Eurasian Journal of Applied Biotechnology

GEOGRAPHIC DISTRIBUTION, ECOLOGICAL ADAPTABILITY AND AGRONOMIC STRATEGIES FOR BLACK PLUM (VITEX DONIANA SWEET) DOMESTICATION IN THE NIGERIAN SAVANNA

2025-08-29T09:01:10+05:00

Vitex doniana Sweet (Lamiaceae), commonly known as black plum, is a versatile tree species vital to Nigeria’s agroforestry systems, recognized for its nutritional, medicinal and ecological benefits. This comprehensive review synthesizes recent advances in the species geographic range, ecological preferences, morphological variation, propagation methods, phytochemical properties and conservation challenges, based on 83 georeferenced occurrence records from the Global Biodiversity Information Facility (GBIF) and studies published since 2010. The analysis shows Vitex doniana’s extensive presence across Nigeria’s savanna and forest zones, especially in Delta, Oyo and Taraba states, demonstrating its adaptability to a variety of habitats. Key findings include notable morphological diversity influenced by climatic and soil factors, promising propagation techniques and a diverse phytochemical profile supporting medical research. In addition to ecological understanding, the review incorporates agronomic considerations for domestication in Nigerian savannas, including soil management, nutrient application, water regulation, spacing, weed control and intercropping practices. These practical insights are essential for guiding large-scale cultivation and agroforestry integration. Challenges such as low seed germination, human activities and climate change continue to threaten the species' survival, calling for integrated strategies. This review recommends combining agroecological zoning, advanced propagation, agronomic standardization and community-led conservation to promote sustainable domestication and biodiversity protection in Nigeria.

EVALUATION OF GENETIC DIVERSITY OF SOYBEANACCESSIONS USING SSR MARKERS

2025-09-04T09:18:02+05:00

Soybean (Glycine max L.) is an important legume crop used to meet the protein and oil demands of many populations worldwide. Assessing the genetic diversity of soybean accessions is crucial for expanding the genetic base and improving the efficiency of breeding programs. The aim of this study was to evaluate the genetic diversity of 188 soybean accessions using 15 polymorphic SSR markers to determine the degree of their genetic differentiation and their potential for use in breeding. As a result of the study, a total of 55 alleles were identified, with an average of 2.68 alleles per locus. The polymorphic information content (PIC) ranged from 0.35 (Satt385) to 0.74 (Satt409), with an average of 0.61. The expected heterozygosity (He) ranged from 0.35 (Satt385) to 0.74 (Satt409), with an average value of 0.60. Furthermore, a phylogenetic Neighbor-Joining (NJ) dendrogram based on the binary allele data revealed three main clusters, each of which was divided into two subclusters. In addition, the analysis of molecular genetic data using the UPGMA method allowed the identification of two and one main clusters at different levels of genetic distance (0.21; 0.22). The obtained clusters reflect various relationships between local and foreign genotypes, indicating both the preservation of the local gene pool and its expansion through the inclusion of foreign cultivars. The information on genetic diversity among soybean genotypes obtained in this study will assist breeders in selecting parental lines for future breeding programs.

GENETIC DIVERSITY ANALYSIS OF ZEA MAYS L. ACCESSIONS BASED ON MICROSATELLITE MARKERS

2025-09-04T09:23:13+05:00

This study aimed to evaluate the genetic diversity of 21 maize (Zea mays L.) accessions cultivated in Kazakhstan using 21 SSR markers. The selected markers revealed substantial polymorphism, with polymorphic information content (PIC) values ranging from 0.557 to 0.962, indicating high marker informativeness. Genetic diversity indices such as the number of alleles (Na), effective number of alleles (Ne), Shannon’s information index (I), and Nei’s gene diversity index (uh) varied significantly among accessions, with the Kazakh accession ZM001 showing the highest diversity. Analysis of Molecular Variance (AMOVA) revealed that 66% of the total genetic variation was attributable to differences among accessions, confirming strong population differentiation (Fst = 0.611) among maize accessions. Cluster analysis, Principal Coordinate Analysis (PCoA), and STRUCTURE analysis consistently grouped accessions according to their geographic origin, distinguishing local, Chinese, and European accessions. These results highlight the effectiveness of SSR markers in revealing genetic structure and demonstrate the existence of untapped allelic variation in the maize germplasm of Kazakhstan.

