Cloprostenol effect with and without male stimulation on oestrus synchronization in Peru guinea pigs.

Cloprostenol, a synthetic derivative of prostaglandin F2α, effectively triggers functional and morphological regression of the corpus luteum (luteolysis). In rural Peru, the guinea pig (Cavia porcellus) holds significance within the local economy and serves as a valuable protein source. Enhancing reproductive efficiency is crucial to achieve uniformity in weight, age, and litter size across commercial systems. Thus, our study aimed to evaluate the effect of cloprostenol with and without male stimulation on the onset, duration, and intensity of oestrus in Peru guinea pigs. A total of 128 guinea pigs (120 females and eight males) between 8 and 12 months of age, weighing between 800 and 1200 g, were distributed in cages of 15 females per treatment. Cloprostenol sodium (0 [control], 0.20, 0.25, and 0.30 mg/animal) was IM administered to the groups with and without male stimulation. Four isolated males in individual cages, different from the one used for the treatment, were considered to detect oestrus. The oestrus intensity was assessed by observing sexual behaviour signs such as restlessness, moaning, attempts to mount, pelvic elevation, loin stretching, and vulvar inflammation. The oestrus was manifested 2 days after the administration of cloprostenol sodium. At a dose of 0.30 mg/animal and with male stimulation, the earliest oestrus was observed at 46.9 h. There was a reduction in the oestrus duration (p < .05) in guinea pigs that received the three doses of cloprostenol sodium compared to the control group. The highest percentages of frank oestrus intensity (66.7%) were strongly associated with the administered doses of cloprostenol sodium (p < .01). In conclusion, the cloprostenol sodium administration was proper for rapid oestrus induction in Peru guinea pigs.

Response to hormonal treatment and conception rates of Sahiwal cows subjected to fixed time artificial insemination in pastoral dairy systems.

This study aimed at determining factors influencing response of Sahiwal cows/heifers to fixed time artificial insemination protocol in pastoral systems in Kenya. Available cows/heifers were inspected for conformity to Sahiwal breed characteristics, parity, body condition score, and subsequently rectal palpation to determine pregnancy status, ovarian structures, and estimated ovarian diameter. Consequently, these animals were injected with 100 µg of gonadotrophin-releasing hormone. On days 7 and 9, only responsive cows/heifers were injected with 500 µg of cloprostenol and 100 µg of gonadorelin Acetate, respectively. On day 10, animals were inseminated and separated from bulls for 45 days and pregnancy diagnosis done after 90 days. Analysis of variance was performed to determine the effects of production system, parity, and ovarian structures on ovary diameters pre- and post-hormonal treatment. Logistic regression was used fitting a logit function to account for the binomial distribution of conception. Overall, 56.2%, 23.1%, and 20.7% of the animals had follicles (F), corpus luteum (CL), and corpus albicans (CA), respectively, at day 0, and 16.6%, 68.6%, and 14.8%, respectively, at day 7. Human and environmental factors had no influence on conception. Among the animal factors, only the ovarian structures at day 7 had a significant effect on conception. Ovaries with CL at this time were about 6 times significantly more likely to conceive than those with F. For higher conception rates, animals with ovaries with CL should be recruited into the FTAI program as they are significantly more likely to conceive than those with other ovarian structures.

Puberty attainment and reproductive performance of yearling Bos indicus-influenced heifers after two sequential treatments with progesterone.

Number of pubertal heifers at time of breeding season initiation is a primary determinant to pregnancy success during the breeding season. It was hypothesized that pre-breeding progesterone (P4) supplementation (induction) would increase the number of heifers pubertal at the time of imposing estrous synchronization treatment regimens and P/AI. Yearling, Bos indicus-influenced (n = 577) or Bos indicus (n = 174) heifers were or were not treated with P4 (CIDR and Non-CIDR, respectively) for 10 d starting on D-23 (D0 = TAI). Presence of a CL on D-33 or D-23 was considered to indicate heifers were pubertal. On D-13, there was a PGF analogue administered. On D-9, there was treatment with GnRH analogue, 6d-CIDR and PGF. There were inseminations based on estrus (D-2 to D0) or TAI on D0 for non-estrous animals. There were 5.2 % and 62.9 % purebred and crossbred heifers pubertal, respectively. Proportion of prepubertal crossbred than purebred heifers with CL on D-3 was greater as a result of imposing the pubertal induction regimen (P < 0.05 and P> 0.10, respectively). Regardless of puberty status, proportion of heifers in estrus prior to AI in the CIDR group was similar to the heifers of the Non-CIDR group for crossbreds and purebreds. Similarly, P/AI of CIDR group was similar to the Non-CIDR group for crossbreds and purebreds. In summary, imposing the pubertal induction regimen hastened attainment of puberty in yearling crossbred, but not purebred heifers. Puberty induction did not affect estrous response, neither fertility after imposing an estrous synchronization treatment regimen.

