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Introduction: Improving ewe longevity is an important breeding and management goal, as death loss and early culling of mature ewes are economic burdens in the sheep industry. Ewe longevity can be improved by selecting for positive reproductive outcomes. However, the breeding approaches for accomplishing this come with the challenge of recording a lifetime trait. Characterizing genetic factors underpinning ewe longevity and related traits could result in the development of genomic selection strategies to improve the stayability of sheep through early, informed selection of replacement ewes. Methods: Towards this aim, a genome-wide association study (GWAS) was performed to identify genetic markers associated with ewe longevity, reproductive, and production traits. Traits evaluated included longevity (i.e., length of time in the flock), parity and the lifetime number of lambs born, lambs born alive, lambs weaned, and weight of lambs weaned. Ewe records from previous studies were used. Specifically, Rambouillet (n = 480), Polypay (n = 404), Suffolk (n = 182), and Columbia (n = 64) breed ewes (N = 1,130) were analyzed against 503,617 SNPs in across-breed and within-breed GWAS conducted with the Bayesian-information and Linkage-disequilibrium Iteratively Nested Keyway (BLINK) model in R. Results: The across-breed GWAS identified 25 significant SNPs and the within-breed GWAS for Rambouillet, Polypay, and Suffolk ewes identified an additional 19 significant SNPs. The most significant markers were rs411309094 (13:22,467,143) associated with longevity in across-breed GWAS ( p -value = 8.3E-13) and rs429525276 (2:148,398,336) associated with both longevity ( p -value = 6.4E-15) and parity ( p -value = 4.8E-15) in Rambouillet GWAS. Significant SNPs were identified within or in proximity (±50 kb) of genes with known or proposed roles in reproduction, dentition, and the immune system. These genes include ALPL, ANOS1, ARHGEF26, ASIC2, ASTN2, ATP8A2, CAMK2D, CEP89, DISC1, ITGB6, KCNH8, MBNL3, MINDY4, MTSS1, PLEKHA7, PRIM2, RNF43, ROBO2, SLCO1A2, TMEM266, TNFRSF21, and ZNF804B . Discussion: This study proposes multiple SNPs as candidates for use in selection indices and suggests genes for further research towards improving understanding of the genetic factors contributing to longevity, reproductive, and production traits of ewes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Smitchger, Taylor, Mousel, Schaub, Thorne, Becker and Murdoch.)

As chronic wasting disease (CWD) continues to spread across North America, the relationship between CWD and host genetics has become of interest. In Rocky Mountain elk (Cervus elaphus nelsoni), one or two copies of a leucine allele at codon 132 of the prion protein gene (132L*) has been shown to prolong the incubation period of CWD. Our study examined the relationship between CWD epidemiology and codon 132 evolution in elk from Wyoming, USA, from 2011 to 2018. Using PCR and Sanger sequencing, we genotyped 997 elk and assessed the relationship between genotype and CWD prevalence estimated from surveillance data. Using logistic regression, we showed that each 1% increase in CWD prevalence is associated with a 9.6% increase in the odds that an elk would have at least one copy of leucine at codon 132. In some regions, however, 132L* variants were found in the absence of CWD, indicating that evolutionary and epidemiologic patterns can be heterogeneous across space and time. We also provide evidence that naturally occurring CWD is not rare in 132L* elk, which merits the study of shedding kinetics in 132L* elk and the influence of genotype on CWD strain diversity. The management implications of cervid adaptations to CWD are difficult to predict. Studies that investigate the degree to which evolutionary outcomes are shaped by host spatial structure can provide useful epidemiologic insight, which can in turn aid management by informing scale and extent of mitigation actions., (© Wildlife Disease Association 2024.)

Despite the benefits of crossbreeding on animal performance, genetic evaluation of sheep in the U.S. does not directly incorporate records from crossbred lambs. Crossbred animals may be raised in different environments as compared to purebreds. Systemic factors such as age of dam and birth and rearing type may, therefore, affect purebred and crossbred performance differently. Furthermore, crossbred performance may benefit from heterozygosity, and genetic and environmental variances may be heterogeneous in different breeds and their crosses. Such issues must be accounted for in a combined (purebred and crossbred) genetic evaluation. The objectives of this study were to i) determine the effect of dam age and birth type on birth weight, and dam age and birth-rearing type on weaning weight, in purebred and crossbred lambs, ii) test for heterogeneous genetic and environmental variances in those weights, and iii) assess the impact of including weights on crossbred progeny on sire estimated breeding values (EBV). Performance records were available on purebred Columbia and Suffolk lambs. Crossbred information was available on lambs sired by Suffolk, Columbia or Texel rams mated to Columbia, Suffolk, or crossbred ewes. A multiple-trait animal model was fitted in which weights from Columbia, Suffolk, or crossbred lambs were considered different traits. At birth, there were 4,160, 2,356, and 5,273 Columbia, Suffolk, and crossbred records, respectively, with means (SD) of 5.14 (1.04), 5.32 (1.14), and 5.43 (1.23) kg, respectively. At weaning, on average at 122 (12) d, there were 2,557, 980, and 3,876 Columbia, Suffolk, and crossbred records, respectively, with corresponding means of 39.8 (7.2), 40.3 (7.9), and 39.6 (8.0) kg. Dam age had a large positive effect on birth and weaning weight in pure and crossbred lambs. At birth, however, the predicted effect was larger in crossbred and Suffolk lambs. While an increase in a number of lambs born and reared had a strong and negative influence on birth and weaning weight, the size of the effect did not differ across-breed types. Environmental variances were similar at birth and weaning, but additive variances differed among breed types for both weights. Combining purebred and crossbred information in the evaluation not only improved predictions of genetic merit in purebred sires but also allowed for direct comparisons of sires of different breeds. Breeders thus can benefit from an additional tool for making selection decisions., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science.)

The tick-transmitted disease bovine babesiosis causes significant economic losses in many countries around the world. Current control methods include modified live-attenuated vaccines that have limited efficacy. Recombinant proteins could provide effective, safe, and low-cost alternative vaccines. We compared the expression of the Babesia bovis thrombospondin-related anonymous protein (TRAP) family from parasites in bovine blood, in vitro induced sexual stages, and kinetes from tick hemolymph. Quantitative PCR showed that in blood and sexual stages, TRAP3 was highly transcribed as compared to the other TRAPs. In contrast, the TRAP1 gene was highly transcribed in kinetes as compared to the other TRAPs. Fixed immunofluorescence assays showed that TRAP2, 3, and 4 proteins were expressed by both blood and sexual stages. Conversely, TRAP1 protein, undetected on blood and induced sexual stages, was the only family member expressed by kinetes. Live IFA revealed that TRAP2, 3, and 4 proteins were expressed on the surface of both B. bovis blood and sexual stages. Modeling of B. bovis TRAP1 and TRAP4 tertiary structure demonstrated both proteins folded the metal-ion-dependent adhesion site (MIDAS) domain structure of Plasmodium TRAP. In conclusion, TRAP proteins may serve as potential vaccine targets to prevent infection of bovine and ticks with B. bovis essential for controlling the spread of bovine babesiosis.

Crossbreeding is a common practice among commercial sheep producers to improve animal performance. However, genetic evaluation of U.S. sheep is performed within breed type (terminal sire, semi-prolific, and western range). While incorporating crossbred records may improve assessment of purebreds, it requires accounting for heterotic and breed effects in the evaluation. The objectives of this study were to: 1) describe the development of a paternal composite (PC) line, 2) determine the effect of direct and maternal heterosis on growth traits of crossbred lambs, 3) estimate (co)variance components for direct and maternal additive, and uncorrelated maternal environmental, effects, and 4) provide an interpretation of the estimates of random effects of genetic groups, and to use those solutions to compare the genetic merit of founding breed subpopulations. Data included purebred and crossbred records on birth weight (BN; n = 14,536), pre-weaning weight measured at 39 or 84 d (WN; n = 9,362) depending on year, weaning weight measured at 123 d (WW; n = 9,297), and post-weaning weight measured at 252 d (PW; n = 1,614). Mean (SD) body weights were 5.3 (1.1), 16.8 (3.9) and 28.0 (7.6), 39.1 (7.2), and 54.2 (8.7) kg for BN, WN (at the two ages), WW, and PW, respectively. In designed experiments, the Siremax, Suffolk, Texel, Polypay, Columbia, Rambouillet, and Targhee breeds were compared within the same environment. Estimates of heterotic effects and covariance components were obtained using a multiple trait animal model. Genetic effects based on founders' breeds were significant and included in the model. Percent estimates of direct heterosis were 2.89 ± 0.61, 2.60 ± 0.65, 4.24 ± 0.56, and 6.09 ± 0.86, and estimates of maternal heterosis were 1.92 ± 0.87, 4.64 ± 0.80, 3.95 ± 0.66, and 4.04 ± 0.91, for BN, WN, WW, and PW, respectively. Correspondingly, direct heritability estimates were 0.17 ± 0.02, 0.13 ± 0.02, 0.17 ± 0.02, and 0.46 ± 0.04 for BN, WN, WW, and PW. Additive maternal effects accounted for trivial variation in PW. For BN, WN, and WW, respectively, maternal heritability estimates were 0.16 ± 0.02, 0.10 ± 0.02, and 0.07 ± 0.01. Uncorrelated maternal environmental effects accounted for little variation in any trait. Direct and maternal heterosis had considerable impact on growth traits, emphasizing the value of crossbreeding and the need to account for heterosis, in addition to breed effects, if crossbred lamb information is included in genetic evaluation., (Published by Oxford University Press on behalf of the American Society of Animal Science 2022.)