STUDY OF BACTERIAL LYSATE IN THE FIGHT AGAINST ANTIBIOTIC RESISTANCE

2025-09-04T11:16:25+05:00

Bacterial lysates are widely used today as immunomodulatory agents that enhance the effectiveness of antibacterial therapy through synergistic interactions with antibiotics. The aim of this study was to evaluate the pharmacological properties of a bacterial lysate obtained from antibiotic-resistant strains of opportunistic microorganisms such as Acinetobacter baumaniі, Klebsiella pneumoniae SCAID PND1-2022 (246), and Pseudomonas aeruginosa SCAID PHRX1-2019 to assess its potential in combating the growing problem of antimicrobial resistance. The study analyzed the properties of the lysate and its effects on the body using in vivo studies at doses of 300.0 and 500.0 mg/kg. No signs of acute toxicity were observed, suggesting that this substance is safe for use. Furthermore, destroyed bacterial cells retain the ability to induce a specific immune response, stimulating immune system activation. Given the inherent resistance of bacterial strains to antibiotics, it can be concluded that the use of bacterial lysate promotes the development of an adaptive immune response directed against highly resistant pathogens, including superbugs.

FORECASTING THE SPATIAL DYNAMICS OF THE EPIZOOTIC PROCESS PLAGUE AMONG THE WILD ANIMALS IN THE DESERT PLAGUE FOCI OF KAZAKHSTAN FOR THE PERIOD 2020-2024. BASED ON GIS TECHNOLOGIES

2025-08-28T15:51:26+05:00

The present study is aimed at spatial and temporal analysis and prediction of the epizootic process of plague infection among wild animals in desert natural plague foci of the Republic of Kazakhstan using geoinformation modeling methods. The data of epizootological monitoring for 2020–2024 were used as the initial base, including information on the abundance and distribution of the main reservoir species (Rhombomys opimus), vector activity (ectoparasites), as well as climatic, landscape and anthropogenic characteristics of the studied territories.

Spatial data was processed and visualized using the ArcGIS and QGIS software platforms. Situational, analytical, and predictive electronic maps of epizootic activity have been constructed.

By the modeling results, it was found that in the future, epizootic activity is most likely to increase in the western parts of the desert zone of Kazakhstan, including in the North Aral, Volga-Ural sandy, Predustyurt and Ustyurt foci. A potential increase in risk is also expected in the Betpakdalinsky, Moyinkumsky and Dariyalyk-Takyr autonomous foci. Under the conditions of predicted climatic changes, the probability of expanding the circulation zones of the plague pathogen may increase, which increases the risk of infection of humans and camels in enzootic territories.

The research results have high applied significance and can be used to optimize epizootological surveillance systems, plan preventive and anti-epidemic measures, and minimize the risk of transmission of plague infection from animals to humans.

DETERMINATION OF THE ANTIOXIDANT ACTIVITY OF MICROCLONES OF WALNUT (JUGLANS REGIA L.)

2025-08-19T18:26:20+05:00

Walnut (Juglans regia L.), a plant belonging to the genus Juglans of the family Juglandaceae, native to the region extending from the Balkans east to the Himalayas. Since ancient times, walnut has been widely used in both folk and traditional medicine for various diseases and is among the medicinal plants of interest due to its rich content of antioxidants such as flavonoids, phenolic acids, melatonin, folate, gamma-tocopherol, selenium and proanthocyanidins in various organs (kernels, shells, roots and leaves). However, until now there have been virtually no studies on how walnut can respond to abiotic stresses in vitro culture and what benefits such a study can provide, so our work is devoted to the analysis of the antioxidant activity of J. regia shoot microclones in vitro after exposure to low positive temperatures. The results of this work show that Juglans regia effigia microclones subjected to cold stress have greater antioxidant potential than microclones grown under more favorable temperature conditions.

EFFECT OF HIGH- AND LOW-TEMPERATURE STRESS ON THE METABOLOME OF N. SIBIRICA PALL. AND NITRARIA SCHOBERI L. MICROCLONES IN VITRO

2025-08-11T10:30:12+05:00

The results of an in vitro study of the effects of high temperature and cold stress on the metabolome of N. sibirica Pall. and Nitraria schoberi L. microclones are presented. Gas chromatography–mass spectrometry (GC–MS) analysis revealed significant stress-induced changes in the metabolome spectra of the studied species. These results are important for the development of metabolomics and for improving our understanding of plant responses to abiotic stress factors. Furthermore, the study's results help determine the role of individual metabolites in these processes, which is important for developing strategies for the targeted induction of valuable secondary metabolites in vitro.

