Cross-sectional study to describe bovine leukemia virus herd and within-herd ELISA prevalence and bovine leukemia virus proviral load of convenience-sampled Kansas beef cow-calf herds.

OBJECTIVE: Determine bovine leukemia virus (BLV) seroprevalence of adult female cattle in Eastern Kansas beef herds and the proviral load (PVL) of those cattle found to be ELISA positive. ANIMALS: Convenience sample of 2,845 cows from 44 beef herds. PROCEDURES: BLV serostatus was determined using an ELISA antibody test (gp-51; IDEXX). BLV quantitative PCR (qPCR) status and PVL were determined utilizing a qPCR test (SS1 qPCR test; CentralStar Laboratories). The association of age, herd size, and body condition score (BCS) category on the probability of being BLV positive was evaluated with a multiple variable logistic regression analysis that used BLV status as a binary outcome, herd nested within ranch as a random effect, and BCS, herd size, and age category as fixed effects. RESULTS: Forty-two of 44 herds had at least 1 BLV ELISA-positive cow (95.5% herd seroprevalence). Overall, 1,564 of the 2,845 cows were BLV ELISA positive (55.0% individual animal prevalence). No association between BLV ELISA status and herd size or BCS was identified. When evaluated by age, the model-adjusted probability of being BLV ELISA positive was lowest for heifers (1 year of age, first parity) and increased until 5 to 6 years of age. Of the 1,564 ELISA-positive animals, 838 were qPCR positive (53.6%). The model-adjusted probability of being qPCR positive was not associated with age, herd size, or BCS category. CLINICAL RELEVANCE: This study indicated that BLV-seropositive status both as a herd classification and individual animal classification was very common in this population. Because the percentage of BLV-seropositive cows varied between herds and by age, this study provides evidence that it is essential for investigators to control for herd and age in any analysis of the association of BLV serostatus and health and production outcomes of interest. Some BLV ELSIA-seropositive cows were classified as BLV negative by qPCR, and risk factors may differ between classification status by ELISA and qPCR.

The effect of bovine leukemia virus on dairy cow longevity.

Bovine leukemia virus (BLV) is a retrovirus of cattle that infects approximately 45% of all US dairy cattle, with about 90% of US dairy herds having at least one infected animal. Studies have found BLV infection to be associated with multiple measures of decreased immune function, which may explain the observed economic losses from milk production, decreased cow longevity, and predisposition to lymphoma and other diseases. Our objective was to measure the association between BLV infection and cow longevity in dairy cow operations. Ninety-one dairy herds from 9 US states volunteered to participate in this study. Milking dairy cows (n = 3,611) were tested for BLV antibodies using an ELISA milk test, and their presence in the herd was monitored for an average of 29 mo. The survival analysis controlled for herd and lactation number. Cows sold for dairy purposes were excluded, and individual cow results were not shared with producers so as not to influence culling decisions. Overall, 47.1% (1,701/3,611) of cows were BLV-positive by ELISA. The significant hazard ratio of 1.30 indicated that positive cows were 30% more likely than their negative herdmates to die or be culled during the monitoring period. These results are consistent with other studies in finding a negative effect of BLV infection on cow lifespan.

Identification of BoLA Alleles Associated with BLV Proviral Load in US Beef Cows.

Bovine leukemia virus (BLV) causes enzootic bovine leukosis, the most common neoplastic disease in cattle. Previous work estimates that 78% of US beef operations and 38% of US beef cattle are seropositive for BLV. Infection by BLV in a herd is an economic concern for producers as evidence suggests that it causes an increase in cost and a subsequent decrease in profit to producers. Studies investigating BLV in dairy cattle have noted disease resistance or susceptibility, measured by a proviral load (PVL) associated with specific alleles of the bovine leukocyte antigen (BoLA) DRB3 gene. This study aims to investigate the associations between BoLA DRB3 alleles and BLV PVL in beef cattle. Samples were collected from 157 Midwest beef cows. BoLA DRB3 alleles were identified and compared with BLV PVL. One BoLA DRB3 allele, *026:01, was found to be associated with high PVL in relation to the average of the sampled population. In contrast, two alleles, *033:01 and *002:01, were found to be associated with low PVL. This study provides evidence of a relationship between BoLA DRB3 alleles and BLV PVL in US beef cows.

Phenotypic Selection of Dairy Cattle Infected with Bovine Leukemia Virus Demonstrates Immunogenetic Resilience through NGS-Based Genotyping of BoLA MHC Class II Genes.