Monocytes are a core component of the immune system that arise from bone marrow and differentiate into cells responsible for phagocytosis and antigen presentation. Their derivatives are often responsible for the initiation of the adaptive immune response. Monocytes and macrophages are central in both controlling and propagating infectious diseases such as infection by Coxiella burnetii and small ruminant lentivirus in sheep. Genotypes from 513 Rambouillet, Polypay, and Columbia sheep (Ovis aries) were generated using the Ovine SNP50 BeadChip. Of these sheep, 222 animals were subsequently genotyped with the Ovine Infinium® HD SNP BeadChip to increase SNP coverage. Data from the 222 HD genotyped sheep were combined with the data from an additional 258 unique sheep to form a 480-sheep reference panel; this panel was used to impute the low-density genotypes to the HD genotyping density. Then, a genome-wide association analysis was conducted to identify loci associated with absolute monocyte counts from blood. The analysis used a single-locus mixed linear model implementing EMMAX with age and ten principal components as fixed effects. Two genome-wide significant peaks (p < 5x10-7) were identified on chromosomes 9 and 1, and ten genome-wide suggestive peaks (p < 1x10-5) were identified on chromosomes 1, 2, 3, 4, 9, 10, 15, and 16. The identified loci were within or near genes including KCNK9, involved into cytokine production, LY6D, a member of a superfamily of genes, some of which subset monocyte lineages, and HMGN1, which encodes a chromatin regulator associated with myeloid cell differentiation. Further investigation of these loci is being conducted to understand their contributions to monocyte counts. Investigating the genetic basis of monocyte lineages and numbers may in turn provide information about pathogens of veterinary importance and elucidate fundamental immunology., Competing Interests: The authors have declared that no competing interests exist.

Arthropod-borne apicomplexan pathogens remain a great concern and challenge for disease control in animals and humans. In order to prevent Babesia infection, the discovery of antigens that elicit protective immunity is essential to establish approaches to stop disease dissemination. In this study, we determined that poly-N-acetylglucosamine (PNAG) is conserved among tick-borne pathogens including B. bovis , B. bigemina , B. divergens , B. microti, and Babesia WA1. Calves immunized with synthetic ß-(1→6)-linked glucosamine oligosaccharides conjugated to tetanus toxoid (5GlcNH 2 -TT) developed antibodies with in vitro opsonophagocytic activity against Staphylococcus aureus . Sera from immunized calves reacted to B. bovis . These results suggest strong immune responses against PNAG. However, 5GlcNH 2 -TT-immunized bovines challenged with B. bovis developed acute babesiosis with the cytoadhesion of infected erythrocytes to brain capillary vessels. While this antigen elicited antibodies that did not prevent disease, we are continuing to explore other antigens that may mitigate these vector-borne diseases for the cattle industry.

Background: Babesia bovis is one of the most significant tick-transmitted pathogens of cattle worldwide. Babesia bovis parasites have a complex lifecycle, including development within the mammalian host and tick vector. Each life stage has developmental forms that differ in morphology and metabolism. Differentiation between these forms is highly regulated in response to changes in the parasite's environment. Understanding the mechanisms by which Babesia parasites respond to environmental changes and the transmission cycle through the biological vector is critically important for developing bovine babesiosis control strategies., Results: In this study, we induced B. bovis sexual stages in vitro using xanthurenic acid and documented changes in morphology and gene expression. In vitro induced B. bovis sexual stages displayed distinctive protrusive structures and surface ruffles. We also demonstrated the upregulation of B. bovis calcium-dependent protein kinase 4 (cdpk4), tubulin-tyrosine ligase (ttl), and methyltransferase (mt) genes by in vitro induced sexual stages and during parasite development within tick midguts., Conclusions: Similar to other apicomplexan parasites, it is likely that B. bovis upregulated genes play a vital role in sexual reproduction and parasite transmission. Herein, we document the upregulation of cdpk4, ttl, and mt genes by both B. bovis in vitro induced sexual stages and parasites developing in the tick vector. Understanding the parasite's biology and identifying target genes essential for sexual reproduction will enable the production of non-transmissible live vaccines to control bovine babesiosis., (© 2021. The Author(s).)

Mycoplasma ovipneumoniae contributes to polymicrobial pneumonia in domestic sheep. Elucidation of host genetic influences of M. ovipneumoniae nasal detection has the potential to reduce the incidence of polymicrobial pneumonia in sheep through implementation of selective breeding strategies. Nasal mucosal secretions were collected from 647 sheep from a large US sheep flock. Ewes of three breeds (Polypay n = 222, Rambouillet n = 321, and Suffolk n = 104) ranging in age from one to seven years, were sampled at three different times in the production cycle (February, April, and September/October) over four years (2015 to 2018). The presence and DNA copy number of M. ovipneumoniae was determined using a newly developed species-specific qPCR. Breed (P<0.001), age (P<0.024), sampling time (P<0.001), and year (P<0.001) of collection affected log10 transformed M. ovipneumoniae DNA copy number, where Rambouillet had the lowest (P<0.0001) compared with both Polypay and Suffolk demonstrating a possible genetic component to detection. Samples from yearlings, April, and 2018 had the highest (P<0.046) detected DNA copy number mean. Sheep genomic DNA was genotyped with the Illumina OvineHD BeadChip. Principal component analysis identified most of the variation in the dataset was associated with breed. Therefore, genome wide association analysis was conducted with a mixed model (EMMAX), with principal components 1 to 6 as fixed and a kinship matrix as random effects. Genome-wide significant (P<9x10-8) SNPs were identified on chromosomes 6 and 7 in the all-breed analysis. Individual breed analysis had genome-wide significant (P<9x10-8) SNPs on chromosomes 3, 4, 7, 9, 10, 15, 17, and 22. Annotated genes near these SNPs are part of immune (ANAPC7, CUL5, TMEM229B, PTPN13), gene translation (PIWIL4), and chromatin organization (KDM2B) pathways. Immune genes are expected to have increased expression when leukocytes encounter M. ovipneumoniae which would lead to chromatin reorganization. Work is underway to narrow the range of these associated regions to identify the underlying causal mutations., Competing Interests: The authors have declared that no competing interests exist.

Small ruminant lentivirus (SRLV) causes Maedi-Visna or Ovine Progressive Pneumonia in sheep and creates insidious livestock production losses. This retrovirus is closely related to human immunodeficiency virus and currently has no vaccines or cure. Genetic marker assisted selection for sheep disease resiliency presents an attractive management solution. Previously, we identified a region containing a cluster of zinc finger genes that had association with ovine SRLV proviral concentration. Trait-association analysis validated a small insertion/deletion variant near ZNF389 (rs397514112) in multiple sheep breeds. In the current study, 543 sheep from two distinct populations were genotyped at 34 additional variants for fine mapping of the regulatory elements within this locus. Variants were selected based on ChIP-seq annotation data from sheep alveolar macrophages that defined active cis-regulatory elements predicted to influence zinc finger gene expression. We present a haplotype block of variants within regulatory elements that have improved associations and larger effect sizes (up to 4.7-fold genotypic difference in proviral concentration) than the previously validated ZNF389 deletion marker. Hypotheses for the underlying causal mutation or mutations are presented based on changes to in silico transcription factor binding sites. These variants offer alternative markers for selective breeding and are targets for future functional mutation assays.

The Ovine Functional Annotation of Animal Genomes (FAANG) project, part of the broader livestock species FAANG initiative, aims to identify and characterize gene regulatory elements in domestic sheep. Regulatory element annotation is essential for identifying genetic variants that affect health and production traits in this important agricultural species, as greater than 90% of variants underlying genetic effects are estimated to lie outside of transcribed regions. Histone modifications that distinguish active or repressed chromatin states, CTCF binding, and DNA methylation were used to characterize regulatory elements in liver, spleen, and cerebellum tissues from four yearling sheep. Chromatin immunoprecipitation with sequencing (ChIP-seq) was performed for H3K4me3, H3K27ac, H3K4me1, H3K27me3, and CTCF. Nine chromatin states including active promoters, active enhancers, poised enhancers, repressed enhancers, and insulators were characterized in each tissue using ChromHMM. Whole-genome bisulfite sequencing (WGBS) was performed to determine the complement of whole-genome DNA methylation with the ChIP-seq data. Hypermethylated and hypomethylated regions were identified across tissues, and these locations were compared with chromatin states to better distinguish and validate regulatory elements in these tissues. Interestingly, chromatin states with the poised enhancer mark H3K4me1 in the spleen and cerebellum and CTCF in the liver displayed the greatest number of hypermethylated sites. Not surprisingly, active enhancers in the liver and spleen, and promoters in the cerebellum, displayed the greatest number of hypomethylated sites. Overall, chromatin states defined by histone marks and CTCF occupied approximately 22% of the genome in all three tissues. Furthermore, the liver and spleen displayed in common the greatest percent of active promoter (65%) and active enhancer (81%) states, and the liver and cerebellum displayed in common the greatest percent of poised enhancer (53%), repressed enhancer (68%), hypermethylated sites (75%), and hypomethylated sites (73%). In addition, both known and de novo CTCF-binding motifs were identified in all three tissues, with the highest number of unique motifs identified in the cerebellum. In summary, this study has identified the regulatory regions of genes in three tissues that play key roles in defining health and economically important traits and has set the precedent for the characterization of regulatory elements in ovine tissues using the Rambouillet reference genome., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Davenport, Massa, Bhattarai, McKay, Mousel, Herndon, White, Cockett, Smith and Murdoch.)

