CIDR Withdrawal Research Articles

Effects of estradiol cypionate dosage and body condition on reproductive performance of Nellore cattle synchronized for timed-artificial insemination

This experiment was designed to evaluate the effects of increasing doses of estradiol cypionate (ECP) and different body condition score (BCS) on reproductive performance of Bos indicus beef females assigned to a timed-artificial insemination (TAI) management. In this experiment, 1683 Bos indicus Nellore cows were blocked by parity and assigned to receive 1) an intravaginal P4 device (1.9 g of P4) and 2.0 mg of estradiol benzoate on day −11, 12.5 mg (i.m.) of dinoprost tromethamine, 300 IU (i.m.) of equine chorionic gonadotrophin, 0.6 mg (i.m.) of estradiol cypionate and CIDR withdrawal on day −2, followed by TAI on day 0 (n = 849; 0.6ECP) or 2) the same synchronization protocol with 1.0 mg of ECP on day −2 (n = 834; 1.0ECP). In both treatments, estrus expression was measured between days −2 and 0. Body condition score (BCS) was evaluated on days −11, 31, and 71 of the experiment and the BCS variation (Δ) was also determined between these timepoints. Transrectal ultrasonography was performed on days 31, 71, and 111 for pregnancy rate determination. All binary data were analyzed using cow as the experimental unit with GLIMMIX, whereas continuous variables were analyzed with the MIXED procedure of SAS. No treatment effects were observed on estrus expression rate. Treatment × BCS interactions were observed for pregnancy rates in all time points (days 31, 71, and 111), as 1.0ECP cows with a LOW BCS also had a greater P/AI than cows assigned to 0.6ECP. In summary, increasing the dose of ECP benefited the reproductive performance of Nellore beef cows with a reduced BCS (≤2.75), whereas no benefits were seen when the BCS was considered adequate (>2.75).

Ovine endometrial estrogen receptor expression is altered following PG-600 administration.

Dysregulation of the estrogen receptor 1 (ESR1) expression during the establishment of pregnancy could contribute to reduce fertility reported in ewes treated with PG-600. The objective of this study was to evaluate the effect of treatment with PG-600 on ESR1 expression in the ovine endometrium during early diestrus. Polypay ewes (n = 24) that had exhibited an oestrous cycle of normal duration (16-18 days) were treated with intravaginal progesterone-releasing device (CIDR) for 9 days. Cloprostenol was administrated two days before CIDR withdrawal. On the day of CIDR withdrawal (day 0), ewes received a 5mL intramuscular injection of PG-600 (400 IU equine chorionic gonadotropin and 200 IU human chorionic gonadotropin) or saline. Blood samples were collected before treatment and on the day of tissue collection to determine serum estradiol-17β and progesterone concentrations using radioimmunoassay. On days 4 and 7, six ewes from each treatment group were anaesthetised and a laparotomy was performed to obtain intercaruncular endometrial samples. Endometrial samples were collected ipsilateral to the ovary bearing the highest number of corpora lutea. An estradiol exchange assay was used to determine ESR1 concentration. Estradiol concentrations did not differ by time or treatment, but progesterone concentrations were significantly higher in the PG-600-treated group on day 7 despite a similar number of corpora lutea. Endometrial ESR1 concentration was significantly reduced on day 7 in the PG-600 group. In conclusion, although progesterone was higher than controls on day 7, ESR1 did not differ from controls suggesting that treatment with PG600 is unlikely to impair reproductive potential.

Luteal Function, Biometrics, and Echotextural Attributes in Santa Inês Ewes Superovulated with Different Total Doses of Porcine Follicle-Stimulating Hormone

