Enzootic Bovine Leukosis Research Articles

Genetic variation in Japanese Holstein cattle for EBL development

BackgroundInfection with bovine leukemia virus (BLV), the causative agent for enzootic bovine leukosis (EBL), is increasing in dairy farms of Japan. The tendency of tumor development following BLV infection in certain cow families and bull lines has previously been described. We therefore hypothesized the existence of a genetic component which differentiates cattle susceptibility to the disease.ResultsWe analyzed routinely collected large-scale data including postmortem inspection data, which were combined with pedigree information and epidemiological data of BLV infection. A total of 6,022 postmortem inspection records of Holstein cattle, raised on 226 farms served by a regional abattoir over 10 years from 2004 to 2015, were analyzed for associations between sire information and EBL development. We then identified statistically the relative susceptibility to EBL development for the progeny of specific sires and paternal grandsires (PGSs). The heritability of EBL development was calculated as 0.19. Similarly, proviral loads (PVLs) of progeny from identified sires and PGSs were analyzed, but no significant differences were found.ConclusionsThese observations suggest that because EBL development in our Holstein population is, at least in part, influenced by genetic factors independent of PVL levels, genetic improvement for lower incidence of EBL development in cattle notwithstanding BLV infection is possible.

Quantitative Risk Assessment for the Introduction of Bovine Leukemia Virus-Infected Cattle Using a Cattle Movement Network Analysis.

The cattle industry is suffering economic losses caused by bovine leukemia virus (BLV) and enzootic bovine leukosis (EBL), the clinical condition associated with BLV infection. This pathogen spreads easily without detection by farmers and veterinarians due to the lack of obvious clinical signs. Cattle movement strongly contributes to the inter-farm transmission of BLV. This study quantified the farm-level risk of BLV introduction using a cattle movement analysis. A generalized linear mixed model predicting the proportion of BLV-infected cattle was constructed based on weighted in-degree centrality. Our results suggest a positive association between weighted in-degree centrality and the estimated number of introduced BLV-infected cattle. Remarkably, the introduction of approximately six cattle allowed at least one BLV-infected animal to be added to the farm in the worst-case scenario. These data suggest a high risk of BLV infection on farms with a high number of cattle being introduced. Our findings indicate the need to strengthen BLV control strategies, especially along the chain of cattle movement.

Development of an in situ hybridization assay using an AS1 probe for detection of bovine leukemia virus in BLV-induced lymphoma tissues.

Enzootic bovine leukosis (EBL) is a malignant B cell lymphoma caused by infection with bovine leukemia virus (BLV). Histopathological examination is commonly used for diagnosis of the disease, but observation of lymphoma alone does not confirm EBL because cattle may be affected by sporadic forms of lymphoma that are not associated with BLV. Detection of BLV in tumor cells can be definitive evidence of EBL, but currently, there is no technique available for such a purpose. In this study, we focused on a viral non-coding RNA, AS1, and developed a novel in situ hybridization assay for the detection of BLV from formalin-fixed paraffin-embedded (FFPE) tissues. RNA-seq analysis revealed that all examined B lymphocytes derived from clinical EBL abundantly expressed AS1 RNA, indicating a possible target for detection. The in situ hybridization assay using an AS1 probe clearly detected AS1 RNA in fetal lamb kidney cells persistently infected with BLV. The utility of this assay in clinical samples was assessed using three EBL-derived lymph node specimens and one BLV-negative specimen, and AS1 RNA was detected specifically in the EBL-derived tissues. These results suggest that AS1 RNA is a useful target for the detection of BLV from FFPE specimens of tumor tissues. This technique is expected to become a powerful tool for EBL diagnosis.

