Variations of blood metabolites in single- and multiple-bearing Ossimi ewes during the transition period.

Evaluation of the metabolic profile indices allows early detection and treatment of various metabolic disorders during the transition period in ewes. This study aimed to determine the variations in the blood metabolites around lambing in Ossimi ewes. The blood metabolites were investigated in ewes with single (n = 27) and multiple (n = 9) lambs at 3- and 1-week pre-lambing and 3-week post-lambing. The plasma concentrations of glucose were higher in single-bearing ewes than those in multiple-bearing ewes (p < .05), moreover, its lowest value was measured at 1-week prepartum in both groups. Throughout the study period, the serum concentrations of non-esterified fatty acids (NEFA) were significantly increased in ewes with multiple lambs compared to ewes with single lambs (p < .05), and the highest value was found at 1-week before parturition in both groups. In addition, the serum level of beta-hydroxybutyric acid (BHBA) was higher at 3-week postpartum, and it was significantly increased in multiple-bearing ewes than that in single-bearing ones (p < .05) at 3-week pre-lambing. In both groups, the lowest values of total proteins were determined 1-week before lambing, and its concentrations, at 3- and 1-week prepartum, were higher in ewes with single lambs than those with multiple lambs (p < .05). In contrast, the serum concentrations of albumin were significantly lowered 1-week postpartum (p < .05), and without significant differences between both groups (p > .05). The serum activities of aspartate aminotransferase (AST) and gamma-glutamyltransferase (GGT) were significantly increased at 1-week after parturition in both groups (p < .05). Furthermore, the serum activities of AST were higher in multiple-bearing ewes than those in single-bearing ones at 3-week pre-lambing and 3-week post-lambing (p < .05). Variable positive and negative correlations were determined among the blood metabolites. In conclusion, physiological adaptations are associated with the fluctuation of the blood metabolites around lambing. The higher the number of foetuses the higher the metabolic variations in Ossimi ewes. Therefore, regular metabolic profiling for health monitoring may be necessary to avoid disease development during the transition period.

Sheep abortions associated with Neospora caninum and Toxoplasma gondii infections in multiple flocks from Southern Brazil.

Neosporosis and toxoplasmosis are important parasitic causes of abortions in small ruminants. This study verified the occurrence of these diseases in sheep fetuses from Santa Catarina State, Southern Brazil from 2015 to 2022. Sheep fetuses were necropsied with organ sampling for histopathology, and polymerase chain reaction (PCR) for Neospora caninum and Toxoplasma gondii using the Nc5 and SAG2 targets, respectively, in frozen brain tissue. Microbiological culture and RT-PCR for Pestivirus were conducted to discard other abortion causes. One positive fetus for toxoplasmosis was genotyped using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) with ten genetic markers. Fifty-five sheep fetuses were evaluated, with 10 (18.2%) cases of neosporosis and 7 (12.7%) cases of toxoplasmosis, comprising six and four flocks, respectively. Macroscopically, neosporosis abortions exhibited fetal mummification, maceration, and arthrogryposis. Toxoplasmosis abortions showed fetal mummification and maceration. The neosporosis abortions included lymphoplasmacytic myositis (70%; 7/10) and myocarditis (60%; 6/10), in addition to necrotizing encephalitis and gliosis (50%; 5/10). Toxoplasmosis abortions included lymphoplasmacytic necrotizing encephalitis (71.4%; 5/7), lymphoplasmacytic myositis (42.8%, 3/7), and myocarditis (14.3%; 1/7). Through PCR, N. caninum and T. gondii were detected in 6 (60%) and 5 (71.4%) fetuses, respectively. In one fetus, T. gondii genotyping was conducted, which was characterized as atypical genotype ToxoDB #98. All of the cases were negative for Pestivirus and bacterial agents. This study establishes the occurrence of these diseases as causes of abortions, malformations, mummification, and fetal maceration in sheep, with the characterization of an atypical T. gondii genotype in one of the fetuses.

Validation and Analysis of COIL, a Gene Associated with Multiple Lambing Traits in Sheep.