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.

Diagnostic Measures of Disease Progression in Cattle Following Natural Infection with Bovine Leukemia Virus.

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.

Natural Infection of Dairy Cows with Bovine Leukemia Virus Affects Immunoglobulin Levels in Saliva and Serum but Not Milk.

Bovine leukemia virus (BLV) is a retroviral infection that disrupts the immune function of infected animals. It is widespread among U.S. dairy cattle. In this pilot study, the average total IgA and IgM concentrations in milk, saliva, and serum samples from BLV ELISA-positive (ELISA+) dairy cows were compared against samples from BLV ELISA-negative (ELISA-) cows using the Kruskal-Wallis test (with ties). The results from ELISA+ cows were also stratified by lymphocyte count (LC) and proviral load (PVL). In milk and saliva from ELISA+ cows, the average total IgA and IgM concentrations were decreased compared to ELISA- cows, although this was only statistically significant for saliva IgM in cows with low PVL (p = 0.0424). Numerically, the average total IgA concentrations were 33.6% lower in milk and 23.7% lower in saliva, and the average total IgM concentrations were 42.4% lower in milk and 15.5% lower in saliva. No significant differences were observed in the total serum IgA concentrations, regardless of PVL and LC. The total serum IgM from ELISA+ cows was significantly decreased (p = 0.0223), with the largest decreases occurring in the highest PVL and LC subgroups. This pilot study is a first step in investigating the impact of BLV on mucosal immunity and will require further exploration in each of the various stages of disease progression.

Current Developments in the Epidemiology and Control of Enzootic Bovine Leukosis as Caused by Bovine Leukemia Virus.

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.

Bovine leukemia virus detection and dynamics following experimental inoculation.

Bovine leukemia virus (BLV) infects more than 40% of the United States cattle population and impacts animal health and production. Control programs aiming to reduce disease prevalence and incidence depend on the ability to detect the BLV provirus, anti-BLV antibodies, and differences in blood lymphocyte counts following infection. These disease parameters also can be indicative of long-term disease progression. The objectives of this study were to determine the timing and to describe early fluctuations of BLV-detection by qPCR, ELISA, and lymphocyte counts. Fifteen Holstein steers were experimentally inoculated with 100 µL of a blood saline inoculum. Three steers served as in-pen negative controls and were housed with the experimentally infected steers to observe the potential for contract transmission. Five additional negative controls were housed separately. Steers were followed for 147 days post-inoculation (DPI). Infections were detected in experimentally infected steers by qPCR and ELISA an average of 24- and 36 DPI, respectively. Significant differences in lymphocyte counts between experimentally infected and control steers were observed from 30 to 45 DPI. Furthermore, a wide variation in peak proviral load and establishment was observed between experimentally infected steers. The results of this study can be used to inform control programs focused on the detection and removal of infectious cattle.

Impact of bovine leukemia virus infection on beef cow longevity.

Bovine leukosis is a chronic lymphoproliferative disorder caused by bovine leukemia virus (BLV). Previous studies estimate that 38 % of cow-calf beef herds and 10.3 % of individual beef cows in the US are BLV seropositive. About 70 % of BLV infected animals are asymptomatic carriers of the virus, while less than 5% develop lymphosarcoma, the leading reason for carcass condemnation at the US slaughterhouses. Studies provide evidence that BLV infection leads to decreased immune function making animals more vulnerable to other diseases, which could shorten their productive lifespan and increase economic losses in the cattle industry. BLV seropositive dairy cows are reportedly more likely to be culled sooner compared with their uninfected herd mates. Beyond simple prevalence studies, little is known about the impact of BLV infection in beef cattle production or specifically on beef cow longevity. Our objective was to determine the association between BLV infection and cow longevity in beef cow-calf operations. Twenty-seven cow-calf herds from the Upper Midwest volunteered to participate in this study. Female beef cattle (n = 3146) were tested for serum BLV antibodies by ELISA. A subsample of 648 cows were also tested for BLV proviral load (PVL). Culling data was collected for the subsequent 24 months. Twenty-one herds (77.7 %) had at least one BLV-infected animal, and 29.2 % (930/3146) of tested animals were BLV seropositive. Of the BLV-positive cows, 33.7 % (318/943) were culled compared with 32.1 % (541/1682) of the seronegative cows. BLV status did not affect cows' longevity within herds (P = 0.062). However, cows with high BLV PVL had decreased survival within the herd compared with ELISA- negative cows (P = 0.01). Overall, infection with BLV did not impact beef cow longevity unless the disease had progressed to a point of high BLV PVL.