Development of a microchip electrophoresis-based, high-throughput PCR-RFLP method to type Tax 233 variants of bovine leukemia virus in Japan.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). We used microchip electrophoresis in combination with automatic image analysis to develop a novel high-throughput PCR-RFLP to type the gene sequences that encode BLV Tax 233. This method revealed that 233L-Tax is more prevalent than 233P-Tax in cattle in Japan. The proportion infected with BLV carrying the gene encoding 233L-Tax was significantly higher in Holstein cattle than in Japanese Black cattle. Holsteins infected with BLV encoding 233L-Tax had higher proviral loads than did Holsteins infected with BLV encoding 233P-Tax and Japanese Blacks infected with BLV encoding 233L-Tax or 233P-Tax. The novel method developed in this study will be a useful tool for identifying cattle harboring BLV with a higher risk of EBL and viral transmission.

Detection and Molecular Characterization of Bovine Leukemia Virus in Egyptian Dairy Cattle

Bovine leukemia virus (BLV) causes enzootic bovine leukosis (EBL), the most common neoplastic disease in cattle worldwide. The first EBL outbreak in Egypt was reported in 1997. To date, there are few studies regarding BLV diagnosis using only serological detection and no studies investigating the distribution of BLV provirus, which is the retroviral genome integrated into the host genome, in Egypt. The genetic characteristics of Egyptian BLV strains are also unknown. Therefore, we aimed to detect BLV provirus and determine BLV genetic variability among dairy cattle in Egypt. We collected 270 blood samples of dairy cattle from 24 farms located in five provinces in Egypt. Out of the 270 samples, 58 (21.5%) were positive for BLV provirus. Phylogenetic analysis based on 18 420-bp selected sequences out of 50 isolates of the BLV env-gp51 gene demonstrated that Egyptian BLV isolates were clustered into genotype-1 and-4, among 11 genotypes detected worldwide. Furthermore, phylogenetic analysis and alignment of the 501-bp sequence of the env-gp51 gene revealed that at least six genetically different strains are present in Egypt. Genotype-1 isolates comprised four different strains (G1-a, G1-b, G1-c, and G1-d) and genotype-4 isolates included two different strains (G4-x and G4-y). Moreover, in one farm with 100% infection rate, we identified three isolates of G1-a strain, 35 isolates of G4-x strain, and two isolates of G4-y strain. Overall, this study provides the new report on molecular prevalence of BLV in Egypt and records the coexistence of BLV genotype-1 and-4 in Egyptian cattle.

Retrospective analysis of the epizootic situation of enzootic bovine leukosis in Ukraine in 1994–2019

In Ukraine, prevention and control measures of bovine leukosis were regulated by relevant legislation, regulations and instructions developed in the period 1960–1992. Currently, the instruction on prevention and rehabilitation of this disease in cattle is used (approved in 2007). According to it, the identification of infected animals is carried out from 6 months of age by serological (first scheme in AGID or another in ELISA) and genomic (PCR) methods. The infected animals are removed from the herd or slaughtered. In general, because of the diagnostic and preventive measures carried out during the analyzed period (26 years) 10 519 farms were rehabilitated from leukosis (2 346 affected farms remained from previous years). At the same time, more 4 million infected cattle were slaughtered. The majority of affected farms (more than 1 000) were registered in the period between 1994 (2 346 farms) and 2003 (1 247 farms). Since 2014, the number of affected farms has remained mostly below 10 and the number of infected cattle has decreased to 2 000 animals per year. At the same time, the number of rehabilitated farms also decreased (from 1 307 farms in 1998 to 4 farms in 2014). The same trend was registered with the dynamics of the number of animals that were removed from the herd due to leukosis. Thus, in 1995 and 1997 their numbers were 321 178 and 558 649 animals, respectively, and in 2014 it was 1 124. The obtained indicators of intensity and extensiveness of the epizootic process show that the incidence rate was maximal during 1998–2000 and amounted to 3.7–4.3%. The maximum indicators of the coefficient of affection were recorded in 1997–2000 and equal 11.8–15.3%. The rate of foci remained on the level of 90–270 throughout the all analyzed period. During 2008–2019, specialists of the Ukrainian veterinary laboratories investigated more than 47 million samples of cattle blood sera for enzootic bovine leukosis by AGID and ELISA. However, despite the significant diagnostic work, the important factor in the decrease of the number of affected farms and infected animals is the decrease in the total number of cattle in Ukraine (almost 22 million animals in 1994 against 3 million in 2019). Graphic trends of these indicators are comparable and agree with the decrease in the number of cattle in our country by analyzed period. After 2014, the number of affected farms ranged 10–17 per year (mostly in private households). However, the full recovery of cattle in Ukraine from bovine leukosis has not taken place, although our country is closer than ever to this.

