Rams are seasonal breeders that have large testes in comparison with other mammalian species. Testicle size is a good indication of a ram׳s sperm producing ability and varies with season, body condition and mating activity. A breeding soundness evaluation prior to ram introduction to ewes is critical to assess a ram's breeding capacity. In a seasonal breeding system, a ram can breed up to 100 ewes per year, whereas in an artificial insemination program, one ram can be used to inseminate thousands of ewes in a breeding season.

The effect of interferon tau (IFNT) on the uterus is critical for maternal recognition of pregnancy in ruminants, while its direct role in luteal function is less well understood. To address this, we performed two experiments. In Experiment 1, cattle received intrauterine infusions of either: bovine serum albumin (BSA; vehicle) or vehicle with IFNT from Days 14 to 16 of the estrous cycle or vehicle with IFNT from Days 14 to 19 or vehicle with IFNT from Days 14 to 19 with pregnancy associated glycoprotein (PAG) from Days 17 to 19. Corpora lutea (CL) were collected on Day 17 or 20 and RNAseq was performed. In Experiment 2, cultured luteal steroidogenic cells from cyclic (Days 10-12) cattle were treated with IFNT and RNAseq was performed. Treatment with IFNT resulted in luteal changes (in vivo: 130 transcripts; in vitro: 2981 transcripts), while addition of PAG resulted in 13 changed transcripts. Only 31% of the genes that changed in the CL during early pregnancy (Hughes et al., 2020) were regulated by IFNT; these were antiviral and immune regulators. In contrast, 50% of the genes that changed during early pregnancy were not regulated by IFNT and were associated with cellular proliferation and extracellular matrix organization. The remaining 19% of genes were not conclusively identified as either IFNT regulated or non-regulated. This suggests that the temporal changes in the CL during early pregnancy are only partially regulated by IFNT, drawing into question identities of other luteal regulators or the effect of age of CL.

This comprehensive review explores the complex processes of reproduction, pregnancy establishment, and pregnancy diagnostic methods in cattle, sheep, goats, swine, horses, and camelids. It provides an overview of the history of pregnancy detection and an in-depth exploration of the physiology of pregnancy in livestock. The detection of conceptus tissue and fluids, conceptus-produced hormones, and maternal responses to conceptus signals, crucial for pregnancy diagnosis, are also discussed in detail, as are emerging methods for pregnancy diagnosis in livestock species. Overall, this review emphasizes the direct impact of pregnancy diagnosis and efficient pregnancy management for profitability of livestock enterprises.

Subfertility remains a major problem in the dairy industry. Only 35% to 40% of high-yielding dairy cows and 55% to 65% of nonlactating heifers become pregnant after their first service. The immune system plays a critical role in the establishment of pregnancy. However, it can also create challenges for embryo survival and contribute to reduced fertility. We conducted 2 separate experiments to characterize changes in subsets of peripheral blood leukocytes (PBL) and their phenotype over the estrous cycle and early pregnancy in heifers and cows. We used flow cytometry and reverse transcription- quantitative PCR to assess protein and mRNA expression of molecules important for immune function. We observed that CD14+ monocytes and CD3+ T cells tended to be affected by pregnancy status in heifers, whereas CD8B+ lymphocytes and NCR1+ natural killer cells were affected during early pregnancy in cows. Changes in expression of immune function proteins appeared to be greater in heifers than cows. To compare the most striking differences between heifers and cows observed in the initial experiments, we conducted a third experiment where PBL sampled from heifers and cows were simultaneously collected and analyzed under the same experimental conditions. Our results indicate that, compared with heifers, cows had greater mRNA expression of proinflammatory cytokines (IFN-gamma and IL6) and AHR protein along with greater percentage of MM20A+ neutrophils and myeloid cells expressing SIRPA, ITGAM, and ITGAX. Moreover, animals that failed to become pregnant showed altered expression of anti-inflammatory molecules compared with cyclic and pregnant animals. Overall, these findings support the hypothesis that early pregnancy signaling alters the phenotype of immune cells in the peripheral blood and that differences in the peripheral immune response to pregnancy between cows and heifers exist. Because cows have lower conception rates than heifers, it is possible that a more proinflammatory immune status in peripheral blood may play a role in embryo loss.

