Bovine Leukemia Virus Research Articles

A clinical case of enzootic bovine leukosis diagnosed by using clonal analysis of peripheral B lymphocytes in a Japanese Black cow

A 7-year-old Japanese Black cow with anorexia was presented at a local veterinarian. As rectal palpation revealed an enlarged induration of the uterus, lymphoma was suspected, and the patient was introduced to the Veterinary Teaching Hospital of Obihiro University of Agriculture and Veterinary Medicine. Although lymphadenopathy and lymphocytosis were not found, a high proviral load of bovine leukemia virus (BLV) with 464 copies/10 ng DNA was recorded, and lymphocytes with constricted or flower-like nuclei were found occasionally in the peripheral blood; accordingly, enzootic bovine leukosis (EBL) was suspected. According to PCR for clonality analysis of B cells based on immunoglobulin heavy chain (IGH) gene rearrangement, monoclonal proliferation of peripheral blood B cells was evident, suggesting the presence of tumorigenic B cells in the peripheral blood and onset of EBL. Pathological examination including necropsy and histopathological examination confirmed the diagnosis of EBL. Clonality analysis for B cells in the peripheral blood is thought to be useful for diagnosing B-cell tumors such as EBL, especially in cases lacking lymphadenopathy and lymphocytosis. This is the first clinical case of EBL diagnosed by demonstrating monoclonal proliferation of peripheral blood B cells using IGH gene rearrangement-based PCR.

Bovine leukemia virus encoded blv-miR-b4-3p microRNA is associated with reduced expression of anti-oncogenic gene in vivo.

Bovine leukemia virus (BLV) is a retrovirus that causes malignant B-cell lymphoma in up to ten-percent of infected cattle. To date, the mechanisms of BLV linked to malignant transformation remain elusive. Although BLV-encoded miRNAs have been associated with the regulation of different genes involved in oncogenic pathways, this association has not been evaluated in cattle naturally infected with BLV. The objective of this study was to determine the relative expression of BLV-encoded miRNA blv-miR-b4-3p, the host analogous miRNA bo-miR-29a and a couple of potential target mRNAs (HBP-1 and PXDN, with anti-tumorigenic function in B-cells), in cattle naturally infected with BLV compared to uninfected animals (control group). We observed that PXDN was significantly downregulated in BLV-infected cattle (P = 0.03). Considering the similar expression of endogenous bo-miR-29a in both animal groups, the downregulation of PXDN in BLV-naturally infected cattle could be linked to blv-miR-b4-3p expression in these animals. Knowing that PXDN is involved in anti-tumoral pathways in B-cells, the results presented here suggest that blv-miR-b4-3p might be involved in BLV tumorigenesis during natural infection with BLV in cattle.

The current situation on bovine leukemia in Dagestan and methods of its diagnosis

The article presents the current situation on bovine leukemia in the municipalities of Dagestan and a comparative analysis of serological and molecular-genetic methods of diagnosis. In municipal districts, urban districts and areas of animal husbandry for the first half of 2022, the number of animals examined by immune diffusion 531111 identified 2871, infected with the leukemia virus, which is 0,5 % from the number diagnosed. The degree of infection of livestock with bovine leukemia virus in rural areas varied from 0,01 to 5,2 %. The average coverage rate of serological studies of animals on leukemia – 57,2 %. Hematological studies of seropositive animals were not carried out during the analyzed period. 79 new leukemia-affected points have been registered, 95 unhealthy points have been transferred since 2021, only 3 points have been rehabilitated. As on 01.07.2022, 171 leukemia-affected points are registered, including 38 in agricultural enterprises, 4 of them in breeding farms, 27 farms and 106 farms. A comparative analysis of the data on the carrier of antibodies and DNA provirus showed that ELISA revealed 7 animals more carriers of antibodies than in AGID, PCR study revealed 14 and 7 animals more carriers of DNA provirus than carriers of antibodies in AGID and ELISA, respectively. To maximize the detection of seropositive BLV of animals, the integrated use of AGID, ELISA and PCR methods as a promising system in anti-leukemia measures is proposed. The introduction of ELISA and PCR diagnostics into the veterinary practice of the Dagestan Republic will make it possible to determine virus transmission in animals from the first day after birth (ELISA) and from the 20th day of the calf's life (PCR), as well as to increase the effectiveness and shorten the duration of anti-leukemia measures the final stages of the recovery of agricultural enterprises from leukemia infection.

