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.

Characterizing the Cattle Gut Microbiome in Farms with a High and Low Prevalence of Shiga Toxin Producing Escherichia coli.

Cattle are the main reservoirs of Shiga toxin producing Escherichia coli (STEC), a major foodborne pathogen associated with acute enteric disease and hemolytic-uremic syndrome in humans. A total of 397 beef and dairy cattle from 5 farms were included in this study, of which 660 samples were collected for 16S rRNA gene sequencing. The microbiota of farms with a high-STEC prevalence (HSP) had greater richness compared to those of farms with a low-STEC prevalence (LSP). Longitudinal analyses showed STEC-shedders from LSP farms had higher microbiome diversity; meanwhile, changes in the microbiome composition in HSP farms were independent of the STEC shedding status. Most of the bacterial genera associated with STEC shedding in dairy farms were also correlated with differences in the percentage of forage in diet and risk factors of STEC carriage such as days in milk, number of lactations, and warm temperatures. Identifying factors that alter the gut microbiota and enable STEC colonization in livestock could lead to novel strategies to prevent fecal shedding and the subsequent transmission to humans.

Genetic and Phenotypic Factors Associated with Persistent Shedding of Shiga Toxin-Producing Escherichia coli by Beef Cattle.

Shiga toxin-producing Escherichia coli (STEC) is a leading cause of foodborne infections. Cattle are an important STEC reservoir, although little is known about specific pathogen traits that impact persistence in the farm environment. Hence, we sought to evaluate STEC isolates recovered from beef cattle in a single herd in Michigan. To do this, we collected fecal grabs from 26 cattle and resampled 13 of these animals at 3 additional visits over a 3-month period. In all, 66 STEC isolates were recovered for genomics and biofilm quantification using crystal violet assays. The STEC population was diverse, representing seven serotypes, including O157:H7, O26:H11, and O103:H2, which are commonly associated with human infections. Although a core genome analysis of 2,933 genes grouped isolates into clusters based on serogroups, some isolates within each cluster had variable biofilm levels and virulence gene profiles. Most (77.8%; n = 49) isolates harbored stx2a, while 38 (57.5%) isolates formed strong biofilms. Isolates belonging to the predominant serogroup O6 (n = 36; 54.5%) were more likely to form strong biofilms, persistently colonize multiple cattle, and be acquired over time. A high-quality single nucleotide polymorphism (SNP) analysis of 33 O6 isolates detected between 0 and 13 single nucleotide polymorphism (SNP) differences between strains, indicating that highly similar strain types were persisting in this herd. Similar findings were observed for other persistent serogroups, although key genes were found to differ among strong and weak biofilm producers. Together, these data highlight the diversity and persistent nature of some STEC types in this important food animal reservoir.IMPORTANCE Food animal reservoirs contribute to Shiga toxin-producing Escherichia coli (STEC) evolution via the acquisition of horizontally acquired elements like Shiga toxin bacteriophages that enhance pathogenicity. In cattle, persistent fecal shedding of STEC contributes to contamination of beef and dairy products and to crops being exposed to contaminated water systems. Hence, identifying factors important for STEC persistence is critical. This longitudinal study enhances our understanding of the genetic diversity of STEC types circulating in a cattle herd and identifies genotypic and phenotypic traits associated with persistence. Key findings demonstrate that multiple STEC types readily persist in and are transmitted across cattle in a shared environment. These dynamics also enhance the persistence of virulence genes that can be transferred between bacterial hosts, resulting in the emergence of novel STEC strain types. Understanding how pathogens persist and diversify in reservoirs is important for guiding new preharvest prevention strategies aimed at reducing foodborne transmission to humans.

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.

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.

Survival of Mycobacterium bovis during forage ensiling.

OBJECTIVE To determine whether Mycobacterium bovis remains viable in ensiled forages. SAMPLE Alfalfa, mixed mostly grass, and corn silages. PROCEDURES For each of 10 sampling days, six 250-g replicate samples of each feedstuff were created and placed in a film pouch that could be vacuum sealed to simulate the ensiling process. Within each set of replicate samples, 4 were inoculated with 10 mL of mycobacterial liquid culture medium containing viable M bovis and 2 were inoculated with 10 mL of sterile mycobacterial liquid culture medium (controls) on day 0. Pouches were vacuum sealed and stored in the dark at room temperature. On the designated sampling day, 1 control pouch was submitted for forage analysis, and the other pouches were opened, and forage samples were obtained for M bovis culture and analysis with a PCR assay immediately and 24 hours later. RESULTS None of the control samples had positive M bovis culture or PCR assay results. Among M bovis-inoculated samples, the organism was not cultured from alfalfa and corn silage for > 2 days but was cultured from mixed mostly grass silage for 28 days after inoculation and ensiling initiation. Mycobacterium bovis DNA was detected by PCR assay in samples of all 3 feedstuffs throughout the 112-day observation period. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that properly ensiled forages would be an unlikely source for M bovis transmission to cattle. Further research is necessary to determine whether ensiling kills M bovis or forces it to become dormant and, if the latter, elucidate the conditions that cause it to revert to an infectious state.

