Low colostrum intake results in potential accumulation of peroxisome lipid substrates in vaginal tissue of 3-week-old gilts.

Low colostrum intake relates to poorer health and infertility in swine. We previously connected vaginal lipid profiles at weaning to fertility of sows. We hypothesized vaginal lipidome varied with colostrum intake. Our objective was to determine whether indicators of colostrum intake, immunocrit (IM) and weight gain 24 h postnatal (PN), related to vaginal lipids at d21 PN. Gilts (n=60) were weighed and blood sampled to measure IM. On d21 PN vaginal swabs were taken and lipids measured using multiple reaction monitoring. Abundance of multiple lipids differed (P<0.05) between gilts categorized as high versus low IM and high versus low 24 h gain. The abundance of multiple lipids correlated with IM and 24 h gain. Phosphatidylcholine PC(36:3), PC(36:2), and arachidonic acid (C20:4) positively (P<0.05) correlated with IM. The ether lipid PCo(38:6) and multiple cholesteryl esters negatively (P<0.05) correlated with IM. ROC analysis indicated arachidonic acid and docosanoic acid (C22:0) may serve as excellent biomarkers that distinguish between high and low IM. Similar to gilts found to be infertile, lipid profiles of low colostrum intake animals had greater abundance of very long chain fatty acids, lipids with high levels of unsaturation, and cholesteryl esters, which are metabolized in peroxisomes indicating their potential dysfunction.

Evaluating the temperature preferences of sexually mature Duroc, Landrace, and Yorkshire boars.

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 m2/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).

Cryoprotectant effects of natural honey on spermatozoa quality of pre-freezing and frozen-thawed boar semen.

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.

To preserve the semen of male pigs for long-term usage, especially for artificial insemination, semen samples are frozen at temperatures below zero degrees. This research study was conducted with the aim of improving the qualities of semen samples from male pigs that are usually negatively impacted by extremely low temperatures during the preservation process using liquid nitrogen. Honey was added to the preservative mixture used because of its known properties that we hypothesized to be beneficial to frozen pig semen. The findings of the experiments conducted revealed that honey improved the movement of sperm cells in semen samples prior to freezing in liquid nitrogen. Qualities of sperm cells of frozen and thawed semen samples of male pigs, such as motion and shape, were better preserved when honey is added to the preservative media.

Processed Meat Characteristics between Commercial Duroc-Sired and Heritage Breed Large Black Pigs.

This study examined the meat processing characteristics between Duroc-sired (DS) and heritage breed Large Black (LB) pigs fed high forage (FIB) or commercial diets (CON). Fifty pigs (DS, n = 25; LB, n = 25) were used in a 2 × 2 factorial design with breed and diet as fixed effects. Processing traits were measured for fresh bellies, bacon slices, and sausage patties. Results showed that DS bellies were longer (p < 0.001) but thinner (p = 0.026). CON bellies were longer (p = 0.005) and thicker (p < 0.001). LB bellies had decreased lean area (p < 0.001) and processing yield (p = 0.001). DS patties were lighter (L*, p < 0.001) and less red (a*, p < 0.001). LB had increased belly firmness (p = 0.053). The CON diet resulted in better particle definition (p = 0.010) in patties as well as a larger slice area (p = 0.048) in bacon slices. A breed × diet interaction was observed for lipid oxidation (p = 0.001). The findings provided novel insight into the comparison between these breeds and diets. While LB pork may have niche market value, the integration of this breed into commercial bacon processing has limitations in composition that need to be further evaluated to improve the product desirability.

Mammary Development in Gilts at One Week Postnatal Is Related to Plasma Lysine Concentration at 24 h after Birth, but Not Colostrum Dose.

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.

Shotgun proteomics of homogenate milk reveals dynamic changes in protein abundances between colostrum, transitional, and mature milk of swine.

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.

Effects of air exposure and agitation on quality of stored boar semen samples.

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.

Biomarkers predictive of long-term fertility found in vaginal lipidome of gilts at weaning.

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.

Effects of induction on the farrowing process and piglet blood parameters at the time of farrowing.

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.

Shotgun proteome analysis of seminal plasma differentiate boars by reproductive performance.

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.

Effects of the number of sperm and site of uterine semen deposition on conception rate and the number of embryos in weaned sows receiving a single fixed-time insemination.

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.

In utero heat stress alters postnatal phenotypes in swine.

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.

Evaluation of on-farm indicators of gilt reproductive performance potential at 21 days of age.

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.

A standardized model to study effects of varying 24-h colostrum dose on postnatal growth and development.

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.

Temporal analysis of vaginal proteome reveals developmental changes in lower reproductive tract of gilts across the first two weeks postnatal.