In a past study, the team used specific-locus amplified fragment sequencing (SLAF sequencing) to detect single-nucleotide polymorphisms (SNPs) contributing to the differences in lambing numbers in Xinjiang sheep. This study verified the correlation between the COIL gene and lambing number characters in sheep and explored its possible mechanism of action. In this study, three SNPs in the COIL gene, namely COILSNP1 (rs7321466), COILSNP2 (rs7314134), and COILSNP3 (rs7321563), were explored in terms of their possible mechanism of action. A tissue expression profiling analysis revealed that the COIL gene was significantly more expressed in the uterus and ovaries than in other tissues (p < 0.05), whereas an association analysis revealed that the number of lambs born was significantly different among individuals with different genotypes of this COILSNP1 (p < 0.05). The Cell Counting Kit-8(CCK-8) revealed that the overexpression of the COIL gene significantly increased the proliferation of mouse ovarian fibroblasts and sheep fibroblasts (p < 0.05). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) revealed that the overexpression of the COIL gene significantly increased the activity of sheep fibroblasts (p < 0.01) and mouse ovarian fibroblasts (p < 0.05). The overexpression of the COIL gene affected the biogenesis pathway of spliceosomal U snRNPs by validating protein network connections. This activity affects ovulation, embryonic development, and changes in lambing size in sheep.

MitoQ preserves the quality and fertility of liquid-preserved ram sperm.

Supplementing the semen extender with some antioxidants may preserve sperm quality following liquid preservation. The aim of the current study was to evaluate the influence of the use of MitoQ in the semen extender on quality parameters and fertility of liquid-preserved ram semen. In this study, diluted semen samples were divided into five parts and supplemented with 0, 1, 10, 100 and 1000 nM MitoQ. The prepared samples were stored at 3-5 °C for up to 50 h. Motility, viability, mitochondrial activity, membrane integrity, and malondialdehyde concentration of the chilled sperm were assessed at 0, 25, and 50 h. To evaluate reproductive performance, artificial insemination was performed with semen liquid-preserved for 25 h. In results, at 0 h, no difference between the groups was observed. The use of 10 and 100 nM MitoQ resulted in higher (P ≤ 0.05) total motility, progressive motility, membrane integrity, mitochondrial activity, viability, and lower malondialdehyde concentration than the other groups after 25- and 50-h storage. Pregnancy, parturition and lambing rates were higher (P ≤ 0.05) when ewes were inseminated with 25-h chilled semen samples containing 10 and 100 nM MitoQ compared to the control. Therefore, supplementing the semen extender with MitoQ (10 and 100 nM) could be an efficient method to improve the quality and fertility rate of liquid-preserved ram semen.

Single-cell sequencing reveals the reproductive variations between primiparous and multiparous Hu ewes.

BACKGROUND: In the modern sheep production systems, the reproductive performance of ewes determines the economic profitability of farming. Revealing the genetic mechanisms underlying differences in the litter size is important for the selection and breeding of highly prolific ewes. Hu sheep, a high-quality Chinese sheep breed, is known for its high fecundity and is often used as a model to study prolificacy traits. In the current study, animals were divided into two groups according to their delivery rates in three consecutive lambing seasons (namely, the high and low reproductive groups with ≥ 3 lambs and one lamb per season, n = 3, respectively). The ewes were slaughtered within 12 h of estrus, and unilateral ovarian tissues were collected and analyzed by 10× Genomics single-cell RNA sequencing. RESULTS: A total of 5 types of somatic cells were identified and corresponding expression profiles were mapped in the ovaries of each group. Noticeably, the differences in the ovary somatic cell expression profiles between the high and low reproductive groups were mainly clustered in the granulosa cells. Furthermore, four granulosa cell subtypes were identified. GeneSwitches analysis revealed that the abundance of JPH1 expression and the reduction of LOC101112291 expression could lead to different evolutionary directions of the granulosa cells. Additionally, the expression levels of FTH1 and FTL in mural granulosa cells of the highly reproductive group were significantly higher. These genes inhibit necroptosis and ferroptosis of mural granulosa cells, which helps prevent follicular atresia. CONCLUSIONS: This study provides insights into the molecular mechanisms underlying the high fecundity of Hu sheep. The differences in gene expression profiles, particularly in the granulosa cells, suggest that these cells play a critical role in female prolificacy. The findings also highlight the importance of genes such as JPH1, LOC101112291, FTH1, and FTL in regulating granulosa cell function and follicular development.

Reproductive performance and productivity of local and Dorper x local crossbred ewes under community-based management system, Ethiopia.