Throughout their life cycle, Babesia parasites alternate between a mammalian host, where they cause babesiosis, and the tick vector. Transition between hosts results in distinct environmental signals that influence patterns of gene expression, consistent with the morphological and functional changes operating in the parasites during their life stages. In addition, comparing differential patterns of gene expression among mammalian and tick parasite stages can provide clues for developing improved methods of control. Hereby, we upgraded the genome assembly of Babesia bovis, a bovine hemoparasite, closing a 139 kbp gap, and used RNA-Seq datasets derived from mammalian blood and tick kinete stages to update the genome annotation. Of the originally annotated genes, 1,254 required structural changes, and 326 new genes were identified, leading to a different predicted proteome compared to the original annotation. Next, the RNA-Seq data was used to identify B. bovis genes that were differentially expressed in the vertebrate and arthropod hosts. In blood stages, 28% of the genes were upregulated up to 300 fold, whereas 26% of the genes in kinetes, a tick stage, were upregulated up to >19,000 fold. We thus discovered differentially expressed genes that may play key biological roles and serve as suitable targets for the development of vaccines to control bovine babesiosis., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Alveolar macrophages function in innate and adaptive immunity, wound healing, and homeostasis in the lungs dependent on tissue-specific gene expression under epigenetic regulation. The functional diversity of tissue resident macrophages, despite their common myeloid lineage, highlights the need to study tissue-specific regulatory elements that control gene expression. Increasing evidence supports the hypothesis that subtle genetic changes alter sheep macrophage response to important production pathogens and zoonoses, for example, viruses like small ruminant lentiviruses and bacteria like Coxiella burnetii . Annotation of transcriptional regulatory elements will aid researchers in identifying genetic mutations of immunological consequence. Here we report the first genome-wide survey of regulatory elements in any sheep immune cell, utilizing alveolar macrophages. We assayed histone modifications and CTCF enrichment by chromatin immunoprecipitation with deep sequencing (ChIP-seq) in two sheep to determine cis- regulatory DNA elements and chromatin domain boundaries that control immunity-related gene expression. Histone modifications included H3K4me3 (denoting active promoters), H3K27ac (active enhancers), H3K4me1 (primed and distal enhancers), and H3K27me3 (broad silencers). In total, we identified 248,674 reproducible regulatory elements, which allowed assignment of putative biological function in macrophages to 12% of the sheep genome. Data exceeded the FAANG and ENCODE standards of 20 million and 45 million useable fragments for narrow and broad marks, respectively. Active elements showed consensus with RNA-seq data and were predictive of gene expression in alveolar macrophages from the publicly available Sheep Gene Expression Atlas. Silencer elements were not enriched for expressed genes, but rather for repressed developmental genes. CTCF enrichment enabled identification of 11,000 chromatin domains with mean size of 258 kb. To our knowledge, this is the first report to use immunoprecipitated CTCF to determine putative topological domains in sheep immune cells. Furthermore, these data will empower phenotype-associated mutation discovery since most causal variants are within regulatory elements., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Massa, Mousel, Herndon, Herndon, Murdoch and White.)

Signature of selection studies have identified many genomic regions with known functional importance and some without verified functional roles. Multiple studies have identified Transmembrane protein 8B ( TMEM8B) rs426272889 as having been recently under extreme selection pressure in domesticated sheep, but no study has provided sheep phenotypic data clarifying a reason for extreme selection. We tested rs426272889 for production trait association in 770 U.S. Rambouillet, Targhee, Polypay, and Suffolk sheep. TMEM8B rs426272889 was associated with mature weight at 3 and 4 years ( p < 0.05). This suggested selection for sheep growth and body size might explain the historical extreme selection pressure in this genomic region. We also tested Sperm-associated antigen 8 ( SPAG8 ) rs160159557 encoding a G493C substitution. While this variant was associated with mature weights at ages 3 and 4, it was not as strongly associated as TMEM8B rs426272889. Transmembrane protein 8B has little functional information except as an inhibitor of cancer cell proliferation. To our knowledge, this is the first study linking TMEM8B to whole organism growth and body size under standard conditions. Additional work will be necessary to identify the underlying functional variant(s). Once identified, such variants could be used to improve sheep production through selective breeding.

Assessment of immune fitness is valuable in many aspects of livestock management and research. Determining immune consequences of selection for increased disease resistance or inhabiting various environments or climates can lead to different management decisions. The ability to measure immune responses due to different diets, pregnancy status, or aging will increase insight about how these factors contribute to overall immune health. The main objective of these experiments was to adapt a methodology used in cattle and pigs to measure both the humoral and cell-mediated immune response in sheep and goats. The route of administration of two antigens, Candida albicans and hen egg white lysozyme, were compared in sheep to determine differences in antibody or cell-mediated immune response. Subcutaneous injection produced a larger (P < 0.001) cell-mediated response compared to intramuscular injection. Inoculation in the axillary space produced a larger (P = 0.0031) antibody response compared to neck region. Finally, methodology was confirmed in goats. Complete blood cell counts were compared and lymphocytes were highest in low cell-mediated responders while eosinophils were highest in average antibody-mediated responders. This work provides a means to measure immune fitness in sheep and goats allowing for future experiments examining environmental or genetic effects on the immune response., (Published by Elsevier B.V.)

Babesia bovis is a hemoprotozoan parasite of cattle that has a complex life cycle within vertebrate and invertebrate hosts. In the mammalian host, B. bovis undergoes asexual reproduction while in the tick midgut, gametes are induced, fuse, and form zygotes. The zygote infects tick gut epithelial cells and transform into kinetes that are released into the hemolymph and invade other tick tissues such as the ovaries, resulting in transovarial transmission to tick offspring. To compare gene regulation between different B. bovis life stages, we collected parasites infecting bovine erythrocytes and tick hemolymph. Total RNA samples were isolated, and multiplexed libraries sequenced using paired-end 100 cycle reads of a HiSeq 2500. The data was normalized using the TMM method and analysed for significant differential expression using the generalized linear model likelihood ratio test (GLM LRT) in edgeR. To validate our datasets, ten genes were selected using NormFinder. Genes that had no significant fold change between the blood and tick stages in the RNA-Seq datasets were tested by quantitative PCR to determine their suitability as "housekeeping" genes. The normalized RNA-Seq data revealed genes upregulated during infection of the mammalian host or tick vector and six upregulated genes were validated by quantitative PCR. These datasets can help identify useful targets for controlling bovine babesiosis., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships which have, or could be perceived to have, influenced the work reported in this article., (© 2020 The Author(s).)

In this study, we describe a new in vitro tick feeding system that facilitates the study of ticks and tick-borne pathogens. To optimize the system, we used Dermacentor andersoni and Anaplasma marginale as a tick-pathogen interaction model. Ticks were fed on bovine blood containing 10-fold dilutions of the pathogen to determine the effect of dose on tick infection rate. After feeding on infected blood, ticks were transferred to uninfected blood to stimulate bacterial replication within the tick vector. During stimulation feeding, blood samples were collected daily to determine if infected ticks secreted viable A. marginale. The results demonstrated similar attachment rates between the first and second tick feeding. Tick midgut and salivary glands were infected with A. marginale. However, salivary gland infection rates decreased as the percentage of parasitized erythrocytes decreased during tick acquisition feeding. Bacteria recovered from the in vitro system were able to infect a naïve bovine host. Using the highly transmissible A. marginale St. Maries strain, we demonstrated that the artificial tick feeding system is a suitable tool to study tick-pathogen interactions and that A. marginale tick salivary gland infection is dose dependent. This work demonstrates the utility of an artificial tick feeding system to directly study the association between the number of acquired pathogens and transmissibility by ticks.

Though lamb sales account for the majority of annual receipts on U.S. sheep operations, wool is an important income source for many Western flocks. Crossing fine-wool sheep with prolific or composite hair sheep breeds can increase lamb production, but fleece quality and marketability may be reduced by a greater content of nonwool fibers (e.g., med and kemp). The objectives of this study were to compare BW and wool characteristics of Rambouillet, Polypay, and Romanov-White Dorper × Rambouillet (RW-RA) ewes under extensive rangeland management conditions. Ewe BW was collected before mating (fall) and 30 d postlambing (spring) each year from 1 yr up to 4 yr of age. In spring and fall, Rambouillet and Polypay ewes were similar in BW (P ≥ 0.94). Spring BW did not differ (P = 0.13) between RW-RA and Polypay, but Rambouillet ewes were heavier than RW-RA in the spring (P = 0.02). Both Rambouillet and Polypay ewes were heavier (P < 0.07) than RW-RA in the fall. Greasy fleece weight and mid-side wool samples were collected from ewes at 1 and 4 yr of age. Clean fleece weights (CFW) were estimated from average laboratory scoured yield of mid-side wool samples composited within-breed. Average fiber diameter (AFD), SD of fiber diameter (SD-FD), and percentage med (%M), kemp (%K), and total medullated fibers (%T) were quantified on individual mid-side wool samples. There was no difference in 1-yr-old CFW among breed types (P ≥ 0.96). Four-yr-old Rambouillet ewes had heavier CFW (2.29 kg; P < 0.001) than 4-yr-old Polypay (1.83 kg) and RW-RA ewes (1.86 kg), which were not different (P > 0.99). Within 1- and 4-yr-olds, AFD differed among breed type (P < 0.001) and was the finest for Rambouillet (20.1 and 21.9 µm, respectively), intermediate for RW-RA (22.8 and 24.8 µm), and coarsest for Polypay (24.2 and 26.7 µm). Also within 1- and 4-yr olds, SD-FD was lowest in Rambouillet, intermediate in Polypay, and greatest in RW-RA (P < 0.01). Wool from RW-RA ewes had greater %M, %K, and %T (P < 0.001) than wool from Rambouillet and Polypay ewes, which were not different (P > 0.99). Results indicated superior wool production for Rambouillet compared with the coarser, more variable wool produced by Polypay and RW-RA. Still, past research reported greater lamb production in Polypay and RW-RA ewes which, under recent market conditions, would be associated with greater annual gross revenue for these breed types than for Rambouillet., (Published by Oxford University Press on behalf of the American Society of Animal Science 2019.)

