Russian Republic of Karelia is located at the Russian-Finnish border. It contains most of the historical Karelia land inhabited with autochthonous Karels and more recently migrated Russians. Although tuberculosis (TB) incidence in Karelia is decreasing, it remains high (45.8/100 000 in 2014) with the rate of multi-drug resistance (MDR) among newly diagnosed TB patients reaching 46.5 %. The study aimed to genetically characterize Mycobacterium tuberculosis isolates obtained at different time points from TB patients from Karelia to gain insight into the phylogeographic specificity of the circulating genotypes and to assess trends in evolution of drug resistant subpopulations. The sample included 150 M. tuberculosis isolates: 78 isolated in 2013–2014 (“new” collection) and 72 isolated in 2006 (“old” collection). Drug susceptibility testing was done by the method of absolute concentrations. DNA was subjected to spoligotyping and analysis of genotype-specific markers of the Latin-American-Mediterranean (LAM) family and its sublineages and Beijing B0/W148-cluster. The largest spoligotypes were SIT1 (Beijing family, n = 42) and SIT40 (T family, n = 5). Beijing family was the largest (n = 43) followed by T (n = 11), Ural (n = 10) and LAM (n = 8). Successful Russian clone, Beijing В0/W148, was identified in 15 (34.9 %) of 43 Beijing isolates; all В0/W148 isolates were drug-resistant. Seven of 8 LAM isolates belonged to the RD115/LAM-RUS branch, 1 - to the LAM RD174/RD-Rio sublineage. MDR was found in Beijing (32/43), Ural (3/10), and LAM (3/8). In contrast, all T isolates were pansusceptible. Comparison of drug resistant subgroups of the new and old collections showed an increasing prevalence of the B0/W148 clonal cluster, from 18.0 % (mainly polyresistant) in 2006 to 32.6 % in 2014 (mainly MDR and pre-XDR). The West–east increasing gradient is observed for the Ural genotype that may be defined a ‘Russian’ strain. In contrast, the spoligotype SIT40 of the T family appears to be a historical Karelian strain. Circulation of the MDR M. tuberculosis isolates of the Beijing genotype and its B0/W148 cluster continues to critically influence the current situation with the MDR-TB control in northwestern Russia including the Republic of Karelia. Revealed phylogeographic patterns of some genotypes reflect a complex demographic history of Karelia within the course of the 20th century.

Cystoviruses have a phospholipid envelope around their nucleocapsid. Such a feature is unique among bacterial viruses (i.e., bacteriophages) and the mechanisms of virion envelopment within a bacterial host are largely unknown. The cystovirus Pseudomonas phage phi6 has an envelope that harbors five viral membrane proteins and phospholipids derived from the cytoplasmic membrane of its Gram-negative host. The phi6 major envelope protein P9 and the non-structural protein P12 are essential for the envelopment of its virions. Co-expression of P9 and P12 in a Pseudomonas host results in the formation of intracellular vesicles that are potential intermediates in the phi6 virion assembly pathway. This study evaluated the minimum requirements for the formation of phi6-specific vesicles and the possibility to localize P9-tagged heterologous proteins into such structures in Escherichia coli. Using transmission electron microscopy, we detected membranous structures in the cytoplasm of E. coli cells expressing P9. The density of the P9-specific membrane fraction was lower (approximately 1.13 g/cm3 in sucrose) than the densities of the bacterial cytoplasmic and outer membrane fractions. A P9-GFP fusion protein was used to study the targeting of heterologous proteins into P9 vesicles. Production of the GFP-tagged P9 vesicles required P12, which protected the fusion protein against proteolytic cleavage. Isolated vesicles contained predominantly P9-GFP, suggesting selective incorporation of P9-tagged fusion proteins into the vesicles. Our results demonstrate that the phi6 major envelope protein P9 can trigger formation of cytoplasmic membrane structures in E. coli in the absence of any other viral protein. Intracellular membrane structures are rare in bacteria, thus making them ideal chasses for cell-based vesicle production. The possibility to locate heterologous proteins into the P9-lipid vesicles facilitates the production of vesicular structures with novel properties. Such products have potential use in biotechnology and biomedicine.

