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Intramuscular adipose tissue (IMAT; marbling) deposition is the primary determination of beef quality grade in the U.S. While finishing diets increase marbling, they also increase subcutaneous adipose tissue (SAT) accumulation, leading to excessive fattening and economic losses due to ‘waste fat’. Our objective was to investigate the IMAT and SAT-specific transcriptional profile and adipocyte function in vitro. A better understanding of the mechanisms underlying IMAT and SAT-specific characteristics will help to identify molecular and cellular molecules for developing targeted strategies to enhance marbling while limiting overall fatness in beef animals. A section of the Longissimus dorsi muscle from 9-11th ribs was collected to dissect IMAT and adjacent SAT (backfat) from six beef animals harvested at the Texas Tech University Meat Lab. The SAT and IMAT RNA was extracted and sequenced in an Illumina NovaSeq 6000 targeting a minimum of 30 million reads per sample. Transcriptome analysis showed more than 1,800 differently expressed genes (DEGs) in IMAT and SAT. There was a downregulation of genes associated with adipogenesis, lipolysis regulation, insulin sensitivity, and fatty acid metabolism (e.g., LEP, INSIG1, and PLIN4) in IMAT vs. SAT, while pathways of insulin resistance and ß-adrenergic signaling were activated. To elucidate whether the differences between IMAT and SAT transcriptional profiles were translated into changes in the adipocyte function, preadipocytes were isolated, cultured, and differentiated into adipocytes in vitro for 14 followed by functional analyses of adipogenic capacity, insulin sensitivity, and lipolytic responses. Statistical analysis was performed in GraphPad 9. Even though preadipocytes were more abundant in IMAT than in SAT (flow cytometry quantification of CD45-CD31- cell), IMAT preadipocyte proliferation rates were decreased compared with SAT. Additionally, SAT adipocytes were larger than IMAT. These findings suggest an enhancement in SAT adipogenesis/lipogenesis compared with IMAT. Metabolically, SAT adipocytes released greater glycerol concentrations compared with IMAT upon ß-adrenergic stimulation of lipolysis with isoproterenol and tended to have greater insulin-stimulated glucose uptake compared with IMAT. Altogether, these findings indicate that IMAT is unable to accumulate lipids as efficiently as SAT, and these differences may be underlined by a decreased preadipocyte hyperplasia, insulin resistance, and potential dysregulation of ß-adrenergic receptors, all of these similarly suggested by the transcriptome analysis. Our study revealed molecular and cellular differences between IMAT and SAT that may partly explain why it is common to observe increased SAT fatness and limited marbling beef cattle.

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.

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.

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.

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.

Improved reproductive management allowed dairy cow pregnancies to be optimized for beef production. The objective of this sire-controlled study was to characterize effects of maternal genetics and the dairy management system on calf growth. Pregnancies were created with a 2 × 2 factorial arrangement of dam breed (Holstein or Jersey) and mating type (artificial insemination or implantation of an in vitro produced embryo from a commercial beef cow oocyte). Resulting calves were reared in a calf ranch. Additionally, commercial beef cows were inseminated and reared resulting calves on range. Therefore, the five treatments were Angus × Holstein (A×H; n = 19), Angus × Jersey (A×J; n = 22), Angus × Beef gestated by Holstein (H-ET; n = 18), Angus × Beef gestated by Jersey (J-ET; n = 8), and Angus × Beef raised by beef (A×B; n = 20). Beginning at birth, calf body weight, cannon circumference, forearm circumference, top width, hip width, and hip height were measured approximately every 28 days until approximately 196 days of age. At birth, A×J calves weighed the least (P < 0.01). At 150 days of age, body weight was greatest (P < 0.05) among A×B calves, intermediate among H-ET and A×H calves, and least among J-ET and A×J calves (P < 0.05). Morphometric differences were detected between treatments (MANOVA P < 0.01). Primary discriminant function scores identified A×B calves having lesser values than A×J or A×H calves (ANOVA P < 0.01); A×B calves had greater cannon circumference, greater top width, and less hip height (standardized loadings of -0.47, -0.48, and 0.63, respectively). The dairy management system limited growth rate of beef genetics compared to the beef management system. In addition, Holstein dams transmitted greater growth potential than Jersey dams. Replacing maternal dairy genetics with beef genetics increased top width and decreased hip height.

Bovine leukemia virus (BLV) is a contagious, oncogenic deltaretrovirus of cattle with a worldwide distribution. In the US, over 40% of dairy cows are infected with the virus, and evidence of its economic impact is growing. This study evaluated the performance of a field-deployable automatic nucleic acid-extraction/insulated isothermal PCR (iiPCR) system for on-site BLV-proviral DNA detection in dairy cows compared with a conventional laboratory real-time PCR (rt-PCR). Assay performance was verified in parallel tests of 36 archived blood samples with 100% agreement (κ = 1.0; n = 36) between the iiPCR and conventional rt-PCR systems, and the limit of detection of the iiPCR assay was estimated to be 4 copies (genome equivalent) per reaction. The field-deployable iiPCR system was subsequently used on-farm to test freshly collected blood samples, and showed 100% agreement (κ = 1.0; n = 32) with the laboratory rt-PCR system. Fresh blood samples were collected on a second farm and tested on both systems, also with 100% agreement (κ = 1.0; n = 34). The field-deployable iiPCR/POCKIT™ combo system performs as well as a conventional laboratory-based rt-PCR system for detection of BLV proviral DNA in whole blood and may be a useful tool for on-farm evaluation of BLV-infection status in dairy cattle.

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.

The objective of this field trial was to reduce bovine leukemia virus (BLV) transmission and prevalence in commercial dairy herds using proviral load (PVL) and lymphocyte count (LC) measurements as indicators of the most infectious animals for culling or segregation. Bovine leukemia virus causes lymphoma in

This chapter reviews adherence to treatment options in pain medicine from the clinician’s point of view. Adherence is discussed in the context of principle-based ethics through the four tenets of autonomy, nonmaleficence, beneficence, and justice. Treatment adherence for pain medicine on the part of the clinician transcends mere compliance with opioid prescriptions and policies. Adherence is addressed as a broad-based approach that can be clinically appropriate when principle-based ethics are followed. The shortage of clinical therapists and the corresponding role of clinical psychologists are discussed. Ways to find a balance between the issues of abuse and underprescribing are also addressed.