Enzootic Bovine Leukosis (EBL) caused by the bovine leukemia virus (BLV) has been eradicated in over 20 countries. In contrast, the U.S. and many other nations are experiencing increasing prevalence in the absence of efforts to control transmission. Recent studies have shown that BLV infection in dairy cattle has a greater impact beyond the long-recognized lymphoma development that occurs in <5% of infected cattle. Like other retroviruses, BLV appears to cause multiple immune system disruptions, affecting both cellular and humoral immunity, which are likely responsible for increasingly documented associations with decreased dairy production and decreased productive lifespan. Realization of these economic losses has increased interest in controlling BLV using technology that was unavailable decades ago, when many nations eradicated BLV via traditional antibody testing and slaughter methods. This traditional control is not economically feasible for many nations where the average herd antibody prevalence is rapidly approaching 50%. The ELISA screening of cattle with follow-up testing via qPCR for proviral load helps prioritize the most infectious cattle for segregation or culling. The efficacy of this approach has been demonstrated in at least four herds. Breeding cattle for resistance to BLV disease progression also appears to hold promise, and several laboratories are working on BLV vaccines. There are many research priorities for a wide variety of disciplines, especially including the need to investigate the reports linking BLV and human breast cancer.

Transport conditions have the potential to alter the physiological responses of animals to the psychological or physical stress of transport. Transportation may introduce multiple physical and psychological stressors to unweaned calves and adult cattle, including noise, overcrowding, food and water deprivation, extreme temperatures, commingling with unfamiliar animals, handling by unfamiliar humans, and being placed in a novel environment upon arrival. Apart from these factors, the type of road and even driving skill may affect the welfare of animals. One of the concerns regarding cattle transport is that the handling and marketing of animals prior to a journey may lengthen the period of feed withdrawal. Furthermore, feed withdrawal can impact animal welfare through hunger and metabolic stress. Transportation is also associated with a decrease in animal performance as well as an increase in the incidence of bovine respiratory disease. It is well established that the transportation of cattle is a stressor that causes a quantifiable response; however, excessive stress during transport resulting in physiological or pathological changes can be reduced with best management practices. The objective of this review was to analyse the available scientific literature pertaining to the transport by road of cattle, including unweaned calves.

This study describes the longitudinal changes in bovine leukemia virus (BLV) ELISA antibodies, proviral load (PVL), and blood lymphocyte counts (LC) observed over a 2.5-year period in naturally infected cattle. The dataset utilized was from a BLV intervention field trial on three Midwestern dairy herds. Our analysis showed ELISA false negatives were more likely to occur in cattle with low PVL and normal LC. On average, negligible changes in LC were observed during six-month intervals. Periods of lymphocytosis, defined as >10,000 lymphocytes per uL of blood, were observed in 31.5% (68/216) of BLV test-positive cattle. In BLV test-positive cows, an average increase of 2900 to 3100 proviral copies per 100,000 cells was observed during each subsequent six-month sampling interval. The difference between the minimum and maximum PVL observed for an ELISA-positive cow with 3 or more observations ranged from 0 to 115,600 copies per 100,000 cells (median: 12,900; mean: 19,200). Therefore, following the identification of ELISA-positive cattle and the assessment of PVL and LC, subsequent semiannual tests to assess disease progression may not be needed. Further work is needed to determine how available diagnostic tests can be optimized to design cost-effective testing schemes for BLV control programs.

