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Understanding species’ mating systems provides important information about their ecology, life history, and behavior. Direct observations of mating behaviors can be challenging, but molecular techniques can reveal information about mating systems and paternal identity in difficult-to-observe species such as sea turtles. Genetic markers can be used to assess the paternity of a clutch and to assign hatchlings to a father. Leatherback turtles Dermochelys coriacea sometimes mate with multiple individuals, resulting in clutches with mixed paternity; however, the effects of multiple paternity on hatchling quality are unclear. Leatherback hatchlings at Sandy Point National Wildlife Refuge, St. Croix, US Virgin Islands, exhibit visible variation in individual body size, sometimes within the same clutch. We collected morphometrics and tissue samples from hatchlings across multiple nesting seasons (2009, 2012, 2013, 2015, and 2016) and found that hatchlings exhibited small but statistically significant differences in morphometrics between years. We used maternal and hatchling microsatellite genotypes to reconstruct paternal genotypes, assigning fathers to each hatchling. We found multiple paternity in 5 of 17 clutches analyzed and compared differences in morphometrics between full-siblings with differences between half-siblings. We found no significant differences between morphometrics of hatchlings from the same mother but different fathers. We compared within-clutch variances in morphometrics for clutches with and without multiple paternity and found no significant difference in morphological variation between them. Therefore, we could not attribute differences in hatchling size within a clutch to paternal contribution. Understanding other factors affecting hatchling morphology, and other possible fitness metrics, may reveal insights into the benefits, or lack thereof, of polyandry in sea turtles.

Background: Autologous venous bypasses effectively treat extensive infrainguinal arterial occlusive disease with excellent long-term patency rates. However, one-third of patients will experience significant vein graft stenosis, particularly within the first-year post-bypass. Current endovascular interventions yield suboptimal results, with reported re-stenosis rates of 20–50 %. This study investigates the efficacy of cutting balloon (‘slash’) followed by a drug-eluting balloon (‘splash’) angioplasty in treating vein graft stenosis. Methods: This single-centre retrospective review examines consecutive patients who underwent the ‘Slash and Splash’ technique for treating significant stenosis (>70 % stenosis and/or PSV >300 cm/s) in infrainguinal autologous bypasses from June 2017 to January 2023. Follow-up duplex ultrasound was conducted at three months, six months, and yearly thereafter. Primary outcomes assessed technical success and graft patency. Secondary outcomes included major adverse limb events (MALEs), and major amputations Results: Twenty-three patients (mean age 67.4 ± 8.1 years, 44 % male) with significant vein graft stenoses underwent the ‘Slash and Splash’ method to salvage their bypasses. Most patients (91 %) had critical limb-threatening ischemia. Nine patients received femoral-popliteal grafts and 14 received femoral-tibial grafts, with18 utilizing saphenous in-situ conduits. The average follow-up duration was 26.1 ± 16.7 months, with no losses to follow-up. The median time from initial bypass to angioplasty was 10 months. Primary technical success was 100 % and primary patency was 95 %, with only one restenosis requiring repeat angioplasty during follow-up. Primary-assisted and secondary patency rates were 100 %. Freedom from MALEs was 90 %, with two patients requiring amputations secondary to diabetic foot infections, although bypasses remained patent at the time of amputation. Conclusions: The ‘Slash and Splash’ technique effectively treats severe infrainguinal vein bypass graft stenosis, offering excellent mid-term patency and freedom from MALEs. Adoption of this technique should be considered in the treatment of hemodynamically significant vein graft stenoses.

Background: The maternal-offspring relationship, such as whether fetal sex influences maternal health, is essential to explore to advance prenatal and maternal health. While associations exist between fetal sex and maternal health outcomes, it is unclear whether these reflect a causal relationship., Objective: To demonstrate that fetal sex can be randomly assigned to test the causal effect of fetal sex on maternal outcomes., Methods: Holstein dairy cows were stratified and randomized using sealed opaque envelopes to be artificially inseminated with either X- or Y-sorted bull semen until 40 cows became pregnant. Monthly body weight measurements were recorded, and an intravenous glucose tolerance test was performed 30 days before the expected calving day. The primary outcome was insulin area under the curve (AUC), and secondary outcomes were clearance rate, half-life, and AUC for glucose, insulin, and non-esterified fatty acid concentrations. An intention-to-treat (ITT) approach using multiple imputation was employed for primary analysis, and an as-treated (AT) approach was used for secondary analysis., Results: We demonstrated that we could successfully randomize the assignment of fetal sex to dams and test for causal effects of fetal sex on glucoregulatory outcomes using dairy cows as a model. Insulin AUC was not statistically different between groups (ITT p = 0.857, AT p = 0.874), and other outcomes were also not statistically different (p > 0.05)., Conclusion: We demonstrated that causal effects of fetal sex on maternal outcomes can be causally tested in dairy cows. Our study did not provide statistical evidence to support an effect of fetal sex on maternal glucose-related outcomes., Competing Interests: Declaration of competing interest In the last thirty-six months, Dr. Allison has received personal payments or promises for same from: Novo Nordisk Fonden; and Zero Longevity Science (as stock options). Donations to a foundation have been made on his behalf by the Northarvest Bean Growers Association. Dr. Allison's institution, Indiana University, and the Indiana University Foundation have received funds or donations to support his research or educational activities from: Eli Lilly and Company; National Cattlemen's Beef Association; Pfizer, Inc.; Soleno Therapeutics; The Morris Animal Foundation; USDA Agricultural Research Service; WW (formerly Weight Watchers); and numerous other for-profit and non-profit organizations to support the work of the School of Public Health and the university more broadly. All other authors report no disclosures in the last 36 months related to the topic of this study., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Fat supplementation has potential to improve reproductive performance and increase pregnancy rates in cattle by increasing the energy density of the diet. However, some of the positive effects of fat seem to be influenced by the type of fatty acid fed. Docosahexaenoic acid (DHA, 22:6n-3) and eicosapentaenoic acid (EPA, 20:5n-3) are long-chain n-3 poly-unsaturated fatty acids (PUFA) that have important biological effects on reproduction through their involvement in hormone and series 3 prostaglandin synthesis. Ruminant tissues are naturally almost devoid of n-3 long-chain PUFA, specifically EPA and DHA. Algae biomass provides a consistent source of DHA and EPA that could be fed to alter hormonal profiles and improve reproduction of beef heifers. Eighty-eight Angus × Simmental heifers (427 ± 1.8 kg body weight [BW]) were blocked by BW and allotted to 2 treatments (44/treatment, 4 pens/treatment, 11 heifers/pen). Control heifers were fed a diet that contained (dry matter [DM] basis) 52.8% mixed grass silage, 32% corn silage, and 15.2% concentrate. DHAgold (49% fat; 21.8% DHA; DSM Inc.) was included in the algae diet at 1.65% DM, replacing equal parts of corn and DDGS. Diets were formulated to contain 12% crude protein and 0.85 Mcal/kg net energy for gain (NEg). Heifers were fed treatment diets from 54 d prior to the breeding season through the first trimester. Follicular fluid was collected on day 47 for hormonal analysis. Artificial insemination (AI) was from days 54 to 77. Heifers that did not get pregnant to AI were removed from the study on day 109 and placed with a bull. The study ended on day 180. Data were analyzed using the MIXED procedure of SAS. During the pre-breeding period, algae heifers had lesser dry matter intake (P = 0.006) compared to control heifers. Heifers supplemented with algae had a greater average daily gain (P = 0.03) during the breeding period and BW tended (P = 0.06) to be greater for algae compared to control heifers on day 98 and was greater on day 180 (P = 0.03). Dominant follicle diameter and follicular estrogen concentration were unaffected by treatment (P ≥ 0.12). Follicular insulin-like growth factor-1 was greater in algae compared to control heifers (P = 0.03). First service conception rate did not differ between treatments (P = 0.67); however, second service and overall conception were lesser (P ≤ 0.03) in algae compared to control heifers. These data suggest supplementing DHA-rich algae improved growth but decreased conception rates of primiparous beef females., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

An accurate understanding of boar temperature preferences may allow the swine industry to design and utilize environmental control systems in boar facilities more precisely. Therefore, the study objective was to determine the temperature preferences of sexually mature Duroc, Landrace, and Yorkshire boars. Eighteen, 8.57 ± 0.10-mo-old boars ( N = 6 Duroc, 6 Landrace, and 6 Yorkshire; 186.25 ± 2.25 kg) were individually tested in thermal apparatuses (12.20 m × 1.52 m × 1.86 m) that allowed free choice of their preferred temperature within a 8.92 to 27.92 ºC range. For analyses, the apparatuses were divided into five thermal zones (3.71 m 2 /thermal zone) with temperature recorded 1.17 m above the floor in the middle of each zone. Target temperatures for thermal zones 1 to 5 were 10, 15, 20, 25, and 30 ºC, respectively. All boars were given a 24-h acclimation phase followed by a 24-h testing phase within the thermal apparatuses. Daily feed allotments (3.63 kg/d) were provided to each boar and all boars were allowed to consume all feed prior to entering the thermal apparatus. Water was provided ad libitum within the thermal apparatuses with 1 waterer per thermal zone. During testing, boars were video recorded continuously to evaluate behavior (inactive, active, or other), posture (lying, standing, or other), and thermal zone the boar occupied. All parameters were recorded in 15 min intervals using instantaneous scan sampling. Data were analyzed using GLM in JMP 15. For the analyses, only time spent lying or inactive were used because they were observed most frequently (lying 80.02%, inactive 77.64%) and were deemed to be associated with comfort based on previous research. Percent time spent active (19.73%) or standing (15.87%) were associated with latrine or drinking activity and were too low to accurately analyze as an indicator of thermal preference. Breed did not affect temperature preference ( P > 0.05). A cubic regression model determined that boars spent the majority of their time inactive at 25.50 ºC ( P < 0.01) and lying (both sternal and lateral) at 25.90 ºC ( P < 0.01). These data suggest that boar thermal preferences did not differ by breed and that boars prefer temperatures at the upper end of current guidelines (10.00 to 25.00 ºC)., (Published by Oxford University Press for the American Society of Animal Science 2023.)