This study investigates the short and long-term effects of IFNT and PAG on the transcriptome of endometrium and blood leukocytes. Holstein heifers received intrauterine infusions of one of the following treatments: 20 mL of a 200 μg/mL bovine serum albumin solution (BSA; vehicle) from day 14 to 16 of the estrous cycle (BSA), vehicle + 10 μg/mL of IFNT from day 14 to 16 (IFNT3), vehicle + 10 μg/mL of IFNT from day 14 to 19 (IFNT6), and vehicle + 10 μg/mL of IFNT from day 14 to 16 followed by vehicle + 10 μg/mL of IFNT + 5 μg/mL of PAG from day 17 to 19 (IFNT+PAG). RNA-seq analysis was performed in endometrial biopsies and blood leukocytes collected after treatments. Acute IFNT signaling in the endometrium (IFNT3 vs BSA), induced differentially expressed genes (DEG) associated with interferon activation, immune response, inflammation, cell death, and inhibited vesicle transport and extracellular matrix remodeling. Prolonged IFNT signaling (IFNT6 vs IFNT3) altered gene expression related to cell invasion, retinoic acid signaling, and embryo implantation. In contrast, PAG induced numerous DEG in blood leukocytes but only 4 DEG in the endometrium. In blood leukocytes, PAG stimulated genes involved in development and TGFB signaling while inhibiting interferon signaling and cell migration. Overall, IFNT is a primary regulator of endometrial gene expression, while PAG predominantly affected the transcriptome of circulating immune cells during early pregnancy. Further research is essential to fully grasp the roles of identified DEG in both the endometrium and blood leukocytes.

Journal Article I Feel Seen: Exposing Students to Minority Speakers Fosters Learning and Community Building Get access Maria J Solares, Maria J Solares Molecular, Cellular, and Integrative Biosciences Graduate Program, Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA, USADepartment of Biomedical Engineering, Pennsylvania State University, University Park, PA, USACenter for Structural Oncology, The Pennsylvania State University, University Park, PA, USA Search for other works by this author on: Oxford Academic Google Scholar Troy L Ott Troy L Ott Department of Animal Science, The Pennsylvania State University, University Park, PA, USAHuck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA, USA Search for other works by this author on: Oxford Academic Google Scholar Microscopy and Microanalysis, Volume 29, Issue Supplement_1, 1 August 2023, Page 2048, https://doi.org/10.1093/micmic/ozad067.1058 Published: 22 July 2023

A better understanding of changes in plasma amino acid (AA) concentrations during the transition period may help optimize nutrition to improve health, welfare, and production in dairy cows. The aim of this study was to assess changes in plasma AA during the transition period in Holstein cows and first-lactation heifers. Blood samples for AA analysis were collected from the tail vein/artery of 9 multi- and 8 primiparous cows 3 weeks prior to and at weeks 1, 2, and 3 post-calving. Concentration of the sum of essential AA (EAA) was similar between primiparous and multiparous cows before calving but was 17.0% lower in multiparous vs. primiparous cows postpartum, which was related to both greater milk production and imbalance in estimated metabolizable protein supply vs. demand in the multiparous group of cows. In both parity groups, EAA concentrations declined by 20.0% from week 1 to week 3 postpartum. The sum of non-essential protein and non-protein AA increased linearly in the weeks after calving and was similar between primi- and multiparous cows. Compared with prepartum, plasma His concentration decreased 13.1% postpartum in both primiparous and multiparous cows but primiparous cows had 7.6% greater plasma His concentration than multiparous cows pre- and postpartum. Methionine and Lys concentrations were not affected by parity, but concentrations of both AA decreased (22.0% and 24.5%, respectively) after calving. Overall, plasma concentration of EAA decreased after calving, indicating a negative EAA balance (supply vs. demand) around parturition. Lower postpartum plasma concentrations of EAA in multiparous cows likely reflect an overall negative EAA balance due to decreased supply and increased demand to sustain greater milk production, compared with primiparous cows.