EFFECT OF DIFFERENT CRYOPROTECTORS ON VIABILITY OF FROZEN-THAWED SPERMATOZOA OF BALKHASH PERCH (PERCA SHRENKII KESSLER, 1874)

2025-08-28T12:19:12+05:00

In recent years, there has been active development of commercial aquaculture against the background of depletion of natural biological resources of water bodies, considering the growing need for food products. Cryopreservation of spermatozoa of rare and endangered fish species is of great importance, which will help preserve their biodiversity. Scientists are interested in perch as a potential commercial fish, but no studies have been conducted on cryopreservation of Balkhash perch spermatozoa. The Balkhash perch (Perca Shrenkii Kessler, 1874) is endemic to the Balkhash-Ili basin, and currently the population of this species is extremely low. In this study, work was carried out to determine the effect of various cryoprotectants on sperm motility before and after thawing for cryopreservation of Balkhash perch sperm. For this purpose, sperm from 10 males of this species was obtained and examined. During freezing, components of a protective solution were used with the addition of cryoprotectors glycerin and dimethyl sulfoxide to the sperm. The results of the study showed that before the addition of cryoprotectants, the motility was 74.3 ± 5.7%. Thawed samples frozen using the cryoprotectant 2.0 M glycerol retained motility significantly longer than those using 2.0 M dimethyl sulfoxide.

VIABILITY OF FIBROBLASTS OF TUGAI DEER (CERVUS HANGLU BACTRIANUS LYDEKKER, 1900) DURING CRYOPRESERVATION BY DIFFERENT METHODS

2025-08-28T12:19:56+05:00

Currently, there is a rapid and continuous decline in wild animal populations, which threatens accelerated loss of biodiversity on a global scale. Along with in situ conservation strategies, ex situ conservation measures such as germplasm cryopreservation should be widely applied. Cryopreservation in combination with assisted reproductive technologies can become an important means of successfully preserving the biodiversity of the animal world. Methods based on obtaining cell cultures are of great importance in scientific research. The resulting cell lines can be frozen and stored in containers with liquid nitrogen for many years. Such cell and tissue biobanks represent a viable source of genetic material and living cells that offer numerous opportunities for further research.

The aim of this study was to investigate the scientific basis for cryopreservation of somatic cells of tugai deer, which is a species in need of special protection. Studies were conducted on the viability of tugai deer fibroblasts using various cryopreservation methods. The viability of frozen-thawed tugai deer fibroblasts was determined, and it was shown that the most effective cryopreservation of this species fibroblasts was the use of a cryoprotectant 1.5 M ethylene glycol and a slow equilibrium freezing mode using a programmed freezer.

SAFETY ASSESSMENT OF AN ORAL RABIES VACCINE BAIT FOR WILD CARNIVORES IN A SERONEGATIVE DOG MODEL

2025-05-21T09:21:12+05:00

Rabies remains one of the most dangerous zoonotic diseases, causing tens of thousands of deaths annually. One of the key challenges in combating rabies is the vaccination of wild carnivores, domestic, and farm animals, which play a crucial role in the circulation of the virus in nature. The aim of the present study was to evaluate the safety and immunogenicity of an oral rabies vaccine in the form of a baited briquette using seronegative dogs as a model. During the experiment, clinical parameters, serological markers, and the behavior of animals following immunization and controlled infection were assessed. The obtained data indicate good vaccine tolerance, high bait acceptance, and the formation of a strong immune response in the vaccinated animals. The oral vaccine did not cause any adverse effects and provided protection against infection in the majority of vaccinated dogs. The results of the study confirm the potential for using this vaccine form in field conditions, both for target and non-target animals.

OPTIMIZATION OF THE MANUFACTURING CONDITIONS OF AN AGID TEST SYSTEM FOR THE DIAGNOSIS OF BOVINE LEUKEMIA

2025-09-03T10:58:30+05:00

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.

