GnRH Immunocastration in Male Xizang Sheep: Impacts on Rumen Microbiome and Metabolite Profiles for Enhanced Health and Productivity.

Castration is a prevalent and indispensable practice in sheep husbandry, aiding in enhancing meat quality, mitigating aggressive behavior, and managing unwanted reproduction. Nevertheless, the conventional surgical castration procedure poses several challenges, including heightened stress and pain, detrimental impacts on animal welfare, and diminished economic efficacy in farming operations. Consequently, immunocastration methods, serving as substitutes for surgical castration, are progressively finding application in livestock. The rumen, an essential and distinctive digestive and absorptive organ in ruminants, has been associated with enhanced meat quality and productive performance following castration in previous research studies, albeit fewer investigations have explored the potential impacts of GnRH immunization on the rumen's internal milieu in sheep post-de-escalation. Hence, the present study delved into evaluating the impact of GnRH immunocastration on the rumen microbiome and metabolomics in male Xizang sheep. This was achieved through the establishment of a GnRH immunocastration animal model and the collection of rumen fluid for microbiological and comprehensive metabolomics investigations. The outcomes of this investigation unveiled that the impact of GnRH immunocastration on body weight gain was more pronounced during the achievement of the castration objective. In addition, the Firmicutes-to-Bacteroidota ratio in the immune male (IM) group exceeded that of the control group (EM), suggesting that GnRH immunodeficiency may enhance the digestion and absorption of feed in male Xizang sheep. At the taxonomic level, the elevated presence of Prevotella and Quinella bacteria in the IM group compared to the EM group indicated that castration influenced a segment of the rumen microbiota in male Xizang sheep, thereby bolstering the digestive and metabolic efficacy of the rumen concerning nutrient utilization, particularly in the breakdown and absorption of proteins, carbohydrates, and lipids, ultimately expediting the fattening process and weight gain in male Xizang sheep following castration. Moreover, analysis of ruminal fluid metabolomics revealed that GnRH immunization had notable impacts on certain metabolites in the ruminal fluid of male Xizang sheep, with metabolites like 5-hydroxyindole acetic acid and 3-hydroxyindole acetic acid showing significant downregulation in the IM group compared to the EM group, while niacin and tyramine exhibited significant upregulation. These findings indicate a profound influence of GnRH immunization on the maintenance of ruminal equilibrium and ruminal health (including the health of ruminal epithelial cells). This study validates that GnRH immunocastration not only achieves the objectives of castration but also enhances ruminal health in male Xizang sheep, thus laying a foundational theoretical basis for the application and dissemination of GnRH immunocastration technology.

Synergistic Effect of Maternal Micronutrient Supplementation on ORFV DNA Vaccine Immune Response in a Pregnant Model.

Contagious ecthyma is a contagious zoonotic disease caused by the Orf virus that can infect farm animals and humans, but no vaccine is available for pregnant mothers. Excessive oxidative stress during pregnancy can suppress the vaccine immune response in pregnant mothers; hence, maternal micronutrient supplementation could effectively improve the immune response, health, and oxidative status during pregnancy. In this study, we employed an 8-week-old pregnant rat model to receive a single intramuscular dose of 200 µg of ORF DNA vaccine with or without vitamin E and selenium supplementation to evaluate their effect on immune responses (specific IgG and IgG isotypes), oxidative stress, liver enzymes, and blood glucose levels in maternal-neonatal serum and milk secretions. Additionally, antioxidant-related gene expressions were analyzed in the maternal placenta and pups' liver. The results showed that supplementation of vitamin E and selenium with ORF DNA vaccination increased the production of specific antibody and IgG isotypes (IgG1 and IgG2a) and reduced the oxidative stress in neonatal-maternal serum and milk compared to both the control group and those vaccinated without supplementation (p < 0.05). Notably, the ORF DNA vaccine did not cause oxidative stress and hepatic damage. However, combined supplementation of vitamin E and selenium with DNA vaccination significantly decreased serum malondialdehyde (MDA) levels and improved the antioxidant-related enzyme activities of glutathione peroxidase (GPX), superoxide dismutase 1 (SOD1), and selenoprotein P (SELP) in the maternal placenta and liver of pups (p < 0.05). In conclusion, maternal supplementation of vitamin E and selenium enhanced the immune responses of the ORF DNA vaccine by mitigating oxidative stress in pregnant rats and could thus be a promising strategy for better health outcomes for both mothers and neonates.