Lack of effect of melatonin on ovarian function and response to estrous synchronization and fixed-time AI during the nonbreeding season in lactating dairy buffalo (Bubalus bubalis).

The present study was conducted to examine whether pretreatment with melatonin would enhance ovarian follicular functions and increase response to estrous synchronization and fixed-time AI (TAI) during the nonbreeding season in lactating dairy buffalo. In Experiment 1, buffalo cows without a detectable corpus luteum (CL) were assigned on Day -20 (D-20) to three groups: control (n = 12); melatonin (n = 13); progesterone (P4) (n = 15). Cows in the melatonin group were implanted with melatonin on D-20. From D0 to D9, there was imposing of an estrous synchronization treatment regimen using either a standard Ovsynch protocol (control, melatonin) or a P4-based Ovsynch treatment regimen (P4). There were no differences (P > 0.05) among groups for the presence of a CL at D0, size of the largest follicle at D0, ovulation to GnRH injection at D0 and D9, or the time to ovulation after injection of GnRH at D9. In Experiment 2, there was imposing of the same treatment regimens as in Experiment 1, with inclusion of TAI. Females of the P4 group had a greater (P = 0.001) pregnancy/AI percentage (60 %) than those in the control (17 %) and melatonin (23 %) groups. Females of the P4 group also had a larger (P = 0.005) CL at D20 compared with those in the control and melatonin groups. Findings indicate treatment with melatonin for 20 days did not affect ovarian functions or the response to an estrous synchronization treatment regimen and TAI during the nonbreeding season in lactating dairy buffalo.

Inducing ovulation with human chorionic gonadotrophin improves the pregnancy rate in lactating dairy cows receiving an in vitro-produced embryo.

While the global use of in vitro-produced embryos in dairy cattle is on the rise, several technical aspects of embryo transfer procedures have not yet been optimized. This study compares the effects of inducing ovulation using human chorionic gonadotropin (hCG) versus gonadotropin-releasing hormone (GnRH) at the end of a 5-day progesterone(P4)-based protocol for oestrous synchronization on the pregnancy rate of lactating dairy cow recipients of in vitro-produced embryos. Fresh embryos were transferred on Day-seven post-oestrus to ovulating cows receiving GnRH or hCG (groups GnRH and hCG, n = 60 each). Pregnancy was diagnosed by ultrasound on Day 28 post-oestrus. Forty-nine cows became pregnant: 16 in GnRH (26.7%) and 33 in hCG (55%). Taking GnRH-treated cows as reference, the odds ratio for pregnancy of hCG-treated cows was 3.3 (p = .002). In conclusion, hCG treatment given at the end of a 5-day P4-based protocol for oestrous synchronization improved the pregnancy rate in lactating dairy cows receiving an in vitro-produced embryo.

Oxytocin increases pregnancy rates after fixed time artificial insemination in Kazak ewes.