Seroprevalence and risk factors associated with bovine leukemia virus (BLV) seropositivity in cattle herds from Ecuador

Background: Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). This disease mainly affects cattle, causing severe economic losses to producers. Objective: To establish individual and herd seroprevalence and determine the risk factors associated with BLV seropositivity for dairy and dual-purpose cattle herds in Ecuador. Methods: A total of 2,668 serum samples from 386 herds were collected. A questionnaire, including variables related to cattle health, management and the environment was completed by each herd. A commercial blocking enzyme-linked immunosorbent assay (ELISA) test was used to determine seropositivity. A generalized estimating equation model (GEE) was developed to determine the factors associated with BLV seropositivity. Results: Individual seroprevalence of BLV infection in Ecuador was 17.3% (CI95% = 15.86-18.74%). Herd prevalence was 37.8% (CI95% = 33.0-42.6%), and intra-herd prevalence ranged between 12.5 and 100% (median: 37.5%). The risk factors associated with BLV seropositivity were artificial insemination (OR: 2,215; CI95% = 1.402-3.501), concrete floors (OR: 2.178; CI95% = 1.217-3.889), presence of wild ruminants (OR: 2.998; CI95% = 1.788-5.027), and sampling season (wet; OR: 1.996; CI95% = 1.140-3.497). Conclusions: Results indicate that BLV is widespread in cattle herds in Ecuador. In addition, the study suggests that a control program to fight BLV infection should focus on controlling the risk factors identified.

Detection of genotype 1 bovine leukemia virus from a C.schultzei pool: Do Culicoides spp. have a role on the transmission of bovine leukemia virus?

Bovine leukemia virus (BLV) is known as the etiological agent of Enzootic bovine leukosis (EBL), which is the most common neoplastic disease of cattle. While the major route of virus transmission is believed to be iatrogenic, BLV proviral DNA has been identified in biological materials, including nasal secretions, saliva, milk, colostrum, and semen, and in several insect species, including horses flies. However, insects' role in the natural transmission of BLV has not been clearly demonstrated. This study assessed the possible role of midges – Culicoides spp. – in BLV transmission. BLVs were genetically characterized and BLV infection seroprevelance was determined in 224 cattle sampled from 27 different small family herds in five different districts in Hatay province, southern Turkey. Out of the 25 Culicoides spp. pools, one (4.0%; 1/25) was a C.schultzei pool while 2.67% (6/224) of the sampled cattle were positive for BLV nucleic acid. The seroprevalance rates for the sampled herds and all sampled cattle were 7.40% (2/27) and 1.33% (3/224), respectively. According to the phylogenetic analysis, the sequences of the BLVs from the cattle (n = 6) and the one BLV-positive C.schultzei pool clustered on genotype 1 (G1) BLVs. Although these results do not reveal the exact role of Culicoides spp. or other midges flies in BLV transmission, the simultaneous presence of same substitions in BLVs from both cattle and a C.schultzei pool is noteworthy. Further studies on the env gene and other BLV gene regions detected from cattle and C.schultzei pools are ongoing to understand the possible epidemiological relationship between cattle and flies.