Artificial neural network in forecasting of the epizootic situation on the leukosis of cattle

In the real work results of possible scenarios of development of an epizooty of the bovine leukosis virus (BLV) in the Omsk region by means of the spectral analysis and the artificial neural network (ANN) are presented. At the initial stage of researches as a result of comparison of numbers of Wolf to a trend of contamination 2 synchronized sites between the analyzed indicators at an interval of 23 years are established. That was the basis for correction of the earlier described working period of the computer mathematical model which is based on properties of a trigonometrical polynom of Fourier. The recalculation of coefficients of model made further, proceeding from two provisions of a trend for 23 years (from 1994 to 2016 and from 1999 to 2021), weak variability of amplitudes of fluctuations was defined and more considerable dispersion of phase fine tuning of harmonicas. On the basis of the made models the short-term forecast of increase in an indicator of contamination of BLV from 2022 to 2023 connected with the expected strengthening of solar activity is made. At the final stage by means of the special NeuroPro program several trained artificial neural networks (ANN) with nonlinear dependence on Wolf's numbers and harmonicas of the spectral model Fourier on an entrance according to which the estimated level of virus infection carrier state in the territory of the Omsk region in 2022 will be from 14.15 to 26.12% are created.

SELECTION OF SPECIFIC PRIMERS FOR PCR IN BOVINE LEUKEMIA VIRUS

the polymerase chain reaction (PCR) method is widely used to solve various problems. PCR is widely used for the detection of bacterial and viral pathogens. Primers are a very important component of PCR, since the specificity of amplification depends primarily on them. They are necessary for the enzyme to work and are specific to the fragment of interest. Based on the results of the selection of nucleotide sequences of the genome or individual fragments of the virus RNA from the international database on the NCBI website, a large number of sequences for the bovine leukemia virus were identified, which are stored in gene banks and updated daily with new data. The construction of primers in compliance with the necessary parameters is carried out using various computer programs, the main of which are MUSCLE, UGENE V.36.0, Primer-BLAST, Oligo Analyzer and others. The designed primers were then synthesized on the Expedite 8909 oligonucleotide synthesizer, according to the instructions attached to the device. As a result of our experiments, we selected and synthesized specific synthetic oligonucleotides env (g51)_1 and env (g51)_2, for setting up PCR for bovine leukemia.

Phylogenetic analysis based on whole genome sequence of bovine leukemia virus in cattle under 3 years old with enzootic bovine leukosis.

Enzootic bovine leukosis (EBL) is one of bovine neoplasms caused by bovine leukemia virus (BLV). Although EBL is typically observed in cattle over 3 years old, several cases of EBL onset in cattle under 3 years old have been reported in Japan. The mechanism for EBL onset in young cattle remains unclear. Although genetic variation of BLV is limited, the variations could affect viral properties relating to BLV pathogenesis. The purpose of this study was to clarify relationship between early onset of EBL and BLV groups. Moreover, we also aimed to characterize BLV that cause early onset of EBL. Whole genome sequences of BLV in 72 EBL cattle under 3 years old and 50 EBL cattle over 3 years old were identified. Phylogenetic analysis showed that BLV was divided into 4 groups (A, B-1, B-2 and Other). The BLV from EBL cattle under 3 years old were mainly classified as group A and B-1, while those from EBL cattle over 3 years old were mainly included in group B-2. Common sequence of group A and B-1 was compared with those of group B-2. Specific sequences in LTRs, gag-pro-pol, env and tax gene regions were identified in these groups. Amino acid substitutions of Pro and Tax protein were predicted in those nucleotide sequences. Those genetic variations might contribute to the early onset of EBL.