In swine the upper reproductive tract undergoes early postnatal development, however little is known about the lower reproductive tract. Our objective was to measure cytology and proteome of vaginal swab samples taken on postnatal day (PND) 2 and 16 in gilts to determine if temporal changes occurred in cell and protein profiles during the first two weeks after birth. The posterior vagina was swabbed using a cytology brush on PND 0, 2 and 16 and slides were prepared. The proportion of anuclear and superficial cells increased and parabasal decreased (P < 0.05) from PND 0 to 16. Proteins isolated from vaginal swabs taken on PND 2 and 16 from six gilts across three litters were measured using LC-MS/MS. Over 1500 proteins were identified, with 881 differentially expressed (P-adj < 0.05) between PND 2 and 16. One-third of proteins upregulated between days were categorized as secreted, including lipocalins. Categories enriched by downregulated proteins included cell-cell adherens junction, translation and ER to Golgi vesicle-mediated transport, and reflected increased cornification of stratified epithelium and thus mirrored changes in cytology. Changes in cytology and proteome over the first two weeks after birth support that the porcine vagina continues to develop postnatal.

Lipidome profiles of postnatal day 2 vaginal swabs reflect fat composition of gilt's postnatal diet.

We hypothesized that postnatal development of the vagina is impacted by early nutritional environment. Our objective was to determine if lipid profiles of vaginal swabs were different between postnatal gilts suckled by sow or fed milk replacer the first 48 h after birth, with or without a lard-based fat supplement. Gilts (>1.3 kg) were selected at birth across 8 litters and assigned to one of four treatments: 1) suckled by sow (S, n = 8); 2) suckled by sow plus administration of a fat supplement (SF, n = 5); 3) bottle-fed solely milk replacer (B, n = 8); or 4) bottle-fed solely milk replacer plus administration of a fat supplement (BF, n = 7). At 48 h postnatal, vaginal swabs of gilts were taken with a cytology brush, and lipids were extracted for analysis using multiple reaction monitoring (MRM)-profiling. Lipids extracted from serum collected at 48 h from gilts, milk collected at 24 h from sows, and milk replacer were also analyzed with MRM-profiling. Receiver operating characteristic curve analysis found 18 lipids recovered from vaginal swabs that highly distinguished between S and B gilts [area-under-the-curve (AUC) > 0.9], including phosphatidylethanolamine with 34 carbons and four unsaturations in the fatty acyl residues [PE (34:4)]. Twelve lipids from vaginal swabs highly correlated (r > 0.6; p < 0.01) with nutrition source. Lipids with greater abundance in milk replacer drove association. For example, mean intensity of PE (34:4) was 149-fold higher in milk replacer than colostrum. Consequently, PE (34:4) was found to have 1.6- and 2.12-fold higher levels in serum and vaginal swab samples (p < 0.001), respectively, of B gilts as compared to S gilts. Findings support that vaginal swabs can be used to noninvasively study effects of perinatal nutrition on tissue composition.

The impact of farrowing room noise on sows' reactivity to piglets.

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.

Auditory brainstem responses in weaning pigs and three ages of sows.

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.

Comparison of Vocalization Patterns in Piglets Which Were Crushed to Those Which Underwent Human Restraint.

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.

Diet Impacts Pre-implantation Histotroph Proteomes in Beef Cattle.

In ruminants, the period from fertilization to implantation is relatively prolonged, and the survival of embryos depends on uterine secretions known as histotroph. Our objective was to determine if the pre-breeding diet affected histotroph proteomes in beef cattle. Cows were assigned to one of four diets: a control diet (CON), a high-protein diet (PROT), a high-fat diet (OIL), or a high-protein and high-fat diet (PROT + OIL). After 185 days on these diets, an intravaginal progesterone implant (CIDR) was inserted for 7 days. At 9 days after CIDR removal, animals with a corpus luteum were selected ( n = 16; 4 per treatment). Proteins were isolated from the histotroph collected by uterine lavage and analyzed with liquid chromatography-tandem mass spectrometry. Over 2000 proteins were expressed ( n ≥ 3 cows per treatment), with 1239 proteins being common among all of the groups. There were 20, 37, 85, and 123 proteins unique to CON, PROT + OIL, PROT, and OIL, respectively. Relative to CON, 23, 14, and 51 proteins were differentially expressed in PROT + OIL, PROT, and OIL, respectively. Functional analysis found that 53% of histotroph proteins were categorized as extracellular exosome, 3.28% as cell-cell adhesion, and 17.4% in KEGG metabolic pathways. Differences in proteomes among treatments support the idea that pre-breeding diet affects histotroph. Understanding the impact of diet on histotroph proteins may help improve conception rates.