Premature regression of corpora lutea (PRCL) may adversely affect the outcome of hormonal ovarian superstimulation in small ruminants, and the total dose of exogenous gonadotropins used may be one of the causes of this condition. There were two major objectives of the present study: (1) to evaluate the effects of different superovulatory doses of porcine follicle-stimulating hormone (pFSH) on the biometry, blood perfusion (Doppler), and echotextural characteristics of luteal structures; and, (2) to determine the usefulness of biometric, vascular, and echotextural luteal variables, as well as measurements of circulating progesterone (P4) concentrations for early detection of PRCL in superovulated Santa Inês ewes. Twenty-seven Santa Inês ewes received an intravaginal P4-releasing device (CIDR) from Days 0 to 8 (Day 0 = random day of the anovulatory period). An IM injection of d-cloprostenol (37.5 μg) was given at the time of the CIDR insertion and withdrawal. On Day 6, all the ewes received 300 IU of eCG IM and were divided into three treatment groups (each n = 9): G100 (100 mg); G133 (133 mg); and G200 (200 mg of pFSH) administered IM every 12 h in eight injections. Transrectal ovarian ultrasonography and jugular blood sampling for serum P4 measurements were performed on Days 11 to 15. On the day of embryo recovery (Day 15), all the ewes underwent diagnostic videolaparoscopy and were classified, based on their luteal characteristics, into three response groups: nCL (ewes with normal CL only); rCL (ewes with regressing CL only); and ewes with both nCL and rCL following the superovulatory regimen. Our present results indicate that the total pFSH doses of 100 mg and 200 mg result in similar ovulatory responses and luteal function/biometrics, although the percentage of donor ewes with nCL was greater (p < 0.05) for G100 compared with the G200 animals. An application of 133 mg of pFSH was associated with diminished luteogenesis. Lastly, circulating P4 concentrations, ultrasonographic estimates of total luteal area, and CL pixel heterogeneity (standard deviation of numerical pixel values) are promising markers of luteal inadequacy in superovulated ewes.

Oral Administration of Spirulina platensis at Early Gestation Modulates Litter Size and the Expression of Inhibin, Insulin, IGF-I, CO Q10, and BMP-15 in Ewes Induced for Twinning.

Gestation in sheep necessitates the support of nutrients to avoid early embryonic mortalities. Therefore, this study investigates the effects of supplementing either L-arginine or Spirulina alga in the first trimester on the pregnancy rate, litter size, inhibin, insulin, IGF-I, CO Q10, and bone morphogenetic peptide 15 (BMP15) in maternal circulation. Animals were offered barley (500 g/head/day, 14% CP), alfalfa hay (1 kg/head/day, 12% CP), clean water, and balanced salt block licks as free choices. Forty Noemi and Najdi ewes were randomly allotted into three groups: control (C, n = 8), L-arginine (ARG, n = 16), and Spirulina (SP, n = 16). All females were implanted with CIDR for ten days. On days 8, 9, and 10, treated ewes were given a protocol comprised of human recombinant FSH at descending doses (50, 50, 40, 40, and 30, 30 IU, A.M, and P.M, respectively). At the fifth dose, animals were given an equivalent dose of hCG (240 IU). After CIDR withdrawal, ewes were exposed to fertile rams for mating. SP-ewes were orally given 50 ml (2%) Spirulina, and ARG-ewes were given 50 ml (35 mg/kg BW) L-arginine daily for 50 days postbreeding. Blood inhibin, insulin, IGF-I, CO Q10, and BMP15 were determined throughout gestation until parturition. The findings indicated that the conception rates were 25, 75, and 87.5% in C, ARG, and SP, respectively (P < 0.05). The percent of ewes giving birth to twins was 0, 25, and 50% in C, ARG, and SP, respectively (P < 0.05). The survival rates were 100, 81.8, and 83.3%, respectively. Birth weight was 5.6, 3.2, and 3.4 kg in C, ARG, and SP, respectively. Weaning weights were 28.3, 25.6, and 27.2 kg in C, ARG, and SP, respectively. BMP-15 was reduced (P < 0.05) in ARG than in C and SP. However, SP decreased (P < 0.05) inhibin more than in C and ARG. ARG and SP increased (P < 0.05) insulin than in C, whereas SP decreased (P < 0.05) IGF-I. SP increased CO Q10 compared with ARG. Ewes bearing twins revealed higher (P < 0.05) IGF-I (8.57 ng/ml) than those bearing singles (4.63 ng/ml); however, BMP-15 was higher in single (796.6 pg/ml) than in twin-bearing (387.5 pg/ml) ewes. In conclusion, providing early-gestating ewes with Spirulina enhances maternal health, productivity, and reproductive outcomes.

Resumption of postpartum reproductive cyclicity and pregnancy rates after treatment with CIDR plus PGF2 α in normally calved and retained fetal membranes affected water buffaloes