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.

Awareness and Practices Relating to Zoonotic Diseases Among Smallholder Farmers in Nepal.

Increasing livestock production to meet growing demands has resulted in greater interactions at the livestock-wildlife-human interface and more opportunities for zoonotic disease spread. Zoonoses impose enormous burdens on low-income countries like Nepal, where populations are largely dependent on livestock production and access to shared grazing lands, often near protected areas, due to population pressures. Several livestock-associated zoonoses have been reported in Nepal; however, little is known regarding Nepali farmers' knowledge of zoonoses and opportunities for disease management. We conducted a cross-sectional study to investigate Nepali farmers' awareness of zoonoses, assess current health challenges, and evaluate disease prevention and control practices. We found that awareness of zoonotic pathogens was limited, especially in informally educated and illiterate farmers; the majority of which were women. Further, farmers' preventive herd health, food safety, and sanitation practices were not associated with their awareness. Several farmers reported high-risk practices despite being aware of zoonotic diseases, suggesting a disconnect between the farmers' awareness and practice. Our study highlights the need for improving Nepali farmers' knowledge of zoonoses and disease prevention measures. Closing these awareness-practice gaps will require an improved understanding of risk and effective drivers of behavior change, alongside engagement of farmers in development of zoonotic disease prevention programs that encourage participation of both male and female farmers across all levels of education.

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.

Bovine Leukemia Virus and Mycobacterium avium subsp. paratuberculosis Are Not Associated with Shiga Toxin-Producing Escherichia coli Shedding in Cattle.

Bovine leukemia virus (BLV) is a retrovirus that causes enzootic bovine leukosis in cattle, and Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in cattle. Both diseases are chronic in nature and can lead to the disruption of normal immunological or physiological processes. Cattle are the major reservoir of Shiga toxin-producing Escherichia coli (STEC), a cause of foodborne illness in humans. We tested the hypothesis that cattle infected with BLV or MAP are more likely to shed STEC. We conducted a cross-sectional study during the summers of 2011 and 2012 in 11 Michigan cattle herds. A fecal sample from each animal was collected for STEC culture, and multiplex PCR for stx1, stx2, and eaeA was used to screen suspect colonies for STEC confirmation. Antibody detection enzyme-linked immunosorbent assays for BLV and MAP were used to screen serum from each animal. Flow cytometry was used to quantify the percentage of lymphocytes, monocytes, and neutrophils in a subsample (n =497) of blood samples. Of the animals sampled, 34.9% were BLV positive, 2.7% were MAP positive, and 16% were shedding STEC. Cattle in the dairy herds had a higher frequency of BLV and MAP than did those in beef herds, but more cattle in beef herds were shedding STEC. Neither BLV nor MAP was associated with STEC shedding (P values of 0.6838 and 0.3341, respectively). We also observed no association between STEC status and the percentage of neutrophils (P value of 0.3565), lymphocytes (P value of 0.8422), or the lymphocyte-to-monocyte ratio (P value of 0.1800). Although controlling both BLV and MAP is important for overall herd health and productivity, we found no evidence that controlling BLV and MAP has an impact on STEC shedding in cattle.

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.

Effect of feeding a direct-fed microbial on total and antimicrobial-resistant fecal coliform counts in preweaned dairy calves.

OBJECTIVE: To determine whether feeding a direct-fed microbial (DFM) to dairy calves would reduce total and antimicrobial-resistant coliform counts in feces and affect average daily gain (ADG). ANIMALS: 21 preweaned Holstein heifer calves. PROCEDURES: The study had a randomized complete block design. Within each block, 3 consecutively born calves were randomly assigned to 1 of 3 treatment groups within 24 hours after birth (day 0). Calves were fed the DFM at 1.0 g (DFM1; n = 7) or 0.5 g (DFM2; 7) twice daily or no DFM (control; 7) from days 0 through 29. A fecal sample was collected from each calf daily on days 0 through 3 and then every other day through day 29. Fecal samples were cultured, and mean numbers of total coliforms and coliforms resistant to ampicillin, ceftiofur, and tetracycline were compared among the 3 treatment groups. Calves were weighed on days 0 and 29 to calculate ADG. RESULTS: Mean total fecal coliform counts did not differ significantly among the 3 treatment groups. Mean ceftiofur-resistant and tetracycline-resistant coliform counts for the control group were significantly lower, compared with those for the DFM1 and DFM2 groups. Mean ADG did not differ significantly between the DFM1 and DFM2 groups; however, the mean ADG for all calves fed the DFM was 0.15 kg less than that for control calves. CONCLUSION AND CLINICAL RELEVANCE: Results suggested that the DFM fed to the preweaned calves of this study did not reduce total or antimicrobial-resistant coliform counts in feces.