PREPARATION OF HYPERIMMUNE SERA FOR SARS-COV-2 VIRUS

2025-09-08T15:20:04+05:00

Hyperimmune sera are a primary component of diagnostic test systems utilising enzyme immunoassay to identify specific antibodies against infectious disease pathogens. During diagnostic assay development, hyperimmune sera function as positive controls and enable the generation of standard curves for quantitative measurement of antibody levels. Furthermore, they can be used to assess the sensitivity and specificity of established test systems and to verify novel diagnostic approaches. The objective of this study was to produce hyperimmune serum against SARS-CoV-2. Purified and concentrated virus, in conjunction with adjuvants such as aluminium hydroxide or AddaS03, was employed to immunise animals. Consequently, specific sera exhibiting activity in ELISA at dilutions of 1:1600- 1:3200 and in the diffusion precipitation reaction (DPR) at 1:8–1:32 were acquired. The research demonstrated that the use of concentrated, pure SARS-CoV-2 for animal immunisation, supplemented with adjuvants, is a viable and efficacious approach for generating highly active antibodies.

EVALUATION OF THE THERAPEUTIC EFFICACY OF PRECONDITIONED MESENCHYMAL STEM CELLS IN A MOUSE MODEL OF PSORIASIS: PASI AND HISTOLOGICAL ANALYSIS

2025-07-01T12:54:20+05:00

Psoriasis is a common immune-mediated inflammatory skin disease that affects 2-4% of the world's population and about 2.5% of the population of Kazakhstan. Mesenchymal stem cells (MSCs) are of particular interest for the treatment of psoriasis due to their strong immunomodulatory and regenerative potential. One of the strategies to increase the therapeutic efficacy of MSCs is their preconditioning with proinflammatory cytokines, which improves their survival and immunoregulatory properties. Aim of the study: Evaluation of the therapeutic effect of human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) preconditioned with combinations of the cytokines interleukin (IL)-17A, IL-22, and tumor necrosis factor-alpha (TNF-α) in a mouse model of imiquimod (IMQ)-induced psoriasis-like skin inflammation. Results: Assessment of skin inflammation severity using the PASI score, along with subsequent histological analysis, demonstrated that both intact and preconditioned hUCB-MSCs effectively reduced IMQ-induced skin inflammation. In addition, hUCB-MSCs preconditioned with a combination of IL-22 and TNF-α significantly reduced erythema and scaling, while hUCB-MSCs preconditioned with a combination of IL-17A, IL-22 and TNF-α significantly reduced skin thickening. Conclusion: Preconditioned hUCB-MSCs exhibit a pronounced anti-inflammatory potential, confirming their therapeutic promise for the treatment of psoriasis.

SEED GERMINATION BIOLOGY OF RARE TULIPA SPECIES IN NORTHERN AND CENTRAL KAZAKHSTAN

2025-06-23T18:00:01+05:00

Kazakhstan has one of the highest diversities of Tulipa species, most of which are threatened. The threat of their extinction necessitates the use of biotechnological approaches, such as in vitro micropropagation, which allows the preservation of valuable genotypes without harming natural populations.

In the process of increasing climate change and anthropogenic impact, these endemic species are at high risk of extinction. In this regard, the study and conservation of biodiversity of endemic plant species is considered a global priority throughout the world.

The aim of the work was to study the effect of temperature and growth regulators on germination of T. auliekolica, T. turgaica seeds for their introduction into in vitro cultures.

In vitro germination was carried out on ½ MS media with and without the addition of GA₃ (13 and 52 mg/l). Germination was recorded for 60 days, germination (%) and T₅₀ were calculated.

According to the results of the tetrazolium test, the viability of T. turgaica and T. auliecolica seeds was 95% and 100%, respectively. The studied species showed different temperature preferences; for T. auliecolica seeds, the optimal temperature was 4°C, and for T. turgaica, 10°C. In addition, T. auliecolica seeds germinated faster than T. turgaica seeds. The results showed that temperature significantly affects seed germination. Seeds of both species germinated only at low temperatures (4 and 10°C), at 20°C, seed germination was absent in both species. The obtained data are of significant practical importance for the creation of effective methods for in vitro cultivation of these rare Tulipa species. Selection of optimal GA₃ concentrations in combination with temperature regimes significantly increases the success of in vitro propagation, which plays a key role in programs for the conservation of biodiversity of endemic plants.