Control of Bovine Leukemia Virus in Three US Dairy Herds by Culling ELISA-Positive Cows.

The objective of this trial was to evaluate a test-and-cull approach to controlling bovine leukemia virus (BLV) in US dairy herds with a low BLV prevalence. Despite worldwide distribution of the virus, 21 nations have eradicated BLV from their dairy cattle and are currently considered 'BLV-free.' In contrast, the US has attempted no industry-wide BLV control programs and has experienced an increase in BLV prevalence among dairy cows to about 40%. This raises concerns about production efficiency, herd health, and sustainability. In a pilot field trial with three Midwestern-US dairy herds, a test-and-cull approach using ELISA screening of milk samples was successful in reducing BLV prevalence in two herds. In the third herd, BLV prevalence increased following the introduction of infected heifers that were raised at an out-of-state calf raising facility. This trial demonstrated that a test-and-cull approach to BLV control can be successful in US dairy herds with low BLV prevalence, but ongoing surveillance is necessary to prevent reintroduction of the virus.

Breeding bulls as a potential source of bovine leukemia virus transmission in beef herds.

OBJECTIVE: To determine the prevalence of bovine leukemia virus (BLV) in beef bulls; evaluate the presence of BLV provirus DNA in blood, smegma, and semen samples; and analyze whether blood BLV proviral load was associated with differential blood cell counts. DESIGN: Observational cross-sectional study. ANIMALS: 121 beef bulls ≥ 2 years old from 39 Michigan herds. PROCEDURES: Blood, smegma, and semen samples were collected from each bull during a routine breeding soundness examination. An ELISA was used to detect serum anti-BLV antibodies. A coordination of common motifs-quantitative PCR assay was used to detect BLV provirus DNA in blood, smegma, and semen samples. Bulls with positive results on both the BLV serum ELISA and coordination of common motifs-quantitative PCR assay were considered infected with BLV. RESULTS: 19 of 39 (48.7%) herds and 54 of 121 (44.6%) bulls were infected with BLV. Provirus DNA was detected in the blood of all 54 and in smegma of 4 BLV-infected bulls but was not detected in any semen sample. Lymphocyte count was significantly greater in BLV-infected bulls than in uninfected bulls. The proportion of BLV-infected bulls with lymphocytosis (16/54 [29.6%]) was greater than the proportion of uninfected bulls with lymphocytosis (6/67 [9%]). Lymphocyte count was positively associated with BLV proviral load in BLV-infected bulls. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that almost half of beef bulls and herds were infected with BLV, and BLV provirus DNA was detected in the smegma of some BLV-infected bulls. Bulls may have an important role in BLV transmission in beef herds.

Lack of Bovine leukemia virus transmission during natural breeding of cattle.

Bovine leukosis is a chronic lymphoproliferative disorder that leads to significant economic losses in the beef and dairy industries. The major route of virus transmission is believed to be iatrogenic through the transfer of blood containing infected lymphocytes. In addition, BLV proviral DNA has been identified in nasal secretions, saliva, milk, colostrum, semen and smegma; however, natural transmission of BLV through these secretions has not been clearly demonstrated. The use of bulls for natural breeding has been identified as a risk factor in BLV infected dairy herds. However, the risk of BLV-infected bulls transmitting the virus is unknown. The objective of this study was to evaluate the potential for BLV transmission during natural breeding between a BLV-infected bull and uninfected heifers. Forty healthy, BLV seronegative, and proviral-negative beef heifers were randomly assigned to one of two groups: control heifers (n = 20) exposed to a BLV seronegative and proviral negative bull and challenged heifers (n = 20) exposed to a BLV seropositive and proviral-positive bull. Each group was housed with the bull for a period of 38 days in a 5-acre pasture to replicate the housing of commercial beef cattle during the breeding season. Blood samples were collected from heifers at -60, -30 and 0 days prior to breeding and day 30, 60 and 90 after the breeding period ended. Blood samples were tested for BLV antibodies by ELISA and BLV proviral DNA by CoCoMo-qPCR. New infection was not detected by ELISA or CoCoMo-qPCR in any of the challenge or control heifers at any time point during the study. Based on these results, BLV infected bulls that are healthy and aleukemic may not be a significant risk of BLV transmission during a defined breeding season.