Relationships of infrared thermography temperature with core temperature in goat.

Rectal temperature is widely used as an indicator of animal health. However, rectal temperature is conventionally measured by an invasive method, which may reduce animal welfare. So, this study aimed to determine the relationships between the deep-body (core) temperature and body surface temperatures in goats and develop a linear regression equation to establish the core temperature based on body surface temperatures. Body surface temperatures (head, eye, muzzle, horn, back, scrotum and groin) of goats were measured by infrared thermography (IRT). Ambient temperatures were measured by digital thermometer. Core temperatures were measured by a digital vet thermometer. Pearson correlation analysis was used to analyze the relationship between body surface temperatures, ambient temperature, and core temperature. Simple linear regression analysis was used to develop core temperature assessment equations. Correlation analysis showed that groin temperature was highly correlated with core temperature, and low correlated with ambient temperature. The body surface temperature of other region was low correlated with core temperature, and highly correlated with ambient temperature. Regression analysis showed that the determination coefficient of core temperature assessment equation based on groin temperature was the highest (P < 0.0001, R2 = 0.55), and those based on surface temperature of other regions were low (P < 0.01, R2 ≤ 0.16). We concluded that body surface temperatures obtained by IRT could be used for the assessment of goat core temperature. The core temperature assessment equations developed by the temperature of the body surface, which is less affected by ambient temperature, was found to have a higher determination coefficient than the equations developed using body surface temperature that is more affected by ambient temperature.

Effects of yeast culture on in vitro ruminal fermentation and microbial community of high concentrate diet in sheep.

This research aimed to investigate effects of different yeast culture (YC) levels on in vitro fermentation characteristics and bacterial and fungal community under high concentrate diet. A total of 5 groups were included in the experiment: control group without YC (CON), YC1 (0.5% YC proportion of substrate dry matter), YC2 (1%), YC3 (1.5%) and YC4 (2%). After 48 h of fermentation, the incubation fluids and residues were collected to analyze the ruminal fermentation parameters and bacterial and fungal community. Results showed that the ruminal fluid pH of YC2 and YC4 groups was higher (P < 0.05) than that of CON group. Compared with CON group, the microbial protein, propionate and butyrate concentrations and cumulative gas production at 48 h of YC2 group were significantly increased (P < 0.05), whereas an opposite trend of ammonia nitrogen and lactate was observed between two groups. Microbial analysis showed that the Chao1 and Shannon indexes of YC2 group were higher (P < 0.05) than those of CON group. Additionally, YC supplementation significantly decreased (P < 0.05) Succinivibrionaceae_UCG-001, Streptococcus bovis and Neosetophoma relative abundances. An opposite tendency of Aspergillus abundance was found between CON and YC treatments. Compared with CON group, the relative abundances of Prevotella, Succiniclasticum, Butyrivibrio and Megasphaera elsdenii were significantly increased (P < 0.05) in YC2 group, while Apiotrichum and unclassified Clostridiales relative abundances were decreased (P < 0.05). In conclusion, high concentrate substrate supplemented with appropriate YC (1%) can improve ruminal fermentation and regulate bacterial and fungal composition.

Biochemical properties of sheep colostrum and its potential benefits for lamb survival: a review.

Colostrum is the initial secretion of the mammary glands following parturition, which offers main food, protection, and biological active substances for the new born. The most threatening episode of neonate's life is the initial two weeks after birth. This period is associated with high neonatal mortality and morbidity. These worthwhile losses lead to a poor prolificacy rate, low profitability, and ultimately poor performance in animal production. Hence, both diseases and mortality cause valuable losses in terms of production and economic losses. The survival of neonate is correlated with their immune status and passive immune transfer (PIT). Colostrum provides the primary source of nutrition and immunity (PIT) that protects neonates against infections. It must be given as soon as possible after birth since its immunoglobulins are absorbed within the first 16-27 hours after birth, ideally within 2-4 hours. As a result, immunoglobulin (PIT) is the most important component of distressing infectious immunity, and a passable concentration of immunoglobulin in the blood of newborn lambs is linked to their health and survival rate. In this review, we summarized the importance of colostrum in early life and its association with neonatal lamb's survival, profitability and productivity of sheep farming.

Economic values of reproductive and growth traits in Chinese Yiling sheep.

This study aimed to calculate the economic value (EV) of reproductive and growth traits for Yiling sheep. A bio-economic model was developed to assess the economic value of litter size (LS), litter size at weaning (LSW), age at first lambing (AFL), lambing interval (LI), birth weight (BW), weaning weight (WW), and 6-month body weight (6MW). The sensitivity of the economic value of traits to changes in market prices was also analyzed. In this study, the trait with the highest EV was LSW (427.97 ¥), followed by LS (419.96 ¥), BW (52.13 ¥), 6MW (14.46 ¥), WW (11.03 ¥), AFL (-0.51 ¥), and LI (-9.09 ¥). LS was the most important trait in the production system with a relative economic weight of 22.81%, followed by 6MW and LSW with relative economic weights of 18.98% and 19.01%, respectively. All traits assessed, except AFL and LI, had positive economic values, indicating that genetic improvement of these traits would have a positive impact on profitability. The results of the sensitivity analysis showed that the economic value of AFL was not sensitive to price changes. All growth traits were unaffected by price changes in labor and medical costs. In addition, the LS, LSW, LI, WW, and 6MW were sensitive to changes in liveweight and feed prices. Generally, as feed prices increased, the economic value of all traits except LI and BW decreased. Except for LI and BW, the economic value of all traits increased due to the rise in liveweight prices. This suggested that liveweight and feed prices significantly affect the profitability of the production system.

RNA-Seq reveals ACTH-induced steroid hormone pathway participating in goat adrenal gland response to castration.

The content of androgen from adrenal is elevated under castration, and the mechanisms of compensatory secretion of adrenal androgen remain unknown. This study was designed to compare the transcript profiles between adrenals from noncastrated, orchiectomized and immunocastrated Yiling goats. Fifteen goats were randomly divided into three groups: pVAX-asd injection (control) group, pVAX-B2L-(G4S)3-kisspeptin-54-asd immunization (PBK-asd) group, and surgical castration (SC) group. Subsequently, serum was collected every two weeks after the initial immunization for hormone assays. At week 14 after immunization, adrenal glands were collected for transcriptome sequencing and qPCR. Serum testosterone concentration was significantly reduced in PBK-asd and SC group, demonstrating the effectiveness of castration. Both surgical and immunized castration resulted in adrenal hyperplasia, and thickness of adrenal cortex elevated. The specific genes involving castration were enriched in many pathways, including Steroid hormone biosynthesis pathway. The adrenocorticotropic hormone (ACTH), which promotes the production of adrenal steroids, and dehydroepiandrosterone (DHEA), a steroid hormone secreted by adrenal glands, both increased after castration. Further construction of co-expression network for transcription genes and traits (including adrenal weight and cortex thickness, ACTH and DHEA concentration) showed that the trait-related genes were enriched in multiple steroid-related pathways. These results showed that adrenal compensatory hyperplasia and androgen secretion caused by castration may involve in ACTH-induced steroid hormone synthesis.

Selection signature analysis reveals RDH5 performed key function in vision during sheep domestication process.

As one of the most successful domesticated animals in the Neolithic age, sheep gradually migrated all over the world with human activities. During the domestication process, remarkable changes have taken place in morphology, physiology, and behavior, resulting in different breeds with different characters via artificial and natural selection. However, the genetic background responsible for these phenotypic variations remains largely unclear. Here, we used whole genome resequencing technology to compare and analyze the genome differences between Asiatic mouflon wild sheep (Ovis orientalis) and Hu sheep (Ovis aries). A total of 755 genes were positively selected in the process of domestication and selection, and the genes related to sensory perception had directional evolution in the autosomal region, such as OPRL1, LEF1, TAS1R3, ATF6, VSX2, MYO1A, RDH5, and some novel genes. A missense mutation of c.T722C/p.M241T in exon 4 of RDH5 existing in sheep were found, and the T allele was completely fixed in Hu sheep. In addition, the mutation with the C allele reduced the retinol dehydrogenase activity encoding by RDH5, which can impair retinoic acid metabolism and further influenced the visual cycle. Overall, our results showed significant enrichment for positively selected genes involved in sensory perception development during sheep domestication; RDH5 and its variants may be related to the retinal degeneration in sheep. We infer that the wild sheep ancestors with weaker visual sensitivity were weeded out by humans, and the mutation was selective, swept by the dual pressures of natural and artificial selection.

The protective role of maternal genetic immunization on maternal-fetal health and welfare.

Pregnancy is a critical period associated with alterations in physiologic, biologic, and immunologic processes, which can affect maternal-fetal health through development of several infectious diseases. At birth, neonates have an immature immune system that makes them more susceptible to severe viral infections and diseases. For this reason, different maternal nutritional and immunization interventions have been used to improve the immune and health status of the mother and her neonate through passive immunity. Here, we reviewed the protective role of maternal immunization with different types of vaccines, especially genetic vaccines, during pregnancy in maternal-fetal health, immune response, colostrum quality, immune response, and anti-oxidative status. For this purpose, we have used different scientific databases (PubMed and Google Scholar) and other official web pages. We customized the search period range from the year 2000 to 2023 using the key words "maternal immunization" OR "gestation period/pregnancy" OR "genetic vaccination" OR "maternal-fetal health" OR "micronutrients" OR "neonatal immunity" "oxidative stress" OR "colostrum quality". The evidence demonstrated that inactivated or killed vaccines produced significant immune protection in the mother and fetus. Furthermore, most recent studies have suggested that the use of genetic vaccines (mRNA and DNA) during pregnancy is efficient at triggering the immune response in mother and neonate without the risk of undesired pregnancy outcomes. However, factors such as maternal redox balance, nutritional status, and the timing of immunization play essential roles in regulating immune response inflammatory status, antioxidant capacity, and the welfare of both the pregnant mother and her newborn.

LAD-RCNN: A Powerful Tool for Livestock Face Detection and Normalization.

With the demand for standardized large-scale livestock farming and the development of artificial intelligence technology, a lot of research in the area of animal face detection and face identification was conducted. However, there are no specialized studies on livestock face normalization, which may significantly reduce the performance of face identification. The keypoint detection technology, which has been widely applied in human face normalization, is not suitable for animal face normalization due to the arbitrary directions of animal face images captured from uncooperative animals. It is necessary to develop a livestock face normalization method that can handle arbitrary face directions. In this study, a lightweight angle detection and region-based convolutional network (LAD-RCNN) was developed, which contains a new rotation angle coding method that can detect the rotation angle and the location of the animal's face in one stage. LAD-RCNN also includes a series of image enhancement methods to improve its performance. LAD-RCNN has been evaluated on multiple datasets, including a goat dataset and infrared images of goats. Evaluation results show that the average precision of face detection was more than 97%, and the deviations between the detected rotation angle and the ground-truth rotation angle were less than 6.42° on all the test datasets. LAD-RCNN runs very fast and only takes 13.7 ms to process a picture on a single RTX 2080Ti GPU. This shows that LAD-RCNN has an excellent performance in livestock face recognition and direction detection, and therefore it is very suitable for livestock face detection and normalization.

Characteristic of the components and the metabolism mechanism of goat colostrum: a review.

Colostrum contains large number of nutrients that promote the growth, differentiation, and biological functions for goat kids early somatic cells, which is crucial to meet the nutritional demands, immune function, and the health of goat kids later growth. Great attention has been given not only to nutritional ingredient differences between colostrum and normal milk, but also to function differences, and their effect on the physical and sensory properties of goat kid's growth performance and health status. This paper reviews the research progress of goat colostrum in recent years, mainly including the colostrum yield, components, i.e., proteins, lactose, and immunoglobulin, as well as the influence factor, i.e., number of lactation and littler size, nutrition during the gestation, and breeding environment. In addition, this review aims to summarize the synthesis and secretion mechanisms, and the digestion and absorption mechanism of goat colostrum. We conclude that even though the composition and physicochemical properties of goat colostrum are highly dynamic and variable, and the digestion and absorption mechanism has not been made fully clear until now, direct feed microbial (DFM) may be a promising alternative for improving the quality of colostrum that should be further explored for their practical usage.

Recombinant HBsAg-S and RFRP-3 DNA vaccine promotes reproduction hormone secretion in sheep.

