Factors influencing pregnancy rate and loss after laparoscopic artificial insemination with frozen-thawed semen in Lohi sheep under sub-tropical conditions.

This study evaluates factors influencing pregnancy rates per artificial insemination (P/AI) and pregnancy loss in Lohi ewes undergoing laparoscopic AI with frozen-thawed semen under sub-tropical conditions. Data from three experiments comprising ewes (n = 358) of mixed parity (nulliparous; NP and parous; P), various body condition score (BCS) and assigned to long-term (LTP, 11 days) and short-term (STP, 5 days) oestrus synchronization regimen across high breeding season (HBS) and low breeding season (LBS) were analysed. Laparoscopic insemination was conducted 54 h post-sponge removal. Pregnancy diagnosis and loss were evaluated on days 35 and 90 post-insemination via ultrasonography. Results showed parity significantly influenced P/AI, with nulliparous ewes achieving higher pregnancy ratios than parous ewes (p = .001). BCS significantly influenced P/AI (p < .05), with a quadratic relationship observed between BCS and season (BCS*BCS*Season; p = .07). Progestin treatment did not significantly influence the ratio of pregnant ewes (p = .07). Pregnancy losses were significantly higher during LBS than HBS (p < .05), irrespective of progestin treatment. In conclusion, parity and BCS significantly influenced P/AI, with BCS demonstrating a quadratic association with season. Ewes bred during LBS experienced higher pregnancy losses than HBS, irrespective of progestin treatment.

Synergistic effect of extracellular adenosine triphosphate and quercetin on post-thaw quality and fertilization potential of Lohi ram sperm.

This study determined the individual and combined effects of extracellular adenosine triphosphate (ATP) and quercetin (QUE) on the quality of post-thawed sperm and the fertilization potential of Lohi rams. In experiment 1, semen samples from four Lohi rams were pooled and extended with different concentrations of ATP or QUE (control; no ATP or QUE, 1 or 2 mM ATP and 10 or 20 µM QUE). In experiment 2, pooled semen samples were extended with various combinations of ATP and QUE (control; no ATP and QUE, 1 mM ATP + 10 µM QUE, 1 mM ATP + 20 µM QUE, 2 mM ATP + 10 µM QUE and 2 mM ATP + 20 µM QUE). All samples in both experiments were cryopreserved and analyzed for post-thawed sperm quality. In experiment 3, the best combination of ATP and QUE from experiment 2 was to extend semen, which was then used for laparoscopic insemination in estrus-synchronized ewes (n = 83). The results of experiment 1 showed that 1 mM ATP and 20 µM QUE treatments resulted in higher total motility, progressive motility, viability, plasma membrane intactness (PMI), and motion kinetics (VCL, VSL, VAP, LIN, and STR) compared to other treatments (p < 0.05). In experiment 2, the 1 mM ATP +10 µM QUE-treated group exhibited significantly higher total and progressive motility, PMI, and motion kinetics (VSL, VCL, VAP, STR, and BCF) compared to the control group (p < 0.05). In experiment 3, the fertilizing potential of sperms treated with 1 mM ATP +10 µM QUE was greater than that of untreated controls (58.1% vs. 27.5%, respectively, p-value = 0.012). In conclusion, the quality of post-thawed ram semen is enhanced when the extender is supplemented with extracellular 1 mM ATP and 20 µM QUE, whether used separately or in combination with 1 mM ATP and 10 µM QUE. Furthermore, the inclusion of 1 mM ATP and 10 µM QUE together in the extender significantly improves in vivo fertility in Lohi ram.

Effects of extender type and storage time on sperm quality parameters of Kail ram semen stored at 5 °C.

The study was designed to evaluate the effect of different extenders and storage times on sperm quality parameters of extended Kail ram semen. Semen was collected from five adult Kail rams using an artificial vagina. Semen samples with >70% total sperm motility were pooled, diluted with Tris (TR), sodium citrate (SC), and skim milk (SM)-based extenders, and stored at 5 °C. Sperm motility and kinematics, viability, and plasma and acrosomal membrane integrity were assessed every 24 hrs for 120 hrs. Sperm longevity was estimated by placing semen in a water bath at 37°C to assess sperm kinematics. Storage time as the main effect had a negative (P < 0.05) impact on sperm quality parameters. The percentages of total motile (TM), progressive motile (PM), rapid, and medium progressive (MP) motile sperm were similar at 24 hrs of storage for the three extenders. However, semen extended in TR had a higher (P < 0.05) percentage of TM, PM, rapid, and MP sperm compared to SM and SC extenders at 48 hrs of storage. The sperm kinematics (VCL, VSL, VAP, and ALH) of progressive sperm were similar for all three extenders up to 72 hrs of storage. In addition, semen extended in TR had a higher (P < 0.05) percentage of sperm with intact plasma and acrosomal membranes at a storage time of 48 hrs. At 37 °C, the percentage of TM and PM sperm was higher (P < 0.05) in the TR extender than in the SM and SC extenders at 60 minutes and beyond. In conclusion, the Kail ram sperm extended in TR and stored at 5 °C maintained better quality after 48 hrs storage than sperm extended in SM and SC extenders. At 37 °C, sperm extended in TR also retained better quality parameters at 60 min and afterward than sperm extended in SM and SC.