Prevalence of Bovine Leukemia Virus Antibodies in US Dairy Cattle.

OBJECTIVE: To estimate current US herd-level and animal-level prevalence of bovine leukemia virus (BLV) in dairy cows and characterize epidemiologic features. DESIGN: Cross-sectional observational study design and survey. ANIMALS: 4120 dairy cows from 103 commercial dairy herds in 11 states across the US. PROCEDURES: Milk samples were collected from dairy cows through routine commercial sampling and tested for anti-BLV antibodies by antibody capture ELISA. Based on the ELISA results of a sample of an average of 40 cows per herd, within-herd apparent prevalence (AP) was estimated by a directly standardized method and by a lactation-weighted method for each herd. Within-herd AP estimates were summarized to give estimates of US herd-level and animal-level AP. Differences in AP by lactation, region, state, breed, and herd size were examined to characterize basic epidemiologic features of BLV infection. RESULTS: 94.2% of herds had at least one BLV antibody positive cow detected. The average within-herd standardized AP was 46.5%. Lactation-specific AP increased with increasing lactation number, from 29.7% in first lactation cows to 58.9% in 4th and greater lactation cows. Significant differences were not observed based on region, state, breed, or herd size. CONCLUSIONS AND CLINICAL RELEVANCE: These results are consistent with a historical trend of increasing prevalence of BLV among US dairy cattle. Given the findings of other studies on the negative impacts of BLV infection on milk production and cow longevity, these findings are clinically relevant for veterinarians counseling dairy clients on the risks of BLV to their herds.

Dairy Cows Naturally Infected with Bovine Leukemia Virus Exhibit Abnormal B- and T-Cell Phenotypes after Primary and Secondary Exposures to Keyhole Limpet Hemocyanin.

Bovine leukemia virus (BLV) is a retrovirus that is highly prevalent in US dairy herds: over 83% are BLV infected and the within-herd infection rate can be almost 50% on average. While BLV is known to cause lymphosarcomas, only 5% or fewer infected cattle will develop lymphoma; this low prevalence of cancer has historically not been a concern to dairy producers. However, more recent research has found that BLV+ cows without lymphoma produce less milk and have shorter lifespans than uninfected herdmates. It has been hypothesized that BLV infection interferes with normal immune function in infected cattle, and this could lead to reduced dairy production. To assess how naturally infected BLV+ cows responded to a primary and secondary immune challenge, 10 BLV+ and 10 BLV- cows were injected subcutaneously with keyhole limpet hemocyanin (KLH) and dimethyldioctadecylammonium bromide. B- and T-cell responses were characterized over the following 28 days. A total of 56 days after primary KLH exposure, cows were re-injected with KLH and B- and T-cell responses were characterized again over the following 28 days. BLV+ cows produced less KLH-specific IgM after primary immune stimulation; demonstrated fewer CD45R0+ B cells, altered proportions of CD5+ B cells, altered expression of CD5 on CD5+ B cells, and reduced MHCII surface expression on B cells ex vivo; exhibited reduced B-cell activation in vitro; and displayed an increase in BLV proviral load after KLH exposure. In addition, BLV+ cows had a reduced CD45R0+γδ+ T-cell population in the periphery and demonstrated a greater prevalence of IL4-producing T cells in vitro. All together, our results demonstrate that both B- and T-cell immunities are disrupted in BLV+ cows and that antigen-specific deficiencies can be detected in BLV+ cows even after a primary immune exposure.

Comparing the Genetic Diversity and Antimicrobial Resistance Profiles of Campylobacter jejuni Recovered from Cattle and Humans.

