Prevalence Of Bovine Leukemia Virus Research Articles

Herd-level prevalence of bovine leukemia virus, Salmonella Dublin, and Neospora caninum in Alberta, Canada, dairy herds using ELISA on bulk tank milk samples

Endemic infectious diseases remain a major challenge for dairy producers worldwide. For effective disease control programs, up-to-date prevalence estimates are of utmost importance. The objective of this study was to estimate the herd-level prevalence of bovine leukemia virus (BLV), Salmonella Dublin, and Neospora caninum in dairy herds in Alberta, Canada using a serial cross-sectional study design. Bulk tank milk samples from all Alberta dairy farms were collected 4 times, in December 2021 (n = 489), April 2022 (n = 487), July 2022 (n = 487), and October 2022 (n = 480), and tested for antibodies against BLV, S. Dublin, and N. caninum using ELISAs. Herd-level apparent prevalence was calculated as positive samples divided by total tested samples at each time point. A mixed effect modified Poisson regression model was employed to assess the association of prevalence with region, herd size, herd type, and type of milking system. Apparent prevalence of BLV was 89.4, 88.7, 86.9 and 86.9% in December, April, July, and October, respectively, whereas for S. Dublin apparent prevalence was 11.2, 6.6, 8.6, and 8.5%, and for N. caninum apparent prevalence was 18.2, 7.4, 7.8, and 15.0%. For BLV, S. Dublin and N. caninum, a total of 91.7, 15.6, and 28.1% of herds, respectively, were positive at least once, whereas 82.5, 3.6, and 3.0% of herds were ELISA-positive at all 4 times. Compared with the north region, central Alberta had a high prevalence (prevalence ratio (PR) = 1.13) of BLV-antibody positive herds, whereas south Alberta had a high prevalence (PR = 2.56) of herds positive for S. Dublin antibodies. Furthermore, central (PR = 0.52) and south regions (PR = 0.46) had low prevalence of N. caninum-positive herds compared with the north. Hutterite colony herds were more frequently BLV-positive (PR = 1.13) but less frequently N. caninum-positive (PR = 0.47). Large herds (>7,200 L/day milk delivered ∼ > 250 cows) were 1.1 times more often BLV-positive, whereas small herds (≤3,600 L/day milk delivered ∼ ≤ 125 cows) were 3.2 times more often N. caninum-positive. For S. Dublin, Hutterite-colony herds were less frequently (PR = 0.07) positive than non-colony herds only in medium and large stratum but not in small stratum. Moreover, larger herds were more frequently (PR = 2.20) S. Dublin-positive than smaller herds only in non-colony stratum but not in colony stratum. Moreover, N. caninum prevalence was 1.6 times higher on farms with conventional milking systems compared with farms with an automated milking system. These results provide up-to-date information of the prevalence of these infections that will inform investigations of within-herd prevalence of these infections and help in devising evidence-based disease control strategies.

Removing bovine leukemia virus–infected animals with high proviral load leads to lower within-herd prevalence and new case reduction

Bovine leukosis is prevalent in the North American dairy industry and its effect on animal health and production is widely documented. However, not all bovine leukemia virus (BLV) infected animals transmit the virus equally. Animals with high BLV proviral loads (HPL) are associated with higher transmission risks and therefore, their removal may reduce transmission and eventually within-herd prevalence. We aimed to evaluate the impact of selectively removing HPL cows on the within-herd BLV prevalence and incidence rate of BLV infection in 10 dairy herds. Annual blood and/or milk samples were collected from adult cows over 3 years. Positivity with BLV were determined by ELISA tests and proviral loads in blood of BLV-positive animals were estimated with BLV SS1 quantitative PCR assays. Herd managers were encouraged to consider the proviral load when making culling decisions and implement BLV control practices. High proviral load cows had the highest relative risk of removal indicating the farmers prioritized HPL cows for culling. The within-herd BLV prevalence decreased significantly in 4 herds whereas BLV incidence rate decreased in 9 herds. Over the 3 years, the proviral load demonstrated a relatively stable level, suggesting a single proviral load test in an adult cow may suffice to make culling decisions.