In ruminants, it has been observed that capsicum oleoresin can alter insulin responses and that high-intensity artificial sweetener can increase glucose absorption from the small intestine. Because glucose metabolism and insulin responses are critical during early lactation, these supplements might have an effect on the metabolic status of dairy cows during the transition period. The objective of this experiment was to evaluate the effects of rumen-protected capsicum oleoresin fed alone or in combination with artificial sweetener during the transition period on lactational performance and susceptibility to subclinical ketosis in dairy cows. Fifteen primiparous and 30 multiparous Holstein cows (a total of 39 cows finished the study) were arranged in a randomized complete block design during d -21 to 60 relative to parturition. Cows within block were randomly assigned to one of the following treatments: no supplement (CON), supplementation with 100 mg of rumen-protected capsicum/cow per day (RPCap), or RPCap plus 2 g of high-intensity artificial sweetener/cow per day (RPCapS). For both the RPCap and RPCapS treatments, only rumen-protected capsicum was fed during the dry period. From d 8 to 11 of lactation, intake was limited to 70% of predicted dry matter intake to induce subclinical ketosis. Production variables were recorded daily, samples for milk composition were collected on wk 2, 4, 6, and 8, and blood samples were collected on wk -2, 1, 2, and 4 of the experiment for analysis of metabolic hormones and blood cell counts. Supplementation with rumen-protected capsicum increased serum insulin and decreased β-hydroxybutyrate concentrations precalving, indicating a decrease in lipolysis. During the lactation period, RPCap was associated with a trend for increased milk production and feed efficiency following the ketosis challenge. Supplementation with RPCapS appeared to negate the response to rumen-protected capsicum. All cows developed subclinical ketosis during the challenge, and this was not affected by treatment. We conclude that treatments did not decrease susceptibility to subclinical ketosis; however, dietary supplementation with rumen-protected capsicum was effective at improving energy status precalving and tended to increased milk production and feed efficiency. The mechanism underlying these responses is unclear.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Mammalian pregnancy creates unique challenges for immune systems highly evolved to detect and eliminate invading pathogens. Recognition of the challenges created by gestating a semi-allogeneic fetus evolved from the discipline of transplantation biology and were informed by studies on the unique natural parabiosis that occurs when female calves are gestated with twin male fetuses. These pregnancies typically result in an intersex female termed a freemartin, which revealed insights into development of the male and female reproductive tracts. However, they also uncovered important clues on immune tolerance with wide-ranging implications to reproductive biology, transplantation biology and autoimmune disease. Many studies focused on identifying mechanisms through which the fetus evades maternal immune detection and elimination. These included studies characterizing immune interactions between the fetus and mother at the nourishing interface of the placenta and uterine endometrium. This immunological forbearance only occurs under high concentrations of circulating progesterone. Beyond the requirement for progesterone, there has been considerable progress towards understanding the effects of conceptus signals on maternal immune function. One common theme is that pregnancy induces a T helper 2 immune bias as shown in several mammalian species, including domestic ruminants. However, a growing body of evidence shows that the fetus not only evades, but also provokes immune responses locally in the uterus and in peripheral tissues. This is perhaps most dramatically illustrated by domestic ruminants where the conceptus secretes a unique interferon in the opening salvo of hormonal communication with the maternal immune system. The role of interferon tau in regulating expression of genes of the innate immune system in the uterus has been extensively studied. More recently, it was determined that these same genes are also induced in peripheral immune cells and other tissues throughout the body. In addition to interferon tau and progesterone, pregnancy associate glycoproteins and chaperonin 10 (aka Early Pregnancy Factor) are implicated in altering immune function both locally and systemically during pregnancy. While it is tempting to speculate that this activation of innate immunity is designed to counteract selective immunosuppression, knowledge of the importance of local and systemic immune activation to the success of pregnancy remains incomplete. This area remains fertile ground for developing better approaches to diagnose and treat infertility in domestic farm species and humans alike.

This study examined the effect of 3-nitrooxypropanol (3-NOP), a substance under investigation, on enteric methane (CH4) emission, rumen fermentation, lactational performance, sensory properties of milk, and the resumption of ovarian cyclicity in early-lactation dairy cows. Fifty-six multi- and primiparous Holstein cows, including 8 that were rumen cannulated, were used in a 15-wk randomized complete block design experiment. Cows were blocked based on parity and previous lactation milk yield (MY) or predicted MY, and within each block were randomly assigned to one of 2 treatments: (1) control (CON), administered no 3-NOP, or (2) 3-NOP applied at 60 mg/kg of feed dry matter (3-NOP). Enteric CH4 emission was measured during experimental wk 2, 6, 9, and 15, using the GreenFeed system. Dry matter intake (DMI) and MY data were collected daily throughout the experiment, and milk composition samples were collected 7 times during the experiment. Milk samples were collected from 14 to 60 (±2) d after calving, 3 d per week, and assayed for progesterone concentration to determine resumption of ovarian activity. Compared with CON, 3-NOP decreased daily CH4 emission by 26%, CH4 yield (CH4 per kg of DMI) by 21%, and CH4 emission intensity [CH4 per kg of MY or energy-corrected milk (ECM)] by 25%. Enteric emission of carbon dioxide was decreased by 5%, and hydrogen emission was increased 48-fold by 3-NOP. Inclusion of 3-NOP decreased concentration of total volatile fatty acids (by 9.3%) and acetate but increased butyrate molar proportion, ethanol, and formate concentrations in ruminal fluid. Dry matter intake was lower for 3-NOP compared with CON, but DMI expressed as a percentage of body weight was not different between treatments. Treatment had no effect on milk and ECM, body weight change, or body condition score. Milk composition and milk fat and protein yields were not affected by treatment, except that concentrations of short-chain fatty acids in milk were increased by 3-NOP. Nutrient digestibility and blood metabolites and hormones were not affected by 3-NOP, except that insulin was decreased by 3-NOP. There was no effect of 3-NOP on postpartum resumption of ovarian activity, including days to first and second luteal phases, length of first and second luteal phases, and interval from first to second luteal phase. Sensory properties of milk from cows fed 3-NOP and cheese made from that milk were not affected by treatment. In this experiment, 3-NOP decreased daily enteric CH4 emission, emission yield, and emission intensity, improved feed efficiency, and did not affect lactational performance or onset of ovarian activity in early-lactation dairy cows.