Development and implementation of a risk assessment and management program for enzootic bovine leukosis in Atlantic Canada

Over the past 30 yr, the prevalence of bovine leukemia virus (BLV) infection has increased in North America, including Atlantic Canada, at both the herd and individual cow levels. This has occurred despite increased awareness of the disease and its deleterious effects and despite implementation of management practices aimed at reducing disease transmission. Our objectives were to identify risk factors associated with the within-herd prevalence of BLV-infected cows by using a risk assessment and management program workbook, as well as to determine the current level of BLV prevalence in the Atlantic Canada region. We hypothesized that previously established risk factors, including management practices associated with calf rearing and fly control, would affect within-herd BLV prevalence. Bulk tank milk (BTM) samples were collected in January and April of 2016 and again during the same months in 2017 and 2018 from all dairy farms shipping milk in the region. Samples were tested with ELISA for levels of anti-BLV antibodies to estimate within-herd prevalence. Regional BLV prevalence at the herd level was 88.39% of dairy herds infected in 2016 and 89.30% in 2018. All dairy farms shipping milk and who had BTM samples collected in 2017 (n = 605) were eligible to participate in the risk assessment and management program questionnaire (RAMP), which was developed and distributed to all bovine veterinarians in Atlantic Canada. One hundred and six RAMP were returned, with representation from all 4 provinces. The RAMP results were combined with the mean BTM ELISA results, and univariable logistic regression followed by multivariable logistic regression was performed to investigate the association between RAMP risk factors and the estimated within-herd BLV prevalence. Factors in the multivariable model significantly associated with the odds of a herd being classified as >25% estimated within-herd prevalence included history of diagnosis of clinical BLV and calves receiving colostrum from cows with unknown BLV status. Differences in within-herd prevalence were not associated with hypodermic needle and injection practices, rectal sleeve practices, or using bulls for natural breeding, based on these 106 dairy farms.

Risk factor analysis of bovine leukemia virus infection in dairy cattle in Egypt

Identification of the risk factors associated with Enzootic bovine leukosis (EBL) is essential for the adoption of potentially prevention strategies. Accordingly, our objectives were to determine the geographic distribution of Bovine Leukemia Virus (BLV) infection and identify the risk factors associated with cow-level BLV infection in the Egyptian dairy cattle. A cross-sectional study was conducted on 1299 mixed breed cows distributed over four provinces in the Nile Delta of Egypt in 2018. The randomly selected cows on each farm were serologically tested for BLV, and the cow’s information was obtained from the farm records. Four variables (geographic location, herd size, number of parities, and age) were used for risk analysis. A total of 230 serum samples (17.7 %) were serologically positive for BLV. The highest prevalence of BLV infection was associated with parity (OR = 3.4, 95 %CI 2.4–4.9) with 80 % probability of being BLV-positive at parity ≥5, followed by herd size (OR = 1.8, 95 %CI 1.4–2.2). However, geographic location seems to have no impact on the prevalence of BLV infection in Egypt. Our findings strongly indicate that the intensive surveillance and effective prevention strategies against BLV infection in Egypt should be provided to multiparous cows with ≥5 parities and live in large farm with more than 200 cows.

Effect of bovine leukemia virus (BLV) infection on bovine mammary epithelial cells RNA-seq transcriptome profile.

Bovine leukemia virus (BLV) is a δ-retrovirus responsible for Enzootic Bovine Leukosis (EBL), a lymphoproliferative disease that affects cattle. The virus causes immune system deregulation, favoring the development of secondary infections. In that context, mastitis incidence is believed to be increased in BLV infected cattle. The aim of this study was to analyze the transcriptome profile of a BLV infected mammary epithelial cell line (MAC-T). Our results show that BLV infected MAC-T cells have an altered expression of IFN I signal pathway and genes involved in defense response to virus, as well as a collagen catabolic process and some protooncogenes and tumor suppressor genes. Our results provide evidence to better understand the effect of BLV on bovine mammary epithelial cell's immune response.