Aim: Myocardial infarction remains the leading cause of heart failure. The adult human heart lacks the capacity to undergo endogenous regeneration. New blood vessel growth is integral to regenerative medicine necessitating a comprehensive understanding of the pathways that regulate vascular regeneration. We sought to define the transcriptomic dynamics of coronary endothelial cells following ischaemic injuries in the developing and adult mouse and human heart and to identify new mechanistic insights and targets for cardiovascular regeneration., Methods and Results: We carried out a comprehensive meta-analysis of integrated single-cell RNA-sequencing data of coronary vascular endothelial cells from the developing and adult mouse and human heart spanning healthy and acute and chronic ischaemic cardiac disease. We identified species-conserved gene regulatory pathways aligned to endogenous neovascularization. We annotated injury-associated temporal shifts of the endothelial transcriptome and validated four genes: VEGF-C, KLF4, EGR1, and ZFP36. Moreover, we showed that ZFP36 regulates human coronary endothelial cell proliferation and defined that VEGF-C administration in vivo enhances clonal expansion of the cardiac vasculature post-myocardial infarction. Finally, we constructed a coronary endothelial cell meta-atlas, CrescENDO, to empower future in-depth research to target pathways associated with coronary neovascularization., Conclusion: We present a high-resolution single-cell meta-atlas of healthy and injured coronary endothelial cells in the mouse and human heart, revealing a suite of novel targets with great potential to promote vascular regeneration, and providing a rich resource for therapeutic development., Competing Interests: Conflict of interest: None declared., (© The Author(s) 2022. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Natural honey has been successfully used in the preservation of mammalian gametes because of its beneficial properties. The objectives of this study were to determine the inclusion level of honey in extender for improving boar semen quality before freezing and to investigate the effects of honey inclusion in extender and freezing media on post-thaw quality of frozen-thawed boar semen samples. Ejaculates from six terminally crossbred boars were collected using the gloved-hand technique for two experiments. Experiment 1 was a randomized block design, evaluating four inclusion levels of honey in boar semen extender [Control (0H)-Androhep Plus or Androhep Plus with 0.25%, 0.50%, and 0.75% honey (0.25H, 0.50H, and 0.75H respectively)]. Ejaculates were pooled, aliquoted according to treatments, and cooled for 24 h at 17 ºC. The results of this experiment were used to determine inclusion levels in exp. 2. Experiment 2 was a 2 x ×3 factorial design, evaluating the inclusion of honey in boar semen extender and freezing media. Semen samples from individual boars were cooled in extender with or without honey (C0: Androhep Plus; C1: Androhep Plus + 0.25% honey). After 24 h, semen samples were evaluated, diluted in lactose-egg yolk (LEY) media, and one of three freezing media types; F0: 93% LEY + 6% glycerol + 1% Equex-STM Paste (ESP); F1: 93% LEY + (3% glycerol and 3% honey) + 1% ESP; and F2: 93% LEY + 6% glycerol + (0.5% ESP and 0.5% honey). Samples were frozen in 0.5 mL straws using a controlled-rate freezer and stored in liquid nitrogen. In exp. 1, 0.25H and 0.50H improved motility (P = 0.033) and progressive motility (P = 0.001) of cooled boar semen. Nevertheless, 0.25H was selected for exp. 2. In exp. 2, post-thaw motility and progressive motility were highest (P < 0.05) in C0F2 but not different from C1F2. Morphologically normal cells and acrosomes were higher with all inclusion levels of honey (P < 0.05). In conclusion, 0.25% and 0.50% inclusion of honey in Androhep Plus improves motility and progressive motility of cooled boar semen samples after 24 h. Supplementing Androhep Plus with 0.25% honey maintains higher normal sperm cells and acrosomes of cryopreserved boar semen. Replacing 50% Equex-STM paste with honey in freezing media improves post-thaw sperm motility, progressive motility, percentage of normal sperm, and acrosome of cryopreserved boar semen., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Carnitine is an amino acid derivative that performs the functions of increasing energy production as well as acting as an antioxidant for sperm cells. This study was conducted to investigate the effects of the inclusion of carnitine in boar diets on semen output and quality. Sixty-four purebred and hybrid boars at a commercial boar stud were blocked by age and semen quality and randomly allotted to receive a daily 30 g top-dress of either soybean meal (CON) or soybean meal and 625 mg of L-Carnitine (CARN). Supplementation lasted for 12 weeks from May to July 2021 during which weekly semen collection was performed. Semen was evaluated in the stud for concentration and motility parameters using computer-assisted semen analysis (CASA). Samples were shipped to Purdue University for detailed morphology, viability, and CASA analysis performed in samples stored at 17 °C for 5 days. PROC Mixed (SAS v 9.4) was used to analyze data, with boar nested within treatment used in repeated measures analysis. Semen quality estimates from the week before supplementation were used as covariates in the statistical model. Tukey-Kramer adjustment was used for means separation. Carnitine supplementation had no effects on total sperm produced ( P = 0.35). Percentage of motile sperm cells ( P = 0.63), morphologically normal sperm ( P = 0.42), viable sperm ( P = 0.43), or sperm with normal acrosomes ( P = 0.61) in the ejaculates were not different among treatments. Sperm kinematics in CARN ejaculates tended to have greater straight-line velocity and distance ( P = 0.06 and P = 0.07, respectively). There were several interactions of treatment and day of storage for the kinematic parameters. However, these interactions do not show observable trends for CARN to improve or depress sperm function. Overall, the inclusion of 625 mg/d of carnitine in the diet of boars for 12 weeks had no effects on sperm output or quality with minor changes to sperm cell kinematics., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Aims: Endothelial cell (EC) dysfunction drives the initiation and pathogenesis of pulmonary arterial hypertension (PAH). We aimed to characterize EC dynamics in PAH at single-cell resolution., Methods and Results: We carried out single-cell RNA sequencing (scRNA-seq) of lung ECs isolated from an EC lineage-tracing mouse model in Control and SU5416/hypoxia-induced PAH conditions. EC populations corresponding to distinct lung vessel types, including two discrete capillary populations, were identified in both Control and PAH mice. Differential gene expression analysis revealed global PAH-induced EC changes that were confirmed by bulk RNA-seq. This included upregulation of the major histocompatibility complex class II pathway, supporting a role for ECs in the inflammatory response in PAH. We also identified a PAH response specific to the second capillary EC population including upregulation of genes involved in cell death, cell motility, and angiogenesis. Interestingly, four genes with genetic variants associated with PAH were dysregulated in mouse ECs in PAH. To compare relevance across PAH models and species, we performed a detailed analysis of EC heterogeneity and response to PAH in rats and humans through whole-lung PAH scRNA-seq datasets, revealing that 51% of up-regulated mouse genes were also up-regulated in rat or human PAH. We identified promising new candidates to target endothelial dysfunction including CD74, the knockdown of which regulates EC proliferation and barrier integrity in vitro. Finally, with an in silico cell ordering approach, we identified zonation-dependent changes across the arteriovenous axis in mouse PAH and showed upregulation of the Serine/threonine-protein kinase Sgk1 at the junction between the macro- and microvasculature., Conclusion: This study uncovers PAH-induced EC transcriptomic changes at a high resolution, revealing novel targets for potential therapeutic candidate development., Competing Interests: Conflict of interest: S.Y.C. has served as a consultant for United Therapeutics, has held research grants from Actelion and Pfizer, and filed patent applications regarding drug development in pulmonary hypertension. S.Y.C. is a director, officer, and shareholder of Synhale Therapeutics. The other authors declare no competing interests. This manuscript was handled by Consulting Editor Henning Morawietz., (© The Author(s) 2021. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Perinatal nutrition affects future milk production. The number of mammary epithelial cells affect milk production capacity. Therefore, it was hypothesized that the level of colostrum intake affects the proliferation rate and the total number of mammary epithelial cells in the gland. The ratio of newly synthesized protein to newly synthesized DNA reflects the relative amount of cellular differentiation to cell division. The study objective was to determine the relationship between the level of colostrum intake and 24 h-level of circulating amino acid, glucose and insulin with mammary parenchyma histological features, cell division and protein synthesis over the first week postnatal. One of two standardized doses of a homogenate colostrum sample, 10% ( n = 8) and 20% ( n = 8) of birth bodyweight, was fed to gilts over the first 24 h postnatal. Gilts were administered deuterium oxide immediately after birth and daily to label newly synthesized DNA and proteins. Gilts were euthanized on postnatal day seven, and DNA and protein were isolated from mammary parenchyma. DNA and protein fractional synthesis (f) and fractional synthetic rate (FSR) were calculated using mass isotopomer distribution analysis. The ratio of protein f and FSR to DNA f and FSR were calculated and used to indicate the relative amounts of differentiation to cell division. Mammary morphological development was also analyzed by measuring the parenchymal epithelial area and the stromal and epithelial proliferation index on postnatal day seven. Colostrum dose was not related to any of the variables used to evaluate mammary development. However, plasma lysine levels at 24 h postnatal were positively related to average daily gain (ADG; r = 0.54, p = 0.05), DNA f ( r = 0.57; p = 0.03) and DNA FSR ( r = 0.57; p = 0.03) in mammary parenchyma. Plasma lysine was inversely related to the ratio of protein to DNA f and FSR ( r = -0.56; p = 0.04). ADG was related to the parenchymal epithelial area and DNA and protein f and FSR ( p < 0.05). These relationships support the idea that the nutritional environment affects early mammary development and that higher lysine levels in the perinatal period favored a greater degree of cell division versus differentiation in mammary of neonatal pigs and thus, warrant further investigations.

