Background/Objectives: Klebsiella pneumoniae is a major Gram-negative pathogen associated with community-acquired pneumonia (CAP) and a critical contributor to antimicrobial resistance (AMR). Culture-based diagnostics remain the clinical standard but may underestimate microbial diversity and resistance gene profiles. This pilot study compared pathogen detection and antimicrobial resistance gene (ARG) repertoires in matched K. pneumoniae pure cultures and primary sputum samples using targeted next-generation sequencing (tNGS). Methods: We analyzed 153 sputum samples from patients with CAP. Among 48 culture-positive cases, 22 (14% overall; 54% culture-positive) yielded K. pneumoniae. MALDI-TOF MS, phenotypic drug susceptibility testing, and tNGS were conducted on both culture isolates and matched sputum specimens. Microbial composition, ARG diversity, and method concordance were evaluated, with focused analysis of discordant and fatal cases. Results: K. pneumoniae was detected in 14.4% of all CAP cases and accounted for 54.2% of culture-positive samples. Identification rates differed across methods: 35% by MALDI-TOF MS, 45% by culture tNGS, and 29% by sputum tNGS. Sputum tNGS revealed substantially higher microbial diversity than cultures (3.04 vs. 1.42 species per sample) and detected more than sixfold unique ARGs (38 vs. 7), including clinically relevant determinants that were absent from culture isolates. Concordance was high between MALDI-TOF MS and culture tNGS (κ = 0.712), but low between sputum and culture tNGS (κ = 0.279). Among twelve K. pneumoniae isolates included in AMR analysis, all showed resistance to β-lactams, and two-thirds exhibited MDR/XDR phenotypes. Genotypic screening identified seven ARGs, but major ESBL and carbapenemase genes were not detected, suggesting the presence of alternative resistance mechanisms. Overall, sputum tNGS provided additional etiological and resistome information not captured by cultivation and complemented classical diagnostics in CAP involving K. pneumoniae. Conclusions: Culture-based diagnostics and tNGS provide complementary insights into the detection and resistance profiling of K. pneumoniae in CAP, with sputum tNGS revealing broader microbial and resistome information than pure cultures, while classical methods remain essential for species confirmation and phenotypic AST. An integrated diagnostic approach combining both methodologies may improve pathogen detection, guide antimicrobial therapy, and enhance AMR surveillance in K. pneumoniae-associated CAP.

Background and Aims:Pneumonia is an inflammatory condition of the lower respiratory tract, commonly caused by infection and associated with substantial morbidity and healthcare utilization. The burden of pneumonia on healthcare is exacerbated by limited data on pathogenic causes and associated antimicrobial resistance (AMR). Here, we conducted a literature review and synthesized data on the microbial causes and AMR in hospitalized pneumonia patients from Kazakhstan. Methods:We searched PubMed, Medline, Embase, Web of Science, the Cochrane Central Register of Controlled Trials, eLIBRARY, and CyberLeninka for observational studies of hospitalized pneumonia patients reporting microbiologically confirmed pathogen and antimicrobial resistance data from Kazakhstan. Exclusion criteria were unclear pneumonia definitions or absence of pathogen-specific AMR data. We assessed study quality using the Joanna Briggs Institute checklist and extracted data on microbial prevalence and resistance rates by pathogen-antibiotic combinations. We synthesized microbial etiology and AMR patterns and performed random-effects meta-analysis of prevalence data. Results:Nine studies represented 1534 isolates from 1474 inpatients (2008-2022). The most prevalent pathogens were Streptococcus pneumoniae (17.8%), Streptococcus spp. (17.3%), Haemophilus influenzae (12.6%), and Klebsiella pneumoniae (10.7%). Hospital-acquired pathogens comprised 48% of isolates. Acinetobacter baumannii had the highest resistance, exceeding 70% across major antibiotic classes. Escherichia coli had high resistance to β-lactams (> 66%) and fluoroquinolones (63%). Pseudomonas aeruginosa had 60% fluoroquinolone and 27% carbapenem resistance. Fluoroquinolone resistance was widespread across multiple pathogens. K. pneumoniae had relatively lower resistance rates. Pathogen distribution varied by clinical setting and patient population. Conclusion:Data on the etiologic causes and AMR in pneumonia are sparse in Kazakhstan and methodologically heterogeneous. Hospital-acquired pneumonia pathogens were highly prevalent and associated with high AMR rates. These findings underscore the need for improved pathogen surveillance and antimicrobial stewardship in Kazakhstan to address the high burden of antibiotic-resistant and hospital-acquired pneumonia.