It is probable that reduced pregnancy rates in ewes after fixed time artificial insemination (FTAI) is attributable, in part, to the reduced number of normal spermatozoa that colonize the oviduct. Administration of oxytocin stimulates both cervical dilation and uterine/oviductal contractility. The hypothesis that oxytocin can enhance sperm transport into the uteri and the oviducts, and thereby increase pregnancy rates, was tested in the present study. Oestrus was synchronized in 199 multiparous Kazak ewes using intravaginal flurogestone-impregnated sponge. The sponge was left in the vagina for 12 days followed with an injection of 330 IU of eCG at sponge removal. Each ewe was intracervically inseminated twice at 50 hr and 62 hr after the removal of sponges using an insemination catheter containing 0.25 ml of diluted semen. Semen was collected from seven Texel rams and all the ejaculates were pooled and diluted in ultra-high temperature-treated commercial skimmed milk without (Control group, 0.05 ml of saline per mL milk, n = 144) or with oxytocin supplement (Oxytocin group, 0.5 U of oxytocin per ml milk, n = 55). Pregnancy status was determined by transabdominal ultrasound examination 45 days after insemination. Lambing performance was recorded at delivery. Significant differences were observed between the Oxytocin group and the Control group in terms of the pregnancy rate and the fecundity rate (85.5% and 92.7% versus 68.8% and 72.9%, respectively). In conclusion, low dose oxytocin supplementation of semen extender significantly increased pregnancy and fecundity rates in oestrus-synchronized Kazak ewes after FTAI.

Progesterone dose during synchronization treatment alters luteinizing hormone receptor and steroidogenic enzyme mRNA abundances in granulosa cells of Nellore heifers.

The objective was to investigate effects of progesterone (P4) dose on abundance of luteinizing hormone receptor (LHCGR), aromatase (CYP19A1), 3ß-hydroxysteroid dehydrogenase (HSD3B1), and other steroidogenic mRNA transcripts in granulosa cells from dominant follicles. Nellore heifers were assigned to one of six groups: new, first-use controlled internal drug release device (CIDR1) inserted for 5 days (Large-P4-dose-D5; n = 7) or 6 days (Large-P4-dose-D6; n = 8), prostaglandin (PG)F2α administered on D0 and 1 previously-used CIDR (CIDR3) inserted for 5 days (Small- P4-dose-D5; n = 8) or 6 days (Small-P4-dose-D6; n = 8), CIDR1 inserted on D0 and removed plus PGF2α on D5 (Large-P4-dose-proestrus (PE); n = 7), and CIDR3 and PGF2α on D0 and 1, CIDR3 removed plus PGF2α on D5 (Small-P4-dose-PE; n = 7). Duration of P4 treatment (D5 compared to D6) affected abundances of CYP19A1 mRNA transcripts, with there being greater abundances on D6 than D5 (P ≤ 0.05). Heifers treated with the large dose of P4 had a smaller dominant follicle, less serum and intra-follicular estradiol (E2) concentrations (P ≤ 0.05) and lesser LHCGR, CYP19A1, and HSD3B1 transcript abundances (P ≤ 0.05). Heifers treated to induce PE had a larger follicle diameter (P = 0.09), greater intra-follicular E2 concentrations and larger abundances of CYP19A1 mRNA transcript (P ≤ 0.05) than heifers of the D6 group. Overall, treatment with larger doses of P4 resulted in lesser abundances of LHCGR, HSD3B1, and CYP19A1 mRNA transcripts; thus, potentially leading to development of smaller dominant follicles and lesser E2 concentrations.

Cholesterol-cyclodextrin complex as a replacement for egg yolk in bull semen extender: sperm characteristics post-thawing and in vivo fertility.

Egg yolk, a major semen extender constituent, lacks a defined composition, therefore, there are biosecurity concerns with use of egg yolk. Cryopreservation of bull semen without inclusion of animal protein in the semen extender, therefore, is an important consideration. Cholesterol may be delivered and incorporated into the sperm plasma membrane by cyclodextrins to protect sperm during cryopreservation. The aim of this study was to determine suitability of a cholesterol-cyclodextrin semen extender, without inclusion of egg yolk, for cryopreservation of bull semen. Bull semen was collected and cryopreserved in either egg yolk or with inclusions of three different concentrations of cholesterol-cyclodextrin complex (0.5, 1 or 2 mg/mL semen) in Tris-glycerol (TG) extender. Sperm motion characteristics examined using the computer-assisted sperm analysis, and plasma membrane and acrosome integrity examined using flow cytometry, were similar for all extenders. The inclusion of the greatest concentration of cholesterol-cyclodextrin complex (2 mg/mL semen) followed by dilution in TG extender resulted in lesser pregnancy rates (P <  0.05). There was a pregnancy rate of as great as 56 % when sperm cryopreserved in 0.5 mg/mL cholesterol-cyclodextrin Tris-glycerol extender were used for artificial insemination following imposing of a hormonal treatment regimen for synchrony of timing of ovarian functions among cows for conducting fixed-time artificial insemination (FTAI). Results indicate cholesterol-cyclodextrin Tris-glycerol extender, with a chemically defined composition and without inclusion of egg yolk, may be used to cryopreserve bull sperm with there being acceptable pregnancy rates when this semen is used for FTAI.