Efeito da infecção pelo vírus da leucose bovina e carga pró-viral sobre o perfil sistêmico de vacas primíparas no período de transição

ABSTRACT: This study aimed to evaluate the effect of bovine leukemia virus (BLV) on the systemic profile of naturally infected dairy heifers during the transition period. Pregnant Holstein and Jersey heifers (n=24) were distributed in pairs into two experimental groups: (BLV+) and (BLV-). Animals in the BLV+ group were divided into two subgroups based on the median BLV proviral load (high and low). The animals were then assessed at weeks -3, -2, -1, calving time (0), +1, +2, and +3. Blood samples were obtained for hematological and biochemical analyses, as well as haptoglobin measurements. Farm BLV screening revealed a herd BLV prevalence of 57.25% and heifer BLV prevalence of 38.7%. Mean corpuscular hemoglobin concentration was the only hematological variable for which group interaction was observed, with BLV+ cattle having higher values (33.29 ± 3.39%) than BLV- cattle (31.08 ± 2.31%). Aspartate aminotransferase activity was higher in the BLV+ heifers. The BLV+ group had greater incidence of inflammation (haptoglobin ≥ 2.0 mg/dL). Fibrinogen concentrations were also higher at weeks 0 and +1 in BLV+ heifers than in BLV- heifers. A high proviral load affected total leukocyte and lymphocyte count; however, this profile was not observed in the low proviral load and paired BLV- heifers. To our knowledge, this is the first study to report the impact of BLV infection on the health of dairy heifers during the transition period, demonstrating the effect of proviral load on white blood cell changes and early inflammation in infected animals.

Prevalence of Bovine leukemia virus in Egyptian women’s breast cancer tissues

ABSTRACT Bovine leukemia virus (BLV) infects cattle around the world and presents in commercial bovine products. Several studies have suggested that BLV may play a role in breast oncogenesis. This study aimed to determine the risk factors for BLV infection and the presence of the BLV gene in breast tissue. A total of 100 formalin-fixed and paraffin-embedded breast tissue specimens were collected (50 confirmed cancer samples and 50 negative samples). DNA was extracted from breast tissues, and nested PCR was done to detect the tax gene of BLV. Immunohistochemical staining for ER, PR, Her2/neu, and Ki-67 was applied. Bovine leukemia virus DNA was detected in 16% of breast cancer tissue samples and in only 4% of tissue samples without cancer. No statistically significant association was reported between the presence of BLV and tumor grade, stage, or LN metastasis. A high percentage of positivity (100%, 88%, and 75%) was reported for ER, PR, and Ki-67 in BLV-positive cancer samples, respectively. There was a significant association between the BLV tax gene and breast cancer in Egyptian women. Thus, there is a need to control BLV transmission to humans from cattle, and further investigations about its risk related to public health are highly required.

Seroprevalence and risk factors associated with bovine Leukemia virus infection in argentine beef cattle

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis, an endemic disease in dairy cattle of Argentina. However, little is known about the seroprevalence of BLV in beef cattle. In this study, we conducted a cross-sectional study including farms from thirteen provinces of Argentina. A total of 5827 bovine serum samples were collected from 76 farms and analyzed using an in-house developed enzyme-linked immunosorbent assay. Information about herd management was collected through a questionnaire, and univariate and multivariate analyses were performed to detect risk factors associated with BLV infection. Herd-level seroprevalence was 71.05%, while the mean animal-level seroprevalence was 7.23% (median = 2.69%; min = 0, max = 75). Only two provinces had no positive BLV samples. The other eleven provinces showed more than 50% of their farms infected with BLV. The multivariate model revealed that BLV prevalence was significantly associated with the use of animals raised in the same farm for cattle replacement (P = 0.005), breeding cows by natural mating with a bull (P < 0.001), and weaning calves after 6 months of age (P = 0.011). This extensive study revealed that BLV seroprevalence in Argentine beef farms has increased during the last years and allowed identifying some management practices associated with BLV prevalence. These data deserve special attention because BLV infection in beef cattle seems to lead to a dissemination pattern similar to that observed during the last decades in dairy cattle, especially considering that Argentina is the sixth beef producer in the world, with about 5% of global beef production.

The influence of risk factors on bovine leukemia virus infection and proviral load in egyptian cattle.