Characterization of the bovine leukocyte antigen (BoLA) DRB3 gene has shown that specific alleles associate with susceptibility or resilience to the progression of bovine leukemia virus (BLV), measured by proviral load (PVL). Through surveillance of multi-farm BLV eradication field trials, we observed differential phenotypes within seropositive cows that persist from months to years. We sought to develop a multiplex next-generation sequencing workflow (NGS-SBT) capable of genotyping 384 samples per run to assess the relationship between BLV phenotype and two BoLA genes. We utilized longitudinal results from milk ELISA screening and subsequent blood collections on seropositive cows for PVL determination using a novel BLV proviral load multiplex qPCR assay to phenotype the cows. Repeated diagnostic observations defined two distinct phenotypes in our study population, ELISA-positive cows that do not harbor detectable levels of provirus and those who do have persistent proviral loads. In total, 565 cows from nine Midwest dairy farms were selected for NGS-SBT, with 558 cows: 168 BLV susceptible (ELISA-positive/PVL-positive) and 390 BLV resilient (ELISA-positive/PVL-negative) successfully genotyped. Three BoLA-DRB3 alleles, including one novel allele, were shown to associate with disease resilience, *009:02, *044:01, and *048:02 were found at rates of 97.5%, 86.5%, and 90.3%, respectively, within the phenotypically resilient population. Alternatively, DRB3*015:01 and *027:03, both known to associate with disease progression, were found at rates of 81.1% and 92.3%, respectively, within the susceptible population. This study helps solidify the immunogenetic relationship between BoLA-DRB3 alleles and BLV infection status of these two phenotypic groupings of US dairy cattle.

Bovine leukemia virus (BLV) causes Enzootic Bovine Leukosis (EBL), a persistent life-long disease resulting in immune dysfunction and shortened lifespan in infected cattle, severely impacting the profitability of the US dairy industry. Our group has found that 94% of dairy farms in the United States are infected with BLV with an average in-herd prevalence of 46%. This is partly due to the lack of clinical presentation during the early stages of primary infection and the elusive nature of BLV transmission. This study sought to validate a near-complete genomic sequencing approach for reliability and accuracy before determining its efficacy in characterizing the sequence identity of BLV proviral genomes collected from a pilot study made up of 14 animals from one commercial dairy herd. These BLV-infected animals were comprised of seven adult dam/daughter pairs that tested positive by ELISA and qPCR. The results demonstrate sequence identity or divergence of the BLV genome from the same samples tested in two independent laboratories, suggesting both vertical and horizontal transmission in this dairy herd. This study supports the use of Oxford Nanopore sequencing for the identification of viral SNPs that can be used for retrospective genetic contact tracing of BLV transmission.

Bovine leukemia virus (BLV) is a deltaretrovirus that establishes lifelong infection in cattle and remains highly prevalent in dairy herds. This study evaluated associations between maternal BLV infection markers, daughter BoLA-DRB3 variation, and BLV outcomes in dairy youngstock. We studied 201 dam–daughter pairs from five dairy farms. BLV status was assessed using ELISA and quantitative PCR (qPCR); maternal qPCR results were available for 126 dams, and daughters were sampled longitudinally during the youngstock period. BoLA-DRB3 genotypes were determined in 199 daughters, and genotype-level epitope breadth was defined as the non-redundant union of predicted BLV Gag epitopes across both alleles. BLV proviral DNA was detected in 33 of 200 daughters (16.5%) in the qPCR outcome used for the main analysis. Among 38 daughters with BLV proviral DNA detected at least once, first detection occurred preweaning in 2, prebreeding in 24, and post-breeding in 12. Maternal ELISA positivity was associated with higher odds of daughter BLV proviral DNA detection (OR 2.74, 95% CI 1.10–6.83), whereas maternal PVL showed a positive but imprecise association (OR 1.94, 95% CI 0.89–4.22). Greater predicted epitope breadth was nominally associated with lower odds of BLV proviral DNA detection (OR 0.76, 95% CI 0.60–0.97; q = 0.162) and BLV seropositivity (OR 0.80, 95% CI 0.65–0.98; q = 0.162), but neither association remained significant after correction for multiple testing. BoLA-DRB3*014:01:01 carriers also had lower observed frequencies of BLV proviral DNA detection and BLV seropositivity. Overall, maternal infection markers were associated with later BLV outcomes in daughters, whereas the BoLA-DRB3 findings provide suggestive evidence of host immunogenetic heterogeneity.