Approaches of estrous synchronization in sheep: developments during the last two decades: a review.

The objective of the current review was to summarize the protocols used for estrous synchronization in ewes during the last two decades. Progesterone (P4) is a major hormone used in most protocols. P4 in the form of a controlled internal drug releasing (CIDR) device, medroxyprogesterone acetate (MAP), and fluorogestone acetates (FGA) has been used for estrous synchronization. Also, gonadotropins such as equine chorionic gonadotropin (eCG) and gonadotropin-releasing hormone (GnRH) are often administered at the end of P4-based protocols to improve fertility. Moreover, the administration of prostaglandins (PG) and ram effects have been used for estrus induction and synchronization of ewes. The findings of previous studies indicate that the outcome of administering various synthetics P4 analogues (CIDR, MAP, and FGA) in ewes is comparable in terms of estrous synchronization/induction. The supplementation of P4-based protocols with eCG, however, improves the estrus response and pregnancy rate during breeding and non-breeding season. On the other hand, PG is effective for successful estrous synchronization during the breeding season only. Often, two injections of PG are administered either 11 or 14 days apart along with P4-based protocols to lyse ovine corpus luteum (CL) when it is receptive to PG i.e., 3 days post-ovulation. Alternatively, the "ram effect" has been shown to improve the efficacy of P4-based protocols and can be used as an alternative to eCG in ewes. The current review describes the methods of synchronization and their outcomes during breeding and a non-breeding season in ewes.

Follicular dynamics and changes in oestradiol-17ß, progesterone and LH profiles following PGF2α induced oestrus in early and late ovulating Beetal goats.

This study compares the factors associated with variable interval to oestrus and ovulation between early versus late ovulating goats following PGF2α administration. The time of ovulation in Beetal goats (n = 38) was monitored through transrectal ultrasound at every 6 hr following a single dose of PGF2α (experiment 1). Variations in oestrus and ovulation times were further explored through the changes in follicular dynamics, endocrine profiles and behaviour in another set of goats (n = 13) following single PGF2α given randomly during the luteal phase (experiment 2). The ovulation time varied between 60 and 96 hr, and 57% of ovulations occurred by 72 hr following PGF2α (experiment 1). Accordingly, the goats (n = 13) in the second experiment were retrospectively divided either into early and/or late ovulating, that is, ≤72 and/or ≥84 hr following PGF2α . The onset of oestrus, peak estradiol-17ß concentration and LH surge after PGF2α was first observed in early than late ovulating goats (p < 0.05). The goats ovulating early had larger follicle and smaller CL in diameter at the time of PGF2α administration than those ovulating late (5.4 ± 0.2 vs. 4.3 ± 0.2 mm and 10 ± 0.6 vs. 11.8 ± 0.3 mm, respectively; p < 0.05). Likewise, plasma progesterone concentration tended to be lower (p = 0.087) in early than late ovulating goats. In conclusion, the size of dominant follicle and CL at the time of PGF2a determines the interval to ovulation following a single dose of PGF2a during the luteal phase.

Stable reference genes in granulosa cells of bovine dominant follicles during follicular growth, FSH stimulation and maternal aging.

The aim of the present study was to determine a set of reference genes in granulosa cells of dominant follicles that are suitable for relative gene expression analyses during maternal and follicular aging. Granulosa cells of growing and preovulatory dominant follicles were collected from aged and young cows (maternal aging study) and from FSH-stimulated follicles developing under different durations of FSH treatment (follicular aging study). The mRNA levels of the two commonly used reference genes (GAPDH, ACTB) and four novel genes (UBE2D2, EIF2B2, SF3A1, RNF20) were analysed using cycle threshold values. Results revealed that mRNA levels of GAPDH, ACTB, EIF2B2, RNF20, SF3A1 and UBE2D2 were similar (P>0.05) between dominant follicle type, age and among follicles obtained after FSH-stimulation, but differed (P=0.005) due to mRNA processing (i.e. with versus without amplification). The stability of reference genes was analysed using GeNorm, DeltaCT and NormFinder programs and comprehensive ranking order was determined using RefFinder. The mRNA levels of GAPDH and ACTB were less stable than those of UBE2D2 and EIF2B2. The geometric mean of multiple genes (UBE2D2, EIF2B2, GAPDH and SF3A1) is a more appropriate reference control than the use of a single reference gene to compare relative gene expression among dominant and FSH-stimulated follicles during maternal and/or follicular aging studies.

Environmental Effects and Genetic Parameters for Growth Traits of Lohi Sheep.