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leucosis (EBL), which affects cattle globally. In Egypt, BLV control strategies have been ignored because of the shortage of BLV research studies and the silent infection in most animals. This study aimed to identify the risk factors associated with the prevalence of BLV among dairy and beef cattle from six different geographic and climatic provinces in Egypt. Additionally, risk factors affecting the BLV proviral load (PVL) among the positive cattle were targeted. The total BLV prevalence in cattle from six investigated Egyptian provinces was 24.2% (105/433), while the mean PVL (8651.6 copies /105 white blood cells) was absolutely high as estimated by the BLV-CoCoMo-quantitative polymerase chain reaction (qPCR)-2 assay. Analysis of the influence of risk factors (age, sex, breed, production type, farm size, and location) on BLV prevalence indicated that the Holstein breed (OR = 1.582, p = 0.007), beef cattle (OR = 1.088, p = 0.0001), large-size farms (OR = 1.26, p = 0.0001), and cattle from Damietta (OR = 1.43, p = 0.0001) and Cairo (OR = 1.16, p = 0.0001) were ultimately proven the most important risks for BLV infection. The risk factors were analyzed considering the BLV PVL levels in the BLV-positive cases. Significantly high PVL (HPVL) levels were observed in cattle > 5 years old (p < 0.0001), females (p = 0.0008), Holstein (p < 0.0001), dairy cows (p = 0.0053), large-size farms (p < 0.0001), and cattle from Damietta (p < 0.0001) compared to other categories. Contrary, no significant differences in PVL levels were reported between the Native and Mixed cattle breeds (p = 0.13). Ultimately, the logistic regression model indicated that the probability of carrying HPVL in cattle > 5 years is 1.27 (95% CI: 1.03-2.09, p < 0.001) times more likely compared to cattle < 2 years old. In conclusion, the findings were valuably correlating the BLV prevalence with PVL as an indicator of the risk of BLV infection.

Effectiveness of integrated bovine leukemia virus eradication strategies utilizing cattle carrying resistant and susceptible major histocompatibility complex class II DRB3 alleles

Bovine leukemia virus (BLV) has spread worldwide and causes serious problems in the cattle industry owing to the lack of effective treatments and vaccines. Bovine leukemia virus is transmitted via horizontal and vertical infection, and cattle with high BLV proviral load (PVL), which is a useful index for estimating disease progression and transmission risk, are considered major infectious sources within herds. The PVL strongly correlates with highly polymorphic bovine lymphocyte antigen (BoLA)-DRB3 alleles. The BoLA-DRB3*015:01 and *012:01 alleles are known susceptibility-associated markers related to high PVL, and cattle with susceptible alleles may be at a high risk of BLV transmission via direct contact with healthy cows. In contrast, the BoLA-DRB3*009:02 and *014:01:01 alleles comprise resistant markers associated with the development of low PVL, and cattle with resistant alleles may be low-risk spreaders for BLV transmission and disrupt the BLV transmission chain. However, whether polymorphisms in BoLA-DRB3 are useful for BLV eradication in farms remains unknown. Here, we conducted a validation trial of the integrated BLV eradication strategy to prevent new infection by resistant cattle and actively eliminate susceptible cattle in addition to conventional BLV eradication strategies to maximally reduce the BLV prevalence and PVL using a total of 342 cattle at 4 stall-barn farms in Japan from 2017 to 2019. First, we placed the resistant milking cattle between the BLV-positive and BLV-negative milking cattle in a stall barn for 3 yr. Interestingly, the resistant cattle proved to be an effective biological barrier to successfully block the new BLV infections in the stall-barn system among all 4 farms. Concomitantly, we actively eliminated cattle with high PVL, especially susceptible cattle. Indeed, 39 of the 60 susceptible cattle (65%), 76 of the 140 neutral cattle (54%), and 20 of the 41 resistant cattle (48.8%) were culled on 4 farms for 3 years. Consequently, BLV prevalence and mean PVL decreased in all 4 farms. In particular, one farm achieved BLV-free status in May 2020. By decreasing the number of BLV-positive animals, the revenue-enhancing effect was estimated to be ¥5,839,262 ($39,292.39) for the 4 farms over 3 yr. Our results suggest that an integrated BLV eradication program utilization of resistant cattle as a biological barrier and the preferential elimination of susceptible cattle are useful for BLV infection control.

Exploring the presence of bovine leukemia virus among breast cancer tumors in a rural state.