PRODUCTION OF MODIFIED RNA-DEPENDENT DNA POLYMERASE OF HUMAN IMMUNODEFICIENCY VIRUS FOR USE AS REVERSE TRANSCRIPTASE

2025-07-08T17:22:02+05:00

Human immunodeficiency virus reverse transcriptase is an RNA-dependent DNA polymerase that allows the production of complementary DNA on an RNA matrix. The original amino acid sequence of the reverse transcriptase was modified by making 18 substitutions and codonoptimized for expression in Escherichia coli cells. A plasmid vector in which the modified HIV reverse transcriptase gene was integrated under the control of the inducible promoter was obtained by restriction-free cloning. Transformation of competent E. coli cells with this vector yielded a strain producing recombinant modified HIV reverse transcriptase with a molecular mass of 62.5 kDa. The recombinant enzyme was chromatographically purified by two-step elution on Ni²⁺ ions and heparin, yielding 420 mg from 1 liter of induced culture. The RNA-dependent DNA polymerase activity of the purified enzyme was confirmed on RNA samples isolated from Secale cereale and human blood by amplification of 18S rRNA and actin B gene fragments, respectively. The recombinant modified reverse transcriptase exhibits polymerase activity in the temperature range of 50-70ºС, indicating its thermostability. Biochemical parameters show the promising application of the enzyme as a reverse transcriptase in scientific research and diagnostic practice.

CLONING OF THE BACTERIAL ESTERASE GENE AND DETERMINATION OF THE BIOCHEMICAL CHARACTERISTICS OF THE RECOMBINANT ENZYME

2025-07-14T17:21:47+05:00

Esterases are hydrolytic enzymes that catalyze the hydrolysis and transesterification of short-chain fatty acid esters. They are widespread in nature and can be found in plants, animals, and microorganisms. Microbial esterases are used in the production of biofuels, and in cosmetic, food, and pharmaceutical industries. Recently, microbial esterases have attracted interest due to their ability to hydrolyze plastics, particularly polyethylene terephthalate (PET). It was established that Bacillus paralicheniformis T7 secretes an enzyme with esterase activity of 18.32 ± 2.38 U/mL. Protein mass spectrometry in combination with proteomic and genomic analysis identified a bacillary esterase with a molecular weight of 26.8 kDa. Based on the complete genome sequence of the strain, oligonucleotides were designed to amplify the esterase gene, which was subsequently cloned into the pET-28c(+) vector. Transformation of Escherichia coli BL-21(DE3) cells with the esterase-containing vector resulted in a strain that produced recombinant esterase with a yield of 614 mg/L. The recombinant esterase was purified using Ni-NTA affinity chromatography on a Ni²⁺ column. Studies revealed that the recombinant esterase exhibits maximum activity at 40 °C and pH 7.0. The enzyme remains active within the temperature range of 30–55 °C and pH 5.0–8.0. Thermostability assays demonstrated that the esterase is stable after 15 minutes of incubation at 30–50 °C and within the pH range of 8.0–10.0. Submerged fermentation of B. paralicheniformis T7 followed by drying of the culture supernatant yielded a preparation with esterase activity of 15,328.1 ± 528.6 U/g. The results indicate that B. paralicheniformis T7 is a promising esterase-producing strain, and the enzyme itself holds potential for use as a hydrolytic biocatalyst in the degradation of fatty acid esters.

COMPARATIVE ANALYSIS OF PHYSIOLOGICAL RESPONSES OF POTATO SEEDLINGS TO OSMOTIC STRESS OF DIFFERENT INTENSITIES

2025-08-26T12:09:02+05:00

Drought is one of the most significant abiotic stresses that limit plant growth and crop productivity. This study analyzed the physiological responses of potato (Solanum tuberosum L.) seedlings to osmotic stress induced by polyethylene glycol (PEG-6000) at concentrations of 0, 2, 4, 6, and 8%. High concentrations of PEG-6000 were found to significantly inhibit seedling growth and reduc chlorophyll content. Physiological analysis revealed decreased in malondialdehyde (MDA) levels and increased catalase (CAT) activity under osmotic stress conditions. These results deepen our understanding of the physiological processes and internal mechanisms of potato drought tolerance, including changes in chlorophyll content. This knowledge can be applied to breeding programs that develop cultivars resistant to abiotic stresses.