This study evaluated the reproductive and productivity of local and Dorper crossbred ewes in a community-based management system. We analyzed data collected from 2013 to 2021, taking into account different factors such as dam-breed, location, type of birth, season, and year of lambing. Lambing was observed all year-round, but the majority (35%) occurred in September, October, and December. This suggests that pasture availability, which is influenced by climatic-factors, may play a role in the seasonality of lambing. Litter-size at birth and weaning did not show any significant difference. Age at first lambing varied between breeds (P < 0.001), while lambing-interval and annual reproductive rates were unaffected by dam-breed (P > 0.05). The annual number of lambing per year significantly varied based on location and lambing seasons (P < 0.001), with a higher frequency during the major rainy-season compared to the dry-season (1.58vs1.42), highlighting the influence of feed availability. Productivity indices of ewes were calculated. Location and season of lambing had a significant impact on annual ewe productivity, while the ewe genotype showed no significant influence on productivity indices, except for the weight of lambs produced per kilogram of metabolic weight (0.84vs0.72 lambs per kg ewe and year; P < 0.01: 2.02vs1.77 kg lamb per kg0.75 ewe and year), where local ewes outperformed Dorper crossbred ewes. The difference in annual-productivity indices between local and Dorper crossbred ewes was more evident when considering the postpartum weight, as the ewes exhibited higher postpartum weights. However, both ewe genotypes produced comparable lamb weights per year (20.91vs20.16 kg lamb weaned per ewe and year for local and Dorper crossbred ewes, respectively). In summary, under low-input conditions, Dorper crossbred ewes demonstrated comparable reproductive performances and productivity traits to local ewes. Nevertheless, breed and environmental factors identified in this study should be taken into account to enhance sheep productivity in both local and Dorper crossbred ewes.

Mutation of the ETS1 3'UTR interacts with miR-216a-3p to regulate granulosa cell apoptosis in sheep.

ETS1, an important member of the ETS transcription factor family, is involved in a variety of physiological processes in living organisms, such as cell development, differentiation, proliferation and apoptosis, and is thought to be associated with embryonic development and reproduction. However, the polymorphism of ETS1 has been rarely studied, and its potential impact on the formation of reproductive traits in sheep remains unclear. Here, we first analyzed polymorphisms of ETS1 in a population of 382 small-tailed Han sheep with a lambing number record using the Kompetitive Allele Specific PCR (KASP) technique. The results showed the presence of a SNP locus rs161611767 (T > C) in the 3'UTR of ETS1. The association analysis showed the lambing number of first, second and third parity in the individuals with the CC genotype (2.51 ± 0.108, 2.51 ± 0.179, 1.27 ± 0.196) was higher than that of individuals with the TT genotype (1.79 ± 0.086, 1.56 ± 0.102, 0.56 ± 0.100) (P < 0.05). Then, molecular biotechnologies were used to investigate the effects of the EST1 rs161611767 mutant locus on host gene expression in sheep and the underlying mechanism of its effect on sheep reproduction. The RT‒qPCR results showed that the expression of ETS1 was higher in individuals with the CC genotype than in those with the TT genotype (P < 0.05). The dual luciferase reporter assay showed that the luciferase activity of ETS1 in sheep with the TT genotype was decreased compared to CC genotype (P < 0.05), confirming the existence of EST1 rs161611767 in the 3'UTR as a functional SNP. Given that the 3'UTR is an important regulatory region of gene transcription and translation, we performed bioinformatics prediction and confirmed that the SNP rs161611767 of ETS1 was a direct functional target of miR-216a-3p using dual luciferase activity assay, and the binding capacity of allele T was stronger than that of allele C. Subsequently, the cell transfection results showed that miR-216a-3p suppressed the endogenous expression of ETS1 in sheep primary granulosa cells (GCs). Finally, CCK-8, EdU, WB detection of marker proteins and flow cytometry were used to detect the effects of miR-216a-3p on GCs viability and proliferation/apoptosis, respectively. The results showed that miR-216a-3p inhibited the proliferation of GCs while promoting apoptosis of GCs. In conclusion, these results demonstrate that the SNP rs161611767 of ETS1 is associated with lambing number in small-tailed Han sheep, and miR-216a-3p can act as a regulatory element binding to the T mutation in rs161611767 to regulate ETS1 expression and affect GCs development, which may indirectly affect the number of lambs in sheep. These studies provide evidence for the involvement of ETS1 polymorphisms in sheep reproduction and are expected to provide new insights to elucidate the molecular genetic mechanisms of lambing traits in sheep.