Effect of Bovine Leukemia Virus Infection and Proviral Load on Health of Dairy Heifers through the Transition Period

Effect of Bovine Leukemia Virus Infection and Proviral Load on Health of Dairy Heifers through the Transition Period

Effect of Bovine Leukemia Virus Infection and Proviral Load on Health of Dairy Heifers through the Transition Period

Effect of Bovine Leukemia Virus Infection and Proviral Load on Health of Dairy Heifers through the Transition Period

Изучение генетической вариабельности выделенных изолятов вируса лейкоза крупного рогатого скота в Белгородской области

Abstract. The purpose of the research Was to study genetic diversity of bovine leukemia virus isolates obtained in the Belgorod Region using restriction fragment length polymorphism method (RFLP). Scientific novelty. Bovine leukosis is one of the most common chronic infectious diseases of cattle in many countries of the world, which causes significant economic losses in the livestock industry. The typing of bovine leukemia virus (BLV), the study of its genetic structure, the evaluation of the mutation vector and a more detailed disclosure of the biological properties of the pathogen represent fundamental and applied value. Methods. The object of the research was 3-4-year-old cows infected with leukemia virus (n = 10), identified by serological methods in disadvantaged dairy farms. The immunodiffusion (ID) test, hematologic studies, PCR, genotyping, statistical processing of obtained data were conducted. Results. Conducted hematological studies determined the stage of the leukemic process in each animal. The target env fragment of the BLV gene (444 bp) was amplificated by 2-stage nested PCR, and this region was genotyped for all studied leukemia virus isolates using the restriction fragment length polymorphism (RFLP) method. In the course of the work, specific regions of the BLV env (gp51) gene, 970 bp long, were also obtained. We have given a primary assessment of the genetic diversity of BLV with the establishment of a genetic group (Belgian genotype according to RFLP). In the course of the work, specific regions of the env gp51 BLV gene, 444 bp long, were obtained. These fragments will be used for further DNA sequencing followed by phylogenetic analysis and determination of amino acid changes in the structure of the surface glycoprotein (gp51) of the bovine leukemia virus. Monitoring studies of BLV genotypes and the study of antigenic changes in the pathogen will allow timely development of the latest means of controlling and restricting the spread of bovine leukosis and improvement of diagnostic serological and PCR test systems.

Diagnosis and phylogenetic analysis of bovine leukemia virus in dairy cattle in northeastern Brazil.

Enzootic bovine leukosis (EBL) is a chronic viral disease of wide distribution in cattle herds and may take several years for the first manifestation of clinical signs. Most animals do not present clinical signs. However, the economic losses are underestimated due to this disease. Thus, this work aimed to detect and characterize BLV in dairy cattle in the Maranhão state, northeastern Brazil. Blood samples were collected from 176 animals from 8 municipalities in the southeastern state of Maranhão. Bovine blood samples were subjected to DNA extraction and molecular diagnosis using nested PCR assays for BLV, targeting gp51 gene. Positive samples were then sequenced and then subjected to phylogenetic inferences. BLV DNA was detected in 16 cattle (16/176, 9.09%) in 4 municipalities. Phylogenetic analyzes showed that the sequence obtained clustered in a clade containing BLV sequences classified as genotype 6, with a high degree of support. Our data shows BLV occurrence in the Northeast of Brazil and the identification of genotype 6 in this region. These findings contribute to the molecular epidemiology of this agent in Brazil.