An experiment was conducted to compare the estrus induction response, estrus pattern, endometrial regeneration and pregnancy rates following treatment with CIDR plus PGF2 α between normally calved (NC) and retained fetal membranes (RFM) affected water buffaloes. A total of 32 water buffaloes including 16 treated for RFM and 16 NC were selected and divided into 4 groups as I and II, and III and IV respectively. Buffaloes of group I and III were dewormed with Albendazole followed by the administration of mineral mixture in feed for 15 days. Buffaloes of all the groups were treated with CIDR intravaginally for 9 days and they received PGF2 α at 24 hours prior to CIDR withdrawal. After CIDR removal, the buffaloes of all the groups were artificially inseminated during induced estrus. Those buffaloes returned to estrus following breeding at induced estrus, were artificially inseminated twice at an interval of 24 hours. The estrus response following CIDR withdrawal was 100%. The RFM affected buffaloes had significantly longer interval to onset of estrus and shorter duration of estrus than NC buffaloes. More pronounced intensity of estrus following CIDR treatment was observed in NC buffaloes than in RFM affected buffaloes. The mean pregnancy rates obtained in NC and RFM affected buffaloes were 81.25% and 50%. Complete loss of epithelial layer, degenerative changes of endometrium and atrophic changes in the endometrial glands were observed in RFM affected buffaloes before estrus induction. After treatment, RFM affected buffaloes showed clear regeneration of endometrium and its glandular acini. It is concluded that estrus induction using CIDR plus PGF2 α might help to augment the fertility in NC and RFM affected buffaloes at field level.

Effects of additional gonadotropin-releasing hormone and prostaglandin F2α treatment to an estradiol/progesterone-based embryo transfer protocol for recipient lactating dairy cows

This study was designed to evaluate whether the utilization of a second PGF2α treatment at the end of an estradiol/progesterone (E2/P4)-based protocol with or without GnRH at the beginning of the protocol would improve pregnancy rates of lactating Holstein cows assigned to timed embryo transfer. A total of 501 lactating Holstein cows in 5 farms were enrolled in the experiment. Within farm, cows were blocked by parity and, within block, were assigned randomly to (1) insertion of an intravaginal P4 device (controlled internal drug-releasing device; CIDR) and estradiol benzoate on d -11, PGF2α on d -4, CIDR withdrawal and an injection of estradiol cypionate on d -2, and timed embryo transfer on d 7 (1-PGF; n = 164); (2) the same treatments as 1-PGF, but with PGF2α administered on d -4 and -2 (2-PGF; n = 171); and (3) 2-PGF with the addition of a GnRH treatment on d -11 (GnRH+2-PGF; n = 166). Ovaries were scanned by transrectal ultrasonography on d -11, -4, and 7, and blood samples were collected on d -11, -4, 0, and 7 for P4 determination. Treatment comparisons were performed using contrasts. The proportion of cows with a new corpus luteum on d -4 was greater in GnRH+2-PGF cows. Cows in 1-PGF had a greater P4 concentration on d 0 but lesser P4 on d 7 compared with cows in the other groups. Cows assigned to receive 2-PGF (2-PGF and GnRH+2-PGF) had greater estrus expression, and a greater proportion of cows ovulated to estradiol cypionate. No further contrast effects were observed for follicle diameter, double ovulation rate, pregnancy per embryo transfer (P/ET) on d 32 and 60, or pregnancy loss. As P4 concentration on d -4 increased, P/ET on d 60 tended to increase. Cows with P4 ≥3.66 ng/mL on d -4 had greater P/ET on d 32 and 60 than those with P4 below that threshold. Regardless of treatment, cows with P4 concentration ≥3.66 ng/mL also had a greater pregnancy per synchronized protocol (P/SP) on d 60. Also, a P4 concentration on d -4 (low or high) × follicle diameter (continuous) interaction tendency was observed when evaluating P/ET. Although P/ET did not differ among cows with different follicles sizes with reduced P4 concentration on d -4 (<3.66 ng/mL), it increased in cows with larger follicles exposed to increased P4 concentration (≥3.66 ng/mL). When P4 on d 0 was evaluated, P/ET on d 32 and 60 was greater for cows with low (≤0.09 ng/mL) versus high (>0.21 ng/mL) P4; as P4 concentration on d 0 increased, P/ET linearly decreased. In summary, cows with increased P4 concentrations during growth of the ovulatory follicular wave had improved P/ET. Administering a second PGF2α dose reduced P4 concentration on d 0 and increased ovulatory response to the protocol, but no benefits were observed on P/ET or P/SP.

Comparison of Estradiol Cypionate, Estradiol Benzoate and Gonadotropin-Releasing Hormone in Fixed-Time Artificial Insemination in Multiparous Murrah Buffaloes

