S. Novak

Nutritional restriction in lactating primiparous sows selectively affects female embryo survival and overall litter development

This study explored the possibility of sex-specific effects on embryonic survival in primiparous sows subjected to restricted feed intake during the last week of lactation and bred after weaning (Restrict; n = 16), compared with control sows fed close to ad libitum feed intakes (Control; n = 17). Restrict sows were in a substantial negative net energy balance at weaning, and lost 13% of estimated protein and 17% of fat mass during lactation, yet the weaning-to-oestrous interval and ovulation rate were not different between treatments. However, embryonic survival at Day 30 of gestation was lower (P < 0.05) in Restrict than Control sows, and selectively reduced the proportion of female embryos surviving (P < 0.01). A decrease in weight and crown-rump length of surviving female (P < 0.05) and male (P < 0.05) embryos was seen in Restrict litters. The mechanisms mediating this sex-specific effect on embryonic loss in feed-restricted sows are unclear. The data presented here indicate that feed-restriction during the last week of lactation in primiparous sows causes a selective decrease in survival of female embryos and limits the growth of all surviving embryos.

Biomarkers of in vivo fertility in sperm and seminal plasma of fertile stallions

The global proteome of sperm and seminal plasma of fertile stallions was investigated to determine whether associations with relative in vivo fertility exist. Seven stallions at stud in a commercial breeding station were collected throughout the breeding season and bred to a total of 164 mares to determine conception rates. On three occasions during the breeding season, raw semen was obtained from a regular collection for proteomic analysis using two-dimensional electrophoresis and also assessed for routine semen quality end points. First cycle conception rate was negatively related to ejaculate volume (r = -0.43, P = 0.05) and total IGF1 content (ng) per ejaculate (r = -0.58, P = 0.006), whereas overall pregnancy rate was positively related to sperm concentration (r = 0.56, P = 0.01). The abundance of three proteins known to be involved in carbohydrate metabolism in sperm was positively related to fertility. Furthermore, the abundance of four seminal plasma proteins were identified as being negatively related to fertility; these were identified as kallikrein-1E2 (KLK2), clusterin, and seminal plasma proteins 1 (SP1) and 2 (SP2). Abundance of cysteine-rich secretory protein 3 (CRISP3) was positively related to first cycle conception rate (r = 0.495, P = 0.027) and may provide a good marker of fertility. Based on stepwise regression analysis, clusterin and SP1 in seminal plasma together with sperm citrate synthase were predictive of fertility (r = 0.77, P < 0.0001). This study identified proteins within sperm and seminal plasma that could serve as biomarkers of semen quality and fertility in stallions.

Seminal Plasma Proteins as Potential Markers of Relative Fertility in Boars

This study investigated whether specific proteins from distinct seminal plasma fractions of boars could be related to in vivo fertility. Nine boars with acceptable sperm motility and morphology for use in artificial insemination demonstrated major differences in total number born and pregnancy rate when low sperm doses (1.5 billion sperm) were used to breed a minimum of 50 gilts per boar. The 2 lowest-fertility and 2 highest-fertility boars were chosen for evaluation of specific seminal plasma proteins. On 4 occasions, semen was collected and separated into 3 fractions based on sperm concentration (Sperm-Peak, Sperm-Rich, and Sperm-Free), and the fractions were analyzed for total protein concentration and abundance of major seminal plasma glycoprotein (PSP-I), AWN-1, and osteopontin protein using Western blotting techniques. The concentrations of these seminal plasma proteins were lower in the Sperm-Peak fractions compared with the Sperm-Free fractions (P < .05). Seminal plasma from the pooled Sperm-Rich fraction used for artificial insemination was also subjected to two-dimensional gel electrophoresis to investigate novel protein markers related to in vivo fertility. Total piglets born (r = -0.76, P = .01) and sperm motility at day 7 (r = -0.74, P = .037) were again negatively correlated with a 22-kDa protein identified by mass spectrometry as PSP-I. However, fertility index and farrowing rate tended to be positively correlated (P < .10) with a 25-kDa protein, identified as glutathione peroxidase (GPX5), an antioxidant enzyme that may protect sperm membranes from oxidative damage. These candidate proteins merit further investigation as markers of fertility in boars.