Mucus plays crucial roles in higher organisms, from aiding fertilization to protecting the female reproductive tract. Here, we investigate how anisotropic organization of mucus affects bacterial motility. We demonstrate by cryo electron micrographs and elongated tracer particles imaging, that mucus anisotropy and heterogeneity depend on how mechanical stress is applied. In shallow mucus films, we observe bacteria reversing their swimming direction without U-turns. During the forward motion, bacteria burrowed tunnels that last for several seconds and enable them to swim back faster, following the same track. We elucidate the physical mechanism of direction reversal by fluorescent visualization of the flagella: when the bacterial body is suddenly stopped by the mucus structure, the compression on the flagellar bundle causes buckling, disassembly and reorganization on the other side of the bacterium. Our results shed light into motility of bacteria in complex visco-elastic fluids and can provide clues in the propagation of bacteria-born diseases in mucus.

The objective of this experiment was to evaluate productive and reproductive effects of replacing solvent-extracted soybean meal (SSBM) with extruded soybean meal (ESBM) in a total mixed ration for early-lactation dairy cows. Thirty-four Holstein cows (12 primiparous and 22 multiparous) were used in a randomized complete block design experiment with 17 cows per treatment. Feeding was ad libitum for 5 to 10% refusals. A fresh-cow diet was fed the first 21 d in milk followed by a lactation diet from 22 to 60 d in milk. Milk and dry matter intake data were collected throughout the experiment, and samples were collected for blood chemistry and amino acid profile, nutrient digestibility, nitrogen utilization, and enteric methane emission using the GreenFeed system (C-Lock Inc., Rapid City, SD). Dry matter intake, milk yield, and feed efficiency were not different between SSBM and ESBM. Energy-corrected milk yield and efficiency were also not different between diets. Diet had no effect on milk composition, except that milk true protein yield was decreased by ESBM. Enteric methane emission, yield, and intensity were not different between SSBM and ESBM. Because of its greater fat content, ESBM triggered expected changes in milk fatty acid (FA) profile: decreased sum of C16, saturated, and odd- and branched-chain FA and increased sum of preformed FA, polyunsaturated, and trans FA. The ESBM diet increased or tended to increase some essential amino acids in plasma. In this study, ESBM did not affect dry matter intake and did not improve lactational performance or onset of ovarian function in early-lactation dairy cows, and it decreased milk protein yield, possibly due to greater unsaturated FA intake compared with SSBM.

Infertility and subfertility reduce the economic viability of dairy production.Inflammation reduces conception rates in dairy cattle, but surprisingly little information exists about the populations and the functions of immune cells at the conceptus-maternal interface during the periattachment period in dairy cattle.Early pregnancy is accompanied by immune stimulation at insemination and conceptus secretion of IFN-τ, pregnancy-associated glycoproteins, prostaglandins, and other molecules whose effects on immune function during early pregnancy have not been determined.Our working hypothesis is that pregnancy induces changes in immune cell populations and functions that are biased toward immunological tolerance, tissue remodeling, and angiogenesis.This review summarizes current knowledge, starting with insemination and proceeding through early pregnancy, as this is the period of maximal embryo loss.Results indicated that early pregnancy is accompanied by a marked increase in the proportion of endometrial immune cells expressing markers for natural killer (CD335) cells and cytotoxic T cells (CD8) along with an increase in cells expressing major histocompatibility class II antigens (macrophages and dendritic cells).This is accompanied by increased abundance of mRNA for IL-15, a natural killer growth factor, and IL-10 in the endometrium during early pregnancy.Furthermore, expression of indoleamine 2,3 dioxygenase was 15-fold greater in pregnant compared with cyclic heifers at d 17, but then declined by d 20.This enzyme converts tryptophan to kynurenine, which alters immune function by creating a localized tryptophan deficiency and by activation of the aryl hydrocarbon receptor and induction of downstream tolerogenic mediators.Expression of the aryl hydrocarbon receptor is abundant in the bovine uterus, but its temporal and spatial regulation during early pregnancy have not been characterized.Pregnancy is also associated with increased expression of proteins known to inhibit immune activation, including programed cell death ligand-1 (CD274), lymphocyte activation gene-3 (CD223), and cytotoxic T-lymphocyte associated protein-4 (CD152).These molecules interact with receptors on antigen-presenting cells and induce lymphocyte tolerance.Current results support the hypothesis that early pregnancy signaling in dairy heifers involves changes in the proportions of immune cells in the endometrium as well as induction of molecules known to mediate tolerance.These changes are likely essential for uterine wall remodeling, placentation, and successful pregnancy.