Changes in movement, habitat use, and response to human disturbance accompany parturition events in bighorn sheep (Ovis canadensis).

Parturition and the early neonatal period are critical life history stages in ungulates with considerable implications for population growth and persistence. Understanding the changes in behaviour induced by ungulate parturition is important for supporting effective population management, but reliably identifying birth sites and dates presents a challenge for managers. Rocky Mountain bighorn sheep (Ovis canadensis canadensis) are one such highly valued and ecologically important species in montane and subalpine ecosystems of Western North America. In the face of changing patterns of anthropogenic land use, wildlife managers increasingly require site-specific knowledge of the movement and habitat selection characteristics of periparturient sheep to better inform land use planning initiatives and ensure adequate protections for lambing habitat. We used movement data from GPS collared parturient (n = 13) and non-parturient (n = 8) bighorn sheep in Banff National Park, Canada to (1) identify lambing events based on changes in key movement metrics, and (2) investigate how resource selection and responses to human use change during the periparturient period. We fit a hidden Markov model (HMM) to a multivariate characterization of sheep movement (step length, daily home range area, residence time) to predict realistic lambing dates for the animals in our study system. Leave-one-out cross validation of our model resulted in a 93% success rate for parturient females. Our model, which we parameterized using data from known parturient females, also predicted lambing events in 25% of known non-parturient ewes in a test dataset. Using a latent selection difference function and resource selection functions, we tested for postpartum changes in habitat use, as well as seasonal differences in habitat selection. Immediately following lambing, ewes preferentially selected high-elevation sites on solar aspects that were more rugged, closer to escape terrain, and further from roads. Within-home range habitat selection was similar between individuals in different reproductive states, but parturient ewes had stronger selection for low snow depth, sites closer to barren ground, and sites further from trails. We propose that movement-based approaches such as HMMs are a valuable tool for identifying critical parturition habitat in species with complex movement patterns and may have particular utility in study areas without access to extensive field observations or vaginal implant transmitters. Furthermore, our results suggest that managers should minimize human disturbance in lambing areas to avoid interfering with maternal behaviour and ensure access to a broad range of suitable habitat in the periparturient period.

Potential role for human menopausal gonadotropin in ovine superovulatory regimens during the breeding season.

Human menopausal gonadotrophin (hMG) has been reported to produce a comparable superovulatory response to that of follicle stimulating hormone (FSH). Furthermore, hMG has a long half-life as compared with FSH. The present study was designed to compare hMG administered once daily and FSH administered twice daily over a 4 - day period on superovulatory response of Suffolk ewes. During the mid-luteal phase, twenty-four Suffolk donor ewes received intravaginal sponges at day 0 for 12 days. The superovulatory regimens in the Control group (n = 12) and the Treatment group (n = 12) consisted of eight injections of FSH given at twice daily and four injections of hMG given at once daily, respectively. At day 13, the donor ewes were subjected to laparoscopic insemination. Embryos were recovered, classified, and transferred to recipient ewes at day 19. Pregnancy status was determined by ultrasound examination 40 days after transfer. Lambing rate was calculated after all the ewes had delivered. No significant differences were observed between the two groups in terms of the structures recovered, transferable embryos, degenerated embryos, unfertilized oocytes, pregnancy rate and lambing rate. The results showed that once daily injection of hMG can produce a comparable superovulatory response and embryo transfer outcomes to those obtained by twice daily injection of FSH over a 4 - day period. It is feasible that hMG is used to replace FSH and reduce the number of injection treatments in ovine superovulatory regimens.

On-station comparative analysis of reproductive and survival performance between Red Maasai, Dorper, and Merino sheep breeds.

The reproductive performance of ewes and the survivability of lambs to weaning have a critical economic impact on sheep farming worldwide. Further, knowledge of major mortality causes allows an opportunity for improved flock management to evade financial losses. The maximum likelihood estimates for generalised linear mixed models and chi-square test methods were used to examine 971 mating records, 839 and 763 lambs born and weaned (singles or twins) from the Naivasha Sheep and Goats station in Kenya for the years 2011 to 2020 consisting of Dorper, Red Maasai (RedM), and Merino breeds. The RedM (P < 0.05) outperformed Dorper and Merino in weaning rate, whereas reproductive performance between the three breeds was not significantly different (P > 0.05) in litter size and multiple lambings per ewe lambing. On the one hand, Dorper significantly (P < 0.05) outperformed the other two breeds only in weaning weight per lamb born. In addition, among all the major causes of death, pneumonia appeared to be the one to which Dorper breeds were most susceptible (chi-square test, P < 0.05). According to the findings of this study, neither the Dorper nor the Merino sheep breeds were reproductively superior to the RedM in an extensive semi-arid production environment. In addition, Dorper's susceptibility to the leading causes of mortality, particularly pneumonia and sheep pox, were relatively high compared to other breeds and could be a precursor to massive economic losses for Dorper sheep producers. In contrast to the indigenous RedM breed, imported sheep breeds appeared to be more susceptible to major mortality-related under an extensive production system. Therefore, regardless of weaning weight, RedM breed production appears to be a more viable investment for small-scale farmers, particularly in semi-arid regions.