Background/Objectives: Staphylococcus aureus, particularly methicillin-resistant strains, is a leading cause of severe pneumonia. Understanding local molecular epidemiology, including virulence gene profiles and antimicrobial resistance (AMR) mechanisms, is crucial for effective infection control. This pilot study aimed to characterize S. aureus isolates from pneumonia patients in Karaganda, Kazakhstan. Methods: We collected 48 respiratory samples from patients with pneumonia across three medical institutions. Bacterial identification was performed using MALDI-TOF MS. Antimicrobial susceptibility testing (AST) was carried out using European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. Whole-genome sequencing of S. aureus isolates was conducted on an Ion Torrent S5 platform. Genomic analysis included multilocus sequence typing (MLST), identification of virulence and AMR genes, and phylogenetic reconstruction. Results: S. aureus was identified in 14.6% (n = 7) of pneumonia cases included in this study. All isolates (100%, n = 7) were phenotypically resistant to benzylpenicillin. The mecA gene was detected in 57.1% of isolates (n = 4), while phenotypic resistance to methicillin was observed in 28.6% (n = 2) of the isolates. Resistance to azithromycin (57.1%, n = 4) and levofloxacin (42.9%, n = 3) was observed among the isolates. Two isolates (28.6%) were multidrug-resistant (MDR). Genomic analysis revealed the prevalence of the ST22 clone (57.1%, n = 4) in the studied cohort. Other sequence types were ST97, ST8, and ST45 (14.3% each). Phylogenetic analysis showed clustering consistent with MLST profiles. All isolates carried a conserved core virulence arsenal, including hemolysin (hla, hlg), biofilm-forming genes (icaADBC), immune evasion genes (sak, scn), and iron acquisition genes (isd). The Panton-Valentine leukocidin (PVL) genes were detected in three isolates. AMR gene analysis revealed the ubiquitous presence of mepA and tetracycline efflux pump genes, along with regulatory genes (arlRS, mepR, mgrA). The blaZ and ermA genes were not detected despite high phenotypic resistance to penicillin and macrolides. Conclusions: This study reports the identification of the virulent and resistant ST22 S. aureus clone in pneumonia cases in Karaganda, Kazakhstan. The discordance between phenotypic and genotypic AMR profiles underscores the necessity for integrated diagnostic approaches.

This study presents the results of analyzing the sputum microbiome of 46 patients diagnosed with pneumonia using targeted NGS sequencing of the 16S rRNA gene. As a result of the study, 114 species of bacteria belonging to 47 genera and 29 families were identified. Based on the obtained data, potential pathogens associated with the development of pneumonia were identified, and a quantitative analysis of their prevalence among patients was conducted. At the species level, Streptococcus pneumoniae was detected in 26 samples (56.5%). Acinetobacter baumannii was found in 10 samples (21.7%). Klebsiella pneumoniae was detected in 9 samples (19.5%), and Pseudomonas aeruginosa was found in 17.4% of samples. Other species that may be involved in the development of pneumonia included Escherichia coli, Haemophilus influenzae, and Staphylococcus aureus in 15.2%, 10.87%, and 8.7% of samples, respectively. Additionally, Neisseria meningitidis was detected in 5 samples (10.8%), and Stenotrophomonas maltophilia was found in two samples. The obtained results can contribute to a deeper understanding of microbiome communities associated with pneumonia and enrich knowledge about the microbiome of the upper respiratory tract. These data may be useful for developing new strategies for the diagnosis and treatment of pneumonia, as well as for optimizing preventive measures.