The bovine leukemia virus (BLV) is a deltaretrovirus that causes malignant lymphoma and lymphosarcomas in cattle globally and has high prevalence among large scale U.S. dairy herds. Associations between presence of BLV DNA in human mammary tissue and human breast cancer incidence have been reported. We sought to estimate the prevalence of BLV DNA in breast cancer tissue samples in a rural state with an active dairy industry. We purified genomic DNA from 56 fresh-frozen breast cancer tissue samples (51 tumor samples, 5 samples representing adjacent normal breast tissue) banked between 2016 and 2019. Using nested PCR assays, multiple BLV tax sequence primers and primers for the long terminal repeat (LTR) were used to detect BLV DNA in tissue samples and known positive control samples, including the permanently infected fetal lamb kidney cell line (FLK-BLV) and blood from BLV positive cattle. The median age of patients from which samples were obtained at the time of treatment was 60 (40-93) and all were female. Ninety percent of patients had invasive ductal carcinoma. The majority were poorly differentiated (60%). On PCR assay, none of the tumor samples tested positive for BLV DNA, despite having consistent signals in positive controls. We did not find BLV DNA in fresh-frozen breast cancer tumors from patients presenting to a hospital in Vermont. Our findings suggest a low prevalence of BLV in our patient population and a need to reevaluate the association between BLV and human breast cancer.

Evaluation of herd and within-herd prevalence of bovine leukemia virus in eastern Kansas beef cattle

The last U.S. national beef BLV herd prevalence study occurred in 1997 by the USDA and found an AGID/ELISA herd prevalence of 36.4% and an individual animal prevalence of 7.73% of ani­mals tested in the northcentral U.S. which included the state of Kansas. Kansas State University and Michigan State University, in a collaborative effort, performed a BLV herd and with-in-herd prevalence study of beef cattle in eastern Kansas in the fall of 2021.

Molecular prevalence of Bovine Leukemia Virus in specialized dairies in the department of Antioquia, Colombia

Dairy production systems are a sensitive sector of the primary economy frequently affected by pathogens that negatively impact production parameters, the bovine leukemia virus (BLV) one of these. In this study, the molecular prevalence of BLV was determined in the specialized dairy sector of Antioquia using the viral marker of the envelope gene (env). Blood samples were taken from 575 bovines from specialized dairies in Antioquia distributed in 53 herds and located in the three specialized milk production areas of Antioquia (north, east, and Valle de Aburrá). DNA extraction was performed by salting out, and a nested PCR was performed to detect the env gene. The products were visualized on a 2% agarose gel with GelRed as an intercalator. A molecular prevalence of BLV of 17.0% in animals and 71.7% in herds were found, being Valle de Aburrá the area where the highest rate of positive animals was obtained (21.1%), unlike the northern area with the lowest rate (15.6%). The molecular prevalence of BLV in this study is lower than that of previous studies in the department, which ranged between 47 and 73%, and this may be associated with factors of breed resistance, the age of the animals, or management practices in the herds. These results can contribute to creating BLV control strategies and optimizing milk production in the department of Antioquia, being relevant to paying attention to the behavior of this pathogen under different production system conditions.

A safe and effective vaccine against bovine leukemia virus.

Previous attempts to develop a vaccine against bovine leukemia virus (BLV) have not been successful because of inadequate or short-lived stimulation of all immunity components. In this study, we designed an approach based on an attenuated BLV provirus by deleting genes dispensable for infectivity but required for efficient replication. The ability of the vaccine to protect from natural BLV infection was investigated in the context of dairy productive conditions in an endemic region. The attenuated vaccine was tested in a farm in which the prevalence rose from 16.7% in young cattle at the beginning of the study to more than 90% in adult individuals. Sterilizing immunity was obtained in 28 out of 29 vaccinated heifers over a period of 48 months, demonstrating the effectiveness of the vaccine. As indicated by the antiviral antibody titers, the humoral response was slightly reduced compared to wild-type infection. After initial post-vaccination bursts, the proviral loads of the attenuated vaccine remained most frequently undetectable. During the first dairy cycle, proviral DNA was not detected by nested-PCR in milk samples from vaccinated cows. During the second dairy cycle, provirus was sporadically detected in milk of two vaccinated cows. Forty-two calves born from vaccinated cows were negative for proviral DNA but had antiviral antibodies in their peripheral blood. The attenuated strain was not transmitted to sentinels, further supporting the safety of the vaccine. Altogether, these data thus demonstrate that the vaccine against BLV is safe and effective in herd conditions characterized by a very high incidence. This cost-effective approach will thus decrease the prevalence of BLV without modification of production practices. After facing a series of challenges pertaining to effectiveness and biosafety, the vaccine is now available for further large-scale delivery. The different challenges and hurdles that were bypassed may be informative for the development of a vaccine against HTLV-1.