Effects of exogenous GnRH administration on lambing performance of oestrus-synchronized Kazak ewes during the breeding season.

The objective of this study was to investigate the effects of GnRH at insemination on lambing performance of Kazak ewes. Oestrus was synchronized in 774 adult Kazak ewes using an intravaginal sponge impregnated with flurogestone acetate. The sponge was left in the vagina for 12 days followed with an injection of 330 IU of eCG at sponge removal. Each ewe was inseminated twice at 48 h and 60 h after sponge removal. The treatment group (n = 387) was intramuscularly injected at the first insemination with a dose of 25 µg GnRH and the control group (n = 387) with saline solution. The results showed that GnRH administration significantly decreased the fertility rate. In addition, significant differences were observed between the two groups in terms of the twin birth rate, the abortion rate and the litter size. In conclusion, GnRH administration at insemination was not recommended in Kazak ewes undergoing the fixed time artificial insemination during the breeding season. The breed may be a critical determinant of the potential for the exploitation of GnRH application in sheep breeding programmes.

Patterns of Reproductive Management in Sheep and Goat Farms in Greece.

This paper reports findings regarding patterns of reproductive management in 325 sheep flocks and 119 goat herds throughout Greece. The objectives were to describe the patterns of reproductive management in sheep and goat farms, to highlight factors that were associated with these management patterns and the clinical application of the various reproductive management approaches in the farms and to establish potential associations with production outcomes. The median months of the start of the mating period for adult sheep and goats were May and June, respectively and for ewe lambs and replacement goats these were August and September, respectively. The median duration of the mating period was 2 months for adults and 1 month for replacements. The median ratio of female to male animals was 22:1 and 25:1 in sheep and goat farms, respectively. Control of reproduction was applied in 33.2% and 16.8% of sheep and goat farms, respectively, mostly by intravaginal progestagen sponges and less often by melatonin implants; multivariable analysis indicated five (availability of milking parlour, number of ewes in a farm, number of daily milkings, daily period spent by farmers at the farm, farming tradition in the family) and two (number of daily milkings, availability of milking parlour) predictors in sheep and goat farms, respectively. Pregnancy diagnosis ultrasonographically was performed in 36.6% and 16.8% of sheep and goat farms, respectively; multivariable analysis indicated as significant three (management system applied in farms, age of farmer, farming tradition in the family) and two (management system applied in farms, availability of milking parlour) predictors in sheep and goat farms, respectively. The control of reproduction and pregnancy diagnosis were associated with a higher number of vaccinations during pregnancy: 2.6 and 2.7, respectively, versus 2.2 and 2.1 in farms where they were not performed. The average per farm number of lambs/kids born per female animal was 1.33 ± 0.01 and 1.30 ± 0.02 for sheep and goat farms, respectively; five (control of reproduction, location of the farm, presence of working staff, collaboration with veterinary practice, breed of ewes) and two (control of reproduction, breed of female goats) predictors were identified in sheep and goat farms, respectively, for high numbers of lambs/kids. Lambs/kids were taken away from their dams at the age of 50 and 65 days, respectively. The average culling age (females, males) was 5.9 and 4.4 years and 6.9 and 4.9 years for sheep and goats, respectively. Farmers sourced animals to be used as replacements for those removed from the farm, mostly from the animals in their own farms, considering criteria related to milk production. Finally, the application of the control of reproduction was associated with increased milk production and the number of newborns, whilst an inverse correlation between the culling age of animals and milk production was seen.

Bio-economic modelling of sheep meat production systems with varying flock litter size using field data.