Background/Objectives: Acinetobacter baumannii is an increasingly significant nosocomial pathogen causing severe infections globally. The emergence of multidrug-resistant A. baumannii strains has raised concerns about the efficacy of current treatment options. This study aimed to investigate the molecular epidemiology and antimicrobial resistance patterns of A. baumannii isolates from Kazakhstan. Methods: We collected nine A. baumannii isolates in 2022–2023 in Karaganda, Kazakhstan, which were then subjected to whole-genome sequencing (WGS) using the IonTorrent platform for genome characterization. Multilocus sequence typing (MLST) was used to classify the isolates into distinct clonal complexes. In addition, antibiotic susceptibility testing was conducted using the standard methods for a range of antibiotics commonly used against A. baumannii. Results: Our results revealed a high degree of genomic diversity among isolates from Kazakhstan, with multiple distinct classes identified: ST78 (n = 4, 44.4%), ST15 (n = 2, 22.2%), ST2 (n = 2, 22.2%), and ST193 (n = 1, 11%). MLST analysis showed that ST78Pas/1104Oxf (harboring blaOXA-72 and blaOXA-90 genes) were prevalent among the multidrug-resistant isolates. Based on the results of MLST, KL, and OCL, the analyzed isolates were assigned to specific international clones: IC2—ST2(Pas)-KL2/168-OCL1, IC4—ST15(Pas)-KL9-OCL7, and IC6—ST78(Pas)-KL49-OCL1. Notably, these isolates exhibited resistance to multiple antibiotics including meropenem, imipenem, gentamicin, amikacin, and ciprofloxacin. Conclusions: This study highlighted the complex molecular epidemiology of A. baumannii in Kazakhstan over a two-year period, underscoring the need for targeted surveillance strategies to monitor antimicrobial resistance patterns. The emergence and dissemination of multidrug-resistant strains within this timeframe emphasizes the importance of whole-genome sequencing as a diagnostic tool and underscores the challenges posed by these infections.

The influenza virus strain A/mute swan/Mangystau/1-S24R-2/2024 (H5N1; clade 2.3.4.4b) was isolated in embryonated chicken eggs from the lung of a dead swan found around Lake Karakol (Kazakhstan) during a highly pathogenic avian influenza outbreak in 2024. The aim of this study was to characterize the genetic profile of the isolated strain.

Мыт - инфекционное заболевание лошадей, вызываемое грамположительной бактерией Streptococcus equi, характеризующееся абсцессами в подчелюстных и глоточных лимфатических узлах, приводящими к обструкции дыхательных путей. В этом исследовании представлены результаты разработки полимеразной цепной реакции (ПЦР) в реальном времени с использованием зондов TaqMan для обнаружения ДНК S. equi, возбудителя удушья у лошадей. Были разработаны специфические праймеры и зонды, нацеленные на ген rimI («метилтрансфераза I большой субъединицы рибосомальной РНК») S. equi. Было продемонстрировано, что ПЦР в реальном времени позволяет обнаружить всего 50 фемтограмм геномной ДНК. Отсутствие перекрестных реакций с другими респираторными вирусными и бактериальными микроорганизмами, использованными в исследовании, указывает на высокую специфичность ПЦР в реальном времени. Разработанная ПЦР в режиме реального времени значительно сокращает время обработки и получения диагностических результатов, тем самым обеспечивая быстрое реагирование на ранней стадии инфекции, что станет основным преимуществом для контроля и сдерживания распространения мыта лошадей.