Campylobacter jejuni, a leading cause of gastroenteritis in humans, is a foodborne pathogen that can reside in chickens, pigs, and cattle. Because resistance to fluoroquinolones and macrolides, which are commonly used to treat human infections, has emerged in C. jejuni, it is imperative to continously monitor resistance patterns and examine the genetic variation in strains from human infections and animal reservoirs. Our previous study of C. jejuni from human campylobacteriosis cases showed a significantly higher rate of tetracycline resistance compared to national trends, and identified multilocus sequence type (ST)-982 and a history of cattle contact to be associated with tetracycline resistance. To further investigate these associations, we conducted a cross-sectional study to determine the frequency of antimicrobial resistance and examine the genetic diversity of C. jejuni recovered from 214 cattle at three Michigan herds. Overall, the prevalence of C. jejuni was 69.2% (range: 58.6-83.8%) for the three farms, and 83.7% (n = 113) of isolates were resistant to one or more antimicrobials. Resistance to only tetracycline predominated among the cattle isolates (n = 89; 65.9%) with most resistant strains belonging to ST-459 (96.5%) or ST-982 (86.4%). Among the 22 STs identified, STs 459 and 982 were more prevalent in one feedlot, which reported the use of chlortetracycline in feed upon arrival of a new herd. PCR-based fingerprinting demonstrated that the ST-982 isolates from cattle and humans had identical banding patterns, suggesting the possibility of interspecies transmission. Resistance to macrolides (1.5%) and ciprofloxacin (16.3%) was also observed; 14 of the 22 ciprofloxacin resistant isolates represented ST-1244. Together, these findings demonstrate a high prevalence of antimicrobial resistant C. jejuni in cattle and identify associations with specific genotypes. Continuous monitoring and identification of risk factors for resistance emergence are imperative to develop novel methods aimed at decreasing pathogen persistence in food animal reservoirs and the frequency of resistant infections in humans.

Reduced humoral immunity and atypical cell-mediated immunity in response to vaccination in cows naturally infected with bovine leukemia virus.

Bovine leukemia virus (BLV) is a retrovirus that is widely distributed across US dairy herds: over 83% of herds are BLV-infected and within-herd infection rates can approach 50%. BLV infection reduces both animal longevity and milk production and can interfere with normal immune health. With such a high prevalence of BLV infection in dairy herds, it is essential to understand the circumstances by which BLV negatively affects the immune system of infected cattle. To address this question, BLV- and BLV+ adult, lactating Holstein dairy cows were vaccinated with Bovi-Shield GOLD® FP® 5 L5 HB and their immune response to vaccination was measured over the course of 28days. On day 0 prior to vaccination and days 7, 14 and 28 post-vaccination, fresh PBMCs were characterized for T and B cell ratios in the periphery. Plasma was collected to measure titers of IgM, IgG1 and IgG2 produced against bovine herpesvirus 1 (BHV1), Leptospira hardjo and L. pomona, as well as to characterize neutralizing antibody titers produced against BHV1 and bovine viral diarrhea virus types 1 and 2. On day 18 post-vaccination, PBMCs were cultured in the presence of BHV1 and flow cytometry was used to determine IFNγ production by CD4+, CD8+ and γδ T cells and to investigate CD25 and MHCII expression on B cells. BLV+ cows produced significantly lower titers of IgM against BHV1, L. hardjo and L. pomona and produced lower titers of IgG2 against BHV1. γδ T cells from BLV+ cows displayed a hyper reactive response to stimulation in vitro, although no differences were observed in CD4+ or CD8+ T cell activation. Finally, B cells from BLV+ cows exhibited higher CD25 expression and reduced MHCII expression in response to stimulation in vitro. All together, data from this study support the hypothesis that BLV+ cows fail to respond to vaccination as strongly as BLV- cows and, consequently, may have reduced protective immunity when compared to healthy BLV- cows.

Factors Associated with Shiga Toxin-Producing Escherichia coli Shedding by Dairy and Beef Cattle.

UNLABELLED: Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen that can cause hemorrhagic colitis and hemolytic-uremic syndrome. Cattle are the primary reservoir for STEC, and food or water contaminated with cattle feces is the most common source of infections in humans. Consequently, we conducted a cross-sectional study of 1,096 cattle in six dairy herds (n = 718 animals) and five beef herds (n = 378 animals) in the summers of 2011 and 2012 to identify epidemiological factors associated with shedding. Fecal samples were obtained from each animal and cultured for STEC. Multivariate analyses were performed to identify risk factors associated with STEC positivity. The prevalence of STEC was higher in beef cattle (21%) than dairy cattle (13%) (odds ratio [OR], 1.76; 95% confidence interval [CI], 1.25, 2.47), with considerable variation occurring across herds (range, 6% to 54%). Dairy cattle were significantly more likely to shed STEC when the average temperature was >28.9°C 1 to 5 days prior to sampling (OR, 2.5; 95% CI, 1.25, 4.91), during their first lactation (OR, 1.8; 95% CI, 1.1, 2.8), and when they were <30 days in milk (OR, 3.9; 95% CI, 2.1, 7.2). These data suggest that the stress or the negative energy balance associated with lactation may result in increased STEC shedding frequencies in Michigan during the warm summer months. Future prevention strategies aimed at reducing stress during lactation or isolating high-risk animals could be implemented to reduce herd-level shedding levels and avoid transmission of STEC to susceptible animals and people. IMPORTANCE: STEC shedding frequencies vary considerably across cattle herds in Michigan, and the shedding frequency of strains belonging to non-O157 serotypes far exceeds the shedding frequency of O157 strains, which is congruent with human infections in the state. Dairy cattle sampled at higher temperatures, in their first lactation, and early in the milk production stage were significantly more likely to shed STEC, which could be due to stress or a negative energy balance. Future studies should focus on the isolation of high-risk animals to decrease herd shedding levels and the potential for contamination of the food supply.