IMMUNOCHROMATOGRAPHIC ANALYSIS: ASSESSMENT AND PROSPECTS FOR APPLICATION IN THE DIAGNOSIS OF ROTAVIRUS AND ADENOVIRUS INFECTIONS

2025-09-03T09:23:42+05:00

The objective of this review was to provide information on immunochromatographic assays (ICA) relevant to the diagnosis of human adenovirus (HAdV) and rotavirus (HRV) infections. HAdVs and HRVs cause respiratory and gastrointestinal diseases in children and are characterized by strong inflammatory responses. Virological diagnostic methods include virus isolation in cell cultures, molecular-biological techniques, and immunological assays. These methods are considered the gold standard; however, obtaining results requires a significant amount of time. The development of reverse transcription polymerase chain reaction (RT-PCR) has greatly facilitated the diagnosis of viral infections due to its high specificity and sensitivity. However, the complexity of the reaction protocol and the need for additional laboratory equipment make RT-PCR unsuitable for point-of-care testing. Immunochromatographic analysis, due to its low cost, simple sample preparation, and ease of use, represents an excellent alternative for bedside diagnostics. In studies on other infections, ICA has demonstrated the absence of cross-reactivity, high reproducibility, and acceptable sensitivity compared to enzyme-linked immunosorbent assay (ELISA) and RT-PCR. In conclusion, ICA test systems may be useful in clinical practice for the rapid detection of rotavirus and adenovirus infections. However, the possibility of false-positive results and differences in subtype identification across various diagnostic methods should be taken into account.

OPTIMIZATION OF IN VITRO MICROPROPAGATION PROTOCOL OF THE ENDEMIC, ENDANGERED SPECIES AMYGDALUS LEDEBOURIANA FOR CONSERVATION

2025-07-30T14:37:35+05:00

The article presents the results of a study on the in vitro conservation of the rare and endangered species Amygdalus ledebouriana. This almond species is noted for its high resistance to drought and low temperatures, as well as the nutritional value of its fruits. Prior to this study, no research had been conducted on the micropropagation of Amygdalus ledebouriana. In the present work, the authors developed an effective protocol for explant sterilization and establishment in vitro culture. A 9% hydrogen peroxide solution applied for 5 minutes was selected as the most suitable sterilizing agent, ensuring up to 75% explant viability. For shoot multiplication, the QL nutrient medium supplemented with 0.25 mg/L 6-benzylaminopurine (BAP) was optimized, resulting in an average of 7.24 shoots per explant. To date, 400 microshoots have been successfully propagated. An in vitro collection has been established from three natural populations of Amygdalus ledebouriana.

SELECTION OF MARKER REGIONS OF GENES FOR THE MOLECULAR IDENTIFICATION OF HELMINTHS OF THE ASCARIDOIDEA SUPERFAMILY FOUND IN COD FISH

2025-07-10T16:44:30+05:00

In recent years, the consumption of imported fish has increased significantly in Kazakhstan, especially representatives of the cod family, which are characterized by a high level of parasitic invasion. Anisakidosis is a parasitic disease caused by nematodes of the Anisakidae family. The main symptoms of this disease are disorders of the gastrointestinal tract and allergic reactions. Humans are random hosts, consuming raw or undercooked fish and seafood. Until recently, it was believed that heat-treated fish was not dangerous, but recent studies show a high degree of allergy to anisakid antigens, which remain active after heat treatment. In addition, existing data indicate differences in the level of allergenicity between different species of nematodes in the family. In this regard, the species identification of anisakids is relevant for taxonomy, monitoring of distribution and minimizing potential risks to humans. Therefore, the purpose of this study is to select marker regions of genes for the identification of nematodes in cod family fish. Of the four types of primers, NC13/NC2 and NC5/NC2 are the most specific and optimal for amplifying the 5.8S and ITS-2 regions of rDNA. The nucleotide sequences obtained by us were identified as nematodes of the species Anisakis simplex and Hysterothylacium aduncum.

RECOMBINANT PRODUCTION OF THE P60 ANTIGEN OF LISTERIA MONOCYTOGENES

2025-09-05T17:27:01+05:00

Listeria monocytogenes is a significant foodborne pathogen characterized by an intracellular life cycle, pronounced adaptive mechanisms, and a broad spectrum of virulence factors. The aim of this study was to obtain the recombinant p60 antigen of L. monocytogenes from isolates circulating in the Republic of Kazakhstan. Using recombinant DNA technology, a producer strain Escherichia coli ArcticExpress(DE3)/Lmp60 was generated, enabling the expression of the protein with a molecular mass of 50.3 kDa. The specificity of the recombinant p60 antigen was confirmed by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The obtained results highlight the diagnostic value of the p60 protein of L. monocytogenes as a target antigen for the development of highly specific ELISA-based test systems aimed at the rapid identification of L. monocytogenes in food products.