Diagnosis of acute respiratory infections (ARIs) is challenging due to the broad diversity of potential microbial causes. We used metagenomic next-generation sequencing (mNGS) to analyze the nasopharyngeal virome of ARI patients, who had undergone testing with a clinical multiplex PCR panel (Amplisens ARVI-screen-FRT). We collected nasopharyngeal swabs from 49 outpatient adults, 32 of whom had ARI symptoms and were PCR-positive, and 4 asymptomatic controls in Kazakhstan during Spring 2021. We assessed the biodiversity of the mNGS-derived virome and concordance with PCR results. PCR identified common ARI viruses in 65% of the symptomatic cases. mNGS revealed viral taxa consisting of human, non-human eukaryotic and bacteriophage groups, comprising 15, 11 and 28 genera, respectively. Notable ARI-associated human viruses included rhinovirus (16.3%), betaherpesvirus 7 (14.3%) and Epstein-Barr virus (8.16%). The primary phage hosts were Streptococcus spp. (32.7%), Pseudomonas aeruginosa (24.5%) and Burkholderia spp. (20.4%). In total, 47% of ARIs were linked solely to bacterial pathogens, a third to viral-bacterial co-infections, and less than 10% to only viral infections by mNGS. PCR showed low concordance with mNGS, except for rhinovirus. These results underscore the importance of broad diagnostic methods and question the effectiveness of commonly used PCR panels in ARI diagnosis.

Background Respiratory infections are a leading cause of morbidity and mortality worldwide, with a significant proportion thought to have a viral aetiology. Traditional diagnostic approaches often rely on targeted assays for “common” respiratory pathogens, leaving a substantial fraction of infections unidentified. Here, we used metagenomic next generation sequencing (mNGS) to characterize the nasopharyngeal virome associated with acute respiratory infection (ARI), with and without a positive PCR test result for a panel of common respiratory viruses. Methods Nasopharyngeal swabs from symptomatic outpatients (n=49), of whom 32 tested positive by a multiplex viral PCR, and asymptomatic controls (n=4) were characterized by mNGS. The virome taxa were stratified into human, non-human eukaryotic host, and bacteriophage sub-groups. We used a phage host classification as a proxy to establish bacterial taxa present in the nasopharynx. We then compared the virome biodiversity and presence of pathogens with known respiratory effects across the participant sub-groups. Results The nasopharyngeal virome exhibited similar diversity across the PCR-positive and - negative subsets. Among the top ARI-associated human viruses were enterovirus (16.3%, human rhinovirus, HRV-A), roseolovirus (14.3%, human betaherpesvirus 7, HBV-7) and lymphocryptovirus (8.16%, Epstein-Barr virus, EBV). The top three ARI-associated phage hosts were Streptococcus spp (32.7%), Pseudomonas aeruginosa (24.5%) and Burkholderia spp. (20.4%). The virome of both asymptomatic and symptomatic subjects was also abundant in the Staphylococcus (60.4%) and Propionibacterium (Cutibacterium) acnes bacteriophages (90.6%). The PCR and mNGS results were relatively concordant for human rhinovirus (HRV), but not for other PCR panel targets, including human parainfluenza (HPIV), adenovirus (HAdV), bocavirus (BoV) and seasonal coronavirus (HCoV). Conclusions mNGS revealed a high diversity of pathogens that could be cause to respiratory symptomatology, either as a single infection or a co-infection between viral and bacterial species. The clinical significance of the mNGS versus multiplex PCR findings warrants further investigation.

Background and Aim: Equine influenza (EI) is a highly contagious disease that causes fever and upper respiratory tract inflammation. It is caused by influenza virus A, belonging to the Orthomyxoviridae family, with subtypes H3N8 and H7N7. This study presents data on the development of a real-time polymerase chain reaction (RT-PCR) assay using TaqMan probes to detect the H3 subtype of EI virus (EIV). Materials and Methods: The evaluation of the developed RT-PCR assay involved five strains of EIV as positive controls and ten nasopharyngeal swab samples collected from horses. RNA was isolated using the GeneJet Viral DNA and RNA Purification Kit, and primers and probes were designed using the Integrated DNA Technology PrimerQuest Tool. The assay was optimized by investigating the annealing temperature, primer and probes concentrations, sensitivity, and specificity. Sequencing was performed using the Thermo Fisher 3130 Genetic Analyzer, and the evolutionary history was inferred using the Neighbor-Joining method. Results: The designed primers and probes, targeting the H3 gene, were found to be specific to the EIV. The RT-PCR assay was capable of detecting as low as 50 femtogram (f) or 3 × 103 copies of genomic RNA. No cross-reactions were observed with other respiratory viral and bacterial pathogens, indicating the high specificity of the assay. To evaluate its effectiveness, ten nasopharyngeal swab samples collected from farms in North Kazakhstan regions during disease monitoring were analyzed. The accuracy of the analysis was confirmed by comparing the results with those obtained from a commercial RT-PCR assay for EI identification. The developed RT-PCR assay exhibited high sensitivity and specificity for detecting the EIV. Conclusion: The results demonstrate that the developed RT-PCR assay is suitable for diagnosing EI. This simple, highly sensitive, and specific assay for detecting H3 EIV can be a reliable tool for diagnosing and surveilling EI. Implementing this RT-PCR assay in veterinary practice will enhance and expedite the timely response to potential outbreaks of EI, thus positively impacting the overall epizootic well-being of EI in Kazakhstan.