PRMT5 Is Required for Bovine Leukemia Virus Infection In Vivo and Regulates BLV Gene Expression, Syncytium Formation, and Glycosylation In Vitro.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis, which is the most common neoplastic disease of cattle and is closely related to human T-cell leukemia viruses. We investigated the role of a new host protein, PRMT5, in BLV infection. We found that PRMT5 is overexpressed only in BLV-infected cattle with a high proviral load, but not in those with a low proviral load. Furthermore, this upregulation continued to the lymphoma stage. PRMT5 expression was upregulated in response to experimental BLV infection; moreover, PRMT5 upregulation began in an early stage of BLV infection rather than after a long period of proviral latency. Second, siRNA-mediated PRMT5 knockdown enhanced BLV gene expression at the transcript and protein levels. Additionally, a selective small-molecule inhibitor of PRMT5 (CMP5) enhanced BLV gene expression. Interestingly, CMP5 treatment, but not siRNA knockdown, altered the gp51 glycosylation pattern and increased the molecular weight of gp51, thereby decreasing BLV-induced syncytium formation. This was supported by the observation that CMP5 treatment enhanced the formation of the complex type of N-glycan more than the high mannose type. In conclusion, PRMT5 overexpression is related to the development of BLV infection with a high proviral load and lymphoma stage and PRMT5 inhibition enhances BLV gene expression. This is the first study to investigate the role of PRMT5 in BLV infection in vivo and in vitro and to reveal a novel function for a small-molecule compound in BLV-gp51 glycosylation processing.

Overexpression of bovine leukemia virus receptor SLC7A1/CAT1 enhances cellular susceptibility to BLV infection on luminescence syncytium induction assay (LuSIA)

Bovine leukemia virus (BLV) causes enzootic bovine leukosis, the most common neoplastic disease in cattle. We previously reported the development and protocol of the luminescence syncytium induction assay (LuSIA), a method for evaluating BLV infectivity based on CC81-GREMG cells. These cells form syncytia expressing enhanced green fluorescent protein when co-cultured with BLV-infected cells. Recently, we confirmed CAT1/SLC7A1 functions as a receptor of BLV. Here, we focused on CAT1/SLC7A1 to increase the sensitivity of LuSIA. We constructed a bovine CAT1-expressing plasmid and established a new CC81-GREMG-derived reporter cell line highly expressing bovine CAT1 (CC81-GREMG-CAT1). The new LuSIA protocol using CC81-GREMG-CAT1 cells measures cell-to-cell infectivity and cell-free infectivity of BLV faster and with greater sensitivity than the previous protocol using CC81-GREMG. The new LuSIA protocol is quantitative and more sensitive than the previous assay based on CC81-GREMG cells and will facilitate the development of several new BLV assays.

Enzootic bovine leukosis accompanied by splenomegaly in an 8-month-old calf

In this report, an 8-month-old calf (crossbred, Holstein × Japanese Black) developed fever and accompanied abomasum displacement. Blood chemical test showed remarkably high values of white blood cell count and heteromorphic lymphocytes. In pathological appraisal, enlarged splenomegaly and swelling of the lymph nodes were observed. Histopathological examination revealed invasion of tumor cells derived from B1 cells into systemic lymph nodes, liver and spleen. The provirus loads of bovine leukemia virus (BLV) was 1,439 copies per 10 ng DNA by using real time PCR. In conclusion, this case was diagnosed as bovine leukemia caused by BLV infection with a huge splenomegaly.

Serological evidence of enzootic bovine leukosis in the periurban dairy cattle production system of Al Ain, United Arab Emirates.