Increasing prolificacy has been proposed to be the most effective way to increase the biological efficiency and profitability of sheep production. However, use of prolific breeds and genes with major effects on ovulation rate can increase prolificacy to levels that may not be desirable or sustainable in extensive rangeland production systems. This study thus evaluated effects of triplet births on ewe productivity and ewe and lamb performance. An initial study used 666 purebred Polypay litters to compare ewes with triplet litters that were required to raise all the lambs (Treatment A) with those whose triplet litters were reduced to 2 lambs (Treatment R). Adult Polypay ewes had an average litter size of 2.35 lambs per litter. The frequency of litters of 3 or more lambs was 43.2%; 56.0% of lambs were born in litters of 3 or more lambs. Ewes that had singles weaned fewer lambs and less body weight (BW) of lambs (P < 0.001; 0.94 lambs and 40.4 kg, respectively) than ewes that had twins or triplets. Ewes with triplet litters in Treatment A weaned more lambs (P < 0.01) and more BW of lambs (P < 0.05) than ewes that had triplets in Treatment R (2.13 lambs and 62.9 kg, respectively, vs. 1.79 lambs and 55.0 kg, respectively), and weaned more lambs than ewes that had twins (1.77 lambs; P < 0.01). However, neither group of triplet-bearing ewes weaned more BW of lambs than ewes that had twins (58.9 kg; P ≥ 0.34). In 2 sets of data involving 442 purebred Polypay litters and 987 litters from Polypay or Romanov‒White Dorper × Rambouillet ewes mated to terminal sires, ewes were required to raise all triplet-born lambs. Death losses for triplets in these studies (39.6 and 31.6%, respectively) were higher than those in Treatment A of the initial study (26.2%), resulting in greater numbers of lambs weaned for triplet, compared to twin, litters (1.79 vs. 1.68, respectively; P = 0.02) but no greater weight of lambs weaned (54.3 vs. 55.4 kg, respectively; P = 0.17). Based on these 3 sets of data, ewes that were required to rear triplet lambs weaned 0.20 more lambs per litter than ewes that had twins but also had 0.75 additional dead lambs per litter, and thus a lamb mortality overhead of 3.75 additional dead lambs for each additional weaned lamb. We conclude that there is an intermediate optimum prolificacy level for extensive rangeland production systems. If optimum prolificacy is exceeded, removal and artificial rearing of surplus lambs are necessary to avoid increased lamb death losses.

Coxiella burnetii is a globally distributed zoonotic bacterial pathogen that causes abortions in ruminant livestock. In humans, an influenza-like illness results with the potential for hospitalization, chronic infection, abortion, and fatal endocarditis. Ruminant livestock, particularly small ruminants, are hypothesized to be the primary transmission source to humans. A recent Netherlands outbreak from 2007-2010 traced to dairy goats resulted in over 4,100 human cases with estimated costs of more than 300 million euros. Smaller human Q fever outbreaks of small ruminant origin have occurred in the United States, and characterizing shedding is important to understand the risk of future outbreaks. In this study, we assessed bacterial shedding and seroprevalence in 100 sheep from an Idaho location associated with a 1984 human Q fever outbreak. We observed 5% seropositivity, which was not significantly different from the national average of 2.7% for the U.S. (P>0.05). Furthermore, C. burnetii was not detected by quantitative PCR from placentas, vaginal swabs, or fecal samples. Specifically, a three-target quantitative PCR of placenta identified 0.0% shedding (exact 95% confidence interval: 0.0%-2.9%). While presence of seropositive individuals demonstrates some historical C. burnetii exposure, the placental sample confidence interval suggests 2016 shedding events were rare or absent. The location maintained the flock with little or no depopulation in 1984 and without C. burnetii vaccination during or since 1984. It is not clear how a zero-shedding rate was achieved in these sheep beyond natural immunity, and more work is required to discover and assess possible factors that may contribute towards achieving zero-shedding status. We provide the first U.S. sheep placental C. burnetii shedding update in over 60 years and demonstrate potential for C. burnetii shedding to reach undetectable levels after an outbreak event even in the absence of targeted interventions, such as vaccination.

Ewe productivity (i.e., total number or weight of lambs weaned per breeding ewe) is a key indicator of lamb production efficiency. This study compared various measures of ewe productivity and ewe and lamb performance among ewes of 3 breed types mated to rams of 4 terminal-sire breed types in an extensive rangeland production system. Purebred Rambouillet ( = 212), purebred Polypay ( = 236), and crossbred Romanov-White Dorper × Rambouillet (RW-RA; = 231) ewes were produced from locally adapted Polypay and Rambouillet ewes and then annually mated to Columbia, Suffolk, Columbia × Suffolk, or Suffolk × Columbia sires for up to 4 yr, beginning at 1 yr of age. The cumulative number and weight of lambs weaned through 4 yr were greater for RW-RA (5.9 lambs and 153 kg, respectively) and Polypay ewes (4.9 lambs and 123 kg, respectively) than for Rambouillet ewes (2.9 lambs and 99 kg, respectively) and also were greater for RW-RA ewes than for Polypay ewes (all < 0.001). Greater productivity of RW-RA and Polypay ewes, compared with Rambouillet ewes, was driven by greater lambing rates (ewes lambing per ewe exposed) as ewe lambs (87 and 77 vs. 31%, respectively; < 0.001) and greater ( < 0.001) litter sizes as ewe lambs (1.3, 1.3, and 1.0, respectively) and adult ewes (2.1, 2.0, and 1.6, respectively). The RW-RA ewes also had greater longevity ( < 0.01) and cumulative lambing rates ( < 0.001) than Polypay and Rambouillet ewes. Lamb BW at birth and weaning in adult ewes favored less-prolific Rambouillet ewes ( < 0.001), but after adjustment for type of birth and rearing and weaning age, differences in weaning BW among ewe breed types were small and not significant ( = 0.08). Effects of sire breed type on measures of cumulative ewe productivity were not significant ( > 0.74), but Suffolk-sired lambs had the heaviest adjusted birth weights ( = 0.01) and Columbia-sired lambs tended to have the lightest adjusted weaning weights ( = 0.12). Combined effects of heterosis and additive breed effects were associated with greater lambing rates in ewe lambs, larger litters at all ages, and substantially greater number and weight of lambs weaned for Polypay and RW-RA ewes than for Rambouillet ewes.

An accurate and simple-to-perform new version of a competitive ELISA (cELISA) kit that became commercially available in 2015 for testing of cattle for antibody to Anaplasma marginale was validated for detection of Anaplasma ovis antibody in domestic sheep. True positives and negatives were identified using nested PCR (nPCR) as the gold standard. Negative bovine control sera supplied with the kit were used to calculate % inhibition (%I), designated bovine control ELISA (BcELISA), and this was compared to %I calculated from negative ovine sera derived from hand-raised, pathogen-free sheep, designated ovine control ELISA (OcELISA). The receiver operating characteristics area under the curve was 1.0 with a p value <0.001 regardless of the source of the control sera. The cutoff values for negative BcELISA and OcELISA were <30%I and <27%I, respectively. Our work confirmed that this Anaplasma antibody cELISA kit version 2 can be used with the serum controls supplied in the kit to test for A. ovis antibody in domestic sheep. Furthermore, this work confirmed the historically high infection prevalence (>93%) at the U.S. Sheep Experiment Station (Dubois, Idaho), in spite of efforts to reduce the possibility for iatrogenic transmission there, suggesting high levels of tick-borne transmission.

Background: The major histocompatibility complex (MHC) is an organized cluster of tightly linked vertebrate genes with immunological and non-immunological functions. While the important MHC gene DRB1 has been examined in regard to many sheep infectious disease traits, only one study, based on microsatellite markers, has previously examined DRB1 and sheep production traits. Furthermore, to our knowledge no studies have examined DRB1 relationship with lifetime ewe prolificacy traits. Therefore, we analyzed association between the presence of DRB1 SNP haplotypes with internationally recognized standard names and production traits including growth and lifetime prolificacy in 370 Rambouillet, Columbia, and Polypay sheep., Results: The DRB1 *2001 haplotype was associated with increased weaning and mature weights, as well as average daily gain (Šidák P<0.05; corrected for the number of haplotypes tested). Interestingly, the *2001 haplotype also showed a trend toward association with increased total number of lifetime lambs born (Šidák P=0.084) and number of lambs born alive (Šidák P=0.084). In contrast, the DRB1 *0301 haplotype was associated with decreased mature weight (Šidák P=0.01)., Conclusions: Since the *2001 haplotype was present in all three breeds, these results suggest there is at least one functional mutation in the region that influences growth and prolificacy traits that may be broadly present across several breeds. Furthermore, combined use of the similar *2001 and *0301 multi-marker haplotypes that nonetheless have opposing directions of production trait associations will enhance mutation discovery in this region. If undesirable alleles for underlying mutations can be identified, selective pressure against one or a small number of undesirable alleles may improve production with limited impact on MHC genetic diversity and infectious disease susceptibility., (Published by Elsevier B.V.)