Comparing microRNA in milk small extracellular vesicles among healthy cattle and cattle at high risk for bovine leukemia virus transmission

Enzootic bovine leukosis (EBL) is a B-cell lymphosarcoma caused by bovine leukemia virus (BLV) infection. In Japan, cattle diagnosed with EBL are not permitted for human consumption by the law, thereby causing serious economic losses to farmers. The prevalence of BLV is high in Japan (40.9% in dairy cattle and 28.7% in beef cattle, respectively), which makes it difficult to perform the test-and-slaughter of BLV-infected cattle. This necessitates preventing the spread of BLV infection in cattle by early detection, segregation, and the removal of BLV-infected cattle with high proviral load, which are considered high risk for BLV transmission. We aimed to identify cattle that were at high risk for BLV transmission by comparing microRNA (miRNA) profiles in milk small extracellular vesicles (sEV). At first, miRNA profiles in sEV were compared among 4 uninfected cattle and 4 BLV-infected cattle with high proviral load by using a microarray containing mixed probes for miRNA of cattle and humans. Significantly lower amounts of hsa-miR-557 and hsa-miR-19b-1-5p, and insignificantly but higher amounts of hsa-miR-424-5p were observed in milk sEV from BLV-infected cattle than those from uninfected cattle. Next, to evaluate the utility of the aforementioned miRNAs for the identification of cattle that were at high risk for BLV transmission, we performed quantitative real-time PCR using milk sEV newly collected from 5 uninfected cattle and 17 BLV-infected cattle with high proviral load. The cycle threshold value of hsa-miR-424-5p was significantly lower in milk sEV from BLV-infected cattle. The PCR detection was unavailable or a significant difference was not observed for hsa-miR-557 and hsa-miR-19b-1-5p, respectively. These results suggest that the amount of hsa-miR-424-5p was higher in milk sEV from BLV-infected cattle and increasing the hsa-miR-424-5p in milk sEV could be one of the characteristic trends in cattle that are high risk for BLV transmission. Moreover, assessing characteristic miRNA amounts in milk sEV, which can be recovered twice a day by milking, could be useful for the routine monitoring of cattle in dairy herds instead of blood collection.

Genetic diversity of BLV, common in the territory of the Kochenevsky district of the Novosibirsk region

This study evaluated bovine leukemia virus (BLV) prevalence in Kochenevsky district, Novosibirsk region,Russia. BLV prevalence data in farmsteads of private farmers and large livestock farms were obtained; phylogenetic analysis was conducted to define BLV genotypes in the study region. PCR analysis was used to detect BLV proviral cDNA; focal virus spread was identified. The BLV prevalence among the cattle of the private farmsteads ranged from 0% to 79% across the study areas. Among the livestocks of the Kochenevsky district, 72% of samples were positive for BLV. The BVL 1000-bp genome fragment coding the env gene region was obtained for 191 samples. The following phylogenetic analysis was performed and 162 sequences were classified as genotype 4 and the remaining 29 sequences were classified as genotype 7. BLV 7 genotype was identified among Proskudnoe private farmsteads only, while BLV genotype 4 was widespread across the region. Both BLV genotype 7 (n=17) and 4 (n=126) were detected in the dairy farm. Phylogenetic analysis revealed distinct clusters (with a bootstrap support value і75) of BLV sequences most likely indicating to the several transmission routes of the infection in the Kochenevsky district. The study results can be used to replenish fundamental knowledge about the BLV evolution and development of novel BLV prevention and elimination approaches.

Alberta dairy farmers’ and veterinarians’ opinion about bovine leukemia virus control measures