Estimating genetic parameters for growth traits is crucial to plan breeding strategies for improving meat production in indigenous sheep breeds. The study first tested the effects of environmental and maternal effects on five growth traits, including birth weight (BWT), weight at 120 days (WT120), weight at 180 days (WT180), weight at 270 days (WT270) and weight at 365 days of age (WT365) and then estimated genetic parameters for these traits using data obtained in 1215 Lohi sheep. The effects of factors, including year (YOB), season (SOB) and type of birth (TOB), age of dam (AOD) and sex on growth traits of Lohi sheep, were examined using analysis of variance (ANOVA) in R software. Sex, TOB and YOB significantly affected all studied traits. The estimates of direct and maternal heritability for BWT and WT120 were 0.15 ± 0.08 and 0.20 ± 0.06, and 0.45 ± 0.16, 0.21 ± 0.08, respectively. The direct heritability estimates for WT180, WT270 and WT365 were 0.20 ± 0.07, 0.21 ± 0.07 and 0.19 ± 0.08, respectively. Due to the high heritability estimate obtained for WT120 compared to other studied traits, and its strong genetic correlation (>0.9) with post-weaning growth traits, it is recommended that selection must be practiced on WT120 to improve the growth performance of Lohi sheep. The results could be used for the development of genetic/genomic selection programs aiming to improve the production performance of the Lohi sheep.

Effect of Prepartum Dietary Energy Level on Production and Reproduction in Nili Ravi Buffaloes.

The objective of this study was to evaluate the effect of prepartum dietary energy level on postpartum production and reproduction in Nili Ravi buffaloes (n = 21). The buffaloes were offered low energy (LE: 1.31 Mcal/kg DM NEL (net energy for lactation)), medium energy (ME: 1.42 Mcal/kg DM NEL) or high energy (HE: 1.54 Mcal/kg DM NEL) diet for 63 days prepartum, and received the same lactation diet (LD: 1.22 Mcal/kg DM NEL) during 14 weeks postpartum. The effects of dietary energy level and week were analyzed with Proc GLIMMIX model. Dry matter intake (DMI) was lower in buffaloes fed the LE diet compared with buffaloes fed the ME or HE diet. Calf birth weight (CBW) was higher in buffaloes fed the HE diet compared with buffaloes fed the ME or LE diet. Milk production was similar in buffaloes fed the HE, ME or LE diet within 14 weeks postpartum and throughout the lactation. Milk fat was higher in buffaloes fed the LE diet compared with buffaloes fed the ME or HE diet. Milk protein and lactose yields was high in buffaloes fed the HE diet compared with buffaloes fed the ME or LE diet. Body condition score was high in HE and was affected by diet × week interactions during pre and postpartum period. The concentrations of ß-hydroxybutyrate (BHBA) and triglycerides in serum was lowest in buffaloes fed the HE diet compared with the buffaloes fed the ME or LE diet. The buffaloes fed the HE diet had early uterine involution (UI), first estrus, short dry days, and calving interval (CI) compared with buffaloes fed the ME or LE diet. None of buffaloes fed the LE diet exhibited estrus during the first 14 weeks postpartum compared with buffaloes fed the ME or HE diet. In conclusion, prepartum feeding of high energy diet can be helpful in improving the postpartum productive and reproductive performance in Nili Ravi buffaloes.

Effect of weaning method on body weight, testicular growth, sexual behavior, age of puberty, and testosterone concentration of Beetal bucks.

The current study evaluates the effects of early (8th week) and late (16th week of age) weaning of male goat kids on their body growth, testicular growth, sexual behavior, plasma testosterone concentration, and pubertal age. Early (n = 6) and late (n = 7) weaned Beetal bucks were weekly monitored from 18th to 38th week for their body weight, scrotal circumference, testicular volume, testicular echogenicity (via ultrasonography), sexual activities, and plasma testosterone concentration. In comparison to early-weaned, late-weaned bucks showed a marked increase (p < .05) in body weight (11.4 ± 0.8 vs. 13.7 ± 0.6kg), testicular volume (44.1 ± 7.2 vs. 79.8 ± 18.7cm3 ), scrotal circumference (10.7 ± 0.6 vs. 12.8 ± 0.7cm), and testicular echogenicity (28.3 ± 2.7 vs. 38.3 ± 2.1) from 18th, 28th, 21st, and 24th week onward, respectively. Sexual activities started earlier in late- than early-weaned bucks (22nd vs. 25th week, respectively). Moreover, the sexual behavior index was better (p < .05) after the 34th week in late than early-weaned bucks. The plasma concentration of testosterone (at 39 weeks of age) was relatively more and the onset of puberty was 2-3 weeks earlier (p < .05) in late than early-weaned bucks. In conclusion, age-based early weaning of male kids impairs their testicular growth, sexual behavior, and age at puberty compared to conventional weaning.

Optimal timing of artificial insemination and changes in vaginal mucous characteristics relative to the onset of standing estrus in Beetal goats.

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.