Sheep meat producers derive the majority of income from sales of weaned lambs, determined by flock conception rates, litter size, and lamb survival. Field data from commercial flocks can inform sensitivity analyses of the effect of litter size on flock productivity, feed demand, and gross margin. This study adapted an established bio-economic model of a flock of breeding ewes informed by statistical relationships (from linear models) between flock litter size (lambs born per ewe lambing) and production factors (such as flock barren rate, litter birth type and lamb birth weight) identified using 156 145 animal records from the Irish national sheep breeding database. Sensitivity analyses were undertaken to investigate the effects of flock litter size on flock production, feed demand, and gross margin. Results showed that as flock litter size increased, the proportion of lambs born as multiples increased, with 14 % of lambs born as singles when flock litter size was 2.2 lambs born per ewe lambing. Flock gross margin increased from €2 205 to €7 730 as litter size increased from 1.0 to 2.0 lambs born per ewe lambing. As litter size increased from 1.0 to 2.2 lambs born per ewe lambing, flock gross margin increased linearly by, on average, €52 per 0.01 increase in litter size. At a litter size of > 2.2 lambs born per ewe lambing, flock gross margin increased on average €12 per 0.01 increase in litter size. At a litter size of 2.2 lambs born per ewe lambing, flock efficiency (at 65.0 kg of lamb weaned per ewe presented for breeding), weaning rate (at 1.5 lambs weaned per ewe presented for breeding; not including excess lambs from large litters sold within a week after birth and thus not weaned on-farm), and gross margin (at €8 500) began to plateau. The results indicate lower marginal returns in gross margin at very high flock litter size, due to the lower value of additional lambs born as triplets and quadruplets compared with single- and twin-born lambs. However, the diminishing economic returns occurred at higher flock litter size than are currently biologically achieved in most flocks. Quantification from this analysis demonstrates how the value of increasing the number of lambs born changes at very high flock litter size, which can inform the priorities and performance benchmarking for international sheep meat production industries.

Optimal time for laparoscopic intrauterine insemination performed on ewes detected in natural heat.

The aim of this study was to optimize the laparoscopic intrauterine insemination (LAI) methodology by testing different time intervals between the natural heat detection and ewe insemination. Three experiments were performed in the breeding conditions of Southern Kazakhstan. Ewes (n = 243) were exposed for one hour to direct contact with the teaser rams (once a day, morning or evening). Ewes expressing behavioral symptoms of heat were detected and inseminated with the use of LAI during 210-1290 min (3.5-21.5 hrs) after heat detection. Reproductive traits of lambing rate (LR) and litter size (LS) were recorded according to the births registered at 137 to 152 days post insemination. Our statistical model showed significance only for the effects of ewe age category and the time interval from heat detection to LAI on the LR attribute. The highest LR (38.8%) was detected in ewes at 2.5-3.5 years of age. Corrected least-square means of LR indicated 18.5 hrs. as an optimal time for LAI of ewes in natural heat. In the present study, the percentage value of lambing rate obtained at 18.5 h interval was 70.7%. Therefore, our study suggested an effective methodology to spread valuable genetic information in the sheep population in the regions of extensive farming where heat cycle synchronization is not usually performed. Importantly, our study is among the first ones that follow the European strategy to eliminate the occurrence of hormones in livestock production and the environment.

CYP19A1 May Influence Lambing Traits in Goats by Regulating the Biological Function of Granulosa Cells.

Abnormal expression of CYP19A1, a gene related to steroid hormone synthesis, causes steroid hormone disruption and leads to abnormal ovulation in granulosa cells. However, the exact mechanism of CYP19A1 regulation is unclear. In this study, we confirmed the localization of CYP19A1 in goat ovarian tissues using immunohistochemistry. Subsequently, we investigated the effects of CYP19A1 on granulosa cell proliferation, steroid hormone secretion, and expression of candidate genes for multiparous traits by overexpressing and silencing CYP19A1 in goat granulosa cells (GCs). The immunohistochemistry results showed that CYP19A1 was expressed in all types of follicular, luteal, and granulosa cells, with subcellular localization results revealing that CYP19A1 protein was mainly localized in the cytoplasm and nucleus. Overexpression of CYP19A1 significantly increased the mRNA levels of CYP19A1, FSHR, and INHBA, which are candidate genes for multiple birth traits in goats. It also promoted cell proliferation, PCNA and Cyclin E mRNA levels in granulosa cells, and secretion of estrogen and progesterone. However, it inhibited the mRNA levels of STAR, CYP11A1, and 3ßSHD, which are genes related to steroid synthesis. Silencing CYP19A1 expression significantly reduced CYP19A1, FSHR, and INHBA mRNA levels in granulosa cells and inhibited granulosa cell proliferation and PCNA and Cyclin E mRNA levels. It also reduced estrogen and progesterone secretion but enhanced the mRNA levels of STAR, CYP11A1, and 3ßSHD. CYP19A1 potentially influenced the lambing traits in goats by affecting granulosa cell proliferation, hormone secretion, and expression of candidate genes associated with traits for multiple births.