Here, we report the full nucleotide sequence of the RvA1B/KZ/2021/87 rhinovirus, identified through metagenomic sequencing of nasopharyngeal swabs collected from patients exhibiting respiratory symptoms in Kazakhstan during 2021.

Background and Aim:Infectious bovine keratoconjunctivitis (IBK) is a prevalent ocular disease that affects livestock, leading to substantial economic losses due to reduced production and culling of infected animals. Moraxella spp. is common bacterial pathogens that can cause keratoconjunctivitis in livestock. Therefore, rapid and accurate diagnosis is crucial for effective treatment and disease control. This study aimed to develop a multiplex real-time polymerase chain reaction (mRT-PCR) assay for the detection and differentiation of Moraxella bovoculi, Moraxella ovis, and Moraxella bovis. Materials and Methods:Three reference strains of Moraxella as positive controls and 36 lacrimal swab samples collected from cattle were used to evaluate the developed mRT-PCR assay DNA extraction that was performed using the RIBO-sorb DNA/RNA extraction kit. Primers and probes were designed using the SpeciesPrimer pipeline. The annealing temperature, primer and probe concentrations, and sensitivity and specificity of the assay were optimized. Results:An mRT-PCR assay was developed to detect pathogens associated with IBK in cattle on the basis of optimized parameters. The specificity and sensitivity of this assay were confirmed using samples containing individual pathogens (O - M. ovis, B - M. bovis, and BO - M. bovoculi), combinations of two pathogens (O-B, B-BO, and O-BO), and when the DNA of all three pathogens was present in a single reaction (O-B-BO). The analytical sensitivity of mRT-PCR for detecting M. ovis and M. bovoculi DNA was 21 copies or 50 fg per reaction, whereas that for M. bovis was 210 copies or 500 fg per reaction. In addition, this assay has been tested on samples isolated from the affected eyes of cattle in the Akmola region of the Republic of Kazakhstan. Conclusion:For the first time in the Republic of Kazakhstan, the proposed mRT-PCR assay for the simultaneous detection of three Moraxella spp. pathogens has been developed. This assay exhibits the required specificity and high sensitivity for m RT-PCR, facilitating the timely implementation of effective measures for disease control and the prevention of economic losses. These losses are linked to a reduction in livestock breeding value, a reduction in meat and milk production, a reduction in the reproductive performance of heifers, resulting in fewer offspring, as well as costs related to the treatment of affected animals.