Background: Classical scrapie is a transmissible spongiform encephalopathy (TSE) that affects sheep and goats. Our previous bioassay studies in lambs revealed that scrapie prions could be detected in association with peripheral blood monocular cells (PBMC), B lymphocytes and platelet-rich plasma fractions. In the present study, bioassay in lambs was again used to determine if scrapie prions are associated with the other two subsets of PBMC, monocytes and T lymphocytes., Results: PBMC, monocytes and T lymphocytes were isolated from two preclinically affected VRQ/VRQ sheep naturally infected with classical ovine scrapie and intravenously transfused into VRQ/VRQ lambs post-weaning. As determined using standard immunohistochemistry for scrapie, abnormal isoforms of prion protein were detected in lymphoid tissues of lambs inoculated with PBMC (4/4 recipient lambs), monocytes (2/5) and T lymphocytes (1/4). Prion protein misfolding activity was detected by serial protein misfolding cyclic amplification (sPMCA) in PBMC from monocyte and T lymphocyte recipient sheep in agreement with antemortem rectal biopsy results, but such prion protein misfolding activity was not detected from other recipients., Conclusions: These findings show that scrapie prions are associated with monocytes and T lymphocytes circulating in the peripheral blood of sheep naturally infected with classical scrapie. Combined with our previous findings, we can now conclude that all three major subsets of PBMC can harbor prions during preclinical disease and thus, present logical targets for development of a sensitive assay to detect scrapie prions. In this regard, we have also demonstrated that sPMCA can be used to detect scrapie prions associated with PBMC.

Entropion is an inward rolling of the eyelid allowing contact between the eyelashes and cornea that may lead to blindness if not corrected. Although many mammalian species, including humans and dogs, are afflicted by congenital entropion, no specific genes or gene regions related to development of entropion have been reported in any mammalian species to date. Entropion in domestic sheep is known to have a genetic component therefore, we used domestic sheep as a model system to identify genomic regions containing genes associated with entropion. A genome-wide association was conducted with congenital entropion in 998 Columbia, Polypay, and Rambouillet sheep genotyped with 50,000 SNP markers. Prevalence of entropion was 6.01%, with all breeds represented. Logistic regression was performed in PLINK with additive allelic, recessive, dominant, and genotypic inheritance models. Two genome-wide significant (empirical P<0.05) SNP were identified, specifically markers in SLC2A9 (empirical P = 0.007; genotypic model) and near NLN (empirical P = 0.026; dominance model). Six additional genome-wide suggestive SNP (nominal P<1x10(-5)) were identified including markers in or near PIK3CB (P = 2.22x10(-6); additive model), KCNB1 (P = 2.93x10(-6); dominance model), ZC3H12C (P = 3.25x10(-6); genotypic model), JPH1 (P = 4.68x20(-6); genotypic model), and MYO3B (P = 5.74x10(-6); recessive model). This is the first report of specific gene regions associated with congenital entropion in any mammalian species, to our knowledge. Further, none of these genes have previously been associated with any eyelid traits. These results represent the first genome-wide analysis of gene regions associated with entropion and provide target regions for the development of sheep genetic markers for marker-assisted selection.

Strain superinfection occurs when a second strain infects a host already infected with and having mounted an immune response to a primary strain. The incidence of superinfection with Anaplasma marginale, a tick-borne rickettsial pathogen of domestic and wild ruminants, has been shown to be higher in tropical versus temperate regions. This has been attributed to the higher prevalence of infection, with consequent immunity against primary strains and thus greater selective pressure for superinfection with antigenically distinct strains. However an alternative explanation would be the differences in the transmitting vector, Dermacentor andersoni in the studied temperate regions and Rhipicephalus microplus in the studied tropical regions. To address this question, we examined two tropical populations sharing the same vector, R. microplus, but with significantly different infection prevalence. Using two separate markers, msp1α (one allele per genome) and msp2 (multiple alleles per genome), there were higher levels of multiple strain infections in the high infection prevalence as compared to the low prevalence population. The association of higher strain diversity with infection prevalence supports the hypothesis that high levels of infection prevalence and consequent population immunity is the predominant driver of strain superinfection.

Small ruminant lentivirus (SRLV), also called ovine progressive pneumonia virus or maedi-visna, is present in 24% of US sheep. Like human immunodeficiency virus, SRLV is a macrophage-tropic lentivirus that causes lifelong infection. The production impacts from SRLV are due to a range of disease symptoms, including pneumonia, arthritis, mastitis, body condition wasting and encephalitis. There is no cure and no effective vaccine for preventing SRLV infection. However, breed differences in prevalence and proviral concentration indicate a genetic basis for susceptibility to SRLV. Animals with high blood proviral concentration show increased tissue lesion severity, so proviral concentration represents a live animal test for control post-infection in terms of proviral replication and disease severity. Recently, it was found that sheep with two copies of TMEM154 haplotype 1 (encoding lysine at position 35) had lower odds of SRLV infection. In this study, we examined the relationship between SRLV control post-infection and variants in two genes, TMEM154 and CCR5, in four flocks containing 1403 SRLV-positive sheep. We found two copies of TMEM154 haplotype 1 were associated with lower SRLV proviral concentration in one flock (P < 0.02). This identified the same favorable diplotype for SRLV control post-infection as for odds of infection. However, frequencies of haplotypes 2 and 3 were too low in the other three flocks to test. The CCR5 promoter deletion did not have consistent association with SRLV proviral concentration. Future work in flocks with more balanced allele frequencies is needed to confirm or refute TMEM154 association with control of SRLV post-infection., (Published 2014. This article is a U.S. Government work and is in the public domain in the USA. Animal Genetics published by John Wiley & Sons Ltd on behalf of Stichting International Foundation for Animal Genetics.)

Quality measures of lamb longissimus dorsi were evaluated in 514 crossbred wether lambs to assess sire breed differences. Wethers were produced over 3 yr from single-sire matings of 22 Columbia, 22 U.S. Meat Animal Research Center (USMARC)-Composite (Composite), 21 Suffolk, and 17 Texel rams to adult Rambouillet ewes. Lambs were reared to weaning in an extensive western rangeland production system and finished in a feedlot on a high-energy finishing diet. One of three harvest groups were randomly assigned to each lamb, and lambs were transported to The Ohio State University abattoir when the mean BW of wethers remaining in the feedlot reached 54.4, 61.2, or 68.0 kg. After harvest, subjective lean quality scores were assigned and LM pH (immediately after and 24 h after harvest), color (quantified as Minolta L*, a*, and b*), intramuscular fat (IMF), cooking loss percentage, and Warner-Bratzler shear force (WBSF) were determined. Statistical models included fixed effects of sire breed, year of birth, and harvest group and random effects of sire (nested within sire breed and year) and maternal grandsire. Year and harvest group were significant (P < 0.04) for all traits, except harvest group was not for WBSF and lean quality (P > 0.28). At comparable numbers of days on feed, Texel-sired wethers had the greatest (more desirable; P < 0.01) subjective lean quality scores while Composite- and Suffolk-sired lambs were intermediate and Columbia-sired lambs had the lowest (less desirable). Minolta L* values were greater (P < 0.02) for Texel- than Columbia-sired lambs, although this difference is not visually discernible by humans. No significant (P > 0.05) sire breed effects were detected for LM pH at or 24 h after harvest, Minolta a* and b*, IMF, percentage of cooking loss, and WBSF at comparable numbers of days on feed. At comparable chilled carcass weight, significant (P < 0.01) sire breed effects were detected for subjective lean quality score. Texel-sired lambs had greater scores than Columbia- and Suffolk-sired lambs, but Composite-sired lambs did not differ from lambs sired by the other sire breeds. Sire breed effects were not detected (P > 0.15) for LM pH at or 24 h after harvest, Minolta L*, a*, and b*, cooking loss percentage, IMF, and WBSF at comparable chilled carcass weight. Therefore, results of the present study indicated that the 4 sire breeds produced crossbred lamb carcasses that had similar and desirable LM quality measurements, meeting consumer expectations.

Use of lamb BW or chilled carcass weights (CCW), live-animal ultrasound or direct carcass measurements of backfat thickness (BF; mm) and LM area (LMA; cm(2)), and carcass body wall thickness (BWall; mm) to predict carcass yield and value was evaluated using 512 crossbred lambs produced over 3 yr by mating Columbia, U.S. Meat Animal Research Center Composite, Suffolk, and Texel rams to adult Rambouillet ewes. Lambs were harvested at 3 BW endpoints within each year. The predictive value of 3 to 5 additional linear measurements of live-animal or carcass size and shape was also evaluated. Residual correlations (adjusted for effects of year, breed, and harvest group) between ultrasound and direct measurements were 0.69 for BF and 0.65 for LMA. Increasing ultrasound or carcass LMA had positive effects (P < 0.001) on yield of chilled carcass (i.e., on dressing percentage) and, at comparable CCW, on weight of high-value cuts (rack, loin, leg, and sirloin) before trimming (HVW), weight of trimmed high-value cuts (trimmed rack and loin and trimmed boneless leg and sirloin; TrHVW), and carcass value before (CVal) and after (TrCVal) trimming of high-value cuts. By contrast, ultrasound and direct measures of BF had positive effects on yields of CCW and on HVW and CVal but large negative effects on TrHVW and TrCVal. After adjusting for BW at scanning, increases of 1 mm in ultrasound BF or 1 cm(2) in ultrasound LMA were associated with changes of US$-0.32 (P < 0.10) and $1.62 (P < 0.001), respectively, in TrCVal. Carcass BWall was generally superior to carcass BF as a predictor of TrHVW and TrCVal. Carcass LMA was superior to ultrasound LMA but carcass BF was inferior to ultrasound BF for prediction of carcass yield and value. Increasing LMA thus would be expected to improve carcass yield and value. Addition of linear measurements of live-animal or carcass size and shape to the prediction model reduced residual SD (RSD) for TrHVW and TrCVal by 0.4 to 2.2%, but subsequent removal of ultrasound or direct measures of BF and LMA from the prediction model increased RSD by 7.4 to 12.2%. Measurements of CCW, LMA, BF, and BWall would thus be appropriate to support programs for value-based marketing of lamb carcasses and are superior to systems based only on measurements of size and shape in unribbed carcasses.