A high herd and within-herd prevalence of Bovine Leukemia Virus (BLV) infections in the dairy herds of North America and the negative effects thereof caused the Alberta dairy industry to initiate the development of an on farm BLV control program. Because BLV control is dependent on the commitment of the farmer, potential barriers were identified and farmers’ and veterinarians’ points of view toward different control options were investigated to inform how the control program might be adjusted. Conversations with these stakeholders were sought and four focus groups with farmers and eleven interviews with veterinarians were conducted. Testing for BLV, the most common BLV control strategies (testing/culling/segregation/management), as well as on farm best management practices (BMP) to prevent the transmission of BLV, were discussed. The thematic analysis of these conversations resulted in the following findings: Testing of animals was considered important for BLV control, but the financial investment was prohibitive for farmers. Test and cull as well as test and segregation approaches of test positive animals were considered efficient BLV control measures, but impractical and not feasible due to the supply managed Alberta dairy industry (i.e. milk is produced based on demand), with a high prevalence. The management of test positive animals with BMP to prevent new infections and thereby decreasing the within-herd prevalence was considered the only realistic BLV control strategy. The most important barriers for suggested BMP were the cost for some BMP, the inconvenience of performing other BMP, as well as difficulties in performing some BMP consistently and well. Additionally, a lack of knowledge about BLV and its control were identified as an important barrier. On the contrary, farmers indicated being inclined to implement BMP they considered feasible or that were considered a standard within the industry. Further, if BMP increased convenience on farm, they were considered easy to implement. Farmers and veterinarians agreed in many, but not all cases. For example, the single use of examination sleeves was met with differing opinions (i.e. considered doable by farmers while veterinarians assumed it to be too costly). In conclusion, stakeholders’ awareness and communication amongst each other (e.g. veterinarians and farmers) about BLV and its control has to be highlighted in order to manage BLV infection successfully. In addition, by communicating and understanding barriers and motivators for specific BMP, important barriers could be identified (e.g. difficulties while changing needles), and solutions found (e.g. tool belt for needles), thereby improving BLV control efforts on farm.

Intercurrence between bovine leukosis virus infection and mastitis in dairy cattle

There are several factors affecting global milk production leading to severe economic losses. Mastitis and leukosis are highly prevalent diseases especially in Brazilian cattle herds. The present study aimed to evaluate the correlation between mastitis and the bovine leukemia virus (BLV) infection in naturally infected cows belonging to dairy herds in Minas Gerais State. Additionally, the prevalence of both diseases, pathogens causing mastitis, and their association with BLV infection and persistent lymphocytosis were evaluated. The prevalence of BLV was determined using the agar gel immunodiffusion test (AGID) in 1,392 lactating cows from ten dairy herds. From the AGID results, 240 cows were selected and divided into two experimental groups: A case group composed of 160 BLV-positive cows (BLV-P) and the control group of 80 BLV-negative cows (BLV-N). Based on serological and leucocytic evaluations, the animals were classified into three subgroups: BLV-N, BLV-P without persistent lymphocytosis (BLV-PWPL), and BLV-positive with persistent lymphocytosis (BLV-PPL). The California Mastitis Test (CMT) was performed on days 0 (D0) and 60 (D60) in the subgroups and the milk from CMT-positive glands were analyzed. BLV infection was present in 100% of herds, with an overall prevalence of 80.9% in cows older than 24 months. Among BLV-P animals, 74% were classified as BLV-PWPL and 26% as BLV-PPL. Coagulase-negative Staphylococcus (73.30%),Corynebacterium spp. (12.04%), Streptococcus agalactiae (8.90%), and Staphylococcus aureus (5.76%) were predominant (61.41%). There were no statistical differences among the scores of clinical or subclinical mastitis with the pathogen type (contagious or environmental) in the BLV-PWPL or BLV-PPL subgroups. However, BLV-P animals showed higher CMT scores than BLV-N animals, suggesting a higher redisposition to subclinical mastitis in cows affected by leukosis.

Specific antiviral effect of violaceoid E on bovine leukemia virus

Bovine leukemia virus (BLV) infection has spread worldwide causing significant economic losses in the livestock industry. In countries with a high prevalence of BLV, minimizing economic losses is challenging; thus, research into various countermeasures is important for improving BLV control. Because anti-BLV drugs have not been developed, the present study explored a promising chemical compound with anti-BLV activity. Initially, screening of a chemical compound library revealed that violaceoid E (vioE), which is isolated from fungus, showed antiviral activity. Further analysis demonstrated that the antiviral effect of vioE inhibited transcriptional activation of BLV. Cellular thermal shift assay and pulldown assays provided evidence for a direct interaction between vioE and the viral transactivator protein, Tax. These data indicate that interference with Tax-dependent transcription could be a novel target for development of anti-BLV drugs. Therefore, it is suggested that vioE is a novel antiviral compound against BLV.

Bayesian Estimation of the True Seroprevalence and Risk Factor Analysis of Bovine Leukemia Virus Infection in Pakistan.