Milk is an easily digestible source of nutrients and bioactive factors, its composition reflects the neonate's needs, and changes from colostrum to transitional and mature milk. Our objective was to measure milk fat, lactose, total carbohydrate, and protein content in parallel with global proteome of homogenate milk samples to characterize changes across the three phases of swine lactation. Milk samples were collected from multiparous sows (n = 9) on postnatal day 0 (D0; colostrum), 3 (D3; early transitional), 7 (D7; late transitional), and 14 (D14; mature). On D3, percent fat (16 ± 2.1) and lactose (3.8 ± 0.3) were higher (P < 0.05) than on D0 (10 ± 3.9 and 1.5 ± 0.3, respectively). Levels of fat and lactose were not different between D3 and D14. Percent total protein decreased (P < 0.05) between D0 (11 ± 2.1) and D3 (5 ± 0.7), but there was no significant change in percent protein between D3 and D14. Total carbohydrates increased (P < 0.05) between D3 (944 ± 353 µg/mL) and D14 (1,150 ± 462 µg/mL). Quantitative proteomic analysis using liquid chromatography tandem mass spectrometry (LC-MS/MS) of homogenate D0, D3, and D14 milk samples (n = 6) identified 772 protein groups which corresponded to 501 individual protein-coding genes. A total of 207 high confidence proteins were detected in n = 3 sows/day. Of the high confidence proteins, 81 proteins were common among all 3 days of lactation. Among the proteins that decreased between the days (false discovery rate; FDR < 0.05) were multiple apolipoproteins and XDH which decreased between D0 to D3. Proteins that increased across the days (FDR < 0.05) were complement factors and 14-3-3 proteins (YWHAQ, YWHAE). Our data provide a good characterization of milk proteome changes that likely reflect mammary function as well as the neonate's phase-specific developmental needs. This data may be useful in developing approaches to enhance the health and welfare of swine., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science.)

The objective of this study was to investigate the effects of semen volume, air contact inside semen dose tubes, daily agitation of semen doses and extender type on semen quality, thermo-resistance and bacteria growth in extended boar semen doses preserved over 7 days of liquid storage. Ejaculates from 4 proven terminal cross-bred boars were collected using the gloved-hand technique for 4 weeks and used in the 3 × 2 × 2 factorial study. The effects of treatment (CON: 80 ml doses sealed at the top of the tube; 40HIGH: 40 ml doses sealed at top of tube, and 40LOW: 40 ml doses sealed at top of the liquid), agitation (agitated versus not agitated) and extender type (long-term versus short-term) were investigated on semen quality, thermo-resistance and bacteria growth in boar semen doses. The results of the study revealed that motility (p = .031) and viability (p = .041) in 40HIGH were lower than CON. pH (p < .001) was higher in 40HIGH compared with CON and 40LOW. Agitation did not impact motility (p = .581), progressive motility (p = .870), viability (p = .509) or morphology (p = .970), while long-term extender maintained higher motility (p = .002), progressive motility (p = .036), viability (p < .001) and normal acrosome (p < .001) than a short-term extender. VAP (p = .039) of 40HIGH was lower than CON in a thermo-resistance test. Neither treatment (p > .798, .766) nor agitation (p > .396, .476) impacted bacterial growth in this study. In conclusion, air contact negatively impacts boar semen pH and consequently sperm motility. Semen doses prepared with 80 or 40 ml volumes of extended boar semen with minimal air contact in the tubes yield more desirable semen quality and agitating boar semen doses daily does not have negative or positive effects on boar semen quality., (© 2021 Wiley-VCH GmbH.)

A marker indicative of the fertility potential of replacement gilts early in development would decrease culling rates in the sow herd, improve sow herd reproductive efficiency, and reduce production costs. The objective of this study was to determine if vaginal lipid profiles at 21 d postnatal (PN) could predict sow reproductive performance. Vaginal swabs of the anterior vagina were taken at 21 ± 4 d PN from gilts born on a commercial sow production facility for lipidomic analysis. Animals were followed prospectively for 2 yr and assigned to reproductive performance categories based on the observation of estrus or piglets weaned per sow per year (PSY) across two farrowings. Lipids were extracted from cellular material collected with swabs taken from high fertility (HF; n = 28; ≥26 PSY) and infertile (IF; n = 34; no estrus, no pregnancy) animals, and multiple reaction monitoring profiling was used for lipidome analysis. The relative abundance of arachidonic acid (C20:4) and docosahexaenoic acid (C22:6) was lower (P < 0.05) in IF gilts than HF gilts, whereas the abundance of the free fatty acids such as cerotic (C26:0), ximenic (C26:1), and nonadecanoic (C19:0) acids was greater (P < 0.05) in IF gilts. Additionally, eicosapentaenoic acid (C20:5), a precursor of prostaglandins, was higher (P < 0.05) in IF gilts. The perspective of having a panel of lipids captured with vaginal swabs at weaning that can predict the reproductive efficiency of gilts shows promise and warrants future research in this area., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Milk fats are vital to neonate survival and development, but vary highly by diet, maternal metabolic state and stage of lactation. To gain a better understanding of changes in lipid composition of sow milk across lactation, milk was collected from nine multiparous sows on days 0, 3, 7, and 14, relative to birth and lipids were profiled using multiple reaction monitoring (MRM) profiling. Percent fat was determined by creamatocrit, and found to be different (P < 0.05) between day 0 (12.36 ± 5.90%) and day 3 (16.22 ± 3.65%) but not between day 7 (13.13 ± 2.19%) and day 14 (12.13 ± 2.45%). Fat was extracted from milk (n = 6/day) using the Bligh-Dyer method and profiled using tandem mass spectrometry MRM to determine the abundance of lipids defined by class and fatty acyl residue composition. Lipid species relative concentration was calculated from internal standards, and data analysis was performed using Metaboanalyst 4.0. Concentration of phosphatidyl-choline, -serine, -ethanolamine, -inositol, cholesteryl ester and sphingomyelin did not vary across lactation days, nor did the distribution of associated fatty acyl residues. The total abundance of triacylglycerides (TGs) and phosphatidylglycerols (PGs) increased (P < 0.05) from colostrum (day 0) to transitional (days 3 and 7) and mature milk (day 14). As lactation days increased from day 0 to day 14, the number of carbons and unsaturation within fatty acyl residues decreased (P < 0.05) in both TGs and PGs. The proportion of TGs and PGs increased (P < 0.05) relative to other lipid classes. Changes in composition of milk triglycerides and phosphatidylglycerols likely reflect the metabolic activity of the mammary gland and developmental needs of neonates., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Although the visual and geomagnetic orientation cues used by sea turtle hatchlings during sea-finding have been well studied, the potential for auditory stimuli to act as an orientation cue has not been explored. We investigated the response of sea turtle hatchlings to natural and anthropogenic noises present on their nesting beaches during sea-finding. The responses of hatchling leatherback sea turtles, Dermochelys coriacea, collected from the Sandy Point National Wildlife Refuge, St. Croix, were measured in the presence of aerial acoustic sounds within hatchlings' hearing range of 50 to 1600 Hz. The highest sound energy produced by beach waves occurs at frequencies 50-1000 Hz, which overlaps with the most sensitive hearing range of hatchling leatherbacks (50-400 Hz). Natural beach wave sounds, which have highest sound energy at frequencies of 50-1000 Hz, may be masked by human conversations (85-650 Hz) and vehicle traffic noise (60-8000 Hz). In the presence of three stimuli, a) beach wave sounds (72.0 dB re: 20 μPa), b) human conversation (72.4 dB re: 20 μPa), and c) vehicle traffic noise (71.1 dB re: 20 μPa), hatchlings exhibited no phonotaxic response (wave sounds: mean angle = 152.1°, p = 0.645; human conversation: mean angle = 67.4°, p = 0.554; traffic noise: mean angle = 125.7°, p = 0.887). These results may be due to the hatchlings being unable to localize sounds in the experimental arena. Visual and auditory cues may also converge to affect sea-finding orientation. Future studies should focus on the localization ability of sea turtles and on the role that sound may play in orientation when combined with other sensory and environmental cues., Competing Interests: The authors have declared that no competing interests exist.