Options for the control of bovine leukemia virus in dairy cattle.

The subclinical impact of bovine leukemia virus (BLV) on the sustainability of the US dairy industry is only now being fully recognized. Findings of recent longitudinal studies conducted in Michigan dairy herds were consistent with the results of previous studies in showing that within-herd prevalence of BLV-infected cattle was negatively associated with milk production and cow longevity. Risk factors relating to routes of hematogenous transmission such as the use of shared hypodermic needles, shared reproductive examination sleeves, and natural breeding were associated with BLV within-herd prevalence. Few US dairy producers know the prevalence of BLV-infected cattle in their herds or are aware of the insidious economic impact of BLV or the options for BLV control. As an increasing number of countries eradicate BLV from their cattle populations, restrictions on the movement of US cattle and cattle products will likely increase. Veterinarians should be aware of recent developments for screening serum and milk samples for antibodies against BLV and the results of research regarding the economic impact of BLV so they can advise their dairy clients of available alternatives for monitoring and controlling BLV infection.

Impact of bovine leukemia virus infection on neutrophil and lymphocyte concentrations in dairy cattle.

OBJECTIVE: To determine the effect of bovine leukemia virus (BLV) infection on absolute neutrophil and lymphocyte concentrations in healthy lactating Holstein dairy cattle. DESIGN: Observational cross-sectional survey. ANIMALS: 311 healthy lactating Holstein dairy cattle from herds in Michigan (n = 2), Wisconsin (1), Iowa (1), and Pennsylvania (1). PROCEDURES: Whole and anticoagulated (EDTA) blood samples were collected. Serum samples were tested for antibody against BLV by use of an ELISA. Absolute neutrophil and lymphocyte concentrations were measured in EDTA blood samples with an automated hematology analyzer and manual differential cell counts. RESULTS: 208 cows tested positive and 103 cows tested negative for anti-BLV antibodies. Neutrophil concentration was not significantly different between BLV-positive versus BLV-negative cattle. The distribution of lymphocyte concentration was positively skewed for the entire cow population (n = 311) and the BLV-positive subset (208). In contrast, lymphocyte concentration distribution was approximately normal for BLV-negative cows (n = 103). Consequently, the presence or absence of BLV infection strongly influenced the calculated neutrophil-to-lymphocyte concentration ratio. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that absolute lymphocyte concentration is significantly affected by BLV infection in dairy cattle. Accordingly, hematologic reference intervals should be derived from healthy animals that are not infected with BLV and patient BLV status must be considered for meaningful interpretation of lymphocyte concentration. We recommend that the calculated neutrophil-to-lymphocyte ratio be abandoned because it does not provide more information than direct comparison of patient absolute leukocyte concentration with updated reference intervals from healthy BLV-negative cattle.

Herd-level determinants of bovine leukaemia virus prevalence in dairy farms.

The prevalence of bovine leukaemia virus (BLV) was determined in 113 Michigan dairy herds by ELISA testing for anti-BLV antibodies in milk. Additionally, an interview regarding management practices with cooperating herd managers identified farm-level variables thought to be associated with prevalence of BLV. Twenty-three risk factors (P ≤ 0·1) were identified on one-way ANOVA or simple linear regression. Multivariate analysis identified several management practices whose predictive value for increased prevalence of BLV may relate to transmission among herd mates, e.g. reuse of hypodermic needles, lack of fly control, gouge dehorning and increased use of injections in dry cows. Additionally, exclusive breeding of heifers with artificial insemination was associated with decreased BLV prevalence, as compared with at least some use of natural service by a bull. Although intervention studies are needed before causal relationships can be concluded, and unaccounted variables related to transmission exist among dairy herds, these findings suggest management practices that may help dairy producers reduce the transmission of BLV within their herds.