Background The epidemiology of respiratory tract infections (RTI) has dramatically changed over the course of the COVID-19 pandemic. A major effort in the clinical management of RTI has been directed toward diagnosing COVID-19, while the causes of other, common community RTI often remain enigmatic. To shed light on the etiological causes of RTI during a low COVID-19 transmission period in 2021, we did a pilot study using molecular testing for virologic causes of upper RTI among adults with respiratory symptoms from Almaty, Kazakhstan. Methods Adults presenting at two public hospitals with respiratory symptoms were screened using SARS-CoV-2 PCR on nasopharyngeal swabs. A subset of RTI+, COVID-19-negative adults ( n = 50) was then tested for the presence of common RTI viruses and influenza A virus (IAV). Next generation virome sequencing was used to further characterize the PCR-detected RTI pathogens. Results Of 1,812 symptomatic adults, 21 (1.2%) tested SARS-CoV-2-positive. Within the COVID-19 negative outpatient subset, 33/50 subjects (66%) had a positive PCR result for a common community RTI virus, consisting of human parainfluenza virus 3-4 (hPIV 3-4) in 25/50 (50%), rhinovirus (hRV) in 2 (4%), hPIV4-hRV co-infection in four (8%) and adenovirus or the OCR43/HKU-1 coronavirus in two (4%) cases; no IAV was detected. Virome sequencing allowed to reconstruct sequences of most PCR-identified rhinoviruses and hPIV-3/human respirovirus-3. Conclusions COVID-19 was cause to a low proportion of symptomatic RTI among adults. Among COVID-negative participants, symptomatic RTI was predominantly associated with hPIV and hRV. Therefore, respiratory viruses other than SARS-CoV-2 should be considered in the clinical management and prevention of adult RTI in the post-pandemic era.

The COVID-19 pandemic and heightened perception of the risk of emerging viral infections have boosted the efforts to better understand the virome or complete repertoire of viruses in health and disease, with a focus on infectious respiratory diseases. Next-generation sequencing (NGS) is widely used to study microorganisms, allowing the elucidation of bacteria and viruses inhabiting different body systems and identifying new pathogens. However, NGS studies suffer from a lack of standardization, in particular, due to various methodological approaches and no single format for processing the results. Here, we review the main methodological approaches and key stages for studies of the human virome, with an emphasis on virome changes during acute respiratory viral infection, with applications for clinical diagnostics and epidemiologic analyses.

Here, we reported the complete coding sequence of the influenza A/equine/Otar/3/2007 (H3N8) equine virus, first isolated in Kazakhstan in 2007. The hemagglutinin (HA) sequences of the Kazakhstan isolates appeared to be closely related to viruses isolated in early 2000 in Asia. Phylogenetic analysis characterized the Kazakhstan isolates as a member of the Florida sublineage clade 2 by the HA protein sequence.

Capripoxviruses with a host range limited to ruminants have the great potential to be used as vaccine vectors. The aim of this work was to evaluate attenuated sheep pox virus (SPPV) vaccine strain NISKHI as a vector expressing several genes. Open reading frames SPPV020 (ribonucleotide kinase) and SPPV066 (thymidine kinase) were selected as sites for the insertion of foreign genes. Two integration plasmids with expression cassette were designed and constructed. Recombinant SPPVs expressing an enhanced green fluorescent protein (EGFP) (rSPPV(RRΔ)EGFP and rSPPV(TKΔ)EGFP), Foot-and-mouth disease virus capsid protein (VP1), and Brucella spp. outer membrane protein 25 (OMP25) (rSPPV(RRΔ)VP1A-(TKΔ)OMP25) were generated under the transient dominant selection method. The insertion of foreign genes into the SPPV020 and SPPV066 open reading frames did not influence the replication of the recombinant viruses in the cells. Successful foreign gene expression in vitro was assessed by luminescent microscopy (EGFP) and Western blot (VP1 and OMP25). Our results have shown that foreign genes were expressed by rSPPV both in permissive (lamb testicles) and non-permissive (bovine kidney, saiga kidney, porcine kidney) cells. Mice immunized with rSPPV(RRΔ)VP1A-(TKΔ)OMP25 elicited specific antibodies to both SPPV and foreign genes VP1 and OMP25. Thus, SPPV NISKHI may be used as a potential safe immunogenic viral vector for the development of polyvalent vaccines.

We report the near-complete genome sequence of an influenza H5N1 virus strain isolated from a dead swan on the southeastern Caspian seashore in 2006. The results of the surface protein HA phylogenetic analysis showed that the A/swan/Mangystau/3/2006 virus belongs to the EA-nonGsGD clade.