Historically, sows have been induced to farrow using prostaglandin followed by an injection of oxytocin 24 h later. Benefits of induction can include decreased rate of stillbirths, dystocia, and postnatal mortality along with increasing the likelihood of farrowings being attended. Several studies have indicated that oxytocin administration may negatively impact fetal oxygen supply during parturition, potentially from umbilical cords breaking prior to birth, resulting in increased preweaning mortality. Therefore, the objective of this study was to determine if various induction protocols impact umbilical cord breakage and fetal blood parameters at birth. Fifty-eight primiparous and multiparous sows were assigned to one of three treatments: no induction (NO; n = 24) or 2 cc prostaglandin administered on day 114 of gestation followed by either 1 cc of oxytocin 24 h later (OXY24; n = 13) or 0.5 cc of oxytocin at 6 and 12 h after prostaglandin (OXY6; n = 21). Details of the farrowing process were recorded, and umbilical cord blood was collected from piglets at birth and evaluated on an iSTAT machine using an Abbott EC8+ test cartridge. There were no differences in total born, number born alive, stillborns, mummies, or assistance needed during farrowing. Induced sows were more likely to farrow by day 115 compared to naturally farrowing sows ( P = 0.02). Sows in the OXY24 treatment tended to have longer farrowings when compared to both NO and OXY6 (4.8 vs. 3.6 vs. 3.9 h; P = 0.09). Colostrum from OXY6 sows tended to have a greater amount of lactose present than NO and OXY24 ( P = 0.05). Colostrum from sows with longer gestation lengths had a higher percentage of fat ( P = 0.03). Piglets born from NO sows had higher base excess, total carbon dioxide, and glucose, which suggests that these piglets had prolonged moments of asphyxiation ( P < 0.01). OXY24 piglets had the lowest blood pH which is indicative of hypoxic birthing conditions ( P < 0.01). Preweaning mortality was driven largely by a low birth weight coupled with low colostrum intake ( P = 0.03). All piglets, regardless of treatment, displayed signs of stress during farrowing. Induction did not influence preweaning mortality but has the potential to decrease the incidence by increasing attended farrowings., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Selection of replacements for the sow herd is one of the most important facets in swine production. Although our current methods of selection are effective, there is still a large amount of variation in sow reproductive performance traits such as pigs per sow per year (PSY). Therefore, the objective of this study was to determine if on-farm phenotypic traits at 21 d postnatal (PN) or perinatal environmental factors could predict sow reproductive performance. Data were prospectively collected from 2,146 gilts born on a commercial sow production facility and included birth and weaning weights, vulva length and width at 21 d PN, birth and nursing litter size, days nursed, average daily gain from birth to weaning, and age at first estrus. Of the initial animals, 400 (17%) were selected for the sow herd, 353 remained after removal of animals culled for non-reproductive reasons. Animals were assigned to 1 of 5 reproductive performance categories based on observation of estrus or pigs per sow per year (PSY) across two farrowings: High Fertility (HF; 23%; n = 82; ≥26 PSY), Middle Fertility (MF2; 12%; n = 43; 20-25 PSY), Low Fertility (MF3; 15%; n = 54; <20 PSY), Infertile-Estrus (IFe; 10%; n = 36; estrus, no pregnancy), and Infertile-No Estrus (IFno; 39%; n = 138; no estrus, no pregnancy). Generalized linear model analysis indicated vulva width ( P = 0.03) was related to PSY, however, it only explained 1.5% of the total variation in PSY. To determine if preweaning variables were predictive of gilt fertility outcome, animals were grouped as those that became pregnant ( n = 179) or not ( n = 174). Vulva width tended to be greater in fertile animals versus infertile ( P = 0.07). Binomial regression analysis revealed a positive relationship between vulva width and gilt fertility, however, this relationship is not strong enough to make sow herd selection decisions., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Survival, feed efficiency, growth, and fertility of swine are dependent on colostrum intake in the first 24 h after birth. This study determined the effects of three doses of a homogeneous colostrum sample on 24-h body weight, rectal temperature (RT), immunocrit, and growth and survival to postnatal day (PND) 7. Three female piglets were selected from eight litters ( n = 24 piglets) at birth, removed from their litter, and bottle-fed 10% (COL10, n = 8), 15% (COL15, n = 8), or 20% (COL20, n = 8) colostrum based on birth weight over 12 bottle feedings every 2 h. At 24 h, piglets were weighed, RT recorded, and blood was collected to measure immunocrit. Piglets were returned to the litter of origin, and weight was measured daily until PND 7. Colostrum dose had an overall effect on weight gain at 24 h, RT, immunocrit, and growth to PND 7 ( P < 0.05). Piglets in the 20% BrW colostrum group had greater weight gain, RT, and immunocrit at 24 h than COL10 piglets ( P < 0.05), but these variables were not different between COL15 and the other treatments. Despite no difference in average daily gain after being returned to their litters, the greater weight ( P < 0.05) in COL20 compared to COL10 and COL15 was sustained over 7 d. Seven piglets in each treatment survived to PND 7. This model using standardized doses of a homogeneous colostrum sample enables controlled studies aimed at understanding the role of 24-h colostrum intake on piglet development., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

There is a need to identify subfertile boars before they enter into the breeding herd. Seminal plasma proteins are essential for normal sperm function and transport and play an important role in fertilization. The objective of this study was to use liquid chromatography tandem mass spectrometry for shotgun proteome analysis to investigate whether differences in boar fertility phenotype can be differentiated by seminal plasma protein abundance. Following 50 breedings, boars were categorized into one of four phenotypes: high farrowing rate and total born (HFHB; n = 9), high farrowing rate with low total born (HFLB; n = 10), low farrowing rate and total born (LFLB; n = 9), and low farrowing rate with high total born (LFHB; n = 4) that were distinct (p < 0.05) from each other by these variables. There were 506 proteins measured in at least one sample across all animals. There were 245 high confidence proteins and 56 were differentially abundant between the high fertility phenotype (HFHB) and at least one of the three subfertile groups. Findings support that seminal plasma protein profiles are distinct between boars with different fertility phenotypes., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The prenatal environment influences offspring health and development, and this is readily apparent when considering the well-described effects of maternal nutrition and stress on the postnatal metabolism, neural function, and stress response of progeny. Moreover, in laboratory species, sheep, and humans, the effects of in utero heat stress on offspring development have been described in detail for >50 years. Despite our extensive knowledge of the postnatal phenotypes elicited by in utero stressors, the carryover effects of in utero heat stress in pigs have only recently begun to be elucidated. The effects of climate change on increasing global temperatures, combined with greater metabolic heat production in modern swine, has increased heat stress susceptibility in pigs. Greater heat stress susceptibility can negatively affect swine welfare and performance and may impact future generations of pigs through in utero heat stress. Pigs exposed to in utero heat stress develop a variety of postnatal phenotypes that prevent profitable production, and compromise health, and welfare in commercial production systems. Specifically, in utero heat stress alters the postnatal stress response, core body temperature, response to an immune challenge, and is teratogenic. In addition, in utero heat stress changes postnatal body composition through reduced lean and increased adipose tissue accretion rates, respectively. Furthermore, in utero heat stress reduces piglet birth weight, body weight gain, and reproductive efficiency. Although the economic impact of in utero heat stress in pigs has yet to be determined, it likely rivals the postnatal consequences of heat stress and is a threat to the global sustainability of swine production., (Published by Elsevier Inc.)

