Vaccination with live attenuated vaccines is a key element in the prevention of lumpy skin disease. The mechanism of virus attenuation by long-term passaging in sensitive systems remains unclear. Targeted inactivation of virulence genes is the most promising way to obtain attenuated viruses. Four virulence genes in the genome of the lumpy skin disease virus (LSDV) Dermatitis nodulares/2016/Atyrau/KZ were sequentially knocked out by homologous recombination under conditions of temporary dominant selection. The recombinant LSDV Atyrau-5BJN(IL18) with a knockout of the LSDV005, LSDV008, LSDV066 and LSDV142 genes remained genetically stable for ten passages and efficiently replicated in cells of lamb testicles, saiga kidney and bovine kidney. In vivo experiments with cattle have shown that injection of the LSDV Atyrau-5BJN(IL18) at a high dose does not cause disease in animals or other deviations from the physiological norm. Immunization of cattle with the LSDV Atyrau-5BJN(IL18) induced the production of virus-neutralizing antibodies in titers of 4–5 log2. The challenge did not cause disease in immunized animals. The knockout of four virulence genes resulted in attenuation of the virulent LSDV without loss of immunogenicity. The recombinant LSDV Atyrau-5BJN(IL18) is safe for clinical use, immunogenic and protects animals from infection with the virulent LSDV.

In the winter of 2023/2024, the mass death of swans was observed on Lake Karakol on the eastern coast of the Caspian Sea. From 21 December 2023 to 25 January 2024, 1132 swan corpses (Cygnus olor, Cygnus cygnus) were collected and disposed of on the coast by veterinary services and ecologists. Biological samples were collected from 18 birds for analysis at different dates of the epizootic. It was found that the influenza outbreak was associated with a high concentration of migrating birds at Lake Karakol as a result of a sharp cold snap in the northern regions. At different dates of the epizootic, three avian influenza A/H5N1 viruses of clade 2.3.4.4.b were isolated from dead birds and identified as highly pathogenic viruses (HPAIs) based on the amino acid sequence of the hemagglutinin multi-base proteolytic cleavage site (PLREKRRRKR/G). A phylogenetic analysis showed that the viruses isolated from the swans had reassortations in the PB2, PB1, and NP genes between highly pathogenic (HP) and low-pathogenic (LP) avian influenza viruses. Avian influenza viruses A/Cygnus cygnus/Karakol lake/01/2024(H5N1) and A/Mute swan/Karakol lake/02/2024(H5N1) isolated on 10 January 2024 received PB2, PB1, and NP from LPAIV, while A/Mute swan/Mangystau/9809/2023(H5N1) isolated on 26 December 2023 received PB1 and NP from LPAIV, indicating that the H5N1 viruses in this study are new reassortants. All viruses showed amino acid substitutions in the PB2, PB1, NP, and NS1 segments, which are critical for enhanced virulence or adaptation in mammals. An analysis of the genomes of the isolated viruses showed that bird deaths during different periods of the epizootic were caused by different reassortant viruses. Kazakhstan is located at the crossroads of several migratory routes of migratory birds, and the possible participation of wild birds in the introduction of various pathogens into the regions of Kazakhstan requires further study.

Research in the field of medicinal products, especially medicinal plants, has been constantly expanding recently. However, to date, health research has not been widely disseminated. This article focus on the analysis of hexane and chloroform extracts of Artemisia scopaeformis by Gas Chromatography and Mass Spectroscopy (GC-MS) technique. Detailed chemical constituents of A. scopaeformis collected in Kazakhstan were investigated for the first time. The quantitative and qualitative analysis of bioactive constituents of the medicinal plant have been made. A. scopaeformis hexane extract contained 4 compounds: methyleugenol (33.87%), hexadecanoic acid, ethyl ester (41.02%), butyl 4,7,10,13,16,19-do cosahexaenoate (11.55%), hexasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11-dodecamethyl- (13.56%). The chloroform extract of A. scopaeformis contained 5 compounds: fluorene, 2,7-bis(1-hydroxyethyl)- (24.28%), p-dimethylaminobenzylidene p-anisidine (14.59%), 3-acetoxy-5-methyl-2-nitro- terephthalic acid, 4-isopropyl ester 1-methyl ester (11.74%), 3,4-diacetyl-2-methyl-4H-thieno [3,2-b] pyrrole-5- carboxylic acid, methyl ester (17.35%), 4H-1,2,4-triazole-3-thiol, 4-(2-fluorophenyl)-5-(1-methylethyl) – (32.04%). The resulting compounds have significant biological properties: antimicrobial, antioxidant, anti-inflammatory.