The serostatus of enzootic bovine leukosis (EBL) was determined at three dairy farms and the Al Ain Livestock Market (AALM), within the Al Ain region of Abu Dhabi, UAE. Of the 957 bovine sera tested by ELISA, 657 were from Holstein-Friesians from three dairy farms, and 300 from Bos indicus cattle at AALM. The chi-square homogeneity test (CSHT) and the Marascuilo multiple comparison procedure (MMCP) assessed the level of significance between the proportions of EBL-seropositive cattle (ESPC) across the study farms and AALM, and between the age groups at farms 1 and 3. Overall, the proportion of ESPC was 25.7% at dairy farms and AALM, 37.0% for farms and 1.0% for AALM. Furthermore, the proportions of ESPC at farms 1, 2 and 3 were 54.7%, 0.0% and 26.3% respectively, and statistically significant differences were seen across the farm/farm and farm/AALM comparisons, and between two age groups at farms 1 and 3. The 37-72-month-old age group showed the highest proportion of ESPC. This is the first serological evidence of EBL in the UAE. As previously reported, the ESPC are comparatively higher in dairy than Bos indicus cattle. Molecular and more extensive serological studies are needed to further corroborate the present data. Meanwhile, the UAE veterinary authorities will need to formulate national EBL control policies.

New evidence of bovine leukemia virus circulating in Myanmar cattle through epidemiological and molecular characterization.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, which is the most common neoplastic disease of cattle. BLV infects cattle worldwide and causes serious problems for the cattle industry. In this study, we examined the prevalence of BLV infection and the distribution of BLV genotypes in cattle in the northern, central, and southern parts of Myanmar. The prevalence of BLV infection among Myanmar cattle (37.04%) in this study was markedly higher than the prevalence (9.1%) observed in our earlier study in which BLV was detected from the limited number of cattle only from a small area of Myanmar. Phylogenetic analysis of partial env-gp51 sequence of the isolated BLV strains revealed that there are at least three BLV genotypes (genotype-1, genotype-6, and genotype-10) in Myanmar, which have also been detected in the neighboring countries. We performed this study to estimate the BLV proviral load, which is a major diagnosis index for determining the virus transmission risk. The cattle of the three test regions with warm, wet, and humid climatic conditions (upper Sagaing, Yangon, and Kayin) exhibited a high mean proviral load, while cattle of three other regions with low annual rainfall and very high temperature (Mandalay, Magway, and upper Bago) exhibited a low mean proviral load. Further, the level of proviral load and the prevalence of BLV infection in Myanmar native cattle (N = 235) were lower than that in the hybrid cattle (Holstein Friesian × Myanmar native) (N = 62). We also observed that the cattle with high risk for BLV transmission, which have high proviral load, may enhance the BLV infection rate. Hence, to control BLV transmission, it is necessary to eliminate these cattle with high-risk for BLV transmission and to diagnose BLV provirus in cattle in the remaining regions/states of Myanmar sharing a boundary with neighboring countries.

Enzootic bovine leucosis in cows on farms in Almaty and Turkestan regions of Kazakhstan

The objective of this study was to evaluate pathomorphological and histopathological characteristics of internal organ tissues and skeletal muscles of Black-Motley cows with bovine laeukemia necropsied between 2013 and 2017. Pathomorphological and histopathological assessment of enzootic bovine leukosis was performed. The viral presence was proved by haematological study, enzyme-linked immunosorbent assay and agar-gel immunodiffusion. There was the pathomorphological manifestation of an immunosuppressive condition in the cattle, as well as leukemic cell accumulations in the brain, kidneys, duodenum, spleen, lymph nodes, cardiac and skeletal muscles. Our study is considered to bear significance concerning the description of microstructural changes involving lymph node trabeculae and myocardial fibres from leukemic cows. This investigation, therefore, provides new knowledge on histopathological features of enzootic bovine leucosis, which potentially may bring new insights into the disease mechanisms.

Detection and genotyping of bovine leukemia virus (BLV) in Vietnamese cattle.