Interactions between Ewes and Rams during Mating Can Be Used to Predict Lambing Dates Accurately, but Not Sire.

Ewes often lamb over extended periods so the level of nutrition during pregnancy and lambing may be suboptimal for ewes that conceived later during mating. Predicting lambing dates would allow cohorts of ewes with similar gestational ages to be managed more precisely to achieve targets for ewe nutrition, feed on offer, mob sizes and access to shelter to improve lamb survival. The interactions between ewes and rams during mating have been used to predict the time of oestrus and lambing dates successfully, but this has not been tested at a commercial scale. In this study, proximity sensors were used to measure interactions between inexperienced Merino ewes (n = 317) and experienced rams (n = 9) during a 27-day mating period under commercial production conditions. When the gestation length was assumed to be 150 days, 91% of lambing dates were predicted within ±6 days of the actual birth date of lambs and 84% of lambing dates were predicted within ±3 days. The use of proximity sensors during mating was an effective means of predicting lambing dates, and there was no significant difference in accuracy for single bearing verses multiple bearing ewes. However, DNA parentage data showed the ram corresponding with the maximum daily interactions ratio was the sire for only 16% of all progeny, suggesting they could not be used to indicate the sire of the progeny.

Prolonging the time of semen deposition increases the pregnancy rates of ewes subjected to fixed time cervical insemination during the breeding season.

The main limiting factor of artificial cervical insemination in ewes is the long and narrow fibrous cervical canal, which impedes the transport of spermatozoa and leads to lower pregnancy rates. The hypothesis that prolonging the time of semen deposition during ovine cervical insemination can increase pregnancy rates was investigated in this study. Estrus was synchronized in 150 multiparous Ujimqin ewes using a polyurethane intravaginal sponge impregnated with 45 mg of flurogestone acetate. The sponge was left in the vagina for 12 days followed by an injection of 330 IU of eCG at sponge removal. After the exclusion of two ewes due to sponge loss, the remaining 148 ewes were divided into the Treatment group (n = 75) and the Control group (n = 73). Each ewe was inseminated once between 56 and 60 h after the removal of sponges, using a new type of insemination device containing 0.25 ml of diluted semen. Semen was collected from eight Black Suffolk rams and all the ejaculates were pooled and diluted in ultra-high temperature-treated commercial skimmed milk. The time of semen deposition was prolonged to 60 s in the Treatment group, while ewes were given traditional insemination in the Control group. Pregnancy status was determined by transabdominal ultrasound examination 45 days after insemination. Lambing performance was calculated after all the ewes had been delivered. Significant differences were observed between the Treatment group and the Control group in terms of the pregnancy rate and the fecundity rate (73.3% and 93.3% vs 56.2% and 71.2%, p < .05 and p < .01, respectively). In conclusion, prolonging the time of semen deposition significantly increased pregnancy and fecundity rates in estrus-synchronized Ujimqin ewes subjected to fixed time cervical insemination.

Genetic analysis of the effects of heat stress before and after lambing on pre-weaning live weight in Spanish Merino lambs.

BACKGROUND: Heat stress (HS) is a major environmental effect on sheep production. Hereby, we estimated the genetic (co)variance component of HS on the pre-weaning performance of 19,022 Merino lambs by analysing the climatological index of temperature and relative humidity (recorded 30 days before lambing and after lambing) using transversal and longitudinal mixed linear models. METHODS AND RESULTS: The global impact of HS during the last 30 days of pregnancy was -17% for birthweight and ranged between -4% and -8% for live weight at 15, 30 days of age (W30), and average daily gain from birth at 30 days. The results from both statistical approaches showed very similar heritabilities (h2 ), ranging from 0.192 to 0.237 for the direct genetic (D) effects and from 0.072 to 0.082 for the maternal genetic (M) effects, but the antagonism between (D) and (M) was higher when a longitudinal model was used. A significant genotype-environmental effect was also found regardless of whether the climatological covariables were considered in the model. In addition, we employed D and M breeding values for W30 as an example to create a new subjacent index by first using a principal component analysis and employing the leading eigenvalues as a weighted factor that provides the information needed to identify those genotypes that maximise the response for both genetic effects over a wide range of climate-environment levels. CONCLUSIONS: Our study revealed that the HS indexes of the mother during the gestation period have a significant effect on the growth of the lambs during the early stages of life.