Reducing the number of sperm needed to produce a litter with artificial insemination (AI) allows greater use of higher genetic merit boars. Induced ovulation with single fixed-time artificial insemination (SFTAI), combined with intrauterine (IUI) or deep uterine insemination (DUI), could improve fertility with low numbers of sperm. The objectives of the study were to determine the fertility effects of sperm numbers and the site of insemination. At weaning (0 h), sows (n = 534) were assigned by parity and estrus induction method (equine chorionic gonadotropin [eCG] or Control) to receive 1,200 × 106 sperm by IUI; 600, 300, or 150 × 106 sperm by IUI or DUI; or 75 × 106 sperm by DUI. At 80 h postweaning, sows received OvuGel and 26 h later a SFTAI using pooled semen. Sows were exposed to boars once daily and ultrasound was performed to determine follicle size and time of ovulation. Following SFTAI, sows were slaughtered 27 d after AI to determine pregnancy and litter traits. Data were analyzed using different models to test for effects of estrus induction, interaction of three levels of sperm (600 to 150) with two levels for site (IUI vs. DUI), and the overall effects of AI method (eight treatments). There was no effect (P > 0.05) of estrus induction on estrus (93%) within 5 d of weaning or on follicle size (6.1 mm) at OvuGel, but wean-to-estrus interval (3.8 vs. 4.0 d) was slightly reduced (P < 0.01) as was AI-to-ovulation interval (15.9 vs. 17.0 h, P = 0.04) for eCG and Control, respectively. There was no effect (P > 0.05) of estrus induction on pregnancy rate (78.6%), number of corpora lutea (CL; 21.7), or number of viable embryos (12.2). There was no effect of number of sperm or site of insemination and no interaction (P > 0.05) on pregnancy rate (range: 80.9% to 70.5%), but AI occurring after ovulation reduced the pregnancy rate (P < 0.02). The total number of embryos (range: 16.5 to 10.3) was not affected by estrus induction, number of sperm, or site of insemination (P > 0.05), but was influenced by AI treatment (P < 0.01). Treatments with a higher number of sperm (1,200 and 600) had more embryos compared with those with a lower number of sperm (300 to 75). The numbers of embryos also increased with the number of CL (P < 0.0001). These results suggest that the lower number of sperm affects litter size more than the pregnancy status. Acceptable fertility can be achieved with low numbers of sperm when using a SFTAI and uterine deposition, but AI-to-ovulation interval and ovulation rate influence final fecundity., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

This article involved a broad search of applied sciences for milestone technologies we deem to be the most significant innovations applied by the North American pork industry, during the past 10 to 12 years. Several innovations shifted the trajectory of improvement or resolved significant production limitations. Each is being integrated into practice, with the exception being gene editing technology, which is undergoing the federal approval process. Advances in molecular genomics have been applied to gene editing for control of porcine reproductive and respiratory syndrome and to identify piglet genome contributions from each parent. Post-cervical artificial insemination technology is not novel, but this technology is now used extensively to accelerate the rate of genetic progress. A milestone was achieved with the discovery that dietary essential fatty acids, during lactation, were limiting reproduction. Their provision resulted in a dose-related response for pregnancy, pregnancy maintenance and litter size, especially in maturing sows and ultimately resolved seasonal infertility. The benefit of segregated early weaning (12 to 14 days of age) was realized for specific pathogen removal for genetic nucleus and multiplication. Application was premature for commercial practice, as piglet mortality and morbidity increased. Early weaning impairs intestinal barrier and mucosal innate immune development, which coincides with diminished resilience to pathogens and viability later in life. Two important milestones were achieved to improve precision nutrition for growing pigs. The first involved the updated publication of the National Research Council nutrient requirements for pigs, a collaboration between scientists from America and Canada. Precision nutrition advanced further when ingredient description, for metabolically available amino acids and net energy (by source plant), became a private sector nutrition product. The past decade also led to fortuitous discoveries of health-improving components in ingredients (xylanase, soybeans). Finally, two technologies converged to facilitate timely detection of multiple pathogens in a population: oral fluids sampling and polymerase chain reaction (PCR) for pathogen analysis. Most critical diseases in North America are now routinely monitored by oral fluid sampling and prepared for analysis using PCR methods.

Heat stress due to increasing extremes in ambient temperature and humidity results in reduced semen quality in boars. This has caused reduced efficiency of the swine industry, requiring more boars to breed the same number of sows. Vitamins such as vitamin C (VC) and E (VE) have been shown to improve semen quality in boars. Recently, vitamin D has been shown to improve semen quality in boars. The purpose of this experiment was to evaluate the effects of increased supplemental vitamins on boar reproduction during the summer season in a commercial boar stud. One hundred and sixty Pig Improvement Company (PIC) terminal line boars (n = 32 per treatment) and 39 maternal, heat-sensitive boars (n = 7 or 8 per treatment) were randomly allocated to treatment and fed a corn and soybean meal-based diet adjusted based on individual boar body condition score. A control (CNT) diet was used that met PIC recommendations for boars. Increased supplementation of specific vitamins was given in the form of a top-dress and consisted of CNT wheat middlings, CNT plus VC (560 mg/day), CNT plus 25-hydroxy vitamin D3 (VD) (125 µg/day), CNT plus VE (275 mg/day) and CNT plus VC, VD and VE (CDE). The experiment was split into three periods based on maximum daily high temperatures in the barn, where period 1 was weeks 1 to 4, period 2 was weeks 5 to 11 and period 3 was weeks 12 to 14. Semen was collected from boars as needed using the stud's normal production schedule and was analyzed for sperm quantity and quality characteristics. There were no dietary effects on semen volume, sperm concentration or total sperm production (P ≥ 0.553). Total motility of sperm was not impacted by diet (P = 0.115); although, VC tended (P = 0.064) to have a greater progressive motility than CDE. Percentages of morphologically normal sperm and normal acrosomes were not affected by dietary supplementation (P ≥ 0.157). Period effects were observed for most semen quality parameters, with quality generally becoming reduced over time. The present study demonstrates that increased supplementation of vitamins beyond PIC recommendations was not beneficial for boar reproduction during the summer.

Piglet crushing is a devastating welfare concern on swine farms; however, some sows appear unresponsive to a piglet's call. Sow hearing ability is rarely considered despite the extensive body of research performed on crushing. In this study, pigs of four age groups (weaning, n = 7; gilts, n = 5; 2nd and 3rd parity, n = 5; 5th parity and up, n = 5) were anesthetized and auditory brainstem responses (ABRs) were performed to measure if pig hearing diminishes with age in a mechanically ventilated barn. Before testing, pigs were placed in a sound dampening box. ABRs were performed on animals using 1,000 clicks at two decibel (dB) levels: 90 and 127 dB sound pressure level. Latencies and amplitudes of waves I-V were measured and interpeak latencies for waves I-III, III-V, and I-V were calculated. Five pigs (three 2nd and 3rd parity, and two 5th parity and above) had no detectable waves at either decibel. Sows in 2nd and 3rd parities had very few distinguishable waves, with only wave I and II present in two sows. Amplitudes of waves I and V increased with increased dB ( P < 0.001). Increasing dB decreased the latency of each of the recorded waves ( P < 0.01). The vast majority of commercial swine are raised in noisy barn environments; it is possible that these environments directly affect the ability for pigs to hear and normal hearing development in this population of animals. Hearing has a significant effect on swine welfare as hearing is integral to successful animal handing and during moments of animal-to-animal communication. Hearing is a considerable welfare issue on farms and ways to decrease pig hearing loss should be considered., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Objective: To elucidate perspectives on opioids and opioid use from hospitalized veterans with comorbid chronic pain using qualitative methods., Design: This was an analysis of individual qualitative interviews. The semistructured interview guide was developed by a hospitalist with clinical expertise in pain treatment with guidance from a medical anthropologist. Interviews aimed to understand participants' experiences of chronic pain., Setting: A Midwestern Veterans Health Administration inpatient hospital unit., Subjects: Nineteen inpatient veterans with a history of chronic pain or antecedent opioid use., Methods: Recently admitted veterans were screened for chronic pain diagnosis on admission and antecedent opioid use. Eligible veterans were approached to participate in an in-person interview during their hospitalization., Results: The following themes were identified in relation to opioid use: other patients as the problem (by misusing opioids resulting in broad limits to opioid access), empathy for providers (perceived to be working under prescribing constraints), and opioids as a last resort., Conclusions: Although participants were not specifically questioned about opioid medications, discussion of opioids was prevalent in discussions of chronic pain. Findings suggest the potential utility of engaging hospitalized veterans in conversations about opioids and alternative pain management strategies., (2018 American Academy of Pain Medicine. This work is written by US Government employees and is in the public domain in the US.)

Advances in high-throughput sequencing (HTS) technologies coupled with increased interdisciplinary collaboration are rapidly expanding capacity in the scope and scale of wildlife genetic studies. While existing HTS methods can be directly applied to address some evolutionary and ecological questions, certain research goals necessitate tailoring methods to specific study organisms, such as high-throughput genotyping of the same loci that are comparable over large spatial and temporal scales. These needs are particularly common for studies of highly mobile species of conservation concern like marine turtles, where life history traits, limited financial resources and other constraints require affordable, adaptable methods for HTS genotyping to meet a variety of study goals. Here, we present a versatile marine turtle HTS targeted enrichment platform adapted from the recently developed Rapture (RAD-Capture) method specifically designed to meet these research needs. Our results demonstrate consistent enrichment of targeted regions throughout the genome and discovery of candidate variants in all species examined for use in various conservation genetics applications. Accurate species identification confirmed the ability of our platform to genotype over 1,000 multiplexed samples and identified areas for future methodological improvement such as optimization for low initial concentration samples. Finally, analyses within green turtles supported the ability of this platform to identify informative SNPs for stock structure, population assignment and other applications over a broad geographic range of interest to management. This platform provides an additional tool for marine turtle genetic studies and broadens capacity for future large-scale initiatives such as collaborative global marine turtle genetic databases., (© 2019 John Wiley & Sons Ltd.)