Видовое разнообразие нетуберкулезных микобактерий у больных микобактериозом на территориях Северо-Западного федерального округа России

Clarithromycin (CLR) is currently a key antibiotic for Helicobacter pylori infection treatment, however, the data on CLR resistance patterns in Russia are missing. Here, we applied WGS-based approach to H. pylori clinical isolates from Russia to comprehensively investigate sequence variation, identify putative markers of CLR resistance and correlate them with phenotypic susceptibility testing. The phenotypic susceptibility of 44 H. pylori isolates (2014–2022) to CLR was determined by disc diffusion method: 23 isolates were CLR-resistant and 21-CLR-susceptible. All isolates were subjected to WGS and submitted to GenBank. Based on complete sequence analysis, we showed that among all sequence variants, the combination of mutations A2146G/A2147G in the 23S rRNA gene is the most reliable for prediction of phenotypic susceptibility. For the first time, the average number of mutations in 106 virulence-associated genes between resistant and susceptible groups were compared. Moreover, this study presents the first WGS insight into genetic diversity of H. pylori in Russia with a particular focus on the molecular basis of drug resistance: the novel mutations were described as potential markers for the resistance development. Of these, the most prominent was a frameshift deletion (252:CGGGT) in HP0820 coding region, which is a good candidate for further investigation.

Background/Objectives: Non-typhoidal Salmonella (NTS) is one of the most common causative agents of food poisoning in Vietnam, and contaminated livestock meat poses a major risk to human health. The present study aims to provide the genetic characteristics of NTS with a particular focus on antimicrobial resistance and determine phylogenetic relationships between isolates from different sources in Southern Vietnam based on whole-genome sequencing (WGS) data. Methods: A total of 49 NTS isolates from pork/broiler meat, pigs, chickens, and humans were collected in Ho Chi Minh City and four provinces of Southern Vietnam. Phenotypic antimicrobial susceptibility testing (AST) and WGS for all isolates were performed. Results: As a result, 14 different serotypes were identified, among which S. Typhimurium and its monophasic variant were the dominant serotypes for human and pig sources. All chicken samples belonged to S. Indiana, whereas S. Infantis predominated in broiler meat. AST results revealed that 98% of isolates were multidrug resistant. NTS strains isolated from poultry and pigs exhibited resistance to the highest priority antimicrobials—quinolones and polymyxin, as well as to β-lactams, aminoglycosides, tetracycline, and sulfonamide, which are considered to be critical for the treatment of severe diseases. Conclusions: The results highlight the utmost importance of issues related to the selection, spreading, and transmission of multi-resistant strains from animals to humans.

Introduction. Helicobacter pylori was proved to be the principal causative agent of gastroduodenal disorders in human. Although Russian Federation is among the countries with a high prevalence of H. pylori infection (6090%), currently there is a very limited number of studies evaluating H. pylori genotypes in Russia. Objective. Based on the assessment of virulence-associated cagA, oipA, and vacA genes, our study was aimed to determine H. pylori genotypes associated with the clinical outcomes in patients with H. pylori infection in St. Petersburg, Northwest Russia. Materials and methods. Using PCR for the detection of cagA, oipA, and vacA s, m, i allelic variants, we analyzed 61 H. pylori isolates isolated and cultured from biopsies collected during endoscopy of patients with chronic gastritis (G), duodenal ulcer (DU), and gastric cancer (GC). Results. The genetic diversity of H. pylori clinical isolates has been revealed (HGDI 0.88): 41 (67%) of 61 H. pylori isolates were cagA-positive, 38 (62%) oipA-positive. The proportions of cagA+ isolates differed in patients with G (56.7%) and DU (80.9%) (p = 0.06). The s, m, and i allelic variants of the vacA gene were detected in all strains, although the vacA s1 allele was significantly dominant in patients with DU (95.2%) rather than with G (64.9%) (p = 0.01). The vacA alleles m1 and i1 in the isolates from patients with G and DU were found in almost equal proportions: 45.9% and 42.8% for m1 allele, 45.9% and 47.6% for i1 allele, respectively. Seven isolates (11.5%) were positive for different mixed combinations of vacA alleles s, m, and i. Noteworthy, all vacA s2 strains were cagA-negative and had the m2 allele. OipA+ strains were found in almost equal proportions in patients with G (62.2%) and DU (57.1%) (p = 0.71). All three cagA- and oipA-positive isolates from patients with GC carried vacA s1/m1/i1 alleles. Different combinations of virulence-associated determinants constituted 17 genetic profiles. The most common combined genotype cagA+/oipA+/vacA s1/m1/i1 comprised 18 (29.5%) H. pylori isolates. Conclusion. We have determined predominant genotypes in the H. pylori population in the Northwest of Russia. The significant association between vacA s1 genotype of the pathogen and clinical manifestations of H. pylori infection has been established in our study.