The present study was aimed to evaluate the efficiency of estradiol cypionate, estradiol benzoate and gonadotropin-releasing hormone during fixed-time artificial insemination (FTAI) protocols in multiparous Murrah buffaloes. A total of 35 buffaloes were used between September and November 2021 and subjected to a synchronization protocol. On day 0, insertion of an intravaginal progesterone device (CIDR, 1.38 g) was made with intramuscular (i/m) injection of estradiol benzoate (EB, 2 mg); On day 9: CIDR removed with i/m injections of Dinoprost (12.5 mg) and eCG (400 IU), and then buffaloes were randomized into three treatment group: 1) EB 1 mg on day 10 (n=13). 2) estradiol cypionate (EC) 1 mg on day 9 (n=12), and 3) Buserelin acetate (GnRH) 0.0105 mg on day 10 (n=10). The estrus signs observed during the FTAI (56 h after CIDR withdrawal) and conception rate at 90 days after FTAI were recorded. The conception rates were 61.5, 58.3 and 60.0% for EB, EC and GnRH treatment groups, respectively. There was no significant difference between the treatment groups for conception rate and signs of estrus. Therefore, it was concluded that all three treatments can be efficiently used during estrus synchronization protocol with FTAI in multiparous Murrah buffaloes.

Effects of estradiol cypionate dose as an ovulatory stimulus on reproductive performance of lactating dairy cows during the summer season

This experiment was designed to evaluate the effects of increasing doses of estradiol cypionate (ECP) on reproductive function of lactating dairy cows during the summer. In Exp. 1, 643 lactating Holstein cows were blocked by parity and assigned to receive 1) an intravaginal P4 device (1.9 g of P4) and 2.0 mg of estradiol benzoate on day −11, 25 mg (i.m.) of dinoprost tromethamine on day −4, 1.0 mg (i.m.) of estradiol cypionate and CIDR withdrawal on day −2, followed by TAI on day 0 (n = 326; ECP-1) or 2) the same synchronization protocol with 2.0 mg of ECP on day −2 (n = 317; ECP-2). In both treatments, cows were TAI on day 0 of the protocol, and cow rectal temperature was measured on days −2, 0, and 7. In Exp. 2, 608 lactating crossbred Holstein × Gir dairy cows were blocked by parity and enrolled to the same treatments as in Exp. 1, but on day 7, cows received one viable embryo into the uterine horn. In Exp. 1, a greater percentage of ECP-2 cows were detected on estrus (81.3 vs. 91.1%, respectively). A treatment × body condition score (BCS) interaction was observed on day 60 pregnancy per AI (P/AI), as ECP-2 cows with a BCS <2.75 had a greater P/AI vs. ECP-1, but an opposite result was observed in cows with a BCS ≥2.75. Regardless of treatment, there were effects of mean rectal temperature and heat stress events on P/AI. Treatment affected the diameter of the ovulatory follicle at TAI (ECP-1 = 15.3 mm vs. ECP-2 = 14.8 mm) and a greater percentage of ECP-1 cows had larger follicles (≥16.5 mm), but ECP-2 resulted in a greater incidence of early ovulatory cows (ovulating before day 0). Therefore, follicle diameter at TAI affected P/AI on day 60 in cows receiving ECP-2 and tended to affect ECP-1 cows. A treatment effect was observed on time to estrus following ECP treatments and a reduced proportion of ECP-2 cows showed estrus at TAI. Regardless of treatment, cows detected on estrus 48 h after ECP administration had a greater P/AI on day 60 vs. cows detected on estrus 24 h. In Exp. 2, a greater percentage of ECP-2 cows were detected on estrus (68.3 vs. 81.4%, respectively). In summary, a greater dose of ECP increased the percentage of animals expressing estrus, but it did not benefit the reproductive function of lactating dairy cows during the summer, regardless if animals were assigned to a TAI or timed-embryo transfer (TET) protocol.

Dynamic changes in gene expression during follicle development and the efficacy of four timed AI protocols in non-suckling female yaks (Bos grunniens)

The objectives of the study were to investigate changes in the mRNA expression levels of five genes during antral follicle development and to assess the efficacy of four timed-artificial insemination (TAI) protocols in female yaks (Bos grunniens). RT-qPCR analysis revealed that expression levels were greater for follicle-stimulating hormone receptor and bone morphogenic protein 15 in the small follicle, luteinizing hormone receptor, and kit ligand in the large follicle, and growth differentiation factor 9 in the medium follicle (p < 0.05). Non-suckling yaks were treated as a 7-d CIDR, and PGF2α + eCG at CIDR withdrawal and TAI with frozen yak semen at 56–58 h after PGF2α (PPe-7d); either a 7-d CIDR (PPG-7d) or a 5-d CIDR (PPG-5d), and PGF2α at CIDR withdrawal and TAI + GnRH at 70–72 h after PGF2α; and GnRH treatment on Day 0, followed by PGF2α on Day 7 and TAI + GnRH on Day 9 (GPG-7d). The results showed that the pregnancy rate (P/AI) was greater in PPG-5d than in GPG-7d (p < 0.05), but the P/AI was not different among the other TAI protocols. In conclusion, the expression levels of these genes in follicles are dynamically changed during antral follicle development in yaks. The PPG-5d protocol achieved a greater P/AI.