Symposium review: Linking activity-sensor data and physiology to improve dairy cow fertility.

Several studies have demonstrated that the intensity of estrous expression is associated with ovulation, ovarian and uterine function, and fertility, and is dependent on social hierarchy and the housing system used. Data from recent studies involving spontaneous and induced estrus have shown that a greater relative increase and longer estrus (captured by different automated activity monitors; AAM) are both associated with improved pregnancy per artificial insemination (AI; around 10 to 14% increase) and decreased pregnancy losses. Intensity and duration of estrus were surprisingly weakly associated with preovulatory follicle diameter and concentrations of plasma estradiol at estrus, whereas ovulation failure was associated with low estrus intensity. Studies have also shown that the display of estrous behavior near AI was associated with the modification of expression of genes related to the immune system, adhesion molecules, and prostaglandin synthesis in the endometrium. Transcripts in leukocytes and in the conceptus tissue associated with maternal recognition of pregnancy as well as conceptus elongation were all associated with differences in the intensity of estrous expression. Most recently, studies from the United States and Canada have demonstrated that reproductive programs emphasizing detection of estrus using AAM can be successful and comparable to intensive timed AI protocol-based programs that incorporate GnRH and PGF2α treatments. Further, one study concluded that the administration of GnRH at AI for spontaneous estrus events greatly improved pregnancy per AI, but only for cows with reduced intensity of estrous expression, showing the potential to use AAM data as a tool in targeted reproductive programs. Quantitative information from estrus events could be used to improve estrus detection and develop decision-making strategies at the farm level. Future studies in this field should aim to better understand ovarian, conceptus, and endometrial mechanisms associated with either the expression or the intensity of estrus, and to refine the identification of phenotypes related to estrus (relative increase, absolute increase, baseline levels, duration, and repeatability within cow) to improve data usage, estrus detection, and possibly genetic selection.

Timed artificial insemination in crossbred mares: Reproductive efficiency and costs.

Timed artificial insemination (TAI) has boosted the use of conventional artificial insemination (CAI) by employing hormonal protocols to synchronize oestrus and ovulation. This study aimed to evaluate the efficiency of a hormonal protocol for TAI in mares, based on a combination of progesterone releasing intravaginal device (PRID), prostaglandin (PGF2α ) and human chorionic gonadotropin (hCG); and compare financial costs between CAI and TAI. Twenty-one mares were divided into two groups: CAI group (CAIG; n = 6 mares; 17 oestrous cycles) and TAI group (TAIG; n = 15 mares; 15 oestrous cycles). The CAIG was subjected to CAI, involving follicular dynamics and uterine oedema monitoring with ultrasound examinations (US), and administration of hCG (1,600 IU) when the dominant follicle (DF) diameter's ≥35 mm + uterine oedema + cervix opening. The AI was performed with fresh semen (500 × 106 cells), and embryo was recovered on day 8 (D8) after ovulation. In TAI, mares received 1.9 g PRID on D0. On D10, PRID was removed and 6.71 mg dinoprost tromethamine was administered. Ovulation was induced on D14 (1,600 IU of hCG) regardless of the DF diameter's, and AI was performed with fresh semen (500 × 106 cells). On D30 after AI, pregnancy was confirmed by US. The pregnancy rate was 80.0% in TAIG and 82.3% in CAIG (p > .05). The TAI protocol resulted in 65% reduction in professional transport costs, and 40% reduction in material costs. The TAI was as efficient as CAI, provided reduction in costs and handlings, and is recommended in mares.

Luteotropic effects of human chorionic gonadotropin administered 7.5 days after synchronous estrous induction in Morada Nova ewes.