Linear measurements on live lambs and carcasses can be used to characterize sheep breeds and may have value for prediction of carcass yield and value. This study used 512 crossbred lambs produced over 3 yr by mating Columbia, U.S. Meat Animal Research Center (USMARC) Composite, Suffolk, and Texel rams to adult Rambouillet ewes to assess sire-breed differences in live-animal and carcass shape and to evaluate the value of shape measurements as predictors of chilled carcass weight (CCW), weight of high-value cuts (rack, loin, leg, and sirloin; HVW), weight of trimmed high-value cuts (trimmed rack and loin and trimmed, boneless leg and sirloin; TrHVW), and estimated carcass value before (CVal) and after trimming of high-value cuts (TrCVal). Lambs were produced under extensive rangeland conditions, weaned at an average age of 132 d, fed a concentrate diet in a drylot, and harvested in each year in 3 groups at target mean BW of 54, 61, and 68 kg. Canonical discriminant analysis indicated that over 93% of variation among sire breeds was accounted for by the contrast between tall, long, less-thickly muscled breeds with greater BW and CCW (i.e., the Columbia and Suffolk) compared with shorter, more thickly muscled breeds with smaller BW and CCW. After correcting for effects of year, harvest group, sire breed, and shipping BW, linear measurements on live lambs contributed little to prediction of CCW. Similarly, after accounting for effects of CCW, linear measurements on live animals further reduced residual SD (RSD) of dependent variables by 0.2 to 5.7%, with generally positive effects of increasing live leg width and generally negative effects of increasing heart girth. Carcass measurements were somewhat more valuable as predictors of carcass merit. After fitting effects of CCW, additional consideration of carcass shape reduced RSD by 2.1, 3.6, 9.5, and 2.2% for HVW, TrHVW, CVal, and TrCVal, respectively. Effects of increasing carcass leg width were positive for HVW, TrHVW, and TrCVal. We also observed positive effects of increasing carcass length on TrCVal and negative effects of increasing cannon bone length on HVW and CVal. Increasing shoulder width had positive effects on CVal but negative effects on TrHVW. Differences in lamb and carcass shape were significantly associated with carcass yield and value, but the additional accuracy associated with use of these measurements was modest relative to that achieved from use of only shipping BW or CCW.

Ovine lentivirus (OvLV) is a macrophage-tropic lentivirus found in many countries that causes interstitial pneumonia, mastitis, arthritis and cachexia in sheep. There is no preventive vaccine and no cure, but breed differences suggest marker-assisted selective breeding might improve odds of infection and control of OvLV post-infection. Although variants in TMEM154 have consistent association with odds of infection, no variant in any gene has been associated with host control of OvLV post-infection in multiple animal sets. Proviral concentration is a live-animal diagnostic measure of OvLV control post-infection related to severity of OvLV-induced lesions. A recent genome-wide association study identified a region including four zinc finger genes associated with proviral concentration in one Rambouillet flock. To refine this region, we tested additional variants and identified a small insertion/deletion variant near ZNF389 that showed consistent association with proviral concentration in three animal sets (P < 0.05). These animal sets contained Rambouillet, Polypay and crossbred sheep from multiple locations and management conditions. Strikingly, one flock had exceptionally high prevalence (>87%, including yearlings) and mean proviral concentration (>950 copies/μg), possibly due to needle sharing. The best estimate of proviral concentration by genotype, obtained from all 1310 OvLV-positive animals tested, showed insertion homozygotes had less than half the proviral concentration of other genotypes (P < 0.0001). Future work will test additional breeds, management conditions and viral subtypes, and identify functional properties of the haplotype this deletion variant tracks. To our knowledge, this is the first genetic variant consistently associated with host control of OvLV post-infection in multiple sheep flocks., (Published 2013. This article is a U.S. Government work and is in the public domain in the USA. Animal Genetics published by John Wiley & Sons Ltd on behalf of Stichting International Foundation for Animal Genetics.)

A genome-wide association study (GWAS) was performed to investigate seven red blood cell (RBC) phenotypes in over 500 domestic sheep (Ovis aries) from three breeds (Columbia, Polypay, and Rambouillet). A single nucleotide polymorphism (SNP) showed genome-wide significant association with increased mean corpuscular hemoglobin concentration (MCHC, P = 6.2×10(-14)) and genome-wide suggestive association with decreased mean corpuscular volume (MCV, P = 2.5×10(-6)). The ovine HapMap project found the same genomic region and the same peak SNP has been under extreme historical selective pressure, demonstrating the importance of this region for survival, reproduction, and/or artificially selected traits. We observed a large (>50 kb) variant haplotype sequence containing a full-length divergent artiodactyl MYADM-like repeat in strong linkage disequilibrium with the associated SNP. MYADM gene family members play roles in membrane organization and formation in myeloid cells. However, to our knowledge, no member of the MYADM gene family has been identified in development of morphologically variant RBCs. The specific RBC differences may be indicative of alterations in morphology. Additionally, erythrocytes with altered morphological structure often exhibit increased structural fragility, leading to increased RBC turnover and energy expenditure. The divergent artiodactyl MYADM-like repeat was also associated with increased ewe lifetime kilograms of lamb weaned (P = 2×10(-4)). This suggests selection for normal RBCs might increase lamb weights, although further validation is required before implementation in marker-assisted selection. These results provide clues to explain the strong selection on the artiodactyl MYADM-like repeat locus in sheep, and suggest MYADM family members may be important for RBC morphology in other mammals.

Sheep are among the major economically important livestock species worldwide because the animals produce milk, wool, skin, and meat. In the present study, the Illumina OvineSNP50 BeadChip was used to investigate genetic diversity and genome selection among Suffolk, Rambouillet, Columbia, Polypay, and Targhee sheep breeds from the United States. After quality-control filtering of SNPs (single nucleotide polymorphisms), we used 48,026 SNPs, including 46,850 SNPs on autosomes that were in Hardy-Weinberg equilibrium and 1,176 SNPs on chromosome × for analysis. Phylogenetic analysis based on all 46,850 SNPs clearly separated Suffolk from Rambouillet, Columbia, Polypay, and Targhee, which was not surprising as Rambouillet contributed to the synthesis of the later three breeds. Based on pair-wise estimates of F(ST), significant genetic differentiation appeared between Suffolk and Rambouillet (F(ST) = 0.1621), while Rambouillet and Targhee had the closest relationship (F(ST) = 0.0681). A scan of the genome revealed 45 and 41 differentially selected regions (DSRs) between Suffolk and Rambouillet and among Rambouillet-related breed populations, respectively. Our data indicated that regions 13 and 24 between Suffolk and Rambouillet might be good candidates for evaluating breed differences. Furthermore, ovine genome v3.1 assembly was used as reference to link functionally known homologous genes to economically important traits covered by these differentially selected regions. In brief, our present study provides a comprehensive genome-wide view on within- and between-breed genetic differentiation, biodiversity, and evolution among Suffolk, Rambouillet, Columbia, Polypay, and Targhee sheep breeds. These results may provide new guidance for the synthesis of new breeds with different breeding objectives.

Postfabrication carcass component weights of 517 crossbred wether lambs were analyzed to evaluate 4 terminal-sire breeds. Wethers were produced over 3 yr from single-sire matings of 22 Columbia, 22 USMARC-Composite (Composite), 21 Suffolk, and 17 Texel rams to adult Rambouillet ewes. Lambs were reared to weaning in an extensive western rangeland production system and finished in a feedlot on a high-energy finishing diet. When wethers reached a mean BW of 54.4, 61.2, or 68.0 kg, they were transported to The Ohio State University abattoir for harvest. After refrigeration for approximately 24 h, chilled carcass weight (CCW) was measured, carcasses were fabricated according to Style A of Institutional Meat Purchase Specifications, and postfabrication weights were recorded. At comparable numbers of days on feed, Suffolk-sired lambs had heavier (P < 0.04) neck, breast, shoulder, foreshank, rack, loin, leg, sirloin, roast-ready rack, trimmed loin, and boneless leg cuts than progeny of the other sire breeds. Boneless sirloins were heavier (P < 0.01) for Suffolk-sired than Composite-sired lambs but did not differ from those for Columbia- or Texel-sired lambs. Columbia- and Suffolk-sired lambs had heavier (P < 0.01) hindshanks than Texel-sired lambs. Suffolk-sired lambs had heavier (P < 0.01) high-value cuts (rack, loin, leg, and sirloin) and trimmed high-value cuts than progeny of the other sire breeds. Cutting loss (CCW - wholesale cut weights) and high-value trimming loss were greatest (P < 0.02) for Suffolk-sired lambs and least for Texel- and Composite-sired lambs. Sire breed did not affect (P > 0.06) flank weight. Data adjusted to comparable CCW reduced the number of significant sire-breed effects and changed sire-breed rankings of carcass component weights, for which sire breeds differed. After adjusting, Suffolk-sired lambs had lighter (P < 0.05) loins than Columbia- and Composite-sired lambs, Composite-sired lambs had heavier (P < 0.05) high-value cuts than Suffolk-sired lambs, and Suffolk- and Columbia-sired lambs had heavier (P < 0.05) necks than Texel-sired lambs. At predicted backfat thickness of 6.6 mm, Composite-sired lambs had a greater (P < 0.05) percentage of high-value cuts than Suffolk-sired lambs before but not after trimming. Producers can use these results to select terminal-sire breeds that will complement their production system and improve lamb value.