Comparison of Five Protocols of Estrous Synchronization on Reproductive Performance of Hu Sheep.

The purpose of this study is to compare five protocols of estrous synchronization for Hu ewes to obtain the most effective and economical protocol, to apply the advantageous scheme in large-scale sheep farming. Healthy multiparous Hu ewes (n = 150) were randomly divided into five groups, and all ewes were administered fluorogestone acetate (FGA, 45 mg) vaginal sponge. The sponges of the first three groups (Groups I, II, and III) were removed on the 11th day, and 0.1 mg of PGF2α was injected intramuscularly on the ninth day. Group I received 6 µg of gonadotropin-releasing hormone (GnRH) by intramuscular injection at 36th h after withdrawal of the sponge. Group II was injected 330 IU of pregnant mare serum gonadotropin (PMSG) on the ninth day. The combination of 6 µg of GnRH and 330 IU of PMSG was treated in Group III at the same time as Group I and Group II. The sponges of the latter two groups (Groups IV and V) were removed on the 13th day, and 330 IU of PMSG was injected intramuscularly simultaneously. PGF2α (0.1 mg) was administered on the 12th day in Group IV. All ewes were detected for estrus at 24, 36, 48, 60, and 72 h after the sponge removal. The loss of sponge and vaginitis was recorded when the sponge was withdrawn. Cervical artificial insemination (AI) was performed with fresh semen of Dorper rams diluted with skimmed milk. After 30 days of insemination, the conception was detected with a veterinary B-ultrasound scanner. The lambing status of all ewes and the cost of drugs for estrous synchronization in each group were recorded. The results showed the following: (1) on the whole, the average percentage of estrous ewes in the period of 24-36 h and 36-48 h after removal was significantly higher than other three periods and that of the period of 60-72 h was significantly lower than the first three periods after removal; (2) there was no significant difference in percentages of estrous ewes in any of the five time periods, sponge loss rate, vaginitis rate, total percentage of estrous ewes, conception rate, single lambing rate, twinning rate, and multiple lambing rate of ewes among five protocols; (3) total percentage of estrous ewes and conception rate were more than or equal to 80% in the Groups II and III, and the twinning lamb rate of the Group II protocol was 70%; (4) there was no difference in lambing rate of ewes among Groups II, III, IV, and V; (5) the Group III had the highest drug cost of 22.5 CNY. In conclusion, considering the lambing rate, twinning lamb rate, and drug cost for estrous synchronization, Group II was the most advisable for application and promotion in large-scale sheep farms among these five protocols of estrus synchronization.

Maternal reactivity of ewes at lambing is genetically linked to their behavioural reactivity in an arena test.

In sheep, the bond between the dam and her lambs is established during the first hours of a lamb's life. Genetic variability for behavioural reactivity of ewes assessed in an arena test performed 24 hr after lambing has already been reported. However, there is no evidence that this reactivity represents the ewe's maternal reactivity at lambing in outdoor conditions. The objective of this study was to investigate whether or not the behavioural reactivity of ewes in the arena test is genetically related to their maternal reactivity measured at lambing. A total of 935 Romane ewes were studied. The maternal reactivity of ewes at the outdoor lambing site was recorded in response to a human approach and to the handling of the lambs. Their behavioural reactivity was also recorded 24 hr post-lambing in the arena test that involved a separation from the litter and a human presence. Flight distance, aggressive reaction, time to restore contact with the litter, maternal behaviour scores and vocalizations recorded at the lambing site were heritable (0.12-0.34). All of these behaviours were genetically correlated with the behavioural reactivity in the arena test. The highest genetic correlations (from 0.60 to 0.90) were found amongst maternal behavioural scores, flight distance and high-pitched bleats. In conclusion, behavioural reactivity in the arena test can be used to assess early maternal reactivity in standardized conditions. Phenotyping of ewes' behavioural reactivity with a simplified arena test can be performed for genetic improvement in maternal behaviour in sheep.