Simple SummaryThe enzootic bovine leucosis is the most common and economically important viral disease of cattle, caused by bovine leukemia virus (BLV). In cattle, infection with BLV leads to decreased milk production and premature culling and also impairs the immune system predisposing animals to other infections and increasing severity of disease. The aim of this study was to estimate the true seropositivity to BLV at the farm and within-farm levels in Pakistan, using a latent class analysis. In addition, some factors influencing BLV seropositivity were analyzed. We tested 1380 dairy cattle from 451 herds and 92 water buffalo. Analysis at the within-herd and herd levels showed 3.8% of cattle and 1.4% of herds were truly seropositive. All 92 serum samples from water buffalo were negative. The study demonstrated strong association between BLV seroprevalence and herd size but not with common housing of cattle representing indigenous breeds with exotic breed or their crossbred and also common housing of cattle and water buffalo.The objective of this study was to determine the true seroprevalence of bovine leukemia virus (BLV) infection in dairy cattle from Pakistan at the animal and herd-level. We tested 1380 dairy cattle from 451 herds and 92 water buffalo. The sera were tested by ELISA and the results were analyzed using Bayesian inference. The median posterior estimate of the herd level true BLV prevalence was 1.4%, with a 95% credible interval (CI) 0.7–3.1, whereas the median posterior estimate of the within-farm true seroprevalence was 3.8% with a 95% CI 2.8–4.8. All 92 sera collected from water buffalo were negative. Several risk factors potentially associated with seropositivity to BLV infections in Pakistan were analyzed using logistic regression model based on calculation of an odds ratio (OR). The study showed an association between seropositivity and medium herd (≥50) size (OR = 23.57, 95% CI: 3.01–103.48). Common housing of indigenous cattle with exotic-breed cattle (OR = 0.67, 95% CI: 06–2.35) or housing indigenous or their crossbred cattle with exotic-breed cattle (OR = 0.95, 95% CI: 0.14–3.01) had no effect on the BLV seroprevalence. Similarly, common housing of cattle and water buffalo was not risk factor for increased BLV seropositivity (OR = 27.10, 95% CI: 0.63–119.34).

ЛЕЙКОЗ КРУПНОГО РОГАТОГО СКОТА — ДИАГНОСТИКА, ОЗДОРОВЛЕНИЕ, АНТРОПОЗООНОЗНЫЙ ПОТЕНЦИАЛ (ИСТОРИЯ ВОПРОСА) (обзор)

Of tumor diseases in the global animal husbandry industry, the greatest danger is bovine leukemia. Since the last century, this neoplastic disease has remained relevant for veterinary medicine and, in addition, more and more questions arise concerning the potential threat of the Bovine leukemia virus (BLV) for humans. The problem is being discussed (G.Yu. Kosovskiy et al., 2016) which additionally stimulates both fundamental research of the pathogen and the pathology, as well as the methods for detection to improve dairy cattle breeding and veterinary safety. A number of publications are devoted to BLV prevalence, peculiarities, the prospects for improving animal health and welfare based on selection and vaccines developed in the world (S.G. Hopkins et al., 1997; M.A. Juliarena et al., 2017). In our report, we retrospectively compare the experience of BLV eradication in the USSR, Russia and abroad, additionally focusing on the possible role of this pathogen in the occurrence of cancer in humans. In veterinary practice, serological tests, i.e., immunodiffusion (ID) assay in agar gel and enzyme-linked immunosorbent assay (ELISA) are mostly used to detect the BLV infection. G.C. Buehring et al. (2019) reported the detection of BLV proviral DNA in blood leukocytes in 38 % of patients examined by PCR and DNA sequencing. IgG antibodies to BLV were detected in 32 %, IgM — in 58 %, and IgA — in 32 % of the samples tested. Accumulated data indicate metabolic changes in the BLV-positive animals, e.g., disorder in the metabolism of tryptophan, a critical essential acid, leading to the accumulation of dangerous endogenous metabolites in the body. Particularly, free tryptophan, indole and anthranilic acid increased in level 4-8-fold are deposited in the organs of the immune and hematopoietic system (lymph nodes, spleen, liver), in the mammary gland, lungs, and kidneys. It has been established that the milk of leukemic animals differs from the milk of healthy cows in terms of physicochemical, bacteriological (lysozyme), technological parameters and mineral composition. In C57 mice fed with pasteurized milk and heat-treated meat from ID-positive cows, the blood cell profile changed. Available data drive to the unambiguous conclusion that the products derived from BLV-infected cattle pose an increased risk to humans. Despite the lack of convincing evidence of the BLV pathogenicity for humans, concerns about the etiological role of this virus in the occurance of cancer in humans additionally necessitates continuing research on the control and eradication of this common oncogenic retro-virus in livestock farms.