This study was conducted to quantify differences in feed efficiency among 4 sire breeds using records from 1,109 crossbred lambs over 3 yr. Lambs were produced from mating Columbia, USMARC-Composite (Composite), Suffolk, and Texel rams to adult Rambouillet ewes. After weaning, lambs were finished on a high-energy diet in group-fed pens. Efficiency was assessed at 45 and 90 d on study, 50 and 60 kg of BW, and 6.6 and 9.1 mm of fat depth between the 12th and 13th ribs. Sire breed affected most of the traits that were measured. At 90 d, Suffolk-sired lambs had gained 13 to 19% more BW (P < 0.001), were 7 to 13% heavier (P < 0.001), and had consumed 4 to 11% more (P < 0.001) ME than the other breed crosses. However, at 90 d, G:F (59.9 vs. mean of 56.2 g BW gain/Mcal of ME; P < 0.01) and residual BW gain (RG; 0.94 vs. -0.33 kg BW; P < 0.05) were greater for Suffolk-sired lambs than for the other 3 sire breeds. Also, RG for Suffolk-sired lambs was greater (P < 0.01) at 6.6 and 9.1 mm of fat depth (0.62 vs. -0.24 kg and 0.87 vs. -0.33 kg of BW, respectively) and at 60 kg of BW (0.73 vs. 0.31 kg of BW) than for the other sire breeds. At 90 d, Columbia-sired lambs had the greatest (P < 0.01) residual feed intake (RFI; 7.01 vs. mean of -2.33 Mcal ME for the 3 other sire breeds). At 60 kg of BW, Texel- and Composite- (-2.34 Mcal ME, combined) had more favorable (P < 0.01) RFI than Columbia-sired lambs (6.42 Mcal ME), and Suffolk-sired lambs (0.49 Mcal ME) were intermediate and not different from the other 3 sire breeds. At 6.6 mm of fat depth, Columbia- and Suffolk- (3.30 Mcal ME, combined) had greater (P < 0.001) RFI than Texel-sired lambs (-4.23 Mcal ME); at 9.1 mm of fat depth, Columbia-sired lambs had the greatest (P < 0.05) RFI (7.35 vs. -2.11 Mcal ME for the other breeds combined). In summary, except for ME intake at 90 d, Suffolk-sired lambs were equal or superior to Columbia-, Composite-, and Texel-sired lambs for BW, BW gain, ME intake, G:F, RFI, and RG to 90 d, 60 kg of BW, and 9.1 mm of fat depth. Breed rankings were consistent across the study period for G:F and RG, but rankings changed over time for RFI.

A 3-yr study was conducted to comprehensively evaluate Columbia, Suffolk, USMARC-Composite (Composite), and Texel breeds as terminal sires in an extensive rangeland production system. The objective was to estimate breed-of-ram effects on ewe fertility, prolificacy, and dystocia, and sire breed effects on lamb survival and growth until weaning at approximately 132 d of age. Data were from 22 Columbia, 22 Composite, 21 Suffolk, and 17 Texel rams with 957 exposures to 574 adult Rambouillet ewes (3- to 7-yr-old at lambing), 908 lambings, and 1,834 lambs. Ram breed did not affect ewe fertility (mean = 94.9%; p = 0.73), total number born per ewe lambing (mean = 2.02 lambs; p = 0.20), number born alive per ewe lambing (mean = 1.90 lambs; p = 0.24), or number weaned per ewe lambing (mean = 1.45 lambs, p = 0.94). Dystocia rates were different (p = 0.01) for ewes mated to Columbia (12.2%), Composite (13.5%), Suffolk (25.7%), and Texel rams (31.9%) during 1 yr of the study, but differences among ram breeds were not repeatable (p ≥ 0.38) during the other 2 yr. Suffolk-sired lambs were heavier (p ≥ 0.02) at birth (5.5 kg) and weaning (40.3 kg) than lambs sired by the other breeds, which did not differ (p ≥ 0.34) for birth weight (mean = 5.3 kg). Texel-sired lambs (37.4 kg) were lighter (p ≥ 0.02) at weaning than Columbia- (38.8 kg) and Composite-sired (38.4 kg) lambs, which did not differ (p = 0.40) for weaning weight. Sire breed effect approached significance (p = 0.06) for lamb survival to weaning; estimated survival probabilities were 0.87 (Columbia), 0.89 (Composite), 0.93 (Suffolk), and 0.86 (Texel) for lambs reared by their birth dam. Interaction between sire breeds and birth weight affected (p < 0.001) lamb survival and revealed that lightweight Columbia- and Suffolk-sired lambs had a greater risk of death than lightweight lambs sired by Composite and Texel rams, but risk of death did not increase substantially for heavyweight lambs from any of the breeds. When mated to adult Rambouillet ewes in an extensive rangeland production system, the use of Suffolk rams is warranted to improve preweaning growth of market lambs and is not predicted to affect ewe fertility, ewe prolificacy, dystocia, or lamb survival compared with the other sire breeds we tested.

Over 3 yr, postweaning growth patterns and changes in ultrasonic measurements of fat depth (FD) and LM area were assessed for 1,049 crossbred ewe and wether lambs (Ovis aries) produced by mating adult Rambouillet ewes to 1 of 22 Columbia, 22 USMARC-Composite (Composite), 21 Suffolk, or 17 Texel rams and raised to weaning under extensive Western rangeland conditions. After weaning, lambs were transitioned to a high-energy finishing diet in a feedlot, weighed weekly for 13 to 16 wk, and scanned using ultrasound at 2-wk intervals to estimate FD and LM area between the 12th and 13th ribs. Lambs sired by Suffolk rams were 5 to 12% heavier (P < 0.05) at start of test, grew 14 to 22% faster (P < 0.001), and were correspondingly 7 to 14% heavier (P < 0.001) after 90 d on test than lambs sired by rams of the other 3 breeds. At this time, lambs sired by Texel rams were 5% lighter (P < 0.01) than Columbia- or Composite-sired lambs, which did not differ (P > 0.20). Columbia-sired lambs had less FD (8.9 mm) but smaller LM areas (15.6 cm(2)) at 90 d on test (both P < 0.01) than lambs sired by rams of the other breeds (average of 9.8 mm for FD and 16.6 cm(2) for LM area), which did not differ for either measurement at this time. At 60 kg, predicted FD differed among sire breeds (P < 0.001) and were similar for Suffolk- (7.6 mm) and Columbia-sired (7.9 mm) lambs, intermediate for Composite-sired lambs (9.1 mm), and largest for Texel-sired (10.1 mm) lambs. At a BW of 60 kg, LM area differed between all pairs of sire breeds (P < 0.05), and was largest for Texel-sired lambs (16.7 cm(2)), followed by Composite-, Suffolk-, and Columbia-sired lambs (15.7, 14.8, and 14.5 cm(2), respectively). At a predicted FD of 9.1 mm (the boundary between U.S. Yield Grades 3 and 4), the predicted LM area for Suffolk-sired lambs (16.2 cm(2)) was similar (P > 0.10) to that of Texel-sired lambs (16.0 cm(2)) and superior (P < 0.01) to that of Composite- (15.7 cm(2)) and Columbia-sired lambs (15.4 cm(2)). Lambs sired by Suffolk rams were thus equal or superior to lambs sired by the other 3 breeds in growth, FD, and LM area at comparable ages, fatness at 60 kg, and LM area at a FD of 9.1 mm, but inferior in LM area to Texel- and Composite-sired lambs at comparable BW.

To evaluate terminal-sire breeds, harvest BW, prefabrication carcass measurements, and organ weights were evaluated over 3 yr for 518 crossbred wether lambs (ovis aries). Lambs were produced by single-sire matings of 22 Columbia, 22 USMARC-Composite (Composite), 21 Suffolk, and 17 Texel rams to adult Rambouillet ewes. Lambs were raised to weaning under an extensive western rangeland production system and finished in a feedlot on a high-energy finishing diet. Wethers were randomly assigned to be harvested at an average BW of 54.4, 61.2, or 68.0 kg and then transported to The Ohio State University abattoir for harvest. Wether BW was recorded before transport (off-test BW) and before harvest. Prefabrication carcass measurements and organ weights were recorded either after harvest or after an approximate 24-h chill. At comparable numbers of days on feed, Suffolk-sired lambs had heavier (P < 0.01) off-test BW, harvest BW, HCW, chilled carcass weight (CCW), and kidney weights than lambs sired by the other breeds. Suffolk-sired lambs had more (P < 0.01) kidney-pelvic fat than did Columbia-sired lambs; Composite- and Texel-sired lambs were intermediate and did not differ (P > 0.06) from the other crossbred lambs. Texel- and suffolk-sired lambs had larger lm area and greater conformation scores than Columbia-sired lambs (P < 0.03). Texel-sired lambs had greater (P < 0.01) body wall thickness, quality grades, and leg scores than Columbia-sired lambs. Composite- and Suffolk-sired lambs did not differ from each other or from lambs sired by any other breed for body wall thickness (P > 0.18) and were intermediate for quality grades and leg scores. Sire breed did not affect (P > 0.05) shipping shrink, dressing percentage, pelt weight, liver weight, and fat depth. Adjusting data to a comparable off-test BW or CCW modified sire breed rankings for some measurements. Texel-sired lambs were equal or superior to lambs sired by other breeds for HCW, CCW, dressing percentage, pelt weight, LM area, quality grade, and leg and conformation scores. However, Texel-sired lambs also had values for kidney-pelvic fat weight, fat depth, and body wall thickness that were equal to or greater than those of lambs sired by the other breeds, indicating increased fatness at comparable BW. Producers can use these results to select terminal-sire sheep breeds that will complement their production system and improve market lamb value.

