Viral diarrhoea is one of the most common and economically important diseases in cattle, causing significant losses in productivity and increased mortality in young livestock. Effective diagnosis of the disease requires the use of standardised sera that ensure high accuracy and reproducibility of laboratory results. Selection of sera to obtain standard sera for the diagnosis of bovine viral diarrhea represents a key challenge in the development of a standard sera panel. During monitoring in some regions of the Republic of Kazakhstan (Almaty, Mangistau, Karaganda, Turkistan oblasts) 399 samples of biomaterial from cattle were collected in private subsidiary farms and serological method of research was applied. In a serological test, specific diarrhea virus antibodies were detected in 310 serum samples. Comparative studies of sensitivity and stability of the positive sera with positive standard serum from the kit (ID Vet) were performed. During selection of sera, several factors were determined: specificity and sensitivity of the sera used, with consideration of the peculiarities of the epizootic situation in the region. This research work culminates in the production of a positive serum for the candidate from field-derived natural BVDV disease infections, and shortens the production cycle and saves costs.

Bovine leukemia virus (BLV) remains a major concern for cattle industries worldwide due to its persistent nature, economic impact, and challenges in control. In this study, we conducted a comprehensive nationwide survey of BLV in Kazakhstan between 2014 and 2024, utilizing serological diagnostics to assess prevalence and characterize viral genotypes (2024). A total of 433,537 serum samples were screened by agar gel immunodiffusion (AGID), revealing an overall seroprevalence of 5.87%, with the highest rates observed in the North Kazakhstan, Kostanay, and East Kazakhstan regions. In 2024, a targeted analysis of 3736 serum and 536 whole blood samples across 17 regions was performed using AGID, ELISA, real-time PCR, and nested PCR. ELISA demonstrated higher sensitivity than AGID (10.4% vs. 8.2%), confirmed by statistical correlation (r = 0.97, p < 0.001) and a Wilcoxon signed-rank test (p = 0.026). Real-time PCR detected BLV DNA in 4.7% of samples, with the highest positivity in the East Kazakhstan and Abai regions, confirming active viral circulation. Validation of a domestically developed AGID diagnostic kit showed full concordance with commercial assays (IDEXX, IDvet), supporting its use in national surveillance programs. These findings highlight the endemic status of BLV in Kazakhstan. Molecular analysis of sequenced isolates revealed the presence of genotype G-7, consistent with strains circulating in neighboring countries. Together, these results underscore the importance of integrated serological and molecular approaches for effective monitoring and control.

The paper presents data on the study of genetic characteristics of the infl uenza virus A/chicken/ Chelyabinsk/30/2019 H9N2 isolated from pathological material (chicken internal organs) in February 2019 and received from the poultry farm in the Chelyabinsk Oblast. The H9N2 subtype of the isolated virus was identifi ed based on virological analysis. Sequencing of the hemagglutinin gene segment revealed that the amino acid sequence at the cleavage site was RSSR/GLF, which is characteristic of a low virulent avian infl uenza virus. Phylogenetic analysis of the obtained nucleotide sequences of the hemagglutinin gene fragment (1–1539 bp open reading frame) showed that the A/chicken/Chelyabinsk/30/2019 H9N2 isolate belongs to the G1 genetic group of the low virulent infl uenza virus A/H9, the representatives of which are widely spread in the Middle Eastern and Central Asian countries. The complete nucleotide genome sequence of the studied pathogen was determined. The comparative analysis of all genomic segments using the GenBank database revealed a close relationship (over 99%) between the A/chicken/Chelyabinsk/30/2019 H9N2 virus and the A/H9 infl uenza virus isolates circulating in Israel in 2006–2012. According to the analysis of the predicted amino acid sequence of the studied isolate, the positions of some molecular markers that determine the biological properties of the virus have been identifi ed. Most amino acid positions of hemagglutinin (according to H3 subtype sequence numbering) suggest affi nity for the ACA2-3Gal-receptors of avian epithelial cells. Amino acid substitutions were detected at the site within the receptor-binding domain as compared to the A/H9N2 infl uenza virus isolates obtained in Russia in 2018. The primary structure of the A/chicken/Chelyabinsk/30/2019 H9N2 isolate demonstrates a very high level of genetic similarity to the infl uenza virus isolate A/chicken/ Israel/215/2007 H9N2 used as a vaccine strain.