Key Points: Type 1 diabetes mellitus increases cardiovascular risk; hypertension amplifies this risk, while pressure natriuresis regulates long-term blood pressure. We induced type 1 diabetes in rats by streptozotocin injection and demonstrated a substantial impairment of pressure natriuresis: acute increases in blood pressure did not increase renal medullary blood flow, tubular sodium reabsorption was not downregulated, and proximal tubule sodium reabsorption, measured by lithium clearance, was unaffected. Insulin reduced blood glucose in diabetic rats, and rescued the pressure natriuresis response without influencing lithium clearance, but did not restore medullary blood flow. Radiotelemetry showed that diastolic blood pressure was increased in diabetic rats, and its diurnal variation was reduced. Increases in medullary blood flow and decreases in distal tubule sodium reabsorption that offset acute rises in BP are impaired in early type 1 diabetes, and this impairment could be a target for preventing hypertension in type 1 diabetes., Abstract: Type 1 diabetes mellitus (T1DM) substantially increases cardiovascular risk, and hypertension amplifies this risk. Blood pressure (BP) and body sodium homeostasis are linked. T1DM patients have increased total exchangeable sodium, correlating directly with BP. Pressure natriuresis is an important physiological regulator of BP. We hypothesised that pressure natriuresis would be impaired, and BP increased, in the early phase of T1DM. Male Sprague-Dawley rats were injected with streptozotocin (30-45 mg/kg) or citrate vehicle. After 3 weeks, pressure natriuresis was induced by serial arterial ligation. In non-diabetic controls, this increased fractional excretion of sodium from ∼1% to ∼25% of the filtered load (P < 0.01); in T1DM rats, the response was significantly blunted, peaking at only ∼3% (P < 0.01). Mechanistically, normal lithium clearance suggested that distal tubule sodium reabsorption was not downregulated with increased BP in T1DM rats. The pressure dependence of renal medullary perfusion, considered a key factor in the integrated response, was abolished. Insulin therapy rescued the natriuretic response in diabetic rats, restoring normal downregulation of tubular sodium reabsorption when BP was increased. However, the pressure dependence of medullary perfusion was not restored, suggesting persistent vascular dysfunction despite glycaemic control. Radiotelemetry showed that T1DM did not affect systolic BP, but mean diastolic BP was ∼5 mmHg higher than in non-diabetic controls (P < 0.01), and normal diurnal variation was reduced. In conclusion, functional impairment of renal sodium and BP homeostasis is an early manifestation of T1DM, preceding hypertension and nephropathy. Early intervention to restore pressure natriuresis in T1DM may complement reductions in cardiovascular risk achieved with glycaemic control., (© 2018 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.)

Despite much interest in sow welfare, the impact of the acoustic environment on sow reactivity to her piglets is rarely considered. The objective of this study was to understand the impact of noise produced by mechanical ventilation and other sows on a sow's reactivity to her piglets. Sows were farrowed in one of three environments: 1) with eight other sows exposed to constant fan noise (GROUP-FAN; n = 10), 2) alone with fan noise present (ISO-FAN; n = 10), and 3) alone without fans running (ISO-QUIET; n = 10). Sows were subjected for 5 min to a piglet removal event (REMOVAL) by an unknown handler twice, at 24 and 48 h postfarrowing. During a REMOVAL, sows were observed via video recording for changes in posture, eating and drinking behavior, and head orientation. Audio was recorded to quantify vocalizations by the sow. Once piglets were returned, sows underwent further behavior observations for 10 min (RETURN), resulting in approximately 15 min of total video observation. Sows were classified as young (second and third parity) and old (fifth parity and older). The YOUNG sows tended to be more Alert (looking toward the handler or their piglets) during REMOVE than OLD sows ( P = 0.07; 2.01 and 1.33, respectively). The ISO-FAN sows vocalized the loudest during REMOVAL ( P < 0.001) with ISO-QUIET sows performing the quietest vocalizations (GROUP-FAN: 72.22 ± 1.06 dB; ISO-FAN: 73.61 ± 1.07 dB; ISO-QUIET: 67.41 ± 0.99 dB). During RETURN, YOUNG sows spent more time sitting than OLD sows ( P < 0.01; 7.48 ± 1.6% and 0.91 ± 1.8%, respectively). The ISO-QUIET sows tended to have more posture changes during the RETURN with ISO-FAN having the least changes ( P = 0.06; GROUP-FAN: 1.23 ± 0.4; ISO-FAN: 0.44 ± 0.3; ISO-QUIET: 1.61 ± 0.4). Finally, sows decreased the amount of time Alert in the second RETURN ( P = 0.03; first: 3.9 ± 0.6%; second: 2.5 ± 0.6%). Overall, sows acclimated to the removal and return events with decreased vocalizations and decreased Alert behaviors in the second REMOVAL and RETURN. Additionally, YOUNG sows performed more active behaviors than OLD sows, indicating that sows may become less interested in or cannot hear their piglets as they age. Finally, there is some indication that ventilation presence has an effect on sow-piglet communication with ISO-FAN sows having the loudest vocalizations when compared with sows without ventilation noise, indicating that ventilation noise may be a possible competitor with a sow's ability to communicate with her piglets., (© The Author(s) 2018. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Background: Providers often prescribe counseling and/or medications for tobacco cessation without considering patients' treatment preferences., Objective: The primary aims of this study are to describe (1) the development of a discrete choice experiment (DCE) questionnaire designed to identify the attributes and levels of tobacco treatment that are most important to veterans; and (2) the decision-making process in choosing between hypothetical tobacco treatments., Methods: We recruited current smokers who were already scheduled for a primary care appointment within a single Veterans Affairs (VA) healthcare system. Subjects were asked to rate the importance of selected treatment attributes and were interviewed during two rounds of pilot testing of initial DCE instruments. Key attributes and levels of the initial instruments were identified by targeted literature review; the instruments were iteratively revised after each round of pilot testing. Using a 'think aloud' approach, subjects were interviewed while completing DCE choice tasks. Constant comparison techniques were used to characterize the issues raised by subjects. Findings from the cognitive interviews were used to revise the initial DCE instruments., Results: Most subjects completed the DCE questionnaire without difficulty and considered two or more attributes in choosing between treatments. Two common patterns of decision-making emerged during the cognitive interviews: (1) counting 'pros' and 'cons' of each treatment alternative; and (2) using a 'rule-out' strategy to eliminate a given treatment choice if it included an undesirable attribute. Subjects routinely discounted the importance of certain attributes and, in a few cases, focused primarily on a single 'must-have' attribute., Conclusion: Cognitive interviews provide valuable insights into the comprehension and interpretation of DCE attributes, the decision processes used by veterans during completion of choice tasks, and underlying reasons for non'-compensatory decision-making.

Though many studies focused on piglet crushing utilizing piglet vocalizations to test sow response, none have verified the properties of test vocalizations against actual crushing events. Ten sows were observed 48 h after parturition, and crushing events were recorded from all sows. When a crushing event occurred, a second piglet within the same litter was used to solicit a vocalization through manual restraint to compare restrained piglets' call properties to those of crushed piglets'. A total of 659 Restrained calls and 631 Crushed calls were collected. Variables were gathered at the loudest point in a call, and as an average across the entire call. Crushed piglets had a lower fundamental frequency ( p < 0.01; Crushed: 523.57 ± 210.6 Hz; Restrained: 1214.86 ± 203.2 Hz) and narrower bandwidth ( p < 0.01; Crushed: 4897.01 ± 587.3 Hz; Restrained: 6674.99 ± 574.0 Hz) when analyzed at the loudest portion of a call. Overall, piglets which were crushed had a lower mean peak frequency than those which were restrained ( p = 0.01; 1497.08 ± 239.4 Hz and 2566.12 ± 235.0 Hz, respectively). Future research should focus on measuring sow reactivity to Crushed and Restrained piglets to continue to improve research practices.

A novel gel formulation was selected for intravaginal delivery of the GnRH agonist (triptorelin) for synchronizing ovulation in pigs. Studies with gilt models were used to assess LH response profiles. The lowest dose of triptorelin that induced the most gilts to show an LH surge was 100 μg in 1.2% methylcellulose gel. This formulation had a similar effect in weaned sows while also advancing ovulation. The timing of administration was evaluated in sows after weaning. Administration at 96 h induced more sows to ovulate (58%) by 48 h compared to treatment at estrus (45%) or for controls (34%), but the desired level of ovulation synchrony was not achieved. As a result, greater doses of triptorelin were tested and 200 μg given at 96 h after weaning, induced 81% of sows to ovulate within 48 h after treatment. The best synchrony of ovulation occurred when given at 96 h after weaning compared to earlier or later intervals. The optimum time to give a single fixed time AI (SFT-AI) after administration of 200 μg of triptorelin in 1.2% gel (OvuGel ® ) at 96 h after weaning was tested. A SFT-AI at 22 ± 2 h after OvuGel achieved the highest fertility and was practical for staff during the normal work day. In field trials, a SFT-AI 22 ± 2 h after all weaned sows were treated with OvuGel improved (P = 0.04) farrowing rate to 82.5% compared to control sows weaned (80.1%), with no effect on numbers of pigs born alive (12.1). Research continues for identifying the advantages for use of OvuGel in different production systems, and potential application for use in gilts., (Copyright © 2017 Elsevier Inc. All rights reserved.)