Ovulation rate (OR) is an important component of litter size, but mutation(s) in gene(s) underlying OR QTL have yet to be identified in pigs. Markers within an OR QTL on SSC3 were genotyped in three white composite lines selected for ten generations for increased OR or uterine capacity (UC), with one line being an unselected control. Numbers of corpora lutea (CL) and UC (number of fully formed fetuses) were collected at approximately 105 days of gestation, as well as ovary weight (OW), uterine length (UL) and uterine weight (UW) measurements at 160 d of age in generation 12 and 13 females from all three lines. Six microsatellites and ten single nucleotide polymorphisms (SNPs; 0-42 cM) were genotyped in pigs from all lines of generations 11 through 13. The allele frequencies of 24269.1, SW2429, 7907.2 and 7637.2 were different (P < 0.01) in the OR line compared to the control line. A significant (P < 0.05) association of CL with 24269.1 (additive effect 0.65 ± 0.32) was detected, and additive genotypic effects approached significance for markers at 28 through 35 cM (16963.2, 27514.1 and SWR1637). Haplotyping of 7637.2 and 16963.2 (31 through 32 cM) identified a significant additive association of haplotype 1 with CL (-0.62 ± 0.30). These markers were also associated with OW (24296.1 and SWR1637), UL (16963.2, 27514.1 and haplotypes of 7637.2/16963.2) and UW (haplotypes of 7637.2/16963.2). This study verifies an OR QTL on SSC3. However, based on the data, it was concluded that there may be two genes, at 13 through 18 cM and 28 through 35 cM, controlling OR on SSC3p., (© 2011 The Authors, Animal Genetics © 2011 Stichting International Foundation for Animal Genetics.)

Background: Like human immunodeficiency virus (HIV), ovine lentivirus (OvLV) is macrophage-tropic and causes lifelong infection. OvLV infects one quarter of U.S. sheep and induces pneumonia and body condition wasting. There is no vaccine to prevent OvLV infection and no cost-effective treatment for infected animals. However, breed differences in prevalence and proviral concentration have indicated a genetic basis for susceptibility to OvLV. A recent study identified TMEM154 variants in OvLV susceptibility. The objective here was to identify additional loci associated with odds and/or control of OvLV infection., Methodology/principal Findings: This genome-wide association study (GWAS) included 964 sheep from Rambouillet, Polypay, and Columbia breeds with serological status and proviral concentration phenotypes. Analytic models accounted for breed and age, as well as genotype. This approach identified TMEM154 (nominal P=9.2×10(-7); empirical P=0.13), provided 12 additional genomic regions associated with odds of infection, and provided 13 regions associated with control of infection (all nominal P<1 × 10(-5)). Rapid decline of linkage disequilibrium with distance suggested many regions included few genes each. Genes in regions associated with odds of infection included DPPA2/DPPA4 (empirical P=0.006), and SYTL3 (P=0.051). Genes in regions associated with control of infection included a zinc finger cluster (ZNF192, ZSCAN16, ZNF389, and ZNF165; P=0.001), C19orf42/TMEM38A (P=0.047), and DLGAP1 (P=0.092)., Conclusions/significance: These associations provide targets for mutation discovery in sheep susceptibility to OvLV. Aside from TMEM154, these genes have not been associated previously with lentiviral infection in any species, to our knowledge. Further, data from other species suggest functional hypotheses for future testing of these genes in OvLV and other lentiviral infections. Specifically, SYTL3 binds and may regulate RAB27A, which is required for enveloped virus assembly of human cytomegalovirus. Zinc finger transcription factors have been associated with positive selection for repression of retroviral replication. DLGAP1 binds and may regulate DLG1, a known regulator of HIV infectivity.

Visna/Maedi, or ovine progressive pneumonia (OPP) as it is known in the United States, is an incurable slow-acting disease of sheep caused by persistent lentivirus infection. This disease affects multiple tissues, including those of the respiratory and central nervous systems. Our aim was to identify ovine genetic risk factors for lentivirus infection. Sixty-nine matched pairs of infected cases and uninfected controls were identified among 736 naturally exposed sheep older than five years of age. These pairs were used in a genome-wide association study with 50,614 markers. A single SNP was identified in the ovine transmembrane protein (TMEM154) that exceeded genome-wide significance (unadjusted p-value 3×10(-9)). Sanger sequencing of the ovine TMEM154 coding region identified six missense and two frameshift deletion mutations in the predicted signal peptide and extracellular domain. Two TMEM154 haplotypes encoding glutamate (E) at position 35 were associated with infection while a third haplotype with lysine (K) at position 35 was not. Haplotypes encoding full-length E35 isoforms were analyzed together as genetic risk factors in a multi-breed, matched case-control design, with 61 pairs of 4-year-old ewes. The odds of infection for ewes with one copy of a full-length TMEM154 E35 allele were 28 times greater than the odds for those without (p-value<0.0001, 95% CI 5-1,100). In a combined analysis of nine cohorts with 2,705 sheep from Nebraska, Idaho, and Iowa, the relative risk of infection was 2.85 times greater for sheep with a full-length TMEM154 E35 allele (p-value<0.0001, 95% CI 2.36-3.43). Although rare, some sheep were homozygous for TMEM154 deletion mutations and remained uninfected despite a lifetime of significant exposure. Together, these findings indicate that TMEM154 may play a central role in ovine lentivirus infection and removing sheep with the most susceptible genotypes may help eradicate OPP and protect flocks from reinfection., Competing Interests: The authors have declared that no competing interests exist.

Transmission of ovine progressive pneumonia virus (OPPV), a lentivirus of sheep, occurs through both maternal and non-maternal means. Currently, the contribution of each route to the overall flock OPPV prevalence is poorly understood since previous serological epidemiologic studies lacked the ability to accurately track routes of transmission within an infected flock. In this study, the amount of maternal OPP transmission was assessed in a naturally infected ewe flock by applying molecular analyses to proviral sequences derived from peripheral blood leukocytes of OPP positive dam-daughter pairs (N=40). Both proviral envelope (env) and long terminal repeat (LTR) sequences, separately and combined, were utilized in the following 2 sequence analysis methods: phylogenetic analysis and pairwise distance calculations. True maternal transmission events were defined as agreement in 2 out of the 2 sequence analysis methods. Using this criterion, proviral env sequences resulted in a 14.3% maternal transmission frequency, and proviral LTR sequences resulted in a 10% maternal transmission frequency. Both proportions of maternal transmission varied significantly from equality (P<0.0001). This indicates that the remaining 85.7-90% of daughters are infected via non-maternal transmission. This is also the first study to calculate the OPP proviral rate of change for the env gene and LTR promoter. Accurately defining the routes of OPPV transmission provides critical epidemiological data supporting management intended to reduce flock transmission and viral dose., (Published by Elsevier B.V.)

A caprine arthritis-encephalitis virus (CAEV)/maedi-visna virus (MVV) indirect enzyme-linked immunosorbent assay (iELISA) was validated with samples from U.S. sheep and by the use of radioimmunoprecipitation as the standard for comparison. The sensitivity and the specificity were 86.0% (+ or - 5.8%) and 95.9% (+ or - 2.9%), respectively. The iELISA format and phylogenetic differences based on the MVV gag sequence contribute to the reduced sensitivity.

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) of Rocky Mountain elk in North America. Recent studies suggest that tissue and blood mineral levels may be valuable in assessing TSE infection in sheep and cattle. The objectives of this study were to examine baseline levels of copper, manganese, magnesium, zinc, selenium, and molybdenum in the brains of Rocky Mountain elk with differing prion genotypes and to assess the association of mineral levels with CWD infection. Elk with leucine at prion position 132 had significantly lower magnesium levels than elk with 2 copies of methionine. Chronic wasting disease-positive elk had significantly lower magnesium than control elk. The incorporation of manganese levels in addition to magnesium significantly refined explanatory ability, even though manganese alone was not significantly associated with CWD. This study demonstrated that mineral analysis may provide an additional disease correlate for assessing CWD risk, particularly in conjunction with genotype.

Chemokine (C-C motif) Receptor 5 (CCR5) is a chemokine receptor that regulates immune cell recruitment in inflammation and serves as a coreceptor for human immunodeficiency virus (HIV). A human CCR5 coding deletion (termed delta-32) results in strong resistance to HIV infection, and sequence variants in CCR5 regulatory regions have been implicated in delayed progression to acquired immune deficiency syndrome. Both ovine progressive pneumonia virus (OPPV), also known as maedi-visna, and HIV are macrophage-tropic lentiviruses, have similar genomic structures, and cause lifelong persistent host infection, suggesting CCR5 may have a role in regulating OPPV provirus levels. Therefore, the ovine CCR5 genomic sequence was determined, and sequence variants were obtained from the open reading frame and surrounding regulatory sites. One CCR5 variant contained a 4-base deletion within a binding site for octamer transcription factors in the promoter region. A test for differential transcription from each allele in heterozygous animals showed a 3.9-fold transcription difference (P < 0.0001). OPPV proviral levels were also measured in 351 naturally exposed Rambouillet, Polypay and Columbia sheep. Deletion homozygotes showed reduced OPPV proviral levels among these animals (P < 0.01). The association of this CCR5 promoter deletion with OPPV levels will need to be validated in additional populations before the deletion can be recommended for widespread use in marker-assisted selection. However, because of the large impact on transcription and because CCR5 has roles in inflammation, recruitment of effector cells, and cell-mediated immunity, this deletion may play a role in the control of infections of many diverse pathogens of sheep.