This study was conducted in ewes to assess effects of human chorionic gonadotropin (hCG) administration after imposing an estrous induction treatment regimen. Ewes (n = 115) were treated with a 60 mg medroxyprogesterone-intravaginal-sponge for 6 d plus 200 IU of equine chorionic gonadotropin (eCG) im and 37.5 µg d-cloprostenol im 36 h before sponge removal (Day 0). After natural mating, ewes having at least one corpus luteum (CL; n = 108) were administered either 1 mL of saline (G-Control; n = 53) or 300 IU of hCG (G-hCG; n = 55) on Day 7.5 after sponge removal (Day 0). Ovarian ultrasonography and blood collection were performed on Days 7.5, 13.5, 17.5, 21.5, and 30.5. Accessory CL (aCL) were observed in 81.5 % (G-hCG) and 0.0 % (G-Control) of ewes (P = 0.0001). Diameter, area, and volume of luteal tissue were greater (P < 0.05) in G-hCG from Day 13.5 to 30.5. Progesterone (P4) concentrations were greater (P < 0.05) on Days 13.5, 17.5, 21.5 and 30.5 for ewes of the G-hCG group. Pregnancy percentage was similar (P = 0.25) between groups [47.1 % (G-control) compared with 60.0 % (G-hCG)], although total number of lambs produced by estrous synchronized ewes was greater (P = 0.005) in ewes of the G-hCG group (90.9 % compared with 66.0 %). In conclusion, hCG administration 7.5 days after sponge removal from Morada Nova ewes during the non-breeding season is an effective treatment to induce aCL formation, improve luteal tissue biometry and P4 concentrations, and to enhance the total number of lambs born.

Size and number of corpora lutea and serum progesterone concentrations when administering two doses of eCG in an estrous synchronization treatment regimen for dairy cattle.

There was investigation of whether there were ovulations from co-dominant follicles following eCG administration. In all experiments, there was GnRH injection and CIDR insertion on day 0 (D0), CIDR withdrawal on D8, and cloprostenol administration on D8 (Exp. I and II) or D7 and D8 (Exp. III). Females in the control group were not administered any further treatment. Females in other group(s) were treated with eCG (500 IU) on Day 2 in Exp. I, Day 2 (eCG-2) or 8 (eCG-8) in Exp. II and Day 2 (eCG-2) or Days 2 and 6 (eCG-2-6) in Exp. III. Ovaries were examined using ultrasonography. In Experiments I and II, females had follicle emergence on Day 2. At the time of CIDR removal, more eCG-treated heifers (8/9; Exp. I) and cows (5/6; eCG-2; Exp. II) had co-dominant follicles compared to those in the control group (P < 0.05). Occurrence of ovulations from co-dominant for individual cows was minimal. In Experiment III, the time period from CIDR removal to estrus in cows treated with eCG-2 (68 ± 13 h) was longer compared to cows in the control (37±2 h) and eCG-2-6-treated group (38 ± 5 h; P < 0.05). There was a greater proportion of heifers having ovulations and thus greater progesterone concentration in the eCG-2-6 than eCG-2 group (P < 0.05). Administering eCG twice 4 days apart with the initial administration being two days after GnRH administration, at the time of follicle wave emergence, could induce growth of and ovulation from co-dominant follicles and enhance progesterone production in cattle.

Practical application of an impractical bovine genotype: creating bilateral twin pregnancies in Trio allele carriers.