Correlation between the Biodistribution of Bovine Leukemia Virus in the Organs and the Proviral Load in the Peripheral Blood during Early Stages of Experimentally Infected Cattle.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis. However, the propagation and distribution of BLV after primary infection still need to be fully elucidated. Here, we experimentally infected seven cattle with BLV and analyzed the BLV proviral load (PVL) in the blood and various organs. BLV was first detected in the blood of the cattle after one week, and the blood PVL increased for three weeks after infection. The PVL was maintained at a high level in five cattle, while it decreased to a low or medium level in two cattle. BLV was distributed in various organs, such as the heart, lung, liver, kidney, abomasum, and thymus, and, notably, in the spleen and lymph nodes. In cattle with a high blood PVL, BLV was detected in organs other than the spleen and lymph nodes, whereas in those with a low blood PVL, BLV was only detected in the spleen and lymph nodes. The amount of BLV in the organs was comparable to that in the blood. Our findings point to the possibility of estimating the distribution of BLV provirus in organs, lymph nodes, and body fluids by measuring the blood PVL, as it was positively correlated with the biodistribution of BLV provirus in the body of BLV infection during early stages.

Identifying Pathogen and Allele Type Simultaneously in a Single Well Using Droplet Digital PCR.

In the transmission control of chronic and untreatable livestock diseases such as bovine leukemia virus (BLV) infection, the removal of viral superspreaders is a fundamental approach. On the other hand, selective breeding of cattle with BLV-resistant capacity is also critical for reducing the viral damage to productivity by keeping infected cattle. To provide a way of measuring BLV proviral load (PVL) and identifying susceptible/resistant cattle simply and rapidly, we developed a fourplex droplet digital PCR method targeting the BLV pol gene, BLV-susceptible bovine major histocompatibility complex (BoLA)-DRB3*016:01 allele, resistant DRB3*009:02 allele, and housekeeping RPP30 gene (IPATS-BLV). IPATS-BLV successfully measured the percentage of BLV-infected cells and determined allele types precisely. Furthermore, it discriminated homozygous from heterozygous carriers. Using this method to determine the impact of carrying these alleles on the BLV PVL, we found DRB3*009:02-carrying cattle could suppress the PVL to a low or undetectable level, even with the presence of a susceptible heterozygous allele. Although the population of DRB3*016:01-carrying cattle showed significantly higher PVLs compared with cattle carrying other alleles, their individual PVLs were highly variable. Because of the simplicity and speed of this single-well assay, our method has the potential of being a suitable platform for the combined diagnosis of pathogen level and host biomarkers in other infectious diseases satisfying the two following characteristics of disease outcomes: (i) pathogen level acts as a critical maker of disease progression; and (ii) impactful disease-related host genetic biomarkers are already identified. IMPORTANCE While pathogen-level quantification is an important diagnostic of disease severity and transmissibility, disease-related host biomarkers are also useful in predicting outcomes in infectious diseases. In this study, we demonstrate that combined proviral load (PVL) and host biomarker diagnostics can be used to detect bovine leukemia virus (BLV) infection, which has a negative economic impact on the cattle industry. We developed a fourplex droplet digital PCR assay for PVL of BLV and susceptible and resistant host genes named IPATS-BLV. IPATS-BLV has inherent merits in measuring PVL and identifying susceptible and resistant cattle with superior simplicity and speed because of a single-well assay. Our new laboratory technique contributes to strengthening risk-based herd management used to control within-herd BLV transmission. Furthermore, this assay design potentially improves the diagnostics of other infectious diseases by combining the pathogen level and disease-related host genetic biomarker to predict disease outcomes.

Combined Immune Checkpoint Blockade Enhances Antiviral Immunity against Bovine Leukemia Virus.