Indoor Confined Feedlots.

Indoor confined feedlots offer advantages that make them desirable in northern climates where high rainfall and snowfall occur. These facilities increase the risk of certain health risks, including lameness and tail injuries. Closed confinement can also facilitate the rapid spread of infectious disease. Veterinarians can help to manage these health risks by implementing management practices to reduce their occurrence.

Detection of bovine respiratory syncytial virus infections in young dairy and beef cattle in Poland.

BACKGROUND: Bovine respiratory syncytial virus (BRSV) is a major contributor to bovine respiratory disease complex in dairy and beef calves, especially during the first year of life. There is a lack of comprehensive information about the prevalence of infection in cattle herds in Poland as well as in European countries outside the European Union. OBJECTIVE: The aim of this study was to estimate the prevalence of BRSV infections in young beef and dairy cattle in southeastern Poland, a region that has direct contact with non-EU countries. Animals & methods: Nasal swabs and sera (n = 120) were obtained from young cattle aged 6-12 months from 45 farms in eastern and southeastern Poland. BRSV antigen detection in the nasal swabs was carried out using a rapid immunomigration assay used in diagnosing human respiratory syncytial virus (hRSV) infections in humans, while antibodies to BRSV were detected in the sera by ELISA antibody detection. RESULTS: The study confirmed the presence of BRSV infections in young cattle under 12 months of age from both dairy and beef herds. BRSV was detected in 27 of the 45 herds (60%) sampled. CONCLUSIONS: Findings from this study indicate a high prevalence of BRSV infections in cattle in Poland, which may have a significant influence on health status and animal performance. The prevalence of infection is similar to that in other parts of Poland and other countries in Europe. Development of strategies to reduce BRSV infections is needed to improve health and productivity.

Comparison between a conductometric biosensor and ELISA in the evaluation of Johne's disease.

Johne's disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is an important gastrointestinal disease of cattle worldwide because of the economic losses encountered in JD-affected herds. These losses include reduction in milk yield in cows, premature culling and reduced carcass weight of culled diseased animals. In the U.S. dairy industry, economic losses from reduced productivity associated with JD are estimated to cost between $200 and $250 million annually. The development of non-laboratory-based assays would support more frequent testing of animals for JD and could improve its control. Conductometric biosensors combine immunomigration technology with electronic signal detection and have been adapted for the detection of IgG antibody against MAP. In the present study, a capture membrane with limited variability in the immunomigration channel and an optimal concentration of the secondary anti-bovine antibody used in a previously developed conductometric biosensor were compared with a commercially available antibody detection ELISA in their evaluation of JD, using samples of serum from cattle whose JD status where unknown. There was a moderate strength of agreement (kappa = 0.41) between the two assays. Findings from this preliminary study support the continued development of conductometric biosensors for use in the diagnosis of JD.

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.

Effect of constant exposure to cattle persistently infected with bovine viral diarrhea virus on morbidity and mortality rates and performance of feedlot cattle.

OBJECTIVE: To determine the effects of constant exposure to cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV) on health and performance of feedlot cattle. DESIGN: 3 controlled trials. ANIMALS: Crossbred feedlot cattle (trial 1, n = 184; trial 2, 138; trial 3, 138). PROCEDURES: Weaned calves were or were not vaccinated against BVDV at feedlot arrival (trial 1) or 2 (trial 2) or 3 (trial 3) weeks before feedlot arrival. During trial 1, half of the calves were commingled with PI cattle throughout the feeding period. During trial 2, 63 calves were exposed to PI cattle before weaning and all calves were exposed to PI cattle throughout the feeding period. During trial 3, all study calves were exposed to PI cattle throughout the feeding period. Morbidity and mortality rates and average daily gain (ADG) data were analyzed. RESULTS: During trial 1, calves maintained with PI cattle had a higher morbidity rate regardless of BVDV vaccination than did calves not exposed to PI cattle; however, for calves maintained with PI cattle, the morbidity rate for those vaccinated against BVDV was less than that for those not vaccinated against BVDV. During trial 2, calves exposed to PI cattle before weaning or vaccinated against BVDV had lower morbidity and mortality rates and increased ADG, compared with those for calves not exposed to PI cattle before weaning or vaccinated against BVDV. During trial 3, health and performance did not vary between calves that were and were not vaccinated against BVDV. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure of cattle to BVDV naturally or through vaccination before or at feedlot arrival mitigated the negative effects of constant exposure to PI cattle.