Bovine leukemia virus (BLV) belongs to the genus, Deltaretrovirus of the family, Retroviridae and it is the causative agent of enzootic bovine leukosis. The prevalence of BLV in three provinces in the Red River Delta Region in the North of Vietnam, Hanoi, Vinhphuc and Bacninh was studied from April 2017 to June 2018. A total of 275 blood samples collected from cattle were used for serum isolation and DNA extraction. Of these samples, 266 sera were subjected to ELISA test for detecting antibody against BLV gp51 protein and 152 DNA samples were used to detect the 444 bp fragment corresponding to a part of the gp51 region of the env by nested PCR. The results showed that 16.5% (n=44) and 21.1% (n=32) of samples were positive for BLV gp51 antibody and BLV proviral DNA, respectively. Phylogenetic analysis of the partial (423 bp) and complete (913 bp) BLV env-gp51 gene indicated that Vietnamese strains were clustered into genotypes 1, 6 and 10 (G1, G6 and G10). Of those genotypes, G1 genotype was dominant; G6 strains were designated as G6e and G6f subgenotypes; the existence of genotype 10 was confirmed for the first time in Vietnam. The present study provides important information regarding the prevalence of BLV infection and genetic characteristics of BLV strains identified in Vietnam, contributing to promote the establishment of disease control and eradication strategies in Vietnam.

Detection and molecular characterization of bovine leukemia virus in beef cattle presented for slaughter in Egypt.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, the most common neoplastic disease of cattle worldwide and a serious problem for the cattle industry. Previous studies have shown the molecular prevalence of BLV and the coexistence of BLV genotype-1 and -4 in Egyptian dairy cattle; however, the molecular characteristics of BLV in Egyptian beef cattle are unknown. Therefore, we collected blood samples of 168 beef cattle from slaughterhouses in three governorates in Egypt. Based on BLV-CoCoMo-qPCR-2 targeting long terminal repeats and nested PCR targeting the env-gp51 gene, the BLV provirus infection rates were found to be 47/168 (28.0%) and 42/168 (25.0%), respectively. Phylogenetic analysis based on 501 bp of the BLV env-gp51 gene from 42 BLV isolates revealed that at least six distinctive strains (b, e, f, g, x, and z) were prevalent in cattle across the examined regions. Furthermore, phylogenetic analysis of the 420 bp sequence of the BLV env-gp51 region of the six strains against 11 known genotypes showed that the strains b, e, f, and g were clustered into genotype-1, and strains x and z were clustered into genotype-4. Our results also indicated that strains b and x exist in both dairy and beef cattle in Egypt. The present study is the first to detect and genotype BLV among beef cattle in Egypt.

Respuesta inmune humoral contra toxoides de Clostridium perfringens en vaquillonas Holando seropositivas al virus de la Leucosis Bovina Enzoótica

Enzootic bovine leukosis is the main virosis that affects dairy cattle in Uruguay. The subclinical infection causes importants dysfunctions in the immune system impacting directly in the animal health. The aim of this study was to evaluate the immune response of Holstein heifers with positive serology against Leukosis bovine virus (BLV) when they were ordinary immunized against Clostridium perfringens. It was evaluated a seropositive group to BLV animals (n=29) and other seronegative group (n=19) during twelve months. The animals recived three immunizations with a commercial polyvalent vaccine against several Clostridios (days 0, 30, 173). Blood samples were taken in days 0, 30, 60, 90, 180 y 360 post vaccination and then the humoral immune response was analyzed through the use of the ELISA technique to quantify total antibodies against C. perfringens toxins alfa, beta and epsilon. Besides it was in vitro analyzed the amount of neutralizing antibodies against epsilon toxin using the reference technique, seroneutralization. The results demonstrated a low response of neutralizing antibodies to epsilon toxin by SN technique showing no correlation with the results obtained by the commercial ELISA kit. Total antibodies response evaluated using commercial ELISA Kit was variable according to the studied toxoid. No significant differences were found in the immune response against Clostridium perfringens vaccination between BLV seropositive and seronegative animals using the techniques mentioned. The commercial ELISA kit use to analyze the response to vaccination against Clostridiosis, must be discussed and analyzed according to the results of this work.