Bovine twin birth is associated with detriments, including increased embryo/fetal losses, malpresentation, and dystocia. Incidence of these is lessened in bilateral compared with unilateral twin pregnancy. This study was undertaken to assess the use of follicular ablation by aspiration to create bilateral twin pregnancies in females with genetic potential for ~3.5 ovulations per cycle (Trio allele carriers). In experiment 1, carriers (n = 30) and noncarriers (n = 10) were synchronized for ovulation and timed artificial insemination (TAI). Follicles (>5 mm) in excess of one per ovary were aspirated ~16 h preceding TAI. Follicle count for females with follicles on only one ovary was reduced to two. Blood was sampled 2 wk post-TAI to assess progesterone (P4) concentrations; embryo count was determined by ultrasound 6 wk post-TAI. Circulating P4 concentration post-TAI was significantly (P < 0.001) associated with both genotype and subsequent pregnancy status (pregnant noncarriers: 7.06 ± 0.68 ng/mL; pregnant carriers: 5.54 ± 0.55 ng/mL; nonpregnant noncarriers: 5.22 ± 1.05 ng/mL; nonpregnant carriers: 3.13 ± 0.42 ng/mL). Experiment 2 was undertaken to offset the negative effects of follicular aspiration on subsequent P4 concentration observed in experiment 1. Carriers (n = 38) and noncarriers (n = 32) were submitted to TAI and follicle ablation as described for experiment 1. Additionally, accessory corpora lutea (CL) were induced in carriers by the administration of human chorionic gonadotropin (carriers) at day 6 post-TAI. Consequently, P4 concentration post-TAI was significantly (P < 0.05) associated with subsequent pregnancy status (pregnant: 8.48 ± 0.61 ng/mL; nonpregnant: 6.70 ± 0.63 ng/mL) but not with genotype (carrier: 8.01 ± 0.59 ng/mL; noncarrier: 7.17 ± 0.64 ng/mL). Embryo number was greater in carriers (exp. 1: 1.64 ± 0.81; exp 2: 1.45 ± 0.09) vs. noncarriers (1.00 ± 0.00, both experiments). Single, twin, and triplet pregnancies occurred in carriers in experiment 1, whereas multiples in experiment 2 were limited to twin pregnancies. Genotype effects on pregnancy rate were not significant (P > 0.10) in either experiment. Results suggest that follicular ablation to create bilateral twin pregnancies in Trio carriers is feasible but requires the induction of accessory CL to offset the negative effects of follicular aspiration on subsequent P4 concentration and associated fertility outcomes.

Effects of human chorionic gonadotropin and intravaginal progesterone device treatment after artificial inseminations on the reproductive performance of normal and repeat breeder lactating dairy cows.

We examined the effects of human chorionic gonadotropin (hCG) treatment on Day 5 (Day 0 = day of artificial insemination: AI) and intravaginal progesterone device (IVPD) treatment from Day 5 to 19 on the conception and detection rates of return to estrus (re-estrus) in lactating dairy cows. A total of 306 cows from a commercial dairy farm were divided into the following three groups on Day 5: non-treatment group (n = 128), untreated; hCG group (n = 71), 3,000 IU hCG was administered (intramuscularly); IVPD group (n = 107), IVPD was inserted into the vagina from Day 5 to 19. Re-estrus detection was performed up to Day 25. Pregnancy was diagnosed by rectal palpation between Day 50 and 60. There was an interaction between treatment and AI number (P < 0.01) on the conception rate of first-AI. For cows with more than three AIs, the IVPD treatment (66.7%) was more effective than the non-treatment (23.1%) (P < 0.05). The re-estrus detection rate was significantly (P < 0.05) higher in the IVPD group (60.7%) than that in the non-treatment group (41.4%) and tended (P < 0.1) to be higher than that in the hCG group (37.8%). Our results suggested that the conception rate can be improved by IVPD treatment, especially in cows with more than three AIs. In addition, IVPD treatment can induce higher estrus expression up to 25 days after AI in non-pregnant cows.

Responses to intra-luteal administration of cloprostenol in dairy cows.

The aim of the study was to determine the luteolytic dose of cloprostenol administered directly into the corpus luteum (CL; intra-luteal treatment, ILT) in dairy cattle. Cows of two control groups were treated with 500 µg of cloprostenol (Estrumate®) intramuscularly (IM-500) or via ILT with 0.2 mL of physiological solution (ILT-0). Cows of four experimental groups were treated by ILT with cloprostenol in doses 5, 25, 50 and 100 µg (ILT-5, -25, -50 and -100 groups). Progesterone concentrations (P4) and size of CL were evaluated to assess luteolysis at 0, 0.5, 1, 2, 4, 8, 24 and 48 h or at 0, 24 and 48 h after ILT, respectively. Cows in the ILT-0 and -5 groups were unaffected by ILT. The P4 concentrations were less in cows of the IM-500, as well as ILT-25, -50 and -100 groups at 48 h subsequent to ILT. The size of the CL was less in cows of IM-500, as well as ILT-25, -50 and -100 groups at 48 h after ILT. There were P4 concentrations of about 1 ng/mL 48 h after ILT in cows of the IM-500, as well as ILT-50 and -100 groups. In conclusion, the cloprostenol dose of 50 µg administered intra-luteally is a luteolytic dose in cows.