The time of ovulation in relation to estrus duration in gilts

The objective of this study was to determine time of ovulation, monitored by transcutaneous ultrasonography, relative to the duration of estrus in gilts. We exposed 92 cyclic gilts, Camborough x Canabrid terminal line, at Day 19 of their third estrous cycle to vasectomized boars every 6 h for the detection estrus. Transcutaneous ultrasonography was performed every 6 h, starting 24 h after the onset estrus, to determine time of ovulation. Estrus duration was, on average, 52.6 h (range: 30 to 72 h), and ovulation occurred between 30 and 60 h after the onset of estrus (mean: 44 h), about 85 % of the way through the estrus period. The time of ovulation during estrus was dependent on the duration of estrus (Time of ovulation = (duration of estrus) x 0.409 + 22.7; r = 0.57, P = 0.0001). Prediction of the time of ovulation in relation to duration of estrus is important for determining the optimal time for inseminating gilts. This knowledge would contribute to an improvement in the fertilization rate and in reproductive efficiency of the breeding herd.

Restricted feed intake in lactating primiparous sows. I. Effects on sow metabolic state and subsequent reproductive performance

The effects of feed restriction (60% of anticipated feed intake; Restrict; n=60) during the last week of a 21-day lactation in primiparous sows compared with feeding at 90% of anticipated feed intake (Control; n=60) on sow metabolic state, litter growth and sow reproductive performance after weaning were compared. Metabolisable energy (ME) derived from feed was lower, ME derived from body tissues was higher and litter growth rate was reduced (all P<0.05) in Restrict sows during the last week of lactation. Treatment did not affect weaning-to-oestrus interval, pregnancy rate, ovulation rate, embryonic survival or the number of live embryos (P>0.05) at Day 30 of gestation: However, embryo weight was greater (P<0.05) in Control than in Restrict sows (1.55±0.04vs 1.44±0.04g, respectively). These data suggest the biology of the commercial sow has changed and reproductive performance of contemporary primiparous sows is increasingly resistant to the negative effects of lactational catabolism. Overall, catabolism negatively affected litter weaning weight and embryonic development of the next litter, but the extent to which individual sows used tissue mobilisation to support these litter outcomes was highly variable.

Biological Markers of Boar Fertility

The semen evaluation techniques used in most commercial artificial insemination centers, which includes sperm motility and morphology measurements, provides a very conservative estimate of the relative fertility of individual boars. As well, differences in relative boar fertility are masked by the widespread use of pooled semen for commercial artificial insemination (AI) in many countries. Furthermore, the relatively high sperm numbers used in commercial AI practice usually compensate for reduced fertility, as can be seen in some boars when lower numbers of sperm are used for AI. The increased efficiency of pork production should involve enhanced use of boars with strong reproductive efficiency and the highest genetic merit for important production traits. Given that the current measures of semen quality are not always indicative of fertility and reproductive performance in boars, accurate and predictive genetic and protein markers are still needed. Recently, significant efforts have been made to identify reliable markers that allow for the identification and exclusion of sires with reduced reproductive efficiency. This paper reviews the current status of proteomic and genomic markers of fertility in boars in relation to other livestock species.

Feed Restriction and Insulin Treatment Affect Subsequent Luteal Function in the Immediate Postovulatory Period in Pigs: Progesterone Production In Vitro and Messenger Ribonucleic Acid Expression for Key Steroidogenic Enzymes

Abstract Progesterone production and release in vitro, and mRNA expression for key steroidogenic enzymes, were studied in luteal tissue recovered in the immediate postovulatory period from cyclic gilts allocated to one of three treatments: moderate feed restriction during the first (RH) or second week of the estrous cycle, with (HR+I) or without (HR) concomitant injections of long-acting insulin. Time of feed restriction affected neither progesterone production or release, nor mRNA expression for several key steroidogenic enzymes. However, luteal tissue from RH but not from HR gilts responded to LH stimulation by increasing progesterone production and release (P < 0.05). Insulin treatment increased progesterone production and release, restored luteal tissue responsiveness to LH, up-regulated steroidogenic enzyme mRNA expression, and down-regulated the tissue inhibitor of metalloproteinase-I mRNA expression in HR+I compared with HR gilts (P < 0.05). In vitro progesterone production and gene expression were affected by time of tissue collection after ovulation in RH and HR gilts but not in HR+I gilts, and were correlated with temporal changes in oviductal and peripheral plasma progesterone concentrations. Inherent differences in luteal function therefore appear to mediate latent effects of nutrition and insulin treatment on circulating progesterone concentrations in the critical postovulatory period in gilts.