Bovine leukemia virus (BLV) is a retrovirus that causes enzootic bovine leukosis (EBL) in cattle and is widespread in many countries, including Japan. Recent studies have revealed that the expression of immunoinhibitory molecules, such as programmed death-1 (PD-1) and PD-ligand 1, plays a critical role in immunosuppression and disease progression during BLV infection. In addition, a preliminary study has suggested that another immunoinhibitory molecule, T-cell immunoglobulin domain and mucin domain-3 (TIM-3), is involved in immunosuppression during BLV infection. Therefore, this study was designed to further elucidate the immunoinhibitory role of immune checkpoint molecules in BLV infection. TIM-3 expression was upregulated on peripheral CD4+ and CD8+ T cells in BLV-infected cattle. Interestingly, in EBL cattle, CD4+ and CD8+ T cells infiltrating lymphomas expressed TIM-3. TIM-3 and PD-1 were upregulated and coexpressed in peripheral CD4+ and CD8+ T cells from BLV-infected cattle. Blockade by anti-bovine TIM-3 monoclonal antibody increased CD69 expression on T cells and gamma interferon (IFN-γ) production from peripheral blood mononuclear cells from BLV-infected cattle. A syncytium formation assay also demonstrated the antiviral effects of TIM-3 blockade against BLV infection. The combined inhibition of TIM-3 and PD-1 pathways significantly enhanced IFN-γ production and antiviral efficacy compared to inhibition alone. In conclusion, the combined blockade of TIM-3 and PD-1 pathways shows strong immune activation and antiviral effects and has potential as a novel therapeutic method for BLV infection. IMPORTANCE Enzootic bovine leukosis caused by bovine leukemia virus (BLV) is an important viral disease in cattle, causing severe economic losses to the cattle industry worldwide. The molecular mechanisms of BLV-host interactions are complex. Previously, it was found that immune checkpoint molecules, such as PD-1, suppress BLV-specific Th1 responses as the disease progresses. To date, most studies have focused only on how PD-1 facilitates escape from host immunity in BLV-infected cattle and the antiviral effects of the PD-1 blockade. In contrast, how T-cell immunoglobulin domain and mucin domain-3 (TIM-3), another immune checkpoint molecule, regulates anti-BLV immune responses is rarely reported. It is also unclear why PD-1 inhibition alone was insufficient to exert anti-BLV effects in previous clinical studies. In this study, the expression profile of TIM-3 in T cells derived from BLV-infected cattle suggested that TIM-3 upregulation is a cause of immunosuppression in infected cattle. Based on these results, anti-TIM-3 antibody was used to experimentally evaluate its function in influencing immunity against BLV. Results indicated that TIM-3 upregulation induced by BLV infection suppressed T-cell activation and antiviral cytokine production. Some T cells coexpressed PD-1 and TIM-3, indicating that simultaneous inhibition of PD-1 and TIM-3 with their respective antibodies synergistically restored antiviral immunity. This study could open new avenues for treating bovine chronic infections.

Bovine leukemia virus: a perspective insight into the infection and immunity.

Bovine leukemia virus (BLV) is a member of the Retroviridae family and belongs to the Deltaretrovirus genus. It has a close relationship with human T-cell leukemia virus type I. BLV is responsible for causing enzootic bovine leukosis (EBL), a contagious disease that affects the bovine lymphatic system. This virus poses challenges for the global cattle industry, as it impacts cattle populations all over the world. Despite being widespread and impactful, BLV often goes unnoticed, with many researchers unaware of its presence and the potential consequences it carries. BLV demonstrates varying levels of pathogenicity. The majority of cattle (around 70%) become seropositive asymptomatic carriers, displaying no noticeable clinical symptoms. However, a smaller proportion of infected animals experience persistent lymphocytosis, characterized by an elevated number of lymphocytes in the bloodstream. If not monitored and managed, a subset of these persistently infected cattle may advance to lymphosarcoma. This condition typically presents as tumors in different lymphoid tissues, impacting various organs and overall health and productivity. Furthermore, recent research has highlighted the potential association between the occurrence of breast and lung cancer in humans and the presence of BLV. This review will delve into the recent discoveries concerning BLV, specifically exploring its epidemiology, the economic impact it has on the global cattle industry, its implications for human medicine, and the association between different alleles of the major histocompatibility complex (MHC) and susceptibility or resistance to BLV. Bovine leukemia virus, Enzootic bovine leukosis, Major histocompatibility complex, Retroviridae.