In utero stress has been shown to negatively affect intact male rats and mice, though very little research has been conducted in boars. The objectives of the present studies were to determine the effects of in utero heat stress (IUHS) on postnatal development and the response to postnatal heat stress of boars. Ten boars were selected at weaning from litters subjected to IUHS or in utero thermoneutral (IUTN) during 30-60 days of gestation. The boars were evaluated for reproductive performance from birth through 57 weeks of age (WOA). Testicular area tended to be smaller for IUHS boars compared to IUTN boars at 24 WOA (P = 0.080). Libido did not differ for IUHS or IUTN (P = 0.818). Total sperm production was reduced in IUHS boars compared to IUTN boars (P ≤ 0.038). Semen volume and semen concentration did not differ (P ≥ 0.469 and P ≥ 0.664, respectively). Total motility and progressive motility did not differ for IUHS and IUTN boars (P ≥ 0.430 and P ≥ 0.652, respectively). In utero heat stressed boars had a greater incidence of sperm with tail abnormalities than IUTN (P ≤ 0.042). In utero heat stressed boars had a lower incidence of sperm with proximal droplets following mild, postnatal heat stress compared with IUTN (P = 0.005). In utero heat stress resulted in significant reductions in sperm production and increased sperm abnormalities in boars. The IUHS boars may be slightly more tolerant to postnatal heat stress, though more research is needed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Two experiments were conducted to evaluate the use of supplemental betaine in commercially available semen extenders. In experiment 1 ( Exp1 ), semen was collected from six mature boars once weekly for 6 wk (3 wk in summer and 3 wk in winter) and diluted into a commercial extender with the following betaine concentrations: 0, 51, 102, and 205 mM. Semen samples were analyzed on the day of collection ( D0 ) and after 72 h of storage ( D3 ). In experiment 2 ( Exp2 ), semen was collected from four mature boars for 3 wk and was diluted into three commercially available semen extenders (short term, ST ; long term with bovine serum albumin, BSA; and long term without BSA, LT ), with and without supplemental betaine (0 and 70 mM), and analyzed on D0 and D3. Semen was analyzed using computer-assisted sperm assessment (Ceros II, IMV) and morphology using phase contrast microscopy. In Exp1, total motility on D0 was less for 0 mM than that for 102 mM ( P = 0.038) and was substantially reduced for 205 mM compared with 102 mM ( P < 0.001). Supplementation with 205 mM betaine resulted in a significant reduction in the percentage of morphologically normal sperm ( P < 0.001). In Exp2, 70 mM betaine reduced the total motility compared with 0 mM ( P = 0.010) but did not impact percentage of normal sperm ( P = 0.942). The use of supplemental betaine may partially alleviate the dilution effect on sperm, though boar genetics may impact its efficacy. Further research is needed to make a definitive conclusion., (© The Author(s) 2018. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Forty-eight primiparous and multiparous cows were fed a silage-based total mixed ration (TMR; CON), a TMR with 2.5 kg/d DM dried distiller's grains plus solubles (DDGS; MID), or a TMR with 4.7 kg/d DM DDGS (HIGH) from 8.5 ± 4 to 116 ± 4 d postpartum (DPP). All diets were formulated to be isocaloric for a postpartum ADG targeted at 0.22 kg, whereas CP exceeded requirements in both diets containing DDGS. Blood samples were collected from dams at 7- and 21-d intervals for progesterone concentration (resumption of cyclicity) and plasma urea nitrogen, respectively. A weigh-suckle-weigh procedure was used to measure milk production, and samples were collected for milk composition and fatty acid profile. A 5-d Co-Synch + controlled internal drug release (CIDR) protocol was used, and cows were bred (97 ± 4 DPP) by timed AI (TAI). Ultrasonography was used for measurement of the largest antral follicle at TAI and pregnancy diagnosis. Nineteen days after TAI, cow and calf pairs were managed as a single group until weaning and exposed to natural mating for a total of a 60-d breeding season. Starting at weaning, blood samples were collected from heifer offspring at 7-d intervals for progesterone concentration used as an indicator of puberty. A 5-d Co-Synch + CIDR protocol was used, and heifers were bred by TAI. Transrectal ultrasonography was used for measurement of the largest antral follicle at TAI and antral follicle count (AFC) at CIDR device insertion as well as pregnancy diagnosis. There were no differences in dam ADG ( = 0.364); however, DMI decreased with increasing DDGS ( < 0.0001), resulting in an increased feed efficiency ( = 0.042). Milk fat and lactose tended to increase with the addition of DDGS in the diet ( ≤ 0.055), and milk urea nitrogen increased ( < 0.001). There were no differences in resumption of cyclicity ( = 1.00) or diameter of the largest antral follicle ( ≥ 0.372). Timed AI pregnancy rates were not different with the addition of DDGS in the diet ( ≥ 0.174). Heifer ADG was reduced with DDGS during late lactation ( = 0.031). Heifer AFC had a tendency to increase with increasing DDGS ( = 0.081), with no impacts on dominant follicle diameter at TAI ( = 0.161). Heifer TAI pregnancy rate also was not different among treatments ( = 0.508). Overall, feeding 4.7 kg/d DM of DDGS to dams during early lactation increases feed efficiency of the dam, with minor statistically significant effects on dam or heifer offspring reproduction.

Forty-eight primiparous and diparous Angus-Simmental cows were fed 1 of 3 diets; 1) total mixed ration (TMR) based of corn silage and corn stalks (CON), 2) TMR with 3.3 kg/d DM of corn gluten feed (CGF; MID), or 3) TMR with 6.7 kg/d DM of CGF (HIGH). From 11 ± 5 days post-partum (DPP) to 105 ± 5 DPP, all diets were formulated to be isocaloric for a post-partum ADG targeted at 0.22 kg, but CP exceeded requirements in both CGF diets. Blood samples were collected from cows starting at trial initiation until estrous synchronization for determination of plasma progesterone concentration (7 d intervals), as an indicator of resumption of cyclicity, as well as for plasma urea nitrogen (PUN; 21 d intervals). Milk production was assessed at 62 ± 5 DPP via a weigh-suckle-weigh procedure, and milk samples were collected at 64 ± 5 DPP for composition analysis. A 5 d Co-Synch + Control Internal Drug Release (CIDR) protocol was used and cows were bred by timed artificial insemination (TAI). Trans-rectal ultrasonography was used for the evaluation of the dominant follicle at TAI, as well as pregnancy diagnosis. Nineteen days post-TAI, cow and calf pairs were managed as a single group until weaning (205 ± 5 DPP) and exposed to natural mating for a total of a 60 d breeding season. Dam ADG was not significantly different among treatments ( P = 0.849), but, DMI decreased with increasing CGF in the diet ( P = 0.049). There were no differences in final BW ( P = 0.779), however, final BCS was lower in the HIGH treatment when compared to the MID ( P = 0.042). Milk production ( P = 0.457), as well as, milk components ( P ≥ 0.188) were not different, with the exception of milk fat, which tended to be greater in the HIGH treatment ( P = 0.059) when compared to the MID. A treatment by week interaction ( P < 0.0001) was found for PUN concentrations. Concentrations were greater in the HIGH treatment compared to the MID treatment at 63, 84, and 105 d, and greater than the CON treatment at all time points except d-42. There were no differences in resumption of cyclicity ( P = 0.419), dominant follicle ( P = 0.648), or TAI conception rates ( P = 0.761). However, season long pregnancy rates were significantly greater in the CGF treatments when compared to the CON ( P = 0.009). In summary, feeding high or intermediate amounts of CGF neither has a positive nor negative effect on TAI conception rates of beef cows, however, it positively affected season long pregnancy.

Purpose: Extension for Community Health Outcomes (ECHO) is a provider-level telemedicine model successfully applied to hepatitis C care, but little is known about its application to Human Immunodeficiency Virus (HIV) care. We performed a mixed-methods evaluation of 3 HIV ECHO programs in the Veterans Health Administration, focusing on uptake by primary care clinics and veterans., Methods: Administrative data were used to assess program uptake, including adoption (ie, proportion of primary care clinics participating) and reach (ie, proportion of eligible veterans participating). Veterans were considered eligible if they had an HIV diagnosis and lived nearer to a primary care clinic than to the HIV specialty clinic. We interviewed 31 HIV specialists, primary care providers (PCPs), and administrators engaged in HIV ECHO, and we analyzed interview transcripts to identify factors that influenced program adoption and reach., Findings: Nine (43%) of 21 primary care clinics adopted HIV ECHO (range 33%-67% across sites). Program reach was limited, with 47 (6.1%) of 776 eligible veterans participating. Reach was similar among rural and urban veterans (5.3% vs 6.3%). In interviews, limited adoption and reach were attributed partly to: (1) a sense of "HIV exceptionalism" that complicated shifting ownership of care from HIV specialists to PCPs, and (2) low HIV prevalence and long treatment cycles that prevented rapid learning loops for PCPs., Conclusions: There was limited uptake of HIV ECHO telemedicine programs in settings where veterans historically traveled to distant specialty clinics. Other telemedicine models should be considered for HIV care., (© 2016 National Rural Health Association.)