266 Effects of Inflammatory States on Ovarian Biology and Oocyte Competence

Abstract Female fertility is dependent on estradiol concentrations which regulate a multitude of ovarian functions including follicle development and oocyte maturation leading to ovulation of a viable oocyte. Estradiol biosynthesis is regulated by coordinated actions of follicle-stimulating hormone and intra-ovarian control mechanisms including the co-transcription factor beta-catenin. Beta-catenin is a multi-faceted protein recognized for its role in granulosa cell steroid production and is shown to be modulated by lipopolysaccharide (LPS), the endotoxin responsible for stimulation of the immune system in infections caused by Gram-negative bacteria. Beef heifers treated with subacute concentrations of LPS during a synchronized follicular wave demonstrate a decline in serum estradiol concentrations 50 h after CIDR withdrawal, corresponding with dominant follicle maturation and preceding ovulation. The endotoxin exposure also resulted in increased LPS concentration and E2:P4 ratios in follicular fluid suggesting that low dose LPS modulates the intra-follicular hormonal milieu. Additionally, in a granulosa cell line, LPS treatment decreased mRNA expression of aromatase and beta-catenin. These data indicate that LPS alters E2 synthesis by modulating beta-catenin and subsequent steroidogenic enzyme expression. To further explore the contribution of naturally occurring LPS exposure on follicular steroid production and developing oocytes, bovine ovary pairs were collected from local abattoirs. Oocytes were aspirated from small follicles and matured in vitro to evaluate meiotic events related to nuclear maturation and spindle morphology. Small follicles from ovarian pairs were separated by the detectable LPS concentrations into high and low LPS groups. Oocytes matured from low LPS follicles demonstrated an increase in the percent of abnormal maturation events. Data indicate that induced or naturally occurring low doses of LPS can alter circulating and follicular estradiol concentrations impairing oocyte maturation. Perturbation to local ovarian signaling cascades from subclinical inflammatory disease may be an unappreciated factor altering fertility and leading to decreased cow retention.

Effects of lipopolysaccharide on beta-catenin, aromatase, and estrogen production in bovine granulosa cells in vivo and in vitro

Diseases resulting from Gram-negative bacterial infection can induce an immune response by releasing a lipopolysaccharide (LPS) endotoxin that may lead to impaired fertility in cows. To evaluate the effects of LPS on follicular dynamics in a subacute inflammatory disease state, 14 Angus heifers (BW = 413 kg±14) were blocked by weight and assigned to vehicle (n = 7) or LPS treated (n = 7) groups. Heifers received subcutaneous injections of saline (CON) or 2.0 μg/kg LPS on d 2, 5, and 8 of a select synch plus controlled internal drug release device (CIDR) follicular wave synchronization protocol. Fifty hours following CIDR withdrawal, ovaries were harvested, and follicular fluid was collected for hormone and LPS analysis. Daily blood samples were collected from d 0 to d 7. Beginning on d 8 blood samples were collected at 0, 16, 24, 32, 40, and 50 h following LPS challenge. Rectal temperatures were recorded prior to treatment and at regular intervals after each LPS challenge. Heifers treated with LPS exhibited mild (+0.5 °C) hyperthermia (P < 0.05) at 3, 4, and 8 h after the initial LPS challenge (d 2) when compared to vehicle-treated controls. Follicular fluid concentrations of estradiol (E2) increased (P = 0.04) in LPS-treated heifers compared to controls (1,595 ng/mL and 808 ng/mL±240, respectively), while follicular fluid progesterone (P4) concentrations did not differ (P = 0.27) between treatment groups. Additionally, LPS concentrations tended to be increased (P = 0.59) in dominant follicles of LPS-treated heifers, but no difference was detected (P = 0.81) in small developing follicles. To further delineate the impact of LPS on ovarian signaling pathways, a granulosa cell line (KGN) was incubated in the presence or absence of LPS (10 μg/mL) for 48 h. Cells were then collected for gene expression and protein analysis. Cells in both treatment groups expressed toll-like receptor 4, myeloid differentiation factor-2 receptor, and CD-14 complex genes required for LPS signaling. Cells treated with LPS exhibited decreased mRNA expression of aromatase (P = 0.03) and beta-catenin (P = 0.02). However, no change (P > 0.10) was detected in abundance of total beta-catenin protein or beta-catenin phosphorylated isoforms at serine 552 or 675. Based on results from this in vivo experiment, these investigators concluded that low doses of LPS can alter E2 concentrations and this effect may be modulated in part through beta-catenin regulation of aromatase transcription.