Economic losses associated with mastitis due to bovine leukemia virus infection.

Bovine leukemia virus (BLV), which causes enzootic bovine leukosis and immunosuppression, is widely prevalent on Japanese dairy farms. However, in the absence of a national eradication scheme with compensation programs, it is important to estimate BLV-associated economic losses to raise farmers' awareness. Mastitis (includes both clinical and subclinical) is a common disease in the dairy industry and the most common reason for culling. We hypothesized that immunosuppression due to BLV predisposes subclinical mastitis. A retrospective cohort study was conducted to trace Holstein cows at 9 commercial dairy farms in the Nemuro and Kushiro regions of Hokkaido Prefecture, Japan, where monitoring of BLV proviral load is routine. Information regarding Dairy Herd Improvement data, parity number, and delivery day was collected at each farm. Cows with no confirmed infection with BLV during lactation were defined as non-infected. Low-proviral-load and high-proviral-load (H-PVL) cows were defined as those in which proviral load was below and over 2,465 copies/50 ng of DNA, respectively, or 56,765 copies/105 cells, respectively, throughout the lactation period. Survival analysis was performed using the frailty model to estimate the hazard ratio of subclinical mastitis for BLV infection status using data from 1,034 dairy cows after adjusting for parity number and delivery season as confounding factors. Kaplan-Meier survivor curves demonstrated that half of the H-PVL cows developed subclinical mastitis within 52 d after calving. The hazard ratio of subclinical mastitis for H-PVL cows was 2.61 times higher than that of non-infected cows. In 2017, there were 264,443 clinical mastitis cases in Hokkaido. Using field and published data, annual economic losses were estimated using Monte Carlo simulation. The economic loss due to mastitis associated with BLV infection per H-PVL cow was $418.59 (¥43,952), with the annual economic loss in Hokkaido Prefecture due to mastitis caused by BLV infection estimated at $6,097,225 (¥640,208,633). In summary, H-PVL cows were more susceptible to subclinical mastitis than non-infected and low-proviral-load cows, and mastitis due to BLV infection was projected to cause significant economic losses.

Detection of bovine leukemia virus in beef cattle kept in the Central Coast Regions of Vietnam.

Bovine leukemia virus (BLV) is the etiologic agent of enzootic bovine leucosis. Our previous study showed the BLV existence in cattle kept in the Red River Delta Region of Vietnam. However, no positive samples were identified in beef cattle. Besides, information related to the BLV circulation in the remained parts of Vietnam is limited. Therefore, we tested the existence of BLV in 48 beef cattle kept in the Central Coast Regions. Nested PCR targeting the BLV-env-gp51 confirmed the prevalence of 14.6% in investigated regions. Phylogenetic analysis suggested the co-existence of genotypes 1 and 10. The close relationship between strains found in Vietnam, Thailand, Myanmar, and China was revealed suggesting the possibility of BLV transmission through the movement of live cattle.

Estimation of the proviral load in Japanese Black cattle infected with bovine leukemia virus by statistical modeling.

Enzootic bovine leukosis (EBL) is B-cell lymphoma in cattle caused by bovine leukemia virus (BLV) infection. The incidence of EBL has been increasing since 1998 in Japan, resulting in significant economic losses for farms. The BLV genome integrates with the host genome as provirus, leading to sustainably infection. Although most of the BLV-infected cattle are aleukemic, some cattle cause persistent lymphocytosis (PL) and subsequently develop EBL. Recent reports suggest the association between the risk for the transmission of BLV and the developing EBL and the proviral load (PVL) in BLV-infected cattle, which cannot measure readily in the field. This study aims to build a statistical model for predicting PVL of BLV-infected asymptomatic or PL cattle based on data accessible in the field. Five negative binomial regression models with different linear predictors were built and compared for the predictability of PVL. Consequently, the model with two explanatory variables (age in months and logarithm of lymphocyte count) was selected as the best model. The model can be used in the field as a cost-beneficial supporting tool to estimate the risk of transmission of BLV and developing EBL in infected cattle.