Uterine crowding in the sow affects litter sex ratio, placental development and embryonic myogenin expression in early gestation

Uterine crowding in the pig results in intrauterine growth restriction (IUGR), and permanently affects fetal muscle fibre development, representing production losses for the commercial pig herd. The present study sought to understand how different levels of uterine crowding in sows affects muscle fibre development in the early embryo at the time of muscle fibre differentiation and proliferation. Sows either underwent surgical, unilateral oviduct ligation (LIG; n = 10) to reduce the number of embryos in the uterus, or remained as intact, relatively-crowded controls (CTR; n = 10). Embryos and placentae were collected at Day 30 of gestation, and myogenic regulatory factor (MRF) transcript abundance was determined using real-time PCR for both myogenin (MYOG) and myoblast differentiation 1 (MYOD1). Unilateral tubal ligation resulted in lower numbers of embryos in utero, higher placental weights and a higher male : female sex ratio (P < 0.05). Relative MYOD1 expression was not different, but MYOG expression was higher (P < 0.05) in the LIG group embryos; predominantly due to effects on the male embryos. Relatively modest uterine crowding therefore affects MRF expression, even at very early stages of embryonic development, and could contribute to reported differences in fetal muscle fibre development, birthweight and thus post-natal growth performance in swine.

Pig preovulatory oocytes modulate cumulus cell protein and gene expression in vitro.

This study investigated the changes in protein and gene expression in oocytectomized cumulus cells (OOX) of medium-sized follicles from gilts, cultured with or without denuded oocytes isolated from large oestrogenic sow follicles. Proteomic analysis identified 14 proteins that were differentially expressed in OOX, of which the protein 14-3-3 eta, a signal transduction pathway modulator, was down-regulated in the presence of oocytes. Oocyte co-culture also down-regulated FSHR mRNA expression in OOX, as measured by real-time PCR, and FSHR and 14-3-3 eta mRNA abundance were positively correlated. The oocyte also up-regulated HSD3B mRNA, suggesting an effect on cumulus cell progesterone synthesis. Together with data on gene expression in granulosa cells during the follicular phase of the sow oestrous cycle, this study suggests that modulation of the expression of steroidogenesis related proteins and genes in cumulus cells by the porcine preovulatory oocyte reflects the specific physiological requirements of the preovulatory follicle.

Restricted feed intake in lactating primiparous sows. II. Effects on subsequent litter sex ratio and embryonic gene expression

Expression of panels of candidate genes controlling myogenesis, angiogenesis and gender-specific imprinting of development were analysed in embryonic, placental and endometrial tissues recovered at Day 30 of gestation from a subset of primiparous sows that were either feed restricted (Restrict; n=17) or fed to appetite (Control; n=15) during the last week of the previous lactation. Embryos were also sex typed to investigate gender bias in response to treatments. Average embryonic weight was lower in the subset of Restrict compared with Control litters (1.38±0.07vs 1.59±0.08g, respectively) and the male:female sex ratio was higher (P<0.05) in embryos (litters) recovered from Restrict sows. Treatment affected (P≤0.05) the expression of embryonic and placental genes involved in insulin-like growth factor (IGF) 2 signalling, including IGF2, INSR and IGF2R. Embryonic expression of ESR1 was also affected by treatment (P<0.03) and sex×treatment interactions were observed for the expression of embryonic ESR1 (P<0.05) and placental ANGPT2 (P<0.03). At the molecular level, these results support the suggestion that changes in placental function are not the primary mechanism mediating detrimental effects of previous sow catabolism on early embryonic development in the feed-restricted lactational sow model. However, perturbations in the IGF2 system are implicated as mediators of these effects.