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.

Investigation of the Effect of Intravaginal Device-type and Treatment-duration on Vaginal Features in Ewes and Determination of Antibiotic Susceptibility

Background: Seasonal anestrus, which is controlled by the photoperiod in ewes, limits the ability to the fertility of ewe every time. Therefore, using hormones by vaginal devices is the most effective way of increasing fertility in order to form oestrus during the anestrus period and obtain a high rate of pregnancy. But, vaginal devices may cause a change in the profile and number of bacteria in the uterus and vagina, and thus create an environment suitable for infection. For this reason, identification of bacteria that are the source of infection can be achieved by performing antibiotic susceptibility tests and interpreting them correctly. The purpose of this study was to characterize the effect of intravaginal device-type and treatment-duration on vaginal bacteria count and type and determine the antibiotic susceptibility of vaginal bacterias in ewes after a program of oestrus synchronization. Methods: A total of 90 clinically healthy Kivircik ewes were selected. In order to provide oestrus synchronization during anestrus season (June), CIDR was inserted intravaginally to 45 ewes, the other 45 ewes were progestogen sponges inserted. In both groups remained inserted for 5, 9 and 13 days. While vaginal devices were withdrawal both groups ewes were exposed to intramuscular 350 IU PMSG injection. The mucous samples were collected from all ewes 0, 5, 9 and 13 days at time the IVS or CIDR withdrawal. In samples, bacterial concentration, bacterial identification, and antibiotic susceptibility were determined. Result: The changes in the vaginal flora caused by the vaginal devices favored the development of opportunistic microbiota that could potentially be pathogenic. In the study, after vaginal device withdrawal, the bacteria species most frequently observed in both IVS and CIDR groups were E. coli. According to the results of this study, the first choice of antibiotics for vaginal infections caused by G- agents in ewes should be any of AXC, CEP and CTZ compared to other antibiotics.

Real-time assessment of the superovulatory effect of FSH and eCG with laparoscopy at different seasons in Akkaraman ewes.

Conventional methods for determining the reproductive performance of sheep bred either after estrus synchronization during the breeding season or after induction of estrus/ovulation during the non-breeding season take a long time and may give misleading results due to the effect of environmental factors. Laparoscopic observations allow real-time monitoring of ovarian activity around estrus or ovulation. This study was aimed at assessing the superovulatory effects of follicle-stimulating hormone (FSH) and equine chorionic gonadotropin (eCG) treatments by laparoscopy during breeding (September-November, n=12) and non-breeding (April-June, n=12) seasons in Akkaraman sheep. In both seasons, after CIDR withdrawal, the ewes were injected either with 600 IU eCG or 300 μl (20 mg/ml) FSH twice at 12 hour intervals. Plasma P4, E2 and LH concentrations were determined at the time of intra-vaginal CIDR insertion (day 0) and then at its withdrawal (day 12), followed by 3 and 6 days of eCG or FSH injections. After 3 (first observation) and 6 (second observation) days of hormone injections, laparoscopy was performed to record ovarian activity in both seasons. The eCG increased (p⟨0.05) the numbers of large follicles (first observation) and CL (first and second observations) in the breeding season compared to FSH treatment. CL, small-moderate and large follicle numbers of eCG treated ewes were higher (p⟨0.05) than those of FSH at both observations in the non-breeding season. In the breeding season, eCG treated ewes had higher (p⟨0.05) plasma P4 (3 and 6 days after hormones injections) and E2 (3 days after hormones injections) concentrations than those of FSH. In conclusion, the results of the present study indicate that treatment with eCG during the non-breeding season can support ovarian activity, and thus increase ovulation rate and plasma hormone concentrations around induced estrus/ovulation in Akkaraman ewes.

The effect of pre-breeding vitamin E and selenium injection on reproductive performance, antioxidant status, and progesterone concentration in estrus-synchronized Mehraban ewes.