Rational design of capripoxvirus-based vaccine vectors can be achieved by knockout of immunomodulatory genes. In this study, the effect of knockout of the immunomodulatory genes LSDV005, LSDV008 and LSDV066 on the replication of Lumpy skin disease virus in cell cultures and the immune response to an integrated foreign antigen were assessed. The knockout of genes was performed by homologous recombination under conditions of temporary dominant selection. It was found that single knockout of the LSDV005 gene and the LSDV008 gene did not affect the replicative activity of recombinant viruses in vitro (Atyrau-5 and Atyrau-B). Both single knockout of the LSDV066 gene and in combination with knockout of LSDV005 or LSDV008 led to a decrease in the replicative activity of recombinant LSDVs. The recombinant Atyrau-5J(IL18) with LSDV005 gene knockout induced production of antibodies to the integrated antigen in mice. Prime-boost vaccination with all studied recombinants increased the level of interferon-γ. In addition, during immunization with the recombinant Atyrau-5J(IL18) secretion of interleukin-2 was significantly increased. The study of the functions of immunomodulatory genes and their effect on the expression of inserted sequences of foreign antigens is promising for the creation of highly effective polyvalent vector vaccines for animals.

Поксвирусы широко используются в качестве векторов при разработке вакцин против инфекционных заболеваний человека и животных. Ограниченный круг восприимчивых животных и непатогенность для человека каприпоксвирусов определяют огромный потенциал их использования в качестве вакцинных векторов. Технология получения рекомбинантных каприпоксвирусов может быть значительно улучшена путем сочетания временной доминантной селекции по гену gpt Escherichia coli (основан на устойчивости к микофеноловой кислоте) с дифференциальной экспрессией флуоресцентного белка. Рекомбинантный вирус генерируется путем замены зеленого флуоресцирующего белка (в акцепторном вирусе) целевым геном в результате двух последовательных рекомбинаций ДНК. В процессе первого кроссинговера плазмида интеграции с целевым геном встраивается в геном акцепторного вируса. В результате образуется нестабильная генетическая конструкция, которая может существовать только под селективным давлением микофеноловой кислоты. После снятия селективного давления происходит внутримолекулярная рекомбинация вирусного генома в участках гомологии, и образуются два типа вирусов: рекомбинант с целевым геном и исходный акцепторный вирус. Вовлеченные вирусы (зеленый исходный, зеленый промежуточный и бесцветный конечный) визуализируются по-разному с помощью флуоресцентной микроскопии, что позволяет использовать простой и эффективный протокол отбора целевых рекомбинантов. Этот метод (замена генов «от зеленого к бесцветному») значительно сокращает время, необходимое для получения безмаркерного рекомбинантного каприпоксвируса. Poxviruses are widely used as vectors in the development of vaccines against infectious diseases in humans and animals. The limited range of susceptible animals and the non-pathogenicity of capripoxviruses to humans determine the great potential for their use as vaccine vectors. The technology for producing recombinant capripoxviruses can be greatly improved by combining transient dominant selection for the gpt gene of Escherichia coli (based on resistance to mycophenolic acid) with differential expression of green fluorescent protein. The recombinant virus is generated by replacing the green fluorescent protein (in the acceptor virus) with the target gene by two consecutive DNA recombinations. During the first crossover, the integration plasmid with the target gene is inserted into the genome of the acceptor virus. As a result, an unstable genetic construct is formed, which can only exist under the selective pressure of mycophenolic acid. After the selective pressure is removed, intramolecular recombination of the viral genome occurs in homology regions, and two types of viruses are formed: a recombinant with a target gene and an initial acceptor virus. The viruses involved (green parent, green intermediate, and colorless final) are visualized differently by fluorescence microscopy, allowing for a simple and efficient protocol for selecting target recombinants. This method (replacement of genes "from green to colorless") significantly reduces the time required to obtain a marker-free recombinant capripoxvirus.

Вирус гриппа птиц (AIV) подтипа H5 остаѐтся одним из наиболее значимых факторов эпизоотической и биологической безопасности. В рамках данного исследования проведѐн молекулярно-генетический и филогенетический анализ изолятов вируса, выделенных от водоплавающих птиц Северо-Казахстанской области в 2024 году. Полные последовательности гена гемагглютинина (HA 1704 н.п.) были определены методом Сэнгера и использованы для построения филогенетического древа с моделью Tamura – Nei. Обнаруженный полибазичный сайт расщепления NVPQRETR↓GLF указывает на низкопатогенный характер вирусов, характерный для циркуляции среди диких птиц. Филогенетический анализ показал принадлежность изолятов к клади 2.3.4.4b и формирование отдельного кластера с высокой поддержкой (bootstrap 98%), близкого к азиатским штаммам из Китая, Монголии и Кореи. Ключевые позиции рецептор-связывающего участка (Gln-226, Gly-228) сохраняют птичий α2,3- сиалогликановый профиль, что свидетельствует об отсутствии адаптации к млекопитающим. Выявленные аминокислотные замены в антигенных сайтах Sa, Ca2 и Cb не нарушают общую антигенную структуру. Полученные результаты указывают на формирование локальной эволюционной линии H5-вирусов, устойчиво циркулирующей среди водоплавающих птиц региона. Работа подчѐркивает роль Северо-Казахстанской области как ключевого звена в миграционных путях и эпидемиологической сети Евразии и обосновывает необходимость дальнейшего молекулярного мониторинга и геномного секвенирования для раннего выявления потенциально патогенных вариантов.