Risk factors associated with enzootic bovine leukosis in Boyacá and Cundinamarca municipalities, Colombia.

Enzootic bovine leukosis (EBL) is a lymphoproliferative disorder caused by the bovine leukemia virus (BLV), a virus of the Retroviridae family. The infection is distributed worldwide, and a high percentage of animals infected by the BLV are asymptomatic and act as carriers of the virus in many cattle populations. To identify the risk factors associated with EBL in the municipalities of Boyacá and Cundinamarca (Colombia). A simple descriptive cross-sectional study with random sampling was conducted. A total of 1,140 blood samples were taken from cattle (females and males) from the municipalities of Chiquinquirá, Ubaté, and San Miguel de Sema of different breeds and age groups. The samples were processed using the commercial ELISA SERELISA® BLV Ab Mono Blocking kit (sensitivity 97%, specificity 98%). The data were processed with the statistical programs WinEpi and Epi Info® version 7.2.4.0, estimating the prevalence ratio, implementing the chi-square test (p ≤ 0.05) and logistic regression. A true prevalence (TP) and apparent prevalence (AP) of 23.61% and 22.7% in Ubaté, 19.22% and 18.1% in Chiquinquirá, and 15.61% and 14.3% in San Miguel de Sema, respectively, were established. Bovines 2-4 years old were the most prevalent in Ubaté and Chiquinquirá (37.5% and 21.21%, respectively), while in San Miguel de Sema individuals >4 years had the highest percentage of antibodies (18.3%). The Holstein breed had a higher prevalence in Ubaté and San Miguel de Sema (26.02% and 19.67%), and crossbreeds were more BLV-seroprevalence in Chiquinquirá (20.20%). In Ubaté, re-use of needles was identified as a risk factor, contaminated blood in needles is considered one of the main routes of transmission. On the other hand, manual milking was identified as a risk factor in San Miguel de Sema. The non-implementation of an individual needle per animal in Ubaté; the Holstein breed and manual milking in San Miguel de Sema were identified as risk factors for the presence of antibodies against the disease. EBL prevention and control plans should be established that focus on the implementation of management and sanitary practices based on herd biosecurity.

Improvement of the technique for the bovine genome editing using the example of knockout of the CD 209 receptor responsible to the bovine leukemia virus

The aim of this work was to evaluate the possibility of editing the CD209 gene associated with bovine leukemia virus (BLV) infection. Two ways of delivering the gene editing system into the zygote were investigated: delivery of the CRISPR/Cas9 system using a viral vector and injection of mRNA and sgRNA into the oocyte cytoplasm. The most effective was the microinjection of spCas9 protein mRNA and sgRNA targeting CD209 at the zygote stage into the cytoplasm.

Динамика В-лимфоцитов и уровень экспрессии CD20-рецептора в процессе экспериментального лейкоза овец

CD20 is a permanent marker of almost all stages of B-lymphocyte differentiation, with the exception of pro-B cells and plasma cells. The role and functions of this protein are not fully understood today, but it is known that it performs the functions of a calcium channel in cooperation with BCR to regulate all-important processes of B-lymphocytes: from proliferation and differentiation to antibody synthesis. Currently, insufficient attention is paid to the study of the CD20 receptor in the process of chronic viral diseases, although in medicine this molecule is used as an important component in the diagnosis of leukemia’s of various types. This study was aimed at studying the dynamics of B- lymphocytes and the expression level of CD20 receptors after experimental infection with bovine leukemia virus. The data obtained showed that the study of CD20 lymphocytes and CD20 receptor expression levels might be important in the diagnosis of leukemia, as well as be an additional criterion in the study of immune status in long-term chronic infections.