Natural killer (NK) cells are essential for establishment of human and rodent pregnancies. The function of these and other cytotoxic T cells (CTL) is controlled by stimulatory and inhibitory signaling. A role for cytotoxic cells during early pregnancy in cattle has not been described, but regulation of their function at the fetal-maternal interface is thought to be critical for conceptus survival. The hypothesis that the relative abundance of CTL and expression of inhibitory signaling molecules is increased by the conceptus during early pregnancy was tested. The proportions of lymphoid lineage cells and expression of inhibitory signaling molecules in the endometrium during early pregnancy in dairy heifers were determined using flow cytometry, immunofluorescence, and real-time PCR on days 17 and 20 of pregnancy and day 17 of the estrous cycle. Results revealed an increased percentage of NKp46+ and CD8+ cells in the uterus of pregnant heifers. Furthermore, a large percentage of uterine immune cells coexpressed these proteins. Compared to cyclic heifers, CD45+ uterine cells from pregnant heifers exhibited greater degranulation. Endometrium from pregnant heifers had greater mRNA abundance for the inhibitory molecules, CD274 and lymphocyte activating gene 3 (LAG3), and greater cytotoxic T lymphocyte-associated protein 4 (CTLA4), molecules that can interact with receptors on antigen-presenting cells and induce lymphocyte tolerance. This study demonstrates a dynamic regulation of both cytotoxic immune cells and tolerogenic molecules during the peri-implantation period that may be required to support establishment of pregnancy and placentation.

The livestock industries are a major contributor to the economy of the northeastern United States. Climate models predict increased average maximum temperatures, days with temperatures exceeding 25 °C, and higher annual precipitation in the Northeast. These environmental changes combined with increased atmospheric CO 2 concentration are expected to either increase or decrease forage productivity depending on the crop, and may decrease protein content and forage digestibility. Winter damage to sensitive forage species may also increase. Predicted temperature increases are expected to reduce fertility in dairy cattle and heat stress-induced inflammation may limit energy available for productive functions. Additional loss in milk production due to decreased feed intake is estimated to be up to 1% of the projected annual milk production through 2100. The effects of climate change on the beef industry in the Northeast are expected to be minimal. Broiler production in the region may benefit from warmer winter and summer temperatures, but future housing will require greater insulation and ventilation fan capacity. Providing adequate housing and ventilation to offset climate changes will also be important for the layer industry and will likely increase the price of eggs. Climate change is expected to have an economic impact on the horse industry in the region through additional management of land and forage resources, building of shelters, and heat abatement at equine events. Increased temperatures and more intense storms will increase nutrient losses and gaseous emissions from animal manure. Uncertainties about how host animals, pathogens, and disease vectors will respond to climate change highlight the need for continued animal health monitoring.

In this analysis we used a spatially explicit, simplified bottom-up approach, based on animal inventories, feed dry matter intake, and feed intake-based emission factors to estimate county-level enteric methane emissions for cattle and manure methane emissions for cattle, swine, and poultry for the contiguous United States. Overall, this analysis yielded total livestock methane emissions (8916 Gg/yr; lower and upper 95% confidence bounds of ±19.3%) for 2012 (last census of agriculture) that are comparable to the current USEPA estimates for 2012 and to estimates from the global gridded Emission Database for Global Atmospheric Research (EDGAR) inventory. However, the spatial distribution of emissions developed in this analysis differed significantly from that of EDGAR and a recent gridded inventory based on USEPA. Combined enteric and manure methane emissions from livestock in Texas and California (highest contributors to the national total) in this study were 36% lesser and 100% greater, respectively, than estimates by EDGAR. The spatial distribution of emissions in gridded inventories (e.g., EDGAR) likely strongly impacts the conclusions of top-down approaches that use them, especially in the source attribution of resulting (posterior) emissions, and hence conclusions from such studies should be interpreted with caution.