Background: The remarkable diversity and mobility of Newcastle disease viruses (NDV) includes virulent viruses of genotype VI. These viruses are often referred to as pigeon paramyxoviruses 1 because they are normally isolated and cause clinical disease in birds from the Columbidae family. Genotype VI viruses occasionally infect, and may also cause clinical disease in poultry. Thus, the evolution, current spread and detection of NDV are relevant to avian health. Methods: Nucleotide sequencing and phylogenetic studies of three Kazakhstan isolations were performed to characterize the complete fusion (F)-protein gene as well as whole genome sequence. Sequence data were compared with 106 Fusion genes representing different NDV genotypes and sub-genotypes, as well as 225 fusion genes and 37 whole genome of class VI NDV strains from different regions of the world at different time periods. Phylogenetic trees were constructed to determine evolutionary relationships among these strains. We analysed fusion (F) protein gene and whole genome sequences, including the cleavage site. Results: The complete genome of these 3 isolates contained 15142bp, 15085bp and 15102bp in length, similar to those of Newcastle disease virus (NDV) strains in genotypes VIg, with the gene order 3’-NP-P-M-F-HN-L-5’. The cleavage site of the fusion protein was 112KRQKR116-F117, a feature generally associated with virulent NDV strains. Phylogenetic analysis, based on genomic sequences, SNP and fusion gene sequences, revealed that three isolates should be classified as class II genotype VIg NDVs. Phylogenetic analysis revealed that the NDV from various sites in Kazakhstan was highly similar genetically and that it clustered together with NDV of genotype VIg. Based on our data analysis, VIg isolates shared highest sequence identity with Russian and Ukraine isolates of the VIg subgenotype, suggests the possible spread of velogenic NDV in this region through cross-border live bird trade. Conclusion: Our study provides baseline information on the genetic characteristics of NDV circulating in Kazakhstan and we propose that the evolutionary and epidemiological study of virulent NDV could help to provide accurate molecular data about variants circulating in this region, thus aiding in the design of more efficient recombinant vaccines.

Coronaviruses are positive-stranded RNA viruses that infect a variety of hosts, resulting in a range of symptoms from gastrointestinal illness to respiratory distress. Bats are reservoirs for a high diversity of coronaviruses, and focused surveillance detected several strains genetically similar to MERS-coronavirus, SARS-coronavirus, and the human coronaviruses 229E and NL63. The bat fauna of central Asia, which link China to eastern Europe, are relatively less studied than other regions of the world. Kazakhstan is the world's ninth largest country; however, little is understood about the prevalence and diversity of bat-borne viruses. In this study, bat guano was collected from bat caves in three different sites of southern Kazakhstan that tested positive for coronaviruses. Our phylogenetic reconstruction indicates these are novel bat coronaviruses that belong to the genus Alphacoronavirus. In addition, two distinct lineages of Kazakhstan bat coronaviruses were detected. Both lineages are closely related to bat coronaviruses from China, France, Spain, and South Africa, suggesting that co-circulation of coronaviruses is common in multiple bat species with overlapping geographical distributions. Our study highlights the need for collaborative efforts in understudied countries to increase integrated surveillance capabilities toward better monitoring and detection of infectious diseases.

In recent years, there has been a noticeable expansion of the range and severity of the tan spot in Kazakhstan. The high adaptive ability of the fungus contributes to the wide spread and increased harmfulness of the pathogen. It is quite difficult to identify the species of fungus by the symptoms of the manifestation of the disease on the leaves of plants and by the morphological features of conidia. The creation of domestic diagnostic tools to identify the causative agent of wheat tan spot Pyrenophora tritici-repensis is an urgent task. Among molecular methods, the most convenient and operational method is PCR diagnostics. PCR makes it possible to detect fungal DNA even before physiological symptoms become visible on plant tissue. The aim of this work is to develop a PCR test system for the diagnosis of wheat tan spot. As a result of the studies, species-specific primers were selected and synthesized to identify the causative agent of wheat tan spot, the conditions for PCR formulation were optimized: the composition of the reaction mixture and the temperature and time regime. The specificity and sensitivity of the test system was determined. The developed test system showed rather high specificity and allows detection of the pathogen nucleic acid in the samples under study in the amount of 2,6 pg. This test system can be used for the rapid detection of pathogen DNA in samples. Keywords: tan spot, PCR, specificity, sensitivity, diagnosis.