Controlled Internal Drug Release Removal Research Articles

Effect of the feeding condition and number of dominant follicles on oestrus synchronization in Swamp buffalo

The research aimed to assess how feeding conditions and the number of the dominant follicles (DF) influence oestrus synchronization in swamp buffaloes. A total of 170 swamp female buffaloes raised in semi-grazing and housekeeping were included in the study. The synchronization used CIDR method and the follicles on the day of CIDR (Controlled Internal Drug Releasing) insert and removal were tracked by sonography. The results indicated that the age of first mating, age at first calving, and caving interval showed no significant differences in the two feeding conditions. The number of follicles at CIDR insertion and removal decreased in semi-grazing, but increased in housekeeping buffaloes. The percentage of buffalo without DF, one DF, and two DFs at CIDR withdrawal did not differ statistically. The percentage of estrus was 100% in both groups. Pregnancy rates were 53.18% in semi- grazing and 52.63% in housekeeping. The onset of oestrus was similar, but the duration of oestrus was longer in the buffaloes without DF than those with one or two DFs. The percentage of pregnant buffaloes in the group without DF was significantly lower than that in the other two groups. In summary, the efficiency of oestrus synchronization did not differ between semi-grazing and housekeeping methods. Buffaloes without DF displayed longer oestrus periods and lower pregnancy rates compared to those with one or two DFs in the ovarian system.

291 Impact of Early Resynchronization in Combination with Color Doppler Ultrasonography on Cumulative Pregnancy Rates in Postpartum Beef Cows

Abstract The objective of this study is to evaluate a GnRH and progesterone-based early resynchronization strategy combined with color Doppