Search

The Protein Phosphatase 6 Catalytic Subunit (PPP6C) is evolutionarily a conserved gene in eukaryotes known to play a significant role in mammalian reproduction. This study aimed to investigate expression patterns of PPP6C and explore its association with litter size in Shaanbei white cashmere (SBWC) goats. Initially, we determined the mRNA expression levels of PPP6C in both male and female goats across multiple tissues. The results showed that PPP6C mRNA was expressed in multiple tissues, with higher levels in the testis and fallopian tubes, suggesting its involvement in goat reproduction. Additionally, we identified a novel 19 bp InDel within the PPP6C gene in a population of 1030 SBWC goats, which exhibited polymorphism. Statistical analysis revealed a significant association between the19 bp InDel mutation and litter size (P < 0.05). Subsequent, bioinformatics analysis, including linkage disequilibrium (LD) block and selective scanning, highlighted the linkage tendency among most InDel loci did not stand out within B-8 block, there were still some InDel loci linked to the 19 bp within a relatively narrow region. Furthermore, comparative analysis with Bezoars, these selective signals all indicated that this gene was under higher selection pressure, implying that the 19 bp InDel locus within the PPP6C is potentially associated with domesticated traits, particularly in relation to litter size. The results of the present study suggest that the PPP6C is a vital candidate gene affecting prolificacy in goats, with implications for selective breeding programs for goat breeds., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest with this reported study. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed at scrutinizing efficiency of incorporating L-carnitine or M. oleifera leaves extract into semen diluent on improving cryopreservation capacity and in vitro fertilization ability of buck spermatozoa. Ejaculates (n=48) were collected by an artificial vagina from six adult Damascus bucks twice weekly during the breeding season (September-October). Following initial evaluation, ejaculates of each collection session from the same bucks were pooled, diluted (1:10) with glycerolized (3 % glycerol, v/v) tris-citric acid egg yolk diluent and were split into three aliquots. The first aliquot served as control, whereas the second and third aliquots were supplemented with 4 μL/mL L-carnitine and 400 μL/mL moringa leaves extract (v/v), respectively. Thereafter, all specimens were processed for cryopreservation and were stored in liquid nitrogen (-196 °C) for 12 months before post-thaw sperm criteria were analyzed by a computer-assisted sperm analysis (CASA) system. Integrity of sperm DNA post thawing was visualized in all semen groups by fluorescence imaging, and in vitro fertilization ability of spermatozoa was also determined. Inclusion of L-carnitine or moringa leaves extract into the diluent improved (P<0.05) post-thaw sperm physical, morphofunctional and kinematic attributes, whilst maintaining (P<0.05) integrity of sperm DNA throughout the freezing and thawing cycle. Consequently, both supplemented groups yielded higher (P<0.05) in vitro fertilization rates compared to control. These results accentuate the protective roles of these antioxidants on buck sperm against consequences of cryopreservation-induced oxidative stress, hence ameliorating post-thaw sperm quality and fertilization competence. This is crucial for successful application of AI and IVF in goat selective breeding programs., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interests, financial or personal, that could have appeared to influence conducting and reporting this study., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to investigate a new implantation site (intra-auricular subcutaneous - IA) compared to intramuscular (IM) in the cervical portion (cervical splenius muscle) of the neck for ovarian transplantation in goats. Morphological aspects of the implant, follicular activation and morphology, and type I and III collagen deposits of the transplanted tissue were evaluated. Four fragments of the ovarian cortex were allotransplanted at the IA and IM sites in all goat recipients and recovered 7 (IA-7; IM-7) or 15 (IA-15; IM-15) days later and submitted to histological analysis. Two fragments/animal were separated for the fresh control (FC) group. There was a higher percentage of normal and developing primordial follicles at the IA-7 site (P < 0.05) compared to the other treatments, with similar values to the fresh control. Type I and III collagen fibers differed between the groups (P < 0.05), showing a considerable decrease in type I collagen fibers at the IA-7 site compared to the FC. However, the IM-7 and IA-15 sites showed higher values of type I collagen fibers, showing similarity to the FC. Therefore, we conclude that the IA site in goats is an effective site for ovarian tissue transplantation, as it is easily accessible, low invasive and has presented satisfactory rates of morphology and follicular activation., Competing Interests: Declaration of Competing Interest The authors confirm that there are no conflicts of interest for any of the authors, (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the effects of replacing egg yolk extender with sesame oil on the quality of sperm in goats following incubation at 37°C, chilling at 4°C, and freezing. Semen samples were collected from four intact male goats. The individual semen sample was divided into six groups consisting of a control group and five treatment groups with different egg yolk-to-sesame oil ratios. Seminal plasma was removed, and the sperm pellet was diluted with experimental semen extenders. The control group contained an extender of 10 % egg yolk (SO0), and the experimental extenders were composed of 8.75 % egg yolk and 1.25 % sesame oil (SO1.25); 7.5 % egg yolk and 2.5 % sesame oil (SO2.5); 5 % egg yolk and 5 % sesame oil (SO5); 2.5 % egg yolk and 7.5 % sesame oil (SO7.5); and 10 % sesame oil (SO10). Each group of semen was divided into three groups, incubated at 37°C for 1 h, chilled at 4°C for 4 h, or frozen for 24 h. Five replicates were performed. Sperm quality was evaluated, including motility, viability, and functional membrane integrity. The SO1.25 group achieved the highest sperm quality rate among the treatment groups, and the extender did not have a negative effect compared to the control. However, the total replacement of egg yolk with sesame oil in an extender resulted in the lowest sperm quality. In conclusion, the ratios of egg yolk and sesame oil that were acceptable for goat semen cryopreservation were 8.75 % and 1.25 %, respectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cryopreservation of small ruminant's semen is an effective strategy for distributing spermatozoa for reproductive programs, but this process decreases the fertility potential of post-thawed spermatozoa. The aim of this research was to assess the effect of different concentrations of CoQ10 in soybean lecithin (SL)-based extender on buck semen quality during cryopreservation process. Semen samples were collected from five bucks, twice a week, then diluted in the SL-based extender containing different concentrations of CoQ10 as follows: extender containing 0 µM (control, Q0), 0.1 µM (Q0.1), 1 µM (Q1), 10 µM (Q10) and 100 µM (Q100) CoQ10. Motion characteristics, membrane functionality, abnormal morphology, mitochondrial activity, acrosome integrity, viability, apoptotic-like changes, lipid peroxidation, DNA fragmentation and ROS concentration were evaluated after freeze-thawing process. The Q10 resulted in greater (P≤0.05) total motility, progressive motility, average path velocity, membrane integrity, mitochondrial activity, acrosome integrity and viability compared to the other groups. Furthermore, supplementation of freezing extender with 10 µM of CoQ10 presented lower (P≤0.05) apoptotic-like changes, lipid peroxidation, DNA fragmentation and ROS concentration compared to the other groups. Regarding to the protective effect of CoQ10 supplement during cryopreservation process, it could be explored as a potent antioxidant for cryopreservation of buck semen as it preserved the post-thawed buck sperm quality., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to disclose., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Pregnancy success is closely related to the molecular mechanisms that control energy metabolism balance. However, the mechanisms have not been fully understood. Uncoupling protein 2 (UCP2) plays a physiological role by regulating energy metabolism in numerous tissues. In this study, we determined the expression and hormone regulation of UCP2 in goat uterus. UCP2 is expressed in the luminal and glandular epithelia of goat uterus during early pregnancy, as revealed by in situ hybridization and immunohistochemistry conducted on pregnant goats. The signals were detected from day 0 (D0) to D30 of pregnancy, though weak on D16 (the adhesion period). The low levels of UCP2 on D16 were confirmed by RT-qPCR and western blot. In goat uterus and endometrial epithelial cells (EECs), UCP2 was up-regulated by progesterone and estrogen. In addition, after goat EECs were treated with genipin (an inhibitor of UCP2), not only UCP2 expression but also cell proliferation was inhibited. Collectively, UCP2 is dynamically expressed in goat uterus and can affect EEC proliferation, suggesting that it may participate in regulating the energy metabolism balance of goat uterus during early pregnancy., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This review is part of the Festschrift in honor of Dr. Duane Garner and provides an overview of current techniques in cooled storage of semen from livestock animals such as camelids, goats, and sheep. Facing worldwide environmental changes and a trend towards more conscious and healthy eating behaviors, the development of a stable animal breeding industry is a significant challenge for the near future. In the present review, factors influencing semen handling in camelids, goats and sheep are described and relevant methods as well as current trends to improve liquid-storage of cooled semen are discussed, including extenders, additives, cooling rates, and storage temperatures. The species-specific physiology and resulting challenges are taken into consideration. While the main problem for camelid semen processing is the relatively greater viscosity as compared with that of some other animals, the deciding factor for successful artificial insemination (AI) in goats and sheep is the site (i.e., cervical or vaginal) of semen placement in the reproductive tract. Due to the type of cervical anatomy, the penetration of the cervix when using AI instruments is rather difficult. Furthermore, the seminal plasma of small ruminants affects the interaction with milk-based extenders and egg yolk which results in species-specific regimens for cooled liquid-preservation. Comparing all three species, the greatest pregnancy rates were obtained by AI with goat semen after cooled liquid-storage for several days., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The TG interaction factor 1 (TGIF1) is of the TALE homologue domain protein family and is considered as a transcriptional repressor of SMAD protein that interacts with DNA through a specific consensus-binding site for TG and recruits mSin3A and histone deacetylases to the SMAD complex. In this study, there is the first detailed description of TGIF1 on steroidogenesis in goat granulosa cells. When there is a relatively greater expression of the TGIF1 gene, there is a lesser abundance of CYP11A1, CYP19A1, and StAR mRNA transcript and protein and 3β-HSD mRNA transcript in granulosa cells of goats. Furthermore, there were lesser concentrations of 17β-estradiol (E 2 ) and progesterone (P 4 ) in culture medium when there was greater TGIF1 gene expression and there were greater concentrations of these hormones in the culture medium when there was lesser TGIF1 gene expression. There may be functions of TGIF1, therefore, in suppression of SMAD-induced E 2 and P 4 production and in decreasing the phosphorylation of SMAD2/3 in granulosa cells of goats and relative abundance of the SMAD2/3 protein transcription factor, SP1. With suppression of TGIF1 gene expression, there was a reversal of SP1-induced suppression of steroidogenesis-related genes. Results of the present study provide insights about the potential mechanism underlying the regulation of granulosa cell steroidogenesis of goats by TGIF1., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Stromal tissue is an essential componenlt of the ovary not only for providing structural support but also for contributing to the early follicular growth with their bi-directional paracrine signaling. Estradiol is a major female hormone mainly secreted by the follicular cells in the ovary. To examine the relationship between 17β-estradiol and the factors involved in androgen production in stromal cells, ovarian stromal cells were cultured in the graded concentrations (50 and 100 ng/mL) of 17β-estradiol for varying time periods (24 and 48 h). The cells were processed for transmission electron microscopy to study the changes in steroidogenic functions of the cells. The effect of estradiol treatment was also evaluated on the quantity of androgen production and abundance of steroidogenic enzymes and proteins. The results indicated 17β-estradiol increased androgen production in ovarian stromal cells. In addition to enhanced androstenedione and testosterone production, estradiol stimulation was also based on the marked increase in abundance of mRNA transcript of steroidogenic enzymes [Star (Steroidogenic Acute Regulatory Protein), Cyp11a1, Cyp17a1, and hsd3b1 (3β-hydroxysteroid dehydrogenase)], as well as abundances of StAR and CYP11A1 protein. Thus, 17β-estradiol enhanced steroidogenesis in ovarian stromal cells. This study provided a basis for further exploration of regulation of steroidogenesis in ovarian stromal cells and the feedback mechanisms in association with estradiol., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Electroejaculation (EE) is frequently used to collect semen, but this procedure is both stressful and painful. In seasonal-breeding species the sensitivity to stressors might vary with season of the year when semen collection occurs, therefore, the present study was conducted to compare the responses when imposing the EE procedure, stress response to EE and fresh semen characteristics in goat bucks throughout the year. Semen was collected using EE from goat bucks once in autumn (mid-breeding season), winter (transition to the non-breeding season), spring (non-breeding season), and summer (beginning of the breeding season). The number of pulses applied during ejaculation was larger in the autumn and spring than winter and summer months. The number of vocalizations/pulse while bucks were ejaculating was greater in the summer than winter and spring months. The increase in heart rate was greater in spring than summer months. Cortisol concentration was least in the autumn, and concentration of creatine kinase (CK) was greatest in summer. Sperm mass motility was greater in autumn than spring months. Spermatozoa concentration, total number of spermatozoa ejaculated, motile spermatozoa ejaculated, and progressively motile spermatozoa were greater in the autumn and spring than in the winter and summer months. The bucks were less stressed by imposing EE procedures in the autumn, and buck responses were greater in the spring-summer period. Bucks had a greater incremental increase in serum CK and number of vocalizations in the summer months, indicating there was a greater muscular damage and quite likely pain associated with lesser semen quality., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The objectives of the study were to determine the dose-dependent effects of active immunization against inhibin α-subunit (AIINHA) on ovarian dynamics, concentrations of progesterone (P 4 ), pregnancy rate (PR), embryonic and fetal losses (EFL), and prolificacy during the non-breeding season when there was imposing of a progestin-based treatment regimen to induce estrus in Beetal goats. Goats (n = 30) were randomly assigned into following groups: 1) saline (G-CON-0 mg; n = 10), 2) small dose (G-AIINHA-0.5 mg; n = 10), and 3) large dose (G-AIINHA-1 mg; n = 10). The primary administration of inhibin immunogen was administered at Day -48, followed by another administration at Day -20, and subsequently there was induction of estrus using a progestin based treatment regimen that included a single administration of progestin-containing sponge and PGF 2α at Day -8. The sponge was removed, and GnRH was administered at Day -3 followed by breeding (Day 0) at standing estrus. Results indicated mean diameter of the follicles, size of pre-ovulatory follicles and corpora lutea, and post-breeding P 4 concentrations were greater (P < 0.05) in the goat does of the G-AIINHA-0.5 than G-CON-0 group. The PR, and EFL, however, did not differ (P> 0.05) among groups, whereas prolificacy rate was greater (P = 0.04) in goat does of the G-AIINHA-0.5 than G-CON-0 groups. The data from this study indicate G-AIINHA-0.5 is the recommended dose of inhibin immunogen to enhance the reproductive performance during non-breeding season in Beetal goats when estrus is induced using a progestin-based treatment regimen., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In the present study, there was evaluation of cryocapacitation-associated changes, apoptotic-like changes, deprotamination, total antioxidant capacity (TAC), and in vitro sperm functional attributes in Barbari bucks after freezing-thawing. The correlation between deprotamination and sperm functional characteristics was established. Using immunoblotting procedures, there was detection of the presence of a single 28-kDa protein band corresponding to protamine-1. The localization in the head region of the spermatozoa was further validated by an immunofluorescence test. Capacitated (B-) and acrosome-reacted (AR-) pattern spermatozoa, spermatozoa with the externalization of phosphatidylserine and a relatively lesser mitochondrial transmembrane potential, and deprotamination and DNA fragmentation was greater (P < 0.05) after freezing-thawing and indicated there were cryocapacitation- and apoptotic-like changes, respectively. Furthermore, the detection of phosphorylation of tyrosine-containing proteins with use of immunoblotting and immunofluorescence procedures confirmed there were cryocapacitation-like changes in the buck spermatozoa after freezing-thawing. Total antioxidant capacity (TAC), in vitro thermal resistance response, Vanguard distance, progesterone sensitivity, and in vitro capacitation response were less (P < 0.05) in the spermatozoa after freezing-thawing compared with spermatozoa after initial dilution and equilibration. Deprotamination (chromomycin A3-positive cells, CMA3+) and DNA fragmentation (TUNEL+ve) were positively correlated with B- and AR-pattern spermatozoa, while other values for other variables were negatively correlated. In conclusion, the results of this study indicated there was protamine-1 in buck spermatozoa and after freezing-thawing there was a loss of protamine-1 combined with cryocapacitation-associated changes and apoptotic-like changes in buck spermatozoa. Spermatozoa deprotamination might be attributed to increased DNA fragmentation, resulting in compromised fertilizing capacity of buck spermatozoa., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The goal of this study was to identify and characterize effects of repeated estrous synchronization (ES) treatments on the regulation of ovarian intergenic long non-coding RNAs (lincRNAs) in dairy goats. Six does were randomly assigned to a group administered three ES treatment regimens separated by 2 weeks or to a group administered only one ES treatment regimen (control) at the same time as the third ES treatment in the does administered the three hormonal regimens for ES. The paired-end RNA Sequencing procedures were used to evaluate lincRNAs of ovarian tissues. A total of 134 lincRNAs were differentially abundant between the two treatment groups. Several target genes were annotated and were related to hormone activity, cellular response to hormone stimulus, response to hormone, female pregnancy, as well as regulation of hormone secretion. These genes were noticeably enriched in MAPK, Hippo, estrogen signaling pathways, oocyte meiosis, progesterone-mediated oocyte maturation, ovarian steroidogenesis as well as GnRH signaling pathways. According to the enriched GO terms and KEGG pathways of regulated genes, 13 differentially abundant lincRNAs could be promising candidates for regulating reproductive functions of female goats. Current results indicate that repeated treatments with gonadotropins affected hormone sensitivity, estrogen synthesis, and ovarian function. The results also indicated that when there was imposing of the three hormonal treatment regimens for ES, there were several lincRNAs that could contribute to dysregulation of several genes that are important for reproduction in dairy goats. Findings provide novel insights for further investigation of lncRNAs biological functions in goats., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study was conducted to assess effects of two hormonal treatments on ovarian follicular status, estrous synchrony and fertility in dairy goats during the non-breeding season when duration of progestogen device use varied by 12 h. In both experiments, does were administered 60 mg of medroxyprogesterone acetate via intravaginal devices, respectively, for 6 and 6.5 d (G6 and G6.5). At 24 or 36 h before device removal, 200 IU of eCG im and 30 μg d-cloprostenol im were administered. In Experiment 1 (n = 24), data related to sexual behavior and that were collected using ovarian ultrasonography were recorded, and in Experiment 2 (n = 83) fertility was assessed after Flexible Time Artificial Insemination (FxTAI). The interval from device removal to estrus was shorter (P < 0.05) after imposing the G6.5 treatment regimen. Diameter of largest and second-largest ovarian follicles and interval from device removal to ovulation were similar (P> 0.05) between groups. The does treated with the G6.5 hormonal regimen had greater estrous synchrony, associated with greater development of largest follicles at the time of device removal, which might have led to a lesser fertility rate (P > 0.05). Conversely, treatment with the G6 hormonal regimen resulted in a greater conception rate. In conclusion, increasing time the intravaginal device is inserted from 6 to 6.5 d resulted in greater estrous synchrony, advanced ovarian follicular development, abnormal CL function and lesser pregnancy rates in artificially inseminated dairy goats when there were treatments during the non-breeding season., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Glucocorticoids have been widely used as anti-inflammatory therapies. The mechanisms of cortisol action in goat does with endometritis, however, have not been reported. The aim of this study was to investigate the mechanism of cortisol in modulation of effects of E. coli-induced endometritis in the does. Does (n = 24) were assigned to four groups (n = 6): control, E. coli, cortisol, and E. coli + cortisol groups. Does in the cortisol and E. coli + cortisol group were treated with cortisol from 3 days before E. coli inoculations occurred to 36 days post-partum. Does in the E. coli and inoculation groups were administered via intrauterine infusion E. coli O 55 (10 9 CFU/mL) at 0 h. Physical indicators, macroscopic and microscopic changes in the endometrium, uterine secretion cytology and bacteriology were evaluated before (0 h) and at 6, 12, 24, 48, and 72 h after E. coli inoculation. The TLR4 and pro-inflammatory cytokine mRNA transcripts were detected using qPCR. The activations of NF-κB and MAPK signaling pathways were detected using Western blot procedures. As a result, cortisol inhibited the inflammatory response of does by reducing the clinical symptoms, morphological endometrial damage, % PMN in uterine secretions, relative abundance of inflammatory gene mRNA transcripts in the endometrium of does. Cortisol inhibited NF-κB activity by reducing MyD88 and IκB phosphorylation. Treatment with cortisol suppressed the phosphorylation of ERK1/2, p38MAPK, and JNK. These results indicate the anti-inflammatory effect of cortisol in the endometrium of does may be regulated by NF-κB and MAPK pathways., Competing Interests: Declaration of Competing Interest None, (Copyright © 2020 Elsevier B.V. All rights reserved.)

The current study was conducted to determine the optimal timing of artificial insemination and vaginal mucous characteristics relative to the onset of estrus in goats. Does (n = 257) were estrous synchronized using hormonal treatments. Intracervical inseminations with fresh semen were performed at 0, 12, 24 and 36 h after the onset of estrus. Characteristics of vaginal mucus (i.e., color, consistency, and volume) were observed and graded at the time of AI to calculate cumulative mucous score. The vaginal electrical resistance (VER) was recorded (Draminski® detector) at the time of AI. Pregnancy rate was less (P < 0.05) when inseminations occurred at 0 h (28.6%) compared with 12 (58.3%) and 24 h (56.4%) after the onset of estrus; however, pregnancy rate at 36 h (54.5%) did not differ (P> 0.05) compared with inseminations at 0, 12 or 24 h after estrous onset. Relative odds for pregnancy rate at 12, 24 and 36 h were 5.24, 5.20 and 3.29 times greater compared with 0 h. Cumulative mucous score varied (P < 0.05) relative to the onset of estrus and correlated well (P < 0.05) with the color and consistency than volume of the mucus. The VER was less (P < 0.05) at 12 than 0 and 36 h after estrous onset. In conclusion, goats can be inseminated between 12-36 h after the onset of estrus; however, the chances of pregnancy are greater with inseminations at the 12 or 24 h time periods., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interests and have no financial or personal relationship(s) which may have inappropriately influenced the research and write-up of this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In seasonally anestrous goat does, ovulations can be induced by combining a treatment regimen including progestagen, eCG and prostaglandins. Nonetheless, ovulations occur only once and then does return to a seasonally anestrous state. This study was performed to determine whether the presence of a sexually active buck can stimulate a second ovulation after induced luteolysis using prostaglandins following the first ovulation. Three groups of seasonally anestrous does were treated to induce ovulations using an intra-vaginally inserted sponge containing a progestin combined with eCG and prostaglandin administrations. Goats that had ovulations were treated with a prostaglandin 11 days after progestin sponge removal. After the prostaglandin injection, does continued to be isolated from bucks (n = 8), were penned with a control buck (n = 9), or were penned with a sexually active buck (n = 10). The proportion of goats having ovulations after imposing the ovulation-induction protocol was greater than 80% and did not differ among treatment groups (P >  0.05). The proportion of does having ovulations after injecting prostaglandins was greater when does were penned with a sexually active buck (8/10) than does penned with a control buck (0/9) or that were isolated from bucks (0/8; P < 0.05). It is concluded that in seasonally anestrous goat does induced to have ovulations using a hormonal treatment regimen, the presence of a sexually active buck can induce a second ovulation when there is an induced luteolysis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study was conducted to characterise pregnancy-associated glycoprotein (caPAG) in peripheral plasma during gestation and postpartum periods of nulliparous and multiparous does with one or two foetuses using a caPAG specific two-step sandwich ELISA system. Earliest time-points for detection of pregnancy and foetal number with appropriate cut-off values were identified. Plasma samples from 15 pregnant (multiparous: n = 8; nulliparous: n = 7; during pregnancy and postpartum period) and six non-pregnant (during oestrous cycle) goats were collected and analysed. Mean caPAG concentration was greater than the threshold for pregnancy detection (S-N = 0.40) on d22, peaked on d45 and remained unchanged until parturition. From d45 until parturition, caPAG concentration in multiparous does with two foetuses was 1.4 to 1.8 fold greater (P < 0.001) than those with one foetus. For the ELISA, 0.83 (S-N) was the most appropriate cut-off to differentiate does with two from those with a single foetus with an overall sensitivity and accuracy of 88.9% and 84.7%, respectively. Circulating caPAG concentration in multiparous goats was greater (P < 0.05) compared with nulliparous goats during the early pregnancy and postpartum periods. After parturition, caPAG concentrations markedly decreased and were basal within 14 days postpartum. In conclusion, using the caPAG specific ELISA, results indicated there were unique gestational and postpartum profiles for caPAG concentrations that are affected by number of foetuses and parity of the doe. The marked decrease in concentration of caPAG following parturition indicates there would not be compromising of the detection of subsequent pregnancies in goats using this technique., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Litter size is one of the most important traits in goat production and breeding. The most common and presumed single nucleotide polymorphism (SNP) detected in the Growth Differentiation Factor 9 gene is c.1189G>A (rs637044681, Ensembl) which results in an altered sequence of the encoded protein. In some studies, there was no effect of this SNP on litter size, while in other studies there was an effect. In the present study there was a meta-analysis conducted by pooling results from 11 eligible published studies to investigate effects of c.1189G>A polymorphism on litter size using four different genetic models including dominant (AA + AG compared with GG), recessive (AA compared with AG + GG), additive (AA compared with GG) and co-dominant (AA + GG compared with AG). Data were analyzed using fixed and random-effect models based on the I-squared value. Results indicate the c.1189G>A polymorphism is positively associated with litter size with use of the dominant model (SMD = 0.093, 95% CI = 0.028 to 0.158, P-Value = 0.005). There, however, was no effect of the c.1189G>A polymorphism using the recessive (SMD = 0.065, 95% CI = -0.164 to 0.295, P-Value = 0.577), additive (SMD = 0.172, 95% CI = -0.169 to 0.513, P-Value = 0.324) and co-dominant (SMD = -0.083, 95% CI = -0.200 to 0.034, P-Value = 0.164) genetic models. Results from use of the sensitivity analysis indicate the GG genotype affect litter size with use of the additive model (P < 0.01). The results from this meta-analysis indicate the GG genotype is associated with litter size in goats., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Uterine receptivity for the embryo is established and maintained through a series of precise cellular and molecular events, such as DNA methylation. There have been no studies to elucidate entire genome DNA methylation changes associated with embryo receptivity development of the endometrium (RE). In the present study, there was development of a complete genome-wide DNA methylome maps of the RE using whole-genome bisulphite sequencing and bioinformatics analysis. As many as 163.06 Gb of sequencing data averaging 81.53 Gb per sample were obtained for genome bisulphite sequencing of endometrium samples. There were distinct genome-wide DNA methylation patterns in pre-receptive endometrium (PE; Day 5 of gestation) and RE (Day 15 of gestation). There were as many as 16,467 differentially methylated regions (DMRs); 21,391 DMRs were less methylated in RE samples compared with PE samples (P-values ≤ 0.05 and |log2 (fold change)| ≥ 2). Compared with PE samples, methylation ratios of IGF2BP2, ACOX2, PTGDS, VEGFB and PTGDR2 genes were markedly less in RE samples (P-value ≤ 0.05 and |log2 (fold change)| ≥ 2). Conversely, in RE samples there was a markedly greater methylation ratio of IGFBP3 and IGF1R genes. The results of KEGG analysis indicated that these genes were involved in the signalling pathways for insulin, mitogen-activated protein kinase, gonadotropin-releasing hormone, vascular endothelial growth factor and progesterone-mediated oocyte maturation, which participated in differential regulation of goat endometrial development during receptive and prereceptive phases. The results of previous and the present study indicate resulting proteins of IGF2BP2, PTGDS, VEGFB, PGR, IGFBP3 and IGF1R gene expression may have important functions in regulating endometrial receptivity for the embryo., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim was to determine effects of maternal feed restriction in dairy goats at gestational different stages on skeletal muscle development and energy metabolism in kids at birth. Six pregnant goats were fed 50% of total digestible nutrients (TDN) and crude protein (CP) (NRC, 2007) recommendations in the first half of gestation and then fed to 100% of the recommendations in the second half of gestation (treatment R-M). In the other group, eight pregnant goats were fed 100% of TDN and CP in the first half of gestation and 50% of a restricted diet the second half of gestation (treatment M-R). Birth weight, blood glucose concentration, muscle fiber number, and size of kids at birth were not affected by maternal feed restriction. The mRNA and protein abundance of myogenic, adipogenic and fibrogenic markers were not affected (P > 0.05) by maternal diet. With regard to values for variables in kid energy metabolism, mRNA abundance of the glycolic enzyme HKII was less (P = 0.03) in the M-R group. In conclusion, maternal feed restriction in the first or second half of gestation had no affect mRNA abundance on myogenic, adipogenic, and fibrogenic markers nor were there changes in skeletal muscle mesenchymal stem cell population of kids at the time of birth. There, however, may be detrimental effects on energy metabolism by reducing HKII gene expression in skeletal muscle of dairy goat kids at the time of birth., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study was designed to investigate plasma profile of pregnancy-associated glycoprotein (PAG), progesterone (P4) and estrone sulfate (E 1 S) during early- and mid-pregnancy. The goal was to explore the relationships with values for reproductive variables, to detect the most reliable predictor variable, and to identify the most desirable time point for blood collection for determining fetal number in goats. After ultrasonographic examination at d35-40 post-mating, blood sampling of 15 pregnant goats (total 18) was continued until d114. The PAG profile was characterized by gradual increase during early pregnancy from d26 to d51 and thereafter concentrations were relatively constant until d114 of gestation. The effect of fetal number on plasma PAG, P4 and E 1 S was first evident on d28, d51 and d26, respectively. During mid-pregnancy, does with twins had a greater (P < 0.05) PAG (S-N = 2.54 ± 0.12 compared with 1.59 ± 0.11), P4 (18.91 ± 0.67 compared with 14.51 ± 0.47 ng/mL) and E 1 S (16.34 ± 0.76 compared with 11.32 ± 0.44 ng/dL) as compared with does with a singleton fetus. Plasma PAG but not P4 and E 1 S was positively correlated with fetal number and birth weight of kids during early pregnancy. Multivariate linear regression and discriminant function analyzes allowed for identification of plasma PAG as the most reliable predictor for fetal number and birth weight of kids. Furthermore, d58 was the most suitable single time point for prediction of fetal number using PAG as a biomarker. In conclusion, plasma profile of PAG, P4 and E 1 S was affected by fetal count. Plasma PAG was identified as the most reliable predictor variable of fetal number and birth weight of kids as compared to plasma P4 and E 1 S., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this study was to evaluate the effect of the stage of seasonal anestrus and breed on ovarian response in non-estrous cycling goats using a progesterone (P4) injection plus human chorionic gonadotropin (hCG)-based protocol. In Experiment 1, non-estrous cycling local Mexican goats were treated with 20 mg of P4 plus 100 IU of hCG injections 24 h apart during April (early anestrus, n = 13) or June (late anestrus, n = 12). The estrous response, interval from hCG-to-estrus, and interval to ovulation were not affected by season (P > 0.05). In addition, the size of the follicle from which ovulation occurred and the size of the corpus luteum were not different between the two stages of seasonal anestrus (P > 0.05). In Experiment 2, the estrous response was compared between multiparous non-estrous cycling local Mexican (n = 18) and Alpine (n = 19) goats in which stage of the estrous cycle was synchronized using the same P4+hCG protocol as in Experiment 1. Neither the onset of estrus nor the time of ovulation differed between breeds, and the estrus-to-ovulation interval also was similar for both breeds (P > 0.05). The diameter of the dominant follicle at the time of ovulation was similar between local and Alpine goats (P > 0.05). In addition, the pregnancy rate was not different for both local and Alpine goats (P > 0.05). In conclusion, results of this study indicate that the stage of seasonal anestrus or breed do not modify estrous and ovarian response in non-estrous cyclic goats synchronized with a P4 injection plus hCG-based protocol., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this study was to examine whether photoperiod-treated bucks have the same capacity as melatonin-treated bucks to induce reproductive responses in female goats during the spring. On 10 April, 38 anoestrous does were placed with: 1) photoperiod-treated bucks (additional light hours for 83 days from the end of the previous November; PHOTO; n = 12); 2) bucks treated with exogenous melatonin at the beginning of March (MEL; n = 13); and 3) bucks that received no treatments (CONTROL; n = 13). The bucks' sexual behaviour was assessed for 10 days, and doe oestrous behaviour was recorded for the next 32 days by checking for harness marks. Ovulation was confirmed from plasma progesterone concentration (measured twice per week) and ovulation rate was assessed by transrectal ultrasonography. Fecundity, fertility, prolificacy and productivity were also determined. The percentage of does in the PHOTO, MEL and CONTROL group: 1) having ovulations was 92%, 100% and 38% respectively; 2) expressing behavioural oestrous associated with ovulation was 92%, 100% and 31%; and 3) that became pregnant was 75%, 69% and 23%, respectively. The kids produced per doe were 1.08 ± 0.23, 1.15 ± 0.25 and 0.31 ± 0.17 for the PHOTO, MEL, and CONTROL groups, respectively with there being no differences between the PHOTO and MEL groups, however, there was a difference (P < 0.05) between PHOTO/MEL and CONTROL groups. Thus, imposing a stimulatory lighting regimen during the preceding winter period is as efficient as using subcutaneous melatonin implants in stimulating reproductive behaviours and physiological functions of bucks., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Changes in anatomical, behavioural and seminal characteristics were measured throughout the year in seven Beni Arouss bucks maintained at a latitude of 35 °N. Testicular size varied significantly. There were the greatest values during the summer and autumn for scrotal circumference and least in the winter for testicular length. Values for most variables related to sexual behavior were not affected by season except the number of mounts before first ejaculation which was least in the summer. No differences were detected between seasons in terms of semen volume. The sperm concentration, viability and percentage of normal sperm, however, were less during the winter period. The size of sperm heads was larger in the autumn and motility of spermatozoa was greater in the summer and autumn. Results from the correlation analysis indicated that the majority of conventional seminal characteristics were correlated with testis measurements (P <  0.05; r < 0.5). Values for conventional and CASA motility variables indicated there were positive correlations between viability and progressive motility (r = 0.4; P <  0.05) and between the percentage of normal sperm and straight line and average path velocity (r = 0.4; P <  0.05). A global reproduction performance score was established for each buck, which allowed for assessment of magnitude of seasonal changes for each individual. These global score values indicated there was a greater reproductive performance for all bucks during summer and autumn. This study described for the first time seasonal variations of reproductive characteristics of Beni Arouss bucks and results indicate that even though there is a maximal capacity for reproductive performance during the summer and autumn, breeding should be possible throughout the year., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Most goats exposed to males in confined conditions have short ovulatory cycles. The frequency of these cycles can be reduced with a progesterone treatment prior to the introduction of males. The objective of this study was to determine whether extensive management conditions modify the frequency of short ovulatory cycles in progesterone-treated does exposed to photostimulated males. One group of does remained in extensive management conditions and grazed daily from 10:00 to 18:00 h; two other groups were confined separately in shaded pens, and fed alfalfa hay. In March, females from the grazing group (n = 45; grazing-P4 group) and those from one confined group (n = 45; confined-P4 group) were treated with 25 mg of progesterone by intra-muscular injections 48 h before joining with photostimulated males (n = 3 per group). The other confined group did not receive the progesterone treatment and was exposed to two photostimulated males (n = 25; confined-control group). Does were exposed to males for 45 consecutive days. Determination of whether ovulations occurre were made by quantifying plasma progesterone concentrations after introduction of males. The proportion of does that had ovulations at least once was not different between groups (≥98%; P > 0.05). The proportion of does that had short ovulatory cycles differed between groups (P < 0.001), and this proportion was greater in the confined-control group (76%) than in confined-P4 (27%) and grazing-P4 groups (25%; P < 0.001). It is concluded that extensive management conditions do not modify the frequency of short ovulatory cycles in progesterone-treated does exposed to the photostimulated males., (Copyright © 2018. Published by Elsevier B.V.)

When the sexual activity of bucks is minimal, there is a minimal male effect on does regardless of their body weight (BW)/body condition (BC) and whether does are treated with melatonin or not. The study examines whether sexually active bucks can induce an adequate male effect in does with an increasing or decreasing trajectory of change in BW/BC when does are or not treated with melatonin. During natural anoestrus, 46 Blanca Andaluza does were assigned to two groups: 1) low BW/low BC group in which does were fed 1.9 times maintenance requirements for dietary energy for gaining BW/BC (LLg group; n = 23); or 2) a high BW/high BC group in which the does were fed 0.4 times maintenance requirements for dietary energy that resulted in a loss of BW/BC (HHl group; n = 23). There were similar numbers of does in each group that were treated or not treated with melatonin (MEL). Following 48 days of isolation from bucks, four sexually active individuals fitted with marking harnesses were transferred to the paddock containing the does of each group. Blood samples were collected by jugular venipuncture (before the distribution of concentrate) twice per week. The effect of the treatments (increasing or decreasing BW/BC and melatonin) on the different variables that were assessed were analysed using an ANOVA or the Fisher-Freeman-Halton exact probability test as necessary. During the 35 days after treatments were applied, the percentage of females expressing oestrous and having an ovulation were greater in the LLg + MEL than HHl-MEL subgroup (P < 0.05). The interaction of nutrition × melatonin treatment had a significant effect on reproduction of does (P < 0.05). This could be explained by the greater plasma glucose and IGF-1 and lesser plasma non-esterified fatty acid concentrations in does with increasing BW/BC (P < 0.01), and the greater IGF-1 concentrations of MEL-treated females (P < 0.01). The LH concentration and pulsatile release of this hormone from the pituitary were also modified by the presence of the males (P < 0.01). Furthermore, the LLg + MEL-treated does were responsive to the presence of bucks (P < 0.05). The present results indicate sexually active males cannot induce an adequate reproductive response in females with decreasing BW/BC even when does are being treated with melatonin. The presence of bucks enhanced the doe reproductive response when does were treated with melatonin and a pattern of increasing BW/BC., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The present study was conducted to characterize the major proteome of ovarian follicular fluid from locally-adapted, "Canindé" goats in the northeast of Brazil. Eight estrous cycling goats received a hormonal treatment consisting of medroxyprogesterone acetate, D-cloprostenol and FSH. Fluid was collected by laparoscopy from small (<3mm), medium (3-4mm) and large (>4mm) follicles and then, proteins were analyzed by 2-D SDS-PAGE and tandem mass spectrometry. Thirty-six proteins were identified in the goat follicular fluid, including albumin, immunoglobulins, ceruloplasmin, complement factor B, alpha-1B-glycoprotein precursor, serotransferrin, complement C3 and serpins, among others. Albumin and immunoglobulins were the most abundant proteins. Protein concentrations were similar in the fluid from small (45.3±3.1mg/mL), medium (44.2±3.3mg/mL) and large follicles (45.1±2.3mg/mL). The intensities of spots identified in 2-D gels as serotransferrin, zinc-alpha-2-glycoprotein-like, complement factor B and complement protein C3 differed (P<0.05) among follicle categories. The amount of serotransferrin was greater in the medium than small follicles (P<0.05). Content of zinc-alpha-2-glycoprotein-like, complement factor B and complement C3 was greater (P<0.05) in the fluid of large follicles than in medium follicles. Based on gene ontology, the major molecular functions associated with goat follicular fluid proteins were binding and catalytic activity, while the main biological processes were related to regulation, cellular processing, location and the immune system. In conclusion, the major proteome of the follicular fluid from goats subjected to hormonal stimulation was elucidated in the present study. Also, molecules associated with follicle development are potential biomarkers of oocyte competence were prevalent., (Copyright © 2017. Published by Elsevier B.V.)

This study evaluated the efficiency of two d-cloprostenol injections at different intervals on the reproductive parameters of dairy goats. Trial 1 comprised 54 goats allocated to receive two 37.5μg d-cloprostenol doses at intervals of seven (T 7 , n=19), 10 (T 10 , n=18), and 11.5 (T 11.5 , n=17) days. Trial 2 comprised 62 goats allocated to receive injections at T 7 (n=30) and T 11.5 (n=32). Ultrasonography was done and blood was collected just before d-cloprostenol injections. After the second dose, goats were artificially inseminated (AI) with frozen-thawed semen at 18-24h (Trial 1) or at 10-24h (adjusted according to the time of estrus onset in Trial 2) after estrus detection. Estrus response rate did not differ (P>0.05) among groups in Trials 1 (T 7 =94.7%; T 10= 88.9%; T 11.5 =88.2%) and 2 (T 7 =90.0%; T 11.5 =96.9). All females showed progesterone concentrations >1ng/mL before both d-cloprostenol injections. The largest follicle diameter present on ovaries was similar (P>0.05) among treatments at the first and second dose. The second largest follicle diameter was superior (P<0.05) to T 7 than to T 10 and T 11.5 goats at first dose only. This possibly resulted in lower interval to estrus (P<0.05) in T 7 -treated goats than other treated goats in both trials. The conception rate was similar among treatment groups in Trials 1 (T 7 =55.6%; T 10 =18.8%; T 11.5 =26.7%) and 2 (T 7 =85.2%; T 11.5 =93.6%). The three treatments efficiently synchronized estrus. T 7 and T 11.5 protocols resulted in high estrus synchrony and conception rates when adjusting the AI time according to interval of estrus., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Understanding the genetic information of related genes is helpful for the selection and breeding course through marker assisted selection. The aim of the current study was to detect polymorphisms of the KiSS1 gene in 137 animals, including Baladi, Zaraibi and Damascus goat breeds by PCR-RFLP, and DNA sequencing and to investigate the association between these variants and reproductive traits. Comparison of the nucleotide sequence indicated the substitution of T with A at position 121 (T121A) in the intron 1 of the KiSS1 gene in all goat breeds. This substitution distorts the restriction site of the XmnI restriction enzyme and consequently two genotypes were detected (TA and TT). The T121A SNP is associated significantly with litter size in Damascus and Zaribi breeds (p=0.025 and 0.001, respectively). The animals with the TT genotype in Damascus and Zaribi breeds had a significantly higher estradiol 17β level than that recorded in TA genotype at estrus phase (p=0.013 and 0.028, respectively) and late-luteal phase (p=0.067 and 0.041, respectively) of the estrus cycle. Furthermore, animals with the TT genotype in Damascus and Zaribi breeds had significant higher progesterone level at mid-luteal (p=0.037 and 0.045, respectively) phase. Meanwhile, there were no significant differences in progesterone level in late-luteal phase between both genotypes in Zaribi breed (p=0.267). The current trial indicated that the prolific TT genotype in both Damascus and Zaribi breeds had superior estradiol 17β level at estrus phase and an eminent progesterone level at both early and mid-luteal phases of the estrous cycle., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The aim of the present study was to evaluate uterine blood flow (UBF) during the postpartum period in small ruminants. The study involved measures of UBF in 5 ewes and 5 goats during the first 4 weeks after parturition. Transrectal quantification of UBF was assessed by determining the diameter (DM), time averaged mean velocity (TAMEAN), blood flow volume (BFV), blood flow acceleration (ACCE) and pulsatility index (PI) in the uterine arteries ipsilateral to the previously gravid uterine horn(s) at Wk 20 of gestation and every 3days from the day of parturition (D 0) until Day 27 postpartum (D 27). The diameters of the uterine arteries decreased (P<0.01) in both species during the postpartum period. The results revealed decreases (P<0.0001) in BFV, ACCE and TAMEAN, while PI increased during the postpartum period in both sheep and goats. Furthermore, there were positive correlations (P<0.05) between blood flow parameters in sheep and goats, respectively (BFV and DM, r=0.62 and 0.58; BFV and ACCE, r=0.32 and 0.26; BFV and TAMEAN, r=0.51 and 0.37). There were negative correlations (P<0.05) between PI and other parameters (PI and BFV, r=-0.39 and -0.36; PI and DM, r=-0.54 and -0.24; PI and ACCE, r=-0.58 and -0.48; and PI and TAMEAN, r=-0.80 and -0.79) in sheep and goats, respectively. Altogether, these results indicate that non-invasive Doppler ultrasound parameters provide important information toward understanding changes in the vasculature and its perfusion of the uterus during the postpartum period in sheep and goats., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Nerve Growth Factor (NGF) promotes the development of pre-antral ovarian follicles through ovarian innervations and regulation of ovarian response to gonadotropins. The present study was conducted to study the tissue gene expression profile, to characterize the genetic variants, find associations of the NGF gene with prolificacy in the prolific Malabari and less prolific Attappady Black goats because NGF has an important role in reproduction by augmenting ovarian folliculogenesis. Relative abundance of NGF mRNA was greatest in reproductive tissues signifying its role in reproduction. The PCR-SSCP analysis of a 251bp fragment of Exon 3 of the NGF gene from the 277 goats revealed four diplotypes (EE, EF, FF and EG) with respective frequencies of 0.76, 0.22, 0.01 and 0.01. Sequencing of the representative samples revealed one synonymous and one novel non synonymous mutations (g.705G>A and g.715C>T). Statistical analysis indicated that the SNP g.705G>A was associated with litter size in Attappady Black goats (P<0.05) and a PCR-RFLP was designed using the restriction enzyme, BpiI, for rapid screening of the SNP. The results of the present study suggest that the NGF gene is a primary candidate gene affecting prolificacy in goats and may be used for Marker Assisted Selection (MAS) in goats, especially in lowly prolific Attappady Black goats., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Two trials were conducted to evaluate the effect of diverse socio-sexual cues upon male sexual behavior and the reproductive performance of anestrous does (AD). Trials were conducted in northern Mexico (26°N) during the natural anestrous season (Feb-Mar) with crossbred dairy bucks. In Experiment 1, sexually inactive bucks (SIB, n=12) were randomly allotted to three groups, four males/group: a) DEE 9novelty stimulation) - daily exchange of estrogenized females (EF) 12&12h, b) NEE (no novelty stimulation) - no-exchange of EF, 24h, or c) CON (saline-treated&#95; -daily exchange of AF 12&12h. Sexually active bucks (SAB) from the DEE, NEE and CON groups were subsequently exposed to AD (n=72; n=24/group) and the reproductive outcomes were recorded. In Experiment 2, SAB (n=12; n=6/group) were randomly divided in: 1) B+EF - males+four-EF exposed to AF (n=36), and 2) B+NEF; males+four-saline-treated AD and exposed to AD (n=36). Prior to the onset of the experimental breeding in both experiments, the investigative (ISB), consummatory (CSB) and resting (RSB) sexual behavior of males were quantified (2h×d×2d). Sexual behaviors considered were: ISB - flehmen, ano-genital sniffing, approaches, vocalizations, kicking, penis extrusion, CSB; mount attempts and mounts, and RSB - isolation, attempted escape, aggression and distractions. While EF were an effective stimulus (P<0.05) for evoking mounting in SIB males, daily exchange of estrous does used to stimulate males promoted an enhanced response (P<0.05) in terms of both ISB and CSB. After being exposed to AD, the B+EF bucks induced an earlier estrous response (P<0.05) as compared with the B+NEF bucks. The untreated females did not induce any sexual activity in males and stimulation of ovarian function did not occur when saline treated (CON) AD were exposed to AD. Also, the B+EF group induced an enhanced increase (P<0.05) of the male ISB and CSB, inducing in turn an increase percentage onset of estrus in does that had previously been anestrus (P<0.05)., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the reproductive response of anoestrous goats that were either hormonally treated and/or supplemented with maize for 9days to determine which treatment combination was the most effective in enhancing follicular development and ovulation rate, and whether these responses were associated with increases in metabolic hormones. The experiment was carried out using 28 does, using a 2×2 factorial design with seven does in each group to test the effect of synchronisation of oestrus, supplementation with maize and their interactions. Synchronisation of oestrous cycles (P<0.001) but not supplementation with maize or the interaction between the two (P>0.05) increased the number of codominant follicles, the diameter of the largest follicle on Day 9 and growth rate of follicles during the period of supplementation. Compared with non-supplemented animals, supplementation with maize increased the total number of follicles observed between Days 7 and 9 (P=0.039). In addition, nutritional supplementation with maize in combination with synchronisation of oestrus increased the ovulation rate by 43% (P=0.074). Interactions between time and supplementation with maize showed that plasma concentrations of insulin, leptin and IGF-1 were greater in does supplemented with maize compared with non-supplemented does (P<0.001). The findings show that hormonal synchronisation had the most influence on modifying follicular development and ovulation in anoestrous goats. Supplementation with maize increased the concentrations of insulin, leptin and IGF-1, which could potentially modify the sensitivity of follicles to gonadotrophins and reduce rates of atresia., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The present study was designed to study the ultrastructure of goat corpora lutea (CL, n=10) and structural changes as related to steroidogenic functions during the estrous cycle. The reproduction status of goats was estimated by analyzing serum progesterone concentrations. The CL at various stages was surgically collected. To characterize ultrastructural features associated with steroidogenesis, tissue and cellular structures were studied. Blood supplies were examined based on features of the endothelial cells and capillary structures in the CL. Activated endothelial cells and developing vessels were observed in the early stage, whereas mature endothelial cells, accumulating extracellular matrix fibers, and stabilized vessels were observed in the middle and late stages of assessment. In the late stage of assessment, shrunken goat luteal cells scattered around the capillaries were detected and formed circular regression areas. Features of autophagy and luteal cell apoptosis were noted. In large luteal cells, steroidogenic organelles were present, including microvillar channels, endoplasmic reticulum, and mitochondria. Conformational changes in the endoplasmic reticulum and increased mitochondria with tubular cristae were observed in the early-middle CL transitions. In contrast, mitochondria swelled and the cristae transformed to the lamellar type in the late stage, suggesting that organelle plasticity could contribute to steroidogenesis in goat CL. In conclusion, results suggest angiogenesis occurs in early developing CL and programmed cell death occurred in the late stage of CL assessment in the present study. Structures and quantiles of steroidogenic organelles are correlated with the steroidogenic functions in goats., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Goats are seasonal breeders and photoperiod is the main cue controlling the onset and offset of the breeding season. Nevertheless introducing a sexually active buck in a group of females during anestrous can stimulate their reproductive function and induce ovulation. This "male-effect" is very efficient under subtropical latitudes, when using sexually active males previously stimulated by a photoperiodic treatment. However, there is less evidence of its feasibility under temperate latitudes where the more important variation in day length could be responsible for a stronger inhibition of female sexual activity. The aim of this study was therefore to determine whether intense sexual activity can be induced in alpine bucks during the non-breeding season by a long-day treatment under temperate latitude and if these males could be used to produce an efficient male-effect. Bucks (n=21) were divided in two groups, one submitted to a photoperiodic treatment from November 1st to January 15th and then switched to natural photoperiod, while the other group remained entirely under the natural photoperiod. The ones submitted to this light treatment exhibit higher testicular volume and testosterone level 6 weeks after the end of the treatment. At the end of March, bucks were used to stimulate anestrous does (n=41) continuously for 15 days. We showed that (a) light treatment was efficient to induce an increase of sexual activity in bucks and (b) that the introduction of stimulated bucks among females induced a significantly higher proportion of ovulation in anestrous does than control bucks (86% vs 5%). Our results indicate that under temperate latitudes induction of ovulation in females during the anestrous season is feasible using bucks treated with long-days during winter., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Semen collection for cryopreservation is a key step for small ruminant conservation programs. While in these species semen is mainly collected via artificial vagina (AV), electroejaculation (EE) provides a viable alternative for untrained males. Herein we investigated the effect of semen collection method on post-thaw sperm quality by comparing two small ruminant species, sheep and goats. Semen from Blanca-Celtibérica bucks and Manchega rams was collected by AV and EE on the same day and cryopreserved using a standard protocol. At thawing, sperm motion parameters were evaluated by CASA, whereas membrane stability (YO-PRO-1), sperm viability (propidium iodide, PI) and mitochondrial activity (Mitotracker Deep Red) were analyzed using flow cytometry. The semen collection method negatively affected post-thaw sperm quality in bucks but not in rams. Thus, in bucks, post-thaw sperm motility was higher for samples collected by AV as compared to those obtained via EE. Similarly, post-thaw sperm parameters evaluated by flow cytometry were worse for buck samples collected by EE than those collected by AV in the same species, or than ram samples regardless of collection method. These results suggest that ovine and caprine spermatozoa have a different response to the cryopreservation process depending upon the semen collection method used. We hypothesize that the EE procedure may lead to changes in the composition of the ejaculate in bucks that would make spermatozoa more susceptible to the cryopreservation process, whereas this procedure would have had no effect on ram spermatozoa. This assumption requires further investigation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The insulin-like growth factor 1 has an important role in reproduction, foetal development and growth. It regulates the secretion of gonadotrophin releasing hormone, stimulates ovarian function and steroidogenesis. The present study was conducted to characterise the 5' flanking region of goat IGF 1 gene, ascertain ovarian expression of the IGF1 gene, detect SNPs and assess the association with prolificacy in the two indigenous goat breeds of South India viz., low prolific Attappady Black and high prolific Malabari. The 5' flanking region of IGF1 gene was PCR amplified, cloned and sequenced from both breeds. Genotyping was performed in 277 goats from the two genetic groups using the PCR-Single Strand Conformational Polymorphism (SSCP) and the expression of the IGF1 gene in the ovary was analysed by quantitative real time PCR. The 5' flanking region of the IGF1 gene was 601 bp long and located at 450 bp upstream of the start codon. Sequence exhibited 97-99% similarity with that of the sheep, cattle and sika deer IGF1 genes. Three genotypes, PP, PQ and QR were observed at this locus with the frequency of 0.62, 0.30 and 0.08, respectively. Sequencing of the representative PCR products from each genotype revealed two SNPs, g.224A>G and g.227C>T. The population was found to be in Hardy-Weinberg disequilibrium at both loci. Statistical results indicated that these loci were associated with litter size (P ≤ 0.05). However, no significant difference was found in the expression of the IGF1 gene in the ovaries of the two goat breeds. These results suggest the significant influence of the IGF1 gene on prolificacy in goats and identified SNPs would benefit the selection of prolific animals in future breeding programs., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This study aimed to determine the possible effects of a single injection of human chorionic gonadotropin (hCG) as a means for estrus induction in acyclic French-Alpine goats during the reproductive transition period at 25°N, 103°W. The potential effects of hCG upon ovarian function and reproductive performance of goats were also assessed. Multiparous acyclic French-Alpine goats (n = 39; 37.4 ± 8 .5 kg) were primed with 20mg progesterone (P4) 1 day prior to hCG administration. Thereafter, does were treated either with saline (hCG-0; n = 10), 50 (hCG-50; n = 9), 100 (hCG-100; n = 10), or 300 IU of hCG (hCG-300; n = 10). Ovarian structures and pregnancy were monitored by transrectal ultrasonography. In addition, after hCG application, goats were monitored twice daily (0800 and 1800 h) to detect estrus signs, with the use of aproned, sexually active bucks treated with testosterone. Goats were bred 12h after the onset of estrus. Two days after hCG administration, the number of large follicles was higher (P < 0.05) in the hCG-50 and hCG-300 groups (1.7 ± 0.1 and 1.8 ± 0.2, respectively) compared with the hCG-100 and hCG-0 groups (1.4 ± 0.2 and 1.1 ± 0.1, respectively). Although none of the hCG-0-goats depicted estrus, the estrus response from the hCG-50, hCG-100, and hCG-300 groups over the 7-d breeding period was 67%, 100%, and 90%, respectively (P > 0.05), being always accompanied by ovulation. Pregnancy rate (67, 100, and 70%), kidding rate (55%, 80%, and 70%), and litter size (1.6 ± 0.5, 1.5 ± 0.5, and 1.5 ± 0.5) for hCG-50, hCG-100, and hCG-300, respectively, did not differ among the hCG-treated does. Therefore, the combined use of P4-priming plus a 100-IU hCG injection is an effective protocol for inducing estrus in non-cycling Alpine goats during the anestrus-to-estrus transition period, which is of key importance for both goat producers and industrializers., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of this study was to evaluate the effect of different combinations of insulin and FSH concentrations in culture media containing GH on the in vitro follicle morphology, antrum formation, growth rates, estradiol (E2) production, oocyte viability and maturation as well as gene expression for FSHR, GHR, INSR, CYP19A1, CYP17, 3ßHSD. Secondary follicles were individually cultured for 18 days in a basic medium containing 50ng/mL GH supplemented with low insulin concentration (INS-LW: 10ng/mL) or high insulin concentration (INS-HG: 10μg/mL) alone or with a fixed FSH concentration (FSH100: 100ng/mL) or with increasing FSH concentrations (FSH-SEQ: 100ng/mL, days 0-6; 500ng/mL, days 6-12; 1000ng/mL days 12-18). In the INS-LW treatment was observed a higher (P<0.05) incidence of normal follicles at day 18 of culture. However, overall higher (P<0.05) follicular growth, oocyte diameter and meiotic resumption rates were obtained using INS-HG+FSH 100. The INS-HG and INS-HG+FSH100 treatments showed higher E2 production and mRNA levels for CYP19A1, CYP17, 3βHSD when compared to INS-LW and INS-LW+FSH100. However, the addition of increasing FSH concentration, regardless of insulin concentration, did not improve the follicular growth, meotic resumption, E2 production or gene expression of steroidogenic enzymes when compared with INS-HG+FSH100. In conclusion, in presence of GH, a basic medium supplemented with 10μg/mL insulin and 100μg/mL FSH throughout the culture period, improves follicular and oocyte growth, oocyte meiotic resumption and E2 production from isolated preantral caprine follicles cultured in vitro., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A specific problem in goat semen preservation is the detrimental effect of seminal plasma on sperm viability in extenders containing yolk or milk. Thus, the use of chemically defined extenders will have obvious advantages. Although previous studies indicate that the initial pH of an extender is crucial to sustain high sperm motility, changes in extender pH during long-term semen storage have not been observed. Monitoring extender pH at different times of semen storage and modeling its variation according to nonlinear models is thus important for protocol optimization for long-term liquid semen preservation. The present results showed that during long-term liquid storage of goat semen, both sperm motility and semen pH decreased gradually, and a strong correlation was observed between the two. Whereas increasing the initial extender pH from 6.04 to 6.25 or storage with stabilized pH improved, storage with artificially lowered pH impaired sperm motility. Extender renewal improved sperm motility by maintaining a stable pH. Sperm coating with chicken (Gallus gallus) egg yolk improved motility by increasing tolerance to pH decline. A new extender (n-mZAP) with a higher buffering capacity was formulated, and n-mZAP maintained higher sperm motility, membrane integrity and acrosome intactness than the currently used mZAP extender did. Goat semen liquid-stored for 12 d in n-mZAP produced pregnancy and kidding rates similar to those obtained with freshly collected semen following artificial insemination. In conclusion, maintenance of a stable pH during liquid semen storage dramatically improved sperm viability and fertilizing potential., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the hormonal and ovarian responses to the administration of a metastin/kisspeptin analog (TAK-683) under the endocrine environments of luteal and follicular phases in goats. Five estrous cycling goats received a prostaglandin F2α injection followed by 10 days of progesterone treatment by CIDR. The TAK-683 (35nmol) was intravenously administered (Hour 0) on 3 days after CIDR insertion (luteal phase condition; LC) and at 12h after CIDR removal (follicular phase condition; FC). Blood samples were collected at 10min (-2 to 6h), 2h (6-24h) or 6h intervals (24-48h). In the LC, small increases in the basal concentrations of LH were observed after TAK-683 administration from 0 to 6h, which were associated with an increase in estradiol concentration, followed by a surge-like release of LH with a peak at 12.5±1.0h (n=4) after TAK-683 administration. In the FC, a surge-like release of LH occurred immediately after TAK-683 administration with a peak at 6.0±3.5h (n=5), which was earlier than that in the LC (P<0.01). The peak concentration of estradiol did not differ between the two conditions, whereas the time interval from TAK-683 treatment to estradiol peak in the LC was longer than that in the FC (12.0±0.0 compared with 6.0±4.2h, P<0.05). These findings suggest that the timing of surge-like release of LH after TAK-683 administration is associated with blood estradiol concentration at the time of treatment., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This paper compares the effects of exposure to exogenous melatonin (MEL), short days (SD, 8h of light) and long days (LD, 16 h of light), on reproductive activity, sperm motility and other reproductive variables, in Blanca Andaluza bucks. Fourteen males were spilt into two groups of seven animals (G1 and G2). They were subjected to five alternations of 2 months of LD followed by 2 months of SD or MEL before the experimental period of three consecutive intervals of: (1) 2 months of SD (G1, N=7) or MEL (G2, N=7); (2) 2 months of LD (G1+G2, N=14); and (3) 2 months of SD (G2, N=7) or MEL (G1, N=7). Plasma testosterone concentration, live weight, testicular weight and fresh semen quality were determined weekly. Semen was also cooled and frozen-thawed every fortnight, and the same quality variables measured as for fresh sperm. When the bucks were under LD treatment, the testosterone concentration was lower than when under MEL or SD treatment (P<0.01); values for the semen concentration and total number of sperm per ejaculate were also higher (P<0.001). No differences were observed between the MEL and SD treatments in terms of fresh, cooled or frozen-thawed sperm quality. Only some quality variables on fresh semen were improved by MEL and SD treatment (P<0.05). In conclusion the results of the present experiment showed that MEL improved the fresh semen motility variables, but this did not improve the motility of frozen-thawed sperm over that recorded for either SD or LD treatment., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study examines the nutritional and metabolic cue-induced modulation of the reproductive performance response of female goats to the male effect. During natural anoestrus, 48 Blanca Andaluza does were isolated from bucks for 45 days and distributed into two groups: (1) low body weight (BW)/low body condition score (BCS) animals (LL-gain group, N=18), which were fed 1.9 times their maintenance requirements; and (2) high BW/high BCS animals (HH-loss group, N=30), which were fed 0.4 times their maintenance requirements. Following isolation, oestrous activity was recorded daily by visual observation of the marks left by harness-equipped males. Weekly blood samples were taken for the determination of progesterone, glucose, insulin, non-esterified fatty acids (NEFAs) and leptin concentrations. Fecundity, fertility, prolificacy and productivity were also determined. Significantly greater ovarian and oestrous responses, and productivity, were observed in the LL-gain group compared to the HH-loss group (P<0.05). After the introduction to the males, no differences in NEFA concentration were seen between the groups; before introduction the values were higher in the HH-loss group. At the moment of detection of oestrus following male introduction, the insulin concentration of the LL-gain animals was higher (P<0.05). The present results show that the reproductive performances of does subjected to the male effect in spring are poorer in those with a decreasing BW and BCS and better in those with increasing scores. This might be explained by the differences between groups in terms of their plasma insulin concentrations. The NEFA concentration was clearly modified by introduction to the males., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Polymorphisms in the promoter region are likely to impact KISS1 gene transcription and reproductive traits. In this study, Guanzhong (GZ, n=350) and Boer (BE, n=196) goats were used to detect polymorphism in the promoter of the goat KISS1 gene by DNA sequencing. In the GZ goats, the g.1384G>A mutation was identified in the promoter of the goat KISS1 gene. Guanzhong goats were in Hardy-Weinberg disequilibrium at g.1384G>A locus (P<0.05). The 1384A allele was predicted to eliminate methylation, AHR-arnt heterodimers and AHR-related factors (AHRR) and myoblast determining factors (MYOD) transcription factor-binding sites. Statistical results indicated that the g.1384G>A SNP was associated with litter size in the GZ goats (P<0.05). Luciferase assay analysis suggested that the 1384A allele increased luciferase activity when compared to the 1384G allele. The RT-qPCR assay also demonstrated that the 1384A allele had greater amounts of KISS1 mRNA than the 1384G allele in homozygous individuals. Functional analysis suggested that this g.1384G>A SNP may be an important genetic regulator of KISS1 gene expression with effects on downstream processes that are modulated by KISS1 gene because of the changes of methylation and transcription factor-binding sites. Therefore, the current study provides evidence in goats for genetic markers that might be used in breeding programs., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Sperm morphology has been identified as one characteristic which can be useful in prediction of fertility in a species. The development of computer automated sperm morphometry analysis allows for objective analysis of sperm head dimensions. The aim of the current study was to develop an optimum sampling procedure to characterize the Iberian ibex (Capra pyrenaica) sperm head morphometrically. Fresh semen from 11 males was collected using transrectal ultrasonic-guided massage of accessory sex glands and electroejaculation and prepared on slides for morphometric analysis to evaluate technical variation and standardize automated sperm morphometry analysis procedures by Sperm-Class Analyzer(®). Three staining methods (Diff-Quik(®), Hemacolor(®), Spermblue(®)), number of sperm cells necessary to sample and repeatability of the staining technique were assessed. There were significant differences in size of sperm head depending on stain used. Hemacolor(®) was stain most suitable for sperm head morphometry evaluation (length=8.42 μm; width=4.21 μm; area=29.37 μm(2); perimeter=21.93 μm; elongation=0.33; elipticity=2.01; regularity=0.95; rugosity=0.77). Morphometric values obtained from samples of 50, 100, 150, 175 and 200 sperm heads were compared. The most efficient method of analyzing sperm morphometry was to evaluate 100 sperm cells at 60× objective magnification. Thus, this study has allowed for description of optimal sample processing to determine morphometric parameters of sperm heads (size and shape) in Iberian ibex by Sperm-Class Analyzer(®) and provides a basis for future studies on the relationship with freezability and fertility in this species., (Copyright © 2015. Published by Elsevier B.V.)

Here we show the effects of repeated electroejaculation (EE) on mean values of motility, mitochondrial functionality, and expression of active caspases on goat sperm obtained by EE. Evaluations were done using CASA and flow cytometry. A strategy for identification of kinematic sperm subpopulations, when individual data of sperm are not provided by the CASA system, is provided. Fifty semen samples, five of each of ten adult creole goats, were obtained by electroejaculation. Mean values of total motility, progressive motility and flow cytometry evaluations were compared among EEs. Relationships among mean values of variables were investigated using Spearman correlation and principal component analysis (PCA). For identification of kinematic sperm subpopulations, PCA followed by hierarchical clustering was applied on data of the intervals provided automatically by the CASA system. Total motility does no change after repeated EE. Mean values of motility parameters and molecular markers were unrelated in multivariate space, but bivariate correlations were found. Values in upper and lower intervals defined clearly the sperm subpopulations, which had motility parameters changing over time. Taken together, our results show that repeated EE does not affect mean values of total motility, that molecular markers are not related with motility parameters, and that it is possible to identify kinematic sperm subpopulations when individual data, of motility parameters, are not provided by the CASA system., (Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.)

In this work, we examined the ovarian development and its relationship with steroid hormone receptors levels and the precocious puberty in JiNing Gray goats by using optical microscopy, immunohistochemistry, quantitative real-time RT-PCR (qPCR) and Western blotting. We found that in the ovaries of neonatal kids, high level of receptors for estrogen (ERα and ERβ) and progesterone (PR) and their mRNA were observed along with growing follicles. From 0 to 30 days of age, the weight and volume of ovaries increased significantly and the boundary between the inner and outer cortex disappeared, while the expression of ERα, ERβ and PR and their mRNA decreased sharply. When 60 days old, the animals began to ovulate; the expression of ERα, ERβ and PR and their mRNA significantly increased, and the animals reached puberty. On day 90, the animals manifested sexual maturity with biggest mature follicles 6.18mm in diameter, the expression of ERβ and PR protein and their mRNA was maintained at a high level, with decreased expression of ERα and its mRNA. Before puberty, the expression of ovarian ERα (prepubertal dominant receptor) and it's mRNA was significantly higher than that of ERβ (dominant receptor after sexual maturity). The results showed that JiNing Grey goats' ovaries had fast development and early maturation, and ERα, ERβ and PR protein and mRNA expression in the ovary had distinct specificity for time and space, which may be closely related to the strain's progenitive characteristics., (Copyright © 2015. Published by Elsevier B.V.)

The reproductive activity of 84 female Blanca Andaluza goats was monitored over 17 months to determine the role of body condition score (BCS) and body weight (BW) in its control. Following a 3×2 factorial experimental design, the animals were allocated to three groups: low BCS (≤2.50, n=24), medium BCS (BCS=2.75-3.00, n=31) and high BCS (≥3.25, n=29). The same animals, irrespective of the BCS group categorization, were also divided into two groups depending on BW: low BW (≤40kg, n=44) and high BW (>40kg, n=40). Oestrus was evaluated daily using vasectomised males. The ovulation rate was assessed by trans-rectal ultrasonography after the identification of oestrus. Ovulations were determined by monitoring the plasma progesterone concentration weekly. The BCS and BW were recorded once a week and nutritional status adjusted to maintain the initial differences in BW and BCS between the groups. Both BCS and BW had a significant (at least P<0.05) influence on the onset, the end, and the duration of the breeding season, with longer periods of reproductive activity recorded in does with a BCS of ≥2.75 and BW of >40kg. No significant interaction between these variables was observed. Some (11.7%) of the does in the groups with animals of BCS≥2.75 had ovulations during seasonal anoestrus. None of the does with a BCS of ≤2.5 had ovulations during seasonal anoestrus. The ovulation rate of the first and last oestrus was influenced by BW (P<0.01). These results demonstrate that Blanca Andaluza goats show marked reproductive seasonality that is clearly and independently modulated by BCS and BW., (Copyright © 2014 Elsevier B.V. All rights reserved.)

KiSS1 is considered to be a key mediator of molecular mechanism of reproduction (puberty and prolificacy) in mammals. Kisspeptins are a family of structurally related peptides, encoded by KiSS1 gene, with ability to regulate gonadotropin-releasing hormone and hence hypothalamic-pituitary-gonadal axis. The present study investigated the polymorphism of caprine KiSS1 gene in 9 Indian goat breeds differing in sexual precocity and prolificacy. Comparison of KiSS1 amplified sequences of indigenous goats resulted in identification of nine SNPs (intron (1) G296C, T455G, T505A, T693C, T950C and intron (2) T1125C, A2510G, C2540T, A2803G) of which four are novel. These loci were not segregating together (r(2)<0.33). Mutations existed in both, sexually precocious and late-maturing goat breeds as well as low and high prolificacy goat breeds. Three loci reported to be associated with goat litter size (G296C, G2510A and C2540T) were identified in Indian goats as well. Association between loci of KiSS1 gene and age of puberty as well as litter size was explored in Black Bengal (N=158), a sexually precocious and prolific goat breed of India by designing PCR-RFLP. None of the mutations were found to be associated with reproductive traits however, difference in litter size as well age of sexual maturity for different genotypes indicates that the study on additional data based on more number of breeds and animals would be interesting to perform. Considering the importance of the reproductive trait in small ruminants, the results extend the limited information on genetic variation of the caprine KiSS1, which might contribute toward molecular breeding to enhance productivity of goat., (Copyright © 2014 Elsevier B.V. All rights reserved.)