RF-amide related peptides (RFRP) have been proposed as critical regulators of gonadotropin secretion in mammals. This study was designed to construct a DNA vaccine and investigate the effect of vaccine encoding RFRP-3 on reproduction physiology in ewe. A recombinant vaccine was constructed using two copies of the RFRP-3 gene and HBsAg-S that generate a fusion protein to induce an immunology response. Results showed this recombinant vaccine could produce a significant antibody titer in the treated animals (P < 0.05). The specific RFRP-3 antibody response induced by the vaccine was detected at week 2 with a peak at week 6 after the initial immunization. Furthermore, we found that ewes inoculated with pVAX-tPA-HBsAg-S-2RFRP-asd vaccine significantly raised the concentration of GnRH, LH and E2 in serum compared to the control group. LH and E2 concentration in the treated ewes (Group T) was significantly higher than that in control ewes (Group C) at weeks 10, 12 and 14 after the initial immunization, respectively (P < 0.05). Therefore, RFRP-3 can be used as a target for DNA immunization to promote reproductive hormone secretion in ewes and RFRP-3 gene immunization might be a candidate tool to regulate mammal reproduction.

Adrenal gland responses surgical castration and immunocastration by different compensatory manners to increase DHEA secretion.

Androgen from the testis and weak androgens from the adrenal cortex may interact with each other and affect their synthesis and secretion due to their similar functions. The purpose of this study was to investigate the compensatory effect of adrenal in rats after immunocastration and surgical castration, and the interaction between the hypothalamic-pituitary-testis (HPT) axis and the hypothalamic-pituitary-adrenal (HPA) axis. 24 male SD rats aged 8 weeks were randomly divided into three groups and accepted treatments: surgical castration group, immunocastration group and control group. In both surgical castration and immunocastration groups, the secretion of adrenocorticotropic hormone (ACTH) and dehydroepiandrosterone (DHEA) hormones was significantly increased compared with the control group (p < 0.05). In the HPT axis of the immunocastration group, the KISS1 expression was up-regulated, whereas GPR54, LH and LHR expression were down-regulated (p < 0.05). The expression levels of CRH, POMC and MC2R genes were also significantly up-regulated (p < 0.05). In addition, in the immunocastration group, the expression of adrenal LHR mRNA expression was decreased (p < 0.05). The expression of HPT axis genes and adrenal LHR were up-regulated in the surgical castration group (p < 0.05). These results show that in both immunocastration and surgical castration, adrenal androgen is increased, suggesting that the adrenal gland plays a compensatory role. Moreover, it also shows that different castration treatments have effects on adrenal steroid secretion through different mechanisms.

Changes in colostrum ingredients of Hu sheep, as well as the missense mutation genes associated with colostrum yield.

This study focused on the changes in the composition and immune evolution in milk from birth to 144 h postpartum and the genes associated with the colostrum yield of Hu sheep. Twelve Hu sheep, which were bred carefully under animal health standards and have a litter size of two kids and similar gestation length (149 ± 1 days), were used. Lambs were transferred into their own cots to avoid interference. The compositional content (i.e., fat, protein, and lactose) and some other properties, including daily colostrum yield, DM, and SNF, were determined. In addition, immunity molecules (IgG, IgA, and IgM concentrations) received remarkable attention. The DM, SNF, fat, and protein contents were higher in the first days postpartum and then dropped quickly from the time of birth to 144 h postpartum. However, the lactose content displayed an increasing pattern and reached normal milk percentage at 48 h. The highest IgG (103.17 mg/mL), IgA (352.82 µg/mL), and IgM (2.79 mg/mL) colostrum concentrations were observed at partum, decreased quickly, and finally stabilized. The change law of concentration of IgA and IgM in colostrum are the same with IgG. Furthermore, the whole-genome resequencing was performed, and a missense variant locus in the SRC gene and two missense locus variants in the HIF1A gene were significantly associated with the colostrum yield of sheep by using the whole-genome selection signal detection analysis. In conclusions, colostrum contains abundant nutrients especially immunoglobulin, and the HIF1A gene may be used as candidate genes for colostrum yield, which has important information as a basic knowledge for the Hu sheep breeding program.

Multiple-Pathway Synergy Alters Steroidogenesis and Spermatogenesis in Response to an Immunocastration Vaccine in Goat.