The present study was conducted to assess the effects of pre-breeding vitamin E and selenium (ESe) injections on the reproductive performance, antioxidant status, and serum progesterone (P4) concentration in estrus-synchronized Mehraban ewes. During the breeding season, 38 ewes (3-4years) were divided into two groups (n = 18), and the synchronization of estrus was achieved by intravaginal insertion of 0.3g progesterone CIDR device for 13days followed by 350IU eCG at CIDR withdrawal. Ewes were kept under pasture conditions and exposed to Mehraban rams 48h following CIDR withdrawal. The experimental treatments were control and ESe injection. The ESe group was received three intramuscular (5mL) injections of Ese (0.5mg/mL of selenium as sodium selenite and 50IU vitamin E as DL-α- tocopheryl) once every 2weeks. Specifically, Ese was administered at 2weeks before CIDR insertion, at the times of CIDR insertion and CIDR withdrawal. Fertility, prolificacy, lambing rate, and birth weight were recorded after parturition. Blood samples were collected at CIDR insertion, CIDR withdrawal, 5, 10, and 15days after ram exposure. Fertility, prolificacy, lambing rate, and birth weight did not improve by Ese treatment, but viabilityof lambs was higher in ESe than control (P < 0.05). Serum total antioxidant capacity at day 5 and P4 at day 10 after ram exposure was higher in ESe than control (P < 0.05). In conclusion, sheep breeders can use ESe at CIDR insertion and withdrawal times to potentiate antioxidant status and progesterone profile of estrus-synchronized Mehraban ewes.

PSIII-33 A new approach to resynchronize beef cows and perform two Timed Artificial Inseminations 24 days apart

Abstract We aimed to compare the pregnancy rate from 1stTAI and 2ndTAI performed 24 days apart and pregnancy between cows that showed or not estrous on the end of 2nd TAI protocol. Were used 190 lactating cows (Angus: 114; Brangus: 53 and Santa Gertrudis: 23). On the first day of synchronization (D0), insertion of a new CIDR® (Zoetis) plus the 100µg of Cystorrelin®(Merial) treatment, on D7 25mg of Lutalyse® (Zoetis) and CIDR withdrawal. 72 hours after CIDR removal 1stTAI was performed plus GnRH treatment. After 14 days from 1stTAI, all the cows received an one time used CIDR. The pregnancy diagnosis was performed 22 days after 1stTAI, using a subjective visual evaluation of CL blood flow (BF). Pregnancy diagnosis was performed by ultrasonography Color Doppler (MicroMaxx®). Cows with an area of CL≥3cm² and/or ≥25% CL BF were diagnosed as pregnant and underwent CIDR removal without further treatment. Those diagnosed as non-pregnant underwent CIDR removal and received one Estrotect™ (Rockway-Inc.), 25mg of Lutalyse treatment and submitted to 2nd TAI 48h after CIDR removal (D24) plus the 100µg of Cystorrelin. On D31 considered pregnant cows on D22 were examined by US B-mode D31 to verify false positive and effective pregnancies. On D55, cows were submitted to US B-mode to verify the 2ndTAI pregnancy rate. Statistical analysis were performed using GLIMMIX procedure from SAS. The pregnancy on D22 was 48.5% (87/190) and the false positives were 14.9% (13/87). There was no difference on pregnancy 31 days after 1stTAI (38.9%; 74/190) vs. 2nd TAI (32.0%; 33/103; P = 0.24). Cows were more likely to be pregnant on D55 compared to cows that: showed Estrous: 42.9% (21/49) vs. No Estrous: 22.2% (12/54; P = 0.03). In conclusion, the pregnancy rates were the same at 1st to 2ndTAI. And cows that showed estrous had more pregnancy at 2ndTAI.

Evaluation of presynchronization and addition of GnRH at the beginning of an estradiol/progesterone protocol on circulating progesterone and fertility of lactating dairy cows

The objectives of this study were to determine if the utilization of a presynchronization strategy would improve fertility at first artificial insemination (AI) during an E2/P4 ovulation synchronization protocol with or without GnRH administration at the beginning of the protocol. This experiment was conducted using cows (n = 665) at their first postpartum service and the following breeding treatment: CIDR insertion and 2.0 mg of estradiol benzoate (EB) on day −11; 25 mg dinoprost tromethamine (PG) on day −4; PG, CIDR withdrawal, and 1.0 mg of estradiol cypionate (ECP) on day −2; timed-AI on day 0. At 31 ± 3 days postpartum, cows were randomly allocated to one of three treatments on a weekly basis: 1) -P + GnRH: cows assigned to the breeding protocol with 100 μg of GnRH on day −11, 2) P + GnRH: cows assigned to a presynchronization protocol using CIDR insertion +2 mg EB on day −28, PG + ECP and CIDR withdrawal on day −21, and beginning of the breeding protocol plus GnRH (100 μg) on day −11, and 3) +P-GnRH: cows assigned to a presynchronization protocol and the breeding treatment without GnRH on day −11. No treatment effects were observed on P/AI at the pregnancy diagnoses on days 32 and 60, or for pregnancy losses between days 32 and 60 of pregnancy whether analyses included all cows or only cows that ovulated near TAI. Moreover, milk yield negatively affected P/AI. Cows with greater circulating P4 concentrations on day −4 had greater P/AI on day 60. Cows without CL on day −11 had a reduced P/AI and this effect was more significant in cows not treated with GnRH. Cows assigned to -P + GnRH had the lowest circulating P4 concentration on day −4 (3.4 ± 0.16 ng/mL), followed by + P-GnRH (4.56 ± 0.17 ng/mL), and +P + GnRH (5.08 ± 0.17 ng/mL) cohorts. The data of the current study suggest that the combination of a Presynch and GnRH administration at the beginning of a TAI protocol was the most effective way to increase the % of cows with a functional CL and with elevated circulating P4 concentrations at the time of PG treatment.