THE ROLE OF TREC AND KREC IN ASSESSING THE IMMUNE STATUS OF NEWBORNS

2025-09-15T15:38:15+05:00

Newborn screening, based on the quantitative assessment of T-cell receptor excision circles (TREC) and kappa-deleting recombination excision circles (KREC) in dried blood spots, allows for the early diagnosis of various types of primary immunodeficiencies. TREC and KREC are circular DNA, markers of new lymphocyte production, formed during the maturation of T- and B-cells. These DNAs do not possess replication capability and are diluted as a result of cell division, making them useful markers for evaluating the appearance of new lymphocytes. PCR analysis of TREC and KREC enables the precise quantitative measurement of their levels, which is crucial for the diagnosis and treatment of immune diseases related to T- and B-cells. The implementation of TREC and KREC assays into neonatal screening programs facilitates the early detection of immunodeficiencies and the identification of novel genetic defects, thereby expanding early diagnostic capabilities.

This review provides a detailed examination of the role of TREC and KREC markers in the diagnosis of immunodeficiencies in newborns. Additionally, it analyzes the global experience of implementing TREC and KREC into national newborn screening programs.

PRECLINICAL DEVELOPMENT OF A CHIMERIC YELLOW FEVER / TICK-BORNE ENCEPHALITIS VIRUS AS A CANDIDATE VACCINE

2025-09-19T13:33:21+05:00

Tick-borne encephalitis virus (TBEV) is a reemerging pathogen in Kazakhstan. Despite the availability of inactivated TBEV vaccines produced abroad, their reliance on complex multi-dose regimens and frequent boosters limits their implementation for routine use in Kazakhstan. New technologies, including chimerization of different Flavivirus species, enable the development of vaccine candidates which require only a single dose to achieve long-lasting immunity. The ChimeriVax platform leverages the efficient replication machinery of the yellow fever virus (YFV) 17D vaccine strain engineered to express the structural proteins of a different flavivirus.

In this work, the ChimeriVax YFV/TBEV virus was created by replacing the prM-E genes in the YFV genome with the prM-E genes of TBEV. Preclinical evaluation demonstrated robust replication (~10^8 focus-forming units, FFU/mL) in cell cultures and genetic stability over multiple passages. In murine models, the chimeric virus elicited transient viremia (peaking at 10^4 FFU/mL) without mortality even at high doses (10^5 FFU). Immunization induced potent neutralizing antibodies (geometric mean titer: 4,076) and robust cellular immunity, marked by production of the cytokines IFN-γ, TNF-α, and IL-2 upon antigen stimulation. These results position the ChimeriVax YFV/TBEV virus as a promising vaccine candidate.

DEVELOPMENT OF A REAGENT KIT FOR THE DETECTION AND TYPING OF THE “WEST” TOPOTYPE OF BLUETONGUE VIRUS SEROTYPE 9 (BTV-9W) IN BIOLOGICAL SAMPLES USING RT-qPCR

2025-08-06T07:59:35+05:00

Bluetongue (BT), also known as sheep catarrhal fever, is a highly pathogenic viral infection caused by the bluetongue virus (BTV). This disease holds significant economic importance as it is characterized by high mortality rates, reduced productivity, deteriorated animal health, and economic losses.

Kazakhstan is considered free from BT, but the primary vectors of BTV, midges of the genus Culicoides, are widely distributed throughout the country. Southern Kazakhstan offers favorable conditions for the reproduction of midges, and the high density of susceptible livestock combined with the import of animals from BTV-endemic regions creates a significant risk of virus spread. This underscores the need for the development of a domestic test system for the detection of BTV via quantitative PCR, capable of differentiating vaccine strains from field strains of the virus.

This study presents the results of development and validation of a PCR test system for the diagnosis of BT. During development, real-time reverse transcription PCR (RT-qPCR) was utilized for the precise detection of BTV, enabling identification of small amounts of viral RNA in biological samples. The kit is based on primers and a fluorescently labeled probe targeting a conserved region of BVT genome (segment 10), ensuring high specificity and ability to detect various virus serotypes. This test system will be in demand in market and will enhance food security in country.