestinal bumblebee tissues. Identification of the parasite was conducted, using PCR with primers specific for the ribosomal RNA gene cluster and methionine aminopeptidase 2 gene of N. bombi followed by sequencing. Seven hundred twenty-seven individual bumblebees belonging to 16 species were tested; 64 specimens revealed presence of the parasite. Prevalence of Nosema bombi infection was different in each region and varied from 4 to 20 %. No infection was found in Bombus agrorum (n = 194) and Bombus equestris (n = 132), both common bumblebees in West Siberia. Three different genetic variants of the same species, N. bombi, were identified. The first variant belonged to N. bombi (AY008373) identified by Fies et al. (J Apicult Res 40:91-96, 2001), second (N. bombi WS2) was identical to the West Siberian variant identified by Szentgyörgyi et al. (Polish Journal of Ecology 59:599-610, 2011), and the last variant, N. bombi WS3, was new. The results led us to suggest that the prevalence of the N. bombi is related to the population structure of bumblebees and distribution of the particular genetic variants of N. bombi.