BACKGROUND: Animal reproduction performance is crucial in husbandry. Immunocastrated animals serve as an ideal animal model for studying testicular function. During androgen suppression, the testis undergoes dramatic developmental and structural changes, including the inhibition of hormone secretion and spermatogenesis. METHODS: To characterize this process, we investigated the effects of castration using a recombinant B2L and KISS1 DNA vaccine, and then identified functional genes in the testes of Yiling goats using RNA-seq and WGS. The experimental animals were divided into three groups: the PVAX-asd group (control), PBK-asd-immunized group, and surgically castrated group. RESULTS: The results demonstrated that the administration of the recombinant PBK-asd vaccine in goats elicited a significant antibody response, and reduced serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH), resulting in smaller scrotal circumferences and decreased sexual desire compared to the control group. In addition, RNA transcriptome sequencing (RNA-seq) analysis of the testes revealed that the biological processes after immunocastration mainly focused on the regulation of cell matrix adhesion, histone acetylation, negative regulation of developmental processes, apoptosis, and activation of the complement system and the thrombin cascade reaction system. Then, we integrated the whole-genome sequencing and testis transcriptome, and identified several candidate genes (FGF9, FST, KIT, TH, TCP1, PLEKHA1, TMEM119, ESR1, TIPARP, LEP) that influence steroidogenesis secretion and spermatogenesis. CONCLUSIONS: Multiple pathways and polygenic co-expression participate in the response to castration vaccines, altering hormone secretion and spermatogenesis. Taken together, our atlas of the immunocastration goat testis provides multiple insights into the developmental changes and key factors accompanying androgen suppression, and thus may contribute to understanding the genetic mechanism of testis function. Joint analysis of whole genome sequencing and RNA-seq enables reliable screening of candidate genes, benefiting future genome-assisted breeding of goats.

New trends in immunocastration and its potential to improve animal welfare: a mini review.

Castration is frequently used to reduce aggressive behavior and improve the meat quality of animals. Traditionally, surgical and mechanical castration are used to sterilize the animals, but these approaches are associated with a high level of pain, stress, long recovery periods, and post-operative infections. Immunocastration is a new animal-friendly, painless alternative castration technique that is used to prevent undesired sexual behavior, reduce aggressive behavior, prevent unwanted pregnancy, control wildlife populations and wandering species, enhance growth performance, improve meat quality, and treat various sex hormone-dependent disorders. The mechanism of immunocastration includes the immunological block of the hypothalamic-pituitary-gonadal axis (HPG axis) which inhibits gonadotropin secretions, causes atrophy of gonadal tissues, and inhibits gametogenesis, resulting in infertility in both female and male mammals. By the mid-1990s, various immunocastration vaccines have been tested in different animal models to achieve successful castration effects. Recently, genetic immunocastration especially DNA vaccine has gained increasing attention due to its safety, being animal-friendly, and being easy to use. This review aims to evaluate the potential of traditional castration methods, as well as the current status of immunocastration vaccines, their effects, and future prospective.

Polygenic co-expression changes the testis growth, hormone secretion and spermatogenesis to prompt puberty in Hu sheep.

In most male mammals, testis undergoes increased proliferation activity and the onset of spermatogenesis during pubertal development. However, their gene expression patterns and roles in sheep remain unclear. Therefore, we used Illumina Hiseq 2000 sequencing to identify differentially expressed genes (DEGs) from 0, 30, 60, 90, 120, 150 and 180 postnatal days and characterize the transcriptional level of sheep testicular development. Among them, the DEGs changed the most in 90-150 stages. a total of 2546 (1454 up and 1092 down) and 6867 (4683 up and 2184 down) DEGs were identified in D120 vs. D90 and D150 vs. D120, respectively. Functional enrichment analysis revealed that in earlier pubertal development, testis showed higher gene expression in organ morphogenesis, vasculogenesis, neurogenesis and hormone secretion, while in later pubertal development, genes with higher expression mainly concentrated in regulating spermatogenesis process. These results indicated that testis development undergoes the transition from organ growth to spermatogenesis and the genes related to hormone secretion were expressed highly earlier than spermatogenesis during pubertal process. In addition, we found several genes such as ZFP36, TNF, HSD3B1, HSD11B2 played key roles in androgen secretion and SPAG family, SYCP family, SPATA family, SPO11, CABYR, TNP1, TNP2 and CFAP43 performed functions during spermatogenesis process. Taken together, multi-genes cooperation prompt testis development in pubertal process. Our transcriptional atlas of sheep testis provides a comprehensive insight about testicular development and pubertal process.