Coffee pulp supplementation prior to breeding improves oxidative status without affecting fertility of primiparous ewes

Background: Coffee pulp has a high content of antioxidants capable of modifying the oxidative status in small ruminants. However, high amounts for a prolonged time can reduce fertility. Objective: To determine the effect of two inclusion levels of coffee pulp during estrous synchronization on reproductive variables and oxidative status of primiparous ewes. Methods: Sixty Suffolk x Dorset primiparous ewes were distributed into three treatments in a completely randomized design; T0: (n=20) 1.5 kg balanced diet, Tl: (n=20) 1.5 kg balanced diet and 5% coffee pulp, T2: (n=20) 1.5 kg balanced diet and 10% coffee pulp. Supplementation was given for 16 days before estrus synchronization and until the beginning of the breeding season. A progestogen (CIDR®) was inserted for ll days and a dose of PGF2a was applied two days prior to its withdrawal. Estrus detection started 12 hours after CIDR withdrawal. Blood samples were obtained during the supplementation period to measure oxidative status, antioxidant capacity, glucose and insulin, and up to 9 days after breeding to determine progesterone concentration. Pregnancy diagnosis was performed at 30 and 60 days post-breeding. An analysis of repeated measures of mixed effects and frequency analysis were carried out. Results: Inclusion of coffee pulp for a short period prior to breeding did not affect reproductive parameters, nor progesterone, glucose or insulin concentrations (p&gt;0.05); however, antioxidant capacity increased, while lipid oxidation showed an opposite trend (p&lt;0.05). Conclusion: Inclusion of up to 10% coffee pulp in the diet of ewe lambs for 16 days prior to breeding improves oxidative status without causing adverse effects on pregnancy, estrus or prolificacy.

Onset of oestrus and periovulatory events in sheep exposed to 5 and 14 days of CIDR treatment with and without eCG

The present study supports that 5-day short-term CIDR treatments without administration of eCG are equally effective for inducing oestrus behaviour, preovulatory LH discharge and ovulation in sheep than classical protocols based on 14-day treatments plus eCG at CIDR withdrawal. However, the implementation of a 5-day protocol without eCG for fixed-time artificial insemination would be adapted to a later timing of ovulation (p<.05).

Melatonin improves the efficiency of super-ovulation and timed artificial insemination in sheep.

It has been well proved that melatonin participates in the regulation of the seasonal reproduction of ewes. However, the effects of short term treatment of melatonin on ewe’s ovulation are still to be clarified. In this study, the effects of melatonin on the number of embryos harvested from superovulation, and the pregnant rate in recipients after embryo transferred have been investigated. Hu sheep with synchronous estrus treatment were given melatonin subcutaneously injection (0, 5, and 10 mg/ewe, respectively). It was found that the estrogen level in the group of 5 mg melatonin was significantly higher than that of other two groups at the time of sperm insemination (p < 0.05). The pregnant rate and number of lambs in the group of 5 mg melatonin treatment was also significantly higher than that of the rests of the groups (P < 0.05). In another study, 31 Suffolk ewes as donors and 103 small-tailed han sheep ewes as recipients were used to produce pronuclear embryo and embryo transfer. Melatonin (5 mg) was given to the donors during estrus. The results showed that, the number of pronuclear embryos and the pregnancy rate were also significantly higher in melatonin group than that in the control group. In addition, 28 donors and 44 recipient ewes were used to produce morula/blastocyst and embryo transferring. Melatonin (5 mg) was given during estrus. The total number of embryos harvested (7.40 ± 1.25/ewe vs. 3.96 ± 0.73/ewe, P < 0.05) and the pregnant rate (72.3 ± 4.6% vs. 54.7 ± 4.0%, P < 0.05) and number of lambs were also increased in melatonin group compared to the control group. Collectively, the results have suggested that melatonin treatment 36 hours after CIDR withdrawal could promote the number and quality of embryos in vivo condition and increased the pregnant rate and number of lambs.