Intrafollicular injection of nonesterified fatty acids impaired dominant follicle growth in cattle.

Dairy cows frequently undergo a state of negative energy balance (NEB) after parturition and some have impaired ovarian functions that result in delayed resumption of estrous cyclicity and development of follicles without ovulation occurring. During the postpartum period, cows undergo body-fat store losses, hormonal changes, fat mobilization and increases in nonesterified fatty acid (NEFAs) concentrations in blood and follicular fluid. The effect of NEFAs on follicular development and function of follicular cells, however, is not fully understood. The aim of this study, therefore, was to study the effect of an intrafollicular injection of a mixture of oleic, stearic and palmitic NEFAs on dominant follicle development and function of granulosa cells in cows that were not in a NEB state. Follicular size was less at 24 and 48 h after administration of NEFAs compared to that of control follicles injected with vehicle only. At 24 h after intrafollicular injection, the relative mRNA transcript abundance for proteins involved in steroidogenesis (CYP19A1, 3BHSD, STAR, FSHR), metabolism (GLUT1, GLUT3, INSR, IRS1, IRS2, SLC27A1, PPARG), and cell proliferation and apoptosis (CCND2; XIAP) in granulosa cells, as well as estradiol concentrations in follicular fluid were similar in control and NEFA-treated follicles. In conclusion, the results of this study indicate increased intrafollicular concentrations of NEFAs in cows that are not in a NEB state has a detrimental effect on follicle development. We propose intrafollicular injection is a useful approach to further investigate the local effects of NEFAs on the function of follicular cells.

Effect of dose and duration of FSH treatment on ovarian response in prepubertal calves.

The objective of the study was to determine relative effects of dose (200 or 350 mg) and duration (4 or 7 days) of superstimulatory treatment on the ovarian response in prepubertal calves. Calves with similar antral follicular counts at wave emergence (n = 24) were given eight doses of either 25 or 44 mg pFSH every 12 h for 4 days or 14 doses of either 14 or 25 mg pFSH for 7 days beginning at the time of follicular wave emergence and 12.5 mg of pLH im 12 h after the last FSH treatment. On Day 4 of pFSH treatment, calves given 14 mg had fewer follicles ≥3 mm than those given 25 mg (15.1 ± 1.9 and 27.9 ± 3.3, respectively; P = 0.04). At the end of treatment (24 h post-LH), number of follicles ≥9 mm was greater in calves of groups treated with 350 than 200 mg (13.5 ± 1.8 and 8.8 ± 1.3, respectively; P = 0.02) and calves of groups treated for 7 than 4 days (13.3 ± 1.8 and 9.0 ± 1.3, respectively; P = 0.03). The number of spontaneous ovulations was greater in calves of groups treated for 7 than 4 days as was the total number of ovulations (9.7 ± 0.9 and 6.9 ± 1.0, respectively; P ≤ 0.05). In summary, a dose of 25 mg of pFSH per treatment given twice daily for 7 days resulted in a greater ovarian response than other superstimulatory treatments in prepubertal calves.

Comparison of long-term progestin-based protocols to synchronize estrus prior to natural service or fixed-time artificial insemination in Bos indicus-influenced beef heifers.

This experiment was designed to evaluate breeding strategies involving natural service or fixed-time artificial insemination (FTAI) in Bos indicus-influenced beef heifers (n = 1456) when there were field-type management conditions. Body weights and reproductive tract scores (RTS; Scale 1-5) were obtained for heifers before assignment to one of five treatments: 1) Non-synchronized control exposed for natural service (NS), n = 299; 2) melengestrol acetate + natural service (MGA + NS; 0.5 mg/heifer/d), n = 295; 3) 14-d controlled internal drug release insert + natural service (CIDR + NS), n = 289; 4) 14-d MGA-prostaglandin F2α (PG) + FTAI, n = 295; or 5) 14-d CIDR-PG + FTAI, n = 278. Fertile bulls were placed in pastures with heifers of the three NS treatment groups for a 65-day period which began 10 days after progestin treatments (MGA or CIDR) ended. Heifers in FTAI treatment groups were administered PG (25 mg, IM) 16 days after CIDR removal or 19 days following MGA withdrawal, respectively, and FTAI was performed at 66 (CIDR-PG) or 72 h (MGA-PG) after PG. Gonadotropin-releasing hormone (GnRH; 100 µg, i.m.) was administered at FTAI. Pregnancy status was determined at the end of a 65-day breeding period. Pregnancy rates on Days 21 and 65 of the breeding period differed among treatment groups based on pre-treatment pubertal status (P ≤ 0.02) and body weight (P ≤ 0.05) but did not differ by group. These data highlight the need for continued research efforts to improve reproductive management of Bos indicus-influenced females.