Review of the Michigan Upper Peninsula bovine viral diarrhea virus eradication project.

OBJECTIVE: To evaluate the effects of a voluntary regional bovine viral diarrhea virus (BVDV) control project implemented in the Upper Peninsula of Michigan. DESIGN: Longitudinal study. Sample-294 cattle producers and 11,917 cattle from the Upper Peninsula. PROCEDURES: Producer participation was assessed to determine the effectiveness of the project's promotional and educational campaigns. Participating herds were screened for cattle persistently infected (PI) with BVDV by real-time reverse transcriptase PCR assay on ear notch specimens from all newborn calves and cattle that did not calve (bulls and young stock) during the year of enrollment. Responses to a survey administered to producers 4 years after project initiation were evaluated to assess the project's effect on BVDV management practices implemented by producers. RESULTS: 294 of 495 (59%) known cattle producers in the Upper Peninsula participated in the project, and 11,917 cattle from 232 herds were tested for BVDV, of which 22 (0.18%) cattle from 9 (3.9%) herds were identified as PI with BVDV and euthanized or slaughtered. Of 140 survey respondents, 85 (61%) indicated they would test all new herd additions for BVDV, 83 (59%) would quarantine new herd additions for 30 days before introducing them to the main herd, and 81 (58%) would use the fact that their herd was free of cattle PI with BVDV for marketing purposes. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the project enhanced producer knowledge about BVDV and led to changes in producer behavior regarding BVDV management. Stakeholder engagement was as critical to project success as was increased BVDV knowledge.

Effect of transportation, time of sampling, and lymphocyte numbers on gamma interferon response to Mycobacterium bovis in cattle at time of slaughter.

Adapting the gamma interferon (IFNγ) assay for tuberculosis screening at points-of-concentration of cattle would improve global efforts to eradicate bovine tuberculosis. Two separate studies were conducted to evaluate if transportation of cattle, the time of blood collection, and total lymphocyte count affects the retention of a positive IFNγ assay result during slaughter of cattle experimentally sensitized with inactivated Mycobacterium bovis. Study 1 evaluated IFNγ assay responses to M. bovis and Mycobacterium avium stimulations in 5 cows (4 sensitized and 1 control) at the housing facility, after a 30-min transport to the slaughter facility, immediately before stunning, at commencement of exsanguination, and at 5 min after exsanguination commenced. Study 2 evaluated IFNγ assay responses to Mycobacterium antigen stimulations and total lymphocyte count in blood collected from 5 steers (4 sensitized and 1 control) at the housing facility, at commencement of exsanguination and at 2 successive 1-min intervals. The results indicated that blood obtained from sensitized cattle at commencement of exsanguination was more likely to remain positive than blood collected at successive time points; hence the time of blood collection is crucial to obtaining a useful IFNγ assay result for bovine tuberculosis at slaughter. The lymphocyte count progressively declined following exsanguination, and this decline might contribute to the reduction in the measured IFNγ. To compensate for the reduction in IFNγ production, a different set of positive cutoff values might be needed for blood collected at exsanguination. The current findings provide useful preliminary information necessary for making changes to the interpretation of the IFNγ assay on blood collected during exsanguination.

Differential gene expression segregates cattle confirmed positive for bovine tuberculosis from antemortem tuberculosis test-false positive cattle originating from herds free of bovine tuberculosis.

Antemortem tests for bovine tuberculosis (bTB) currently used in the US measure cell-mediated immune responses against Mycobacterium bovis. Postmortem tests for bTB rely on observation of gross and histologic lesions of bTB, followed by bacterial isolation or molecular diagnostics. Cumulative data from the state of Michigan indicates that 98 to 99% of cattle that react positively in antemortem tests are not confirmed positive for bTB at postmortem examination. Understanding the fundamental differences in gene regulation between antemortem test-false positive cattle and cattle that have bTB may allow identification of molecular markers that can be exploited to better separate infected from noninfected cattle. An immunospecific cDNA microarray was used to identify altered gene expression (P ≤ 0.01) of 122 gene features between antemortem test-false positive cattle and bTB-infected cattle following a 4-hour stimulation of whole blood with tuberculin. Further analysis using quantitative real-time PCR assays validated altered expression of 8 genes that had differential power (adj P ≤ 0.05) to segregate cattle confirmed positive for bovine tuberculosis from antemortem tuberculosis test-false positive cattle originating from herds free of bovine tuberculosis.