The c.612A>G mutation of MC4R affects constitutive activity and signaling in domestic goats.

As a key gene for balancing energy and regulating feeding behavior, MC4R is relevant to the growth of ruminants. In this presentation, a highly conserved c.612A>G site in the coding sequence (CDS) of MC4R has been selected during a selective sweep analysis of 35 Yiling goats and 20 other wild goats. This site mutation results in an amino acid change from Ile to Met. The genotyping analysis of the c.612A>G site revealed that the A allele was the dominant allele in the domestic goat populations, while the wild goat individuals only had the G allele. For a better understanding of the biological significance of this site, we examined the protein localization and signal detection to explain the function of the two MC4R receptors. The results showed that both the M204 and I204 receptors can normally localize on the membrane. When stimulating the M204 type without α-MSH, it was defective at the level of basal cAMP and decreased significantly against the I204 type. In contrast, the signaling capacity of the M204 receptor was also lower than that of I204 under the stimulation of α-MSH. In the ERK1/2 pathway, stimulating MC4R with NDP-α-MSH, both the M204 and I204 receptors had normal pERK1/2 levels. These results indicate that the p.I204M mutation may change the function by damaging the constitutive activity and signaling, and thus may regulate goats' appetite. This study has potential application for rearing domestic goats.

Immunocastration with gene vaccine (KISS1) induces a cell-mediated immune response in ram testis: A transcriptome evaluation.

Immunocastration vaccines achieve their effects through neutralization of the endogenous hormone by the humoral antibody produced against the immunized genes, but there is little information regarding cell-mediated immune response on the gonadal function of the immunized model is available. In this study, we used ram as a model animal to identify the cellular immune response in testicular tissues of rams immunized with intranasal KISS1 gene vaccine. The immune castration model was evaluated by sexual behaviours, spermatogenesis and serum hormone profiles after the KISS1 gene immunization. Transcriptome analysis of testicular tissues was carried out to identify the expressions of protein-coding genes involved in cellular immunity. The results showed that we successfully constructed the KISS1 immune castration ram model, in which testicular growth and development, testosterone and kisspeptin-54 levels, and sexual function were suppressed in immunized rams (p < .05). Using Hiseq™ 2000 high sequencing for ram testicular, we identified 21 differentially expressed genes (DEGs) related to cellular immunity, of which, 14 genes were upregulated and seven genes were downregulated in the testis of the immunized group (p < .05). The Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment showed that these differentially expressed genes were enriched in the antigen presentation process mediated by MHC class I and the cytotoxic pathway mediated by natural killer cells. It is concluded that KISS1 gene vaccine induced the cell-mediated immune response in testicular tissue to suppress reproductive activities in rams.

An ancient positively selected BMPRIB missense variant increases litter size of Mongolian sheep populations following latitudinal gradient.

New gene mutation origination is a driving force for the evolution of organisms. The effect of FecB mutation in BMPRIB gene on the litter size of sheep has been well known for a long time, each copy of the mutant allele increases litter size by 0.4-0.5. However, the origin and adaptive evolution mechanism of FecB mutation are still unclear. Here we carried on the thorough analysis on evolutionary features of BMPRIB gene and found that 150 species as a whole is under purifying selection while sheep lineage shows evidence of positive selection. The results of allele age estimation revealed that the FecB mutation in Mongolian sheep of China originated in Mongolian Plateau at about 5000 years ago. Due the two shape drops in temperature subsequently, Mongolian sheep migrated from north to south following the northern nomadic people. Accordingly, the FecB mutant allele frequency increased, with the lowest in sheep locating at Mongolian plateau (0.01) and the highest in sheep locating at Yangtze River valley (0.96). In conclusion, the FecB mutation in Mongolian sheep of China originated in Mongolian Plateau at about 5000 years ago, and the differentiated litter size of Mongolian sheep might be the result of adaptation to various environments during the migration following latitudinal gradient. This study may well exemplify selection on an ancient variation triggered by drastic ecological shifts, and is also helpful to analyze the adaptive evolution mechanism of economic traits of domestic animals and identify major genes and molecular markers.