Use of two cloprostenol administrations 11.5 days apart efficiently synchronizes oestrus in photostimulated multiparous dairy goats in the non-breeding season.

This study assessed the efficiency of synchronous oestrous induction by light programme followed by two doses of cloprostenol in acyclic Saanen goats of different parity orders. Primiparous (n = 22) and multiparous (n = 33) goats were subjected to 16 hr of light and 8 hr of darkness for 60 days (D0-D60), starting 10 days after the winter solstice. All goats received 120 µg cloprostenol doses on D130 (morning) and D141.5 (afternoon) (11.5 days apart). Oestrus behaviour, ovarian follicular dynamics and serum progesterone (P4) analyses were recorded from D0 to D174 at different intervals. Animals in oestrus after D141.5 were randomly assigned into two groups: assisted natural mating (NM) or artificial insemination (AI; 10-24 hr after oestrus onset with frozen-thawed semen). From D57 to D120, 89.0% of goats presented large follicles (5-8 mm) and P4 concentrations were subluteal from D0 to D120. More multiparous compared to primiparous goats (54.5%, 18/33 vs. 18.2%, 4/22) exhibited oestrus after both injections. More primiparous compared to multiparous goats (54.5%, 12/22 vs. 12.1%, 4/33) did not exhibit oestrus after any injection. A total of 35 goats (64%) were in oestrus after the second prostaglandin injection and were subjected to NM or AI. The conception rate was similar among primiparous (70.0%, 7/10) and multiparous (68.0%, 17/25) goats but the pregnancy rate differed, being 31.8% (7/22) and 51.5% (17/33), respectively. No interaction was found between parity order and P4 concentrations in does that became pregnant or not. Thus, the association between light programme (60 days, starting at the beginning of winter) and two cloprostenol administrations 11.5 days apart (starting 70 days after the end of the light treatment) resulted in sufficient synchronous oestrous response in multiparous acyclic Saanen goats to reach satisfactory fertility levels after both NM and AI.

Influence of ipsilateral coexistence of the first wave dominant follicle and corpus luteum on ovarian dynamics and plasma sex steroid hormone concentrations in lactating dairy cows treated with human chorionic gonadotropin.

We examined the effect of human chorionic gonadotropin (hCG) treatment 5 days after estrus on ovarian dynamics and plasma progesterone (P4) and estradiol (E2) concentrations when the first-wave dominant follicle (DF) was ipsilateral or contralateral to the corpus luteum (CL) in lactating dairy cows. Seventy cows were divided into two groups: (1) ipsilateral group (IG; n = 37), in which the first-wave DF was ipsilateral to the CL, and (2) contralateral group (CG; n = 33), in which the first-wave DF was contralateral to the CL. IG and CG were further subdivided into two groups: non-treatment group (IG, n = 18; CG, n = 19), and hCG treatment group: administrated 1500 IU of hCG 5 days after estrus (IG, n = 19; CG, n = 14). Blood sampling and ovarian examination were performed at 3, 5, 7, 9, 11, and 13 days after estrus. Mean diameter of the first-wave DF on Day 9 tended (P = 0.067) to be larger in IG than in CG in the non-treatment group. Mean diameter of CL and plasma P4 and E2 concentrations did not differ between IG and CG in the non-treatment and hCG treatment groups. Accessory CL development did not differ between IG and CG in the hCG treatment group. Our findings indicate that CL development and plasma P4 and E2 concentrations were not affected by the existence of the first-wave DF; however, first-wave DF development was affected by the existence of a CL in the same ovary.