Bovine leukemia virus INFECTION — DIAGNOSTICS, ERADICATION, AND ANTHROPOZOONOTIC POTENTIAL (BACKGROUND) (review)

Of tumor diseases in the global animal husbandry industry, the greatest danger is bovine leukemia. Since the last century, this neoplastic disease has remained relevant for veterinary medicine and, in addition, more and more questions arise concerning the potential threat of the Bovine leukemia virus (BLV) for humans. The problem is being discussed (G.Yu. Kosovskiy et al., 2016) which additionally stimulates both fundamental research of the pathogen and the pathology, as well as the methods for detection to improve dairy cattle breeding and veterinary safety. A number of publications are devoted to BLV prevalence, peculiarities, the prospects for improving animal health and welfare based on selection and vaccines developed in the world (S.G. Hopkins et al., 1997; M.A. Juliarena et al., 2017). In our report, we retrospectively compare the experience of BLV eradication in the USSR, Russia and abroad, additionally focusing on the possible role of this pathogen in the occurrence of cancer in humans. In veterinary practice, serological tests, i.e., immunodiffusion (ID) assay in agar gel and enzyme-linked immunosorbent assay (ELISA) are mostly used to detect the BLV infection. G.C. Buehring et al. (2019) reported the detection of BLV proviral DNA in blood leukocytes in 38 % of patients examined by PCR and DNA sequencing. IgG antibodies to BLV were detected in 32 %, IgM — in 58 %, and IgA — in 32 % of the samples tested. Accumulated data indicate metabolic changes in the BLV-positive animals, e.g., disorder in the metabolism of tryptophan, a critical essential acid, leading to the accumulation of dangerous endogenous metabolites in the body. Particularly, free tryptophan, indole and anthranilic acid increased in level 4-8-fold are deposited in the organs of the immune and hematopoietic system (lymph nodes, spleen, liver), in the mammary gland, lungs, and kidneys. It has been established that the milk of leukemic animals differs from the milk of healthy cows in terms of physicochemical, bacteriological (lysozyme), technological parameters and mineral composition. In C57 mice fed with pasteurized milk and heat-treated meat from ID-positive cows, the blood cell profile changed. Available data drive to the unambiguous conclusion that the products derived from BLV-infected cattle pose an increased risk to humans. Despite the lack of convincing evidence of the BLV pathogenicity for humans, concerns about the etiological role of this virus in the occurance of cancer in humans additionally necessitates continuing research on the control and eradication of this common oncogenic retro-virus in livestock farms.

Pilot implementation of a newly developed bovine leukemia virus control program on 11 Alberta dairy farms

We developed a custom bovine leukemia virus (BLV) control program for the Alberta dairy industry, consisting of a risk assessment and a comprehensive list of best management practices (BMP) aimed at prevention of BLV transmission between cattle. This control program was implemented on 11 farms for approximately 1 yr. Blood samples were collected from all cattle ≥12 mo old, and serum was tested with a commercial ELISA. Risk assessments were performed on each farm, risk-connected on-farm management was identified, and management changes expected to prevent transmission of BLV between cattle were suggested by the first author and agreed upon with each farmer. Throughout the following year, all participating farmers were visited multiple times to identify and overcome barriers to implementation and to monitor progress. After approximately 1 yr of implementing BLV control, all cattle ≥12 mo old on farm with a negative or no previous test result were sampled, and the within-herd prevalence was determined. The median number of cattle on farm that were ≥12 mo was 195 (range 110-524). The initial prevalence averaged 39% (13-66%). On average, 5 BMP (3-7) were suggested to each farmer. On average, 4 BMP (1-7) were implemented. At the second sampling, the average within-herd prevalence of all animals that tested positive (including the previous sampling) was 36% (12-62%). Eight farms reduced their within-herd BLV prevalence, within-herd prevalence stayed constant on 1 farm, and it increased on 1 farm. The remaining farm terminated their participation before the second sampling. The number of seroconversions per farm ranged from 3 to 109, highlighting the success of some producers to minimize new infections. The risk assessment was proven to be a valuable tool to identify flaws in on-farm management, although risk assessment score was unrelated to the within-herd BLV prevalence. Finally, it appeared that implementation of BMP aimed at prevention of BLV transmission between cattle could reduce within-herd BLV prevalence when farmers committed to their implementation.