Avian infectious bronchitis is one of the most common viral infections causing enormous economic losses in the global poultry industry. Due to the lack of mechanisms to correct errors during genome replication, the virus can quickly mutate and generate new strains. This is facilitated by widespread use of live vaccines, simultaneous circulation of field viruses belonging to different serotypes in one flock and rapid spread of the virus. Previous studies of avian infectious bronchitis virus strains and isolates identified in the Russian Federation poultry farms showed that 50% of samples tested positive for the 4-91, D274, H-120, Ma5 vaccine strains, and the other half of samples tested positive for the field viruses belonging to eight GI genetic lineages, while the G1-19 (QX) lineage was dominant. The paper presents identification and genotyping results of the avian infectious bronchitis virus in one of the poultry farms in the Saratov Oblast (the Russian Federation) in 2018–2019. The samples of internal organs and blood, as well as oropharyngeal and cloacal swabs were taken from chicks and layers of different ages in the parent and replacement flocks. The vaccine strain, GI-19 field isolates and variant isolates that do not belong to any of the known genetic lineages were detected. Analysis of test results within a two-year period showed that it is important to study samples taken from birds of different ages. The virus undergoes modification and adaptation inducing new genetic forms by infecting several poultry generations, due to which the heterogeneity of the virus population is ob­served not only in the poultry farm as a whole or in a separate department, but also within one organism. The identified isolates showed tropism for the tissues of intestine, reproductive organs, and, in rare cases, trachea and lungs. The data obtained indicate that, despite the vaccination used, a genetically diverse population of the infectious bronchitis virus circulates in the poultry farm, while the infection may not manifest itself at an early age, but may affect the flock productivity in the future due to pathological changes in the reproductive organs of laying chickens.

Background. Enzootic bovine leukosis (EBL) remains a significant problem in cattle production. Agar gel immunodiffusion (AGID) is widely used for screening; however, diagnostic performance depends on test-kit manufacturing parameters. Aim. To develop and optimize manufacturing conditions for an AGID test system for serological diagnosis of EBL, ensuring reproducibility and compliance with World Organisation for Animal Health (WOAH) recommendations. Methods. BLV antigen was produced on a persistently infected FLK cell line and standardized to gp51 using WOAH reference serum E05. We optimized agarose concentration (0.8–2.0% in 0.2 M Tris, pH 7.2, with 8.5% NaCl), antigen dilutions (1:2–1:64), and control sera dilutions. Plates were incubated in a humid chamber at 20–27 °C with readings at 24–72 h. Validation was performed on bovine sera (positive/weak-positive/negative) and compared with commercial kits. Results. An agarose concentration of 1.0–1.2% provided clear, stable precipitin lines within 24–72 h, balancing diffusion rate and line quality. Optimal antigen dilutions were 1:4 (eliminating nonspecific lines with negative serum while maintaining high sensitivity) and 1:8. Positive control serum was optimal at 1:4–1:8; weak-positive control at 1:32 yielded a reproducible faint line suitable for sensitivity assessment. Negative serum produced no lines. Stable performance was achieved under specified storage and operating conditions; diagnostic characteristics were comparable to those of commercial tests. Conclusions. The optimized AGID test system demonstrates high reproducibility, specificity, and adequate sensitivity, aligns with international requirements, and is suitable for broad implementation in veterinary laboratories in Kazakhstan. Accounting for regional features further enhances its practical value.

Bovine Viral Diarrhea Virus (BVDV) is a globally important cattle pathogen causing substantial economic losses. In Kazakhstan, BVDV’s epidemiological status remains poorly characterized due to the absence of systematic surveillance. We carried out a cross-sectional study of cattle herds across Kazakhstan, using ELISA to detect anti-BVDV antibodies and RT-PCR to identify active infections. Positive samples underwent sequencing for phylogenetic analysis of circulating strains. Additionally, a standard reference serum panel was developed to measure virus neutralization titers (ND₅₀) and to evaluate cross-neutralization with Border Disease virus (BDV). Antibodies against BVDV were prevalent, with seropositivity ranging from 28.89% to 96.13% across surveyed regions. Active BVDV infection was confirmed by RT-PCR in 17 animals. Phylogenetic analysis with 2 samples from Mangystau region classified the virus as BVDV2 genotype. The reference serum panel exhibited high neutralizing titers ND₅₀ up to 1:286 against the local BVDV-1 isolate. Notably, these sera also neutralized BDV, albeit at lower titers ND₅₀ 1:45. These findings provide crucial baseline epidemiological data and enhanced diagnostic tools for BVDV in Kazakhstan. They highlight the need for improved surveillance and will inform strategic control measures against this economically significant cattle disease.