International implementation of the patient-centered medical home (PCMH) model for delivering primary care has dramatically increased in the last decade. A majority of research on PCMH's impact has emphasized the care provided by clinically trained staff. In this article, we report our ethnographic analysis of data collected from Department of Veterans Affairs staff implementing PACT, the VA version of PCMH. Teams were trained to use within-team delegation, largely accomplished through attention to clinical licensure, to differentiate staff in providing efficient, patient-centered care. In doing so, PACT may reinforce a clinically defined culture of care that countermands PCMH ideals. Such competing rubrics for care are brought into relief through a focus on the care work performed by clerks. Ethnographic analysis identifies clerks' care as a kind of emotional dirty work, signaling important areas for future anthropological study of the relationships among patient-centered care, stigma, and clinical authority., (© 2016 by the American Anthropological Association.)

The objective of this study was to evaluate relationships between common semen quality estimates including sperm motility, sperm morphology, spontaneous capacitation status and seminal plasma proteins and boar fertility using heterospermic inseminations and subsequent paternity testing. All boars (n=12) used in the study had excellent semen quality (≥70% normal sperm) that resulted in average farrowing rates and litter sizes of 88.9±0.7% and 11.7±0.1 pigs, respectively. Their ejaculates were combined to make heterospermic insemination doses in such a way that each boar was tested against all of his contemporaries. The proportion of piglets sired by each individual was used to separate boars into three fertility groups: High (71.6±4.8%; n=3); Medium (51.6±3.8%; n=6); and Low (25.2%±5.3%; n=3). Ejaculates from High fertility boars had more motile sperm with normal acrosomes that moved faster in a straight-line and were more likely to undergo an acrosome reaction (p≤0.05) compared with their counterparts in the Low fertility group. Ejaculates from High fertility boars contained the greatest concentrations of three seminal plasma proteins (25.9kD/5.9pI; 55.1kD/4.8pI; and 70.1kD/5.2pI; p≤0.05), whereas concentrations of a 19.1kD/6.8pI were highest in semen from Low fertility boars (p≤0.05). Multiple regression analyses indicated that concentrations of the 25.9kD/5.9pI seminal plasma protein explained 66% of the variation observed in the proportion of pigs sired within a litter among boars (p≤0.00001). These results demonstrate that heterospermic inseminations and subsequent paternity testing is an effective technique for defining relationships between common semen quality tests and fertility, especially in situations where reproductive performance of all the boars is high. Motility, normal acrosome morphology, average linear velocity of motile sperm, and the proportion of sperm capable of an acrosome reaction were all positively associated with boar fertility. However, concentrations of a 25.9kD/5.9pI seminal plasma protein were the best single semen characteristic for ranking boars in terms of their fertility., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Rapidly cooling pigs after heat stress (HS) results in a pathophysiological condition, and because rapid temperature fluctuations may be associated with reduced reproductive success in sows, it lends itself to the hypothesis that these conditions may be linked. Objectives were to determine the effects of rapid cooling on thermal response and future reproductive success in pigs. Thirty-six replacement gilts (137.8±0.9kg BW) were estrus synchronized and then 14.1±0.4 d after estrus confirmation, pigs were exposed to thermoneutral conditions (TN; n=12; 19.7±0.9°C) for 6h, or HS (36.3±0.5°C) for 3h, followed by 3h of rapid cooling (HSRC; n=12; immediate TN exposure and water dousing) or gradual cooling (HSGC; n=12; gradual decrease to TN conditions) repeated over 2 d. Vaginal (T V ) and gastrointestinal tract temperatures (T GI ) were obtained every 15min, and blood was collected on d 1 and d 2 during the HS and recovery periods at 180 and 60min, respectively. Pigs were bred 8.3±0.8 d after thermal treatments over 2 d. Reproductive tracts were collected and total fetus number and viability were recorded 28.0±0.8 d after insemination. HS increased T V and T GI (P=0.01; 0.98°C) in HSRC and HSGC compared to TN pigs. During recovery, T V was reduced from 15 to 105min (P=0.01; 0.33°C) in HSRC compared to HSGC pigs, but no overall differences in T GI were detected (P<0.05; 39.67°C). Rapid cooling increased (P<0.05) TNFα compared to HSGC and TN pigs during recovery-d 1 (55.2%), HS-d 2 (35.1%), and recovery-d 2 (64.9%). Viable fetuses tended to be reduced (P=0.08; 10.5%) and moribund fetuses tended to be increased (P=0.09; 159.3%) in HSRC compared to HSGC and TN pigs. In summary, rapid cooling prior to breeding may contribute to reduced fetal viability and reproductive success in pigs., (Published by Elsevier Ltd.)

Epigenetic modifications established during gametogenesis regulate transcription and other nuclear processes in gametes, but also have influences in the zygote, embryo and postnatal life. This is best understood for DNA methylation which, established at discrete regions of the oocyte and sperm genomes, governs genomic imprinting. In this review, we describe how imprinting has informed our understanding of de novo DNA methylation mechanisms, highlight how recent genome-wide profiling studies have provided unprecedented insights into establishment of the sperm and oocyte methylomes and consider the fate and function of gametic methylation and other epigenetic modifications after fertilization., Competing Interests: Financial & competing interests disclosure KR Stewart was supported by a studentship from the Cambridge Overseas Trusts. L Veselovska was supported by a Babraham Institute: Cambridge University studentship. Work in G Kelsey's laboratory is funded by the Biotechnology and Biological Sciences Research Council and the Medical Research Council. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.

This study evaluated the effect of supplemental dietary betaine at three concentrations (0.0%, 0.63% and 1.26%) on semen characteristics, quality and quality after storage on boars. The trial was conducted between 22 July and 1 October 2014 in a boar stud located in Oklahoma. Boars were blocked by age within genetic line and randomly allotted to receive 0% (CON, n (line T)=22, n (line L)=10), 0.63% (BET-0.63%, n (line T)=21, n (line L)=6) or 1.26% (BET-1.26%, n (line T)=23, n (line L)=7). The diets containing betaine were fed over 10 weeks, to ensure supplemental betaine product (96% betaine) daily intakes of 16.34 and 32.68g, for the BET-0.63% and BET-1.26% diets, respectively. Serum homocysteine concentrations were less for animals with betaine treatments (P=0.016). Rectal temperatures of the boars were unaffected by betaine diets. Betaine tended to increase total sperm in the ejaculates when collectively compared with data of the control animals (P=0.093). Sperm morphology analysis indicated there was a greater percent of sperm with distal midpiece reflex (P=0.009) and tail (P=0.035) abnormalities in boars fed the BET-1.26% than boars fed the BET-0.63% diet. Betaine concentration in the seminal plasma was greater in boars with betaine treatments, with animals being fed the 0.63% and 1.26% diets having 59.2% and 54.5% greater betaine concentrations in seminal plasma as compared with boars of the control group (P=0.046). In conclusion, betaine supplementation at 0.63% and 1.26% tended to increase sperm concentration in the ejaculates by 6% and 13%, respectively, with no negative impacts on semen quality when 0.63% of betaine was included in the diet., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Erasure and subsequent reinstatement of DNA methylation in the germline, especially at imprinted CpG islands (CGIs), is crucial to embryogenesis in mammals. The mechanisms underlying DNA methylation establishment remain poorly understood, but a number of post-translational modifications of histones are implicated in antagonizing or recruiting the de novo DNA methylation complex. In mouse oogenesis, DNA methylation establishment occurs on a largely unmethylated genome and in nondividing cells, making it a highly informative model for examining how histone modifications can shape the DNA methylome. Using a chromatin immunoprecipitation (ChIP) and genome-wide sequencing (ChIP-seq) protocol optimized for low cell numbers and novel techniques for isolating primary and growing oocytes, profiles were generated for histone modifications implicated in promoting or inhibiting DNA methylation. CGIs destined for DNA methylation show reduced protective H3K4 dimethylation (H3K4me2) and trimethylation (H3K4me3) in both primary and growing oocytes, while permissive H3K36me3 increases specifically at these CGIs in growing oocytes. Methylome profiling of oocytes deficient in H3K4 demethylase KDM1A or KDM1B indicated that removal of H3K4 methylation is necessary for proper methylation establishment at CGIs. This work represents the first systematic study performing ChIP-seq in oocytes and shows that histone remodeling in the mammalian oocyte helps direct de novo DNA methylation events., (© 2015 Stewart et al.; Published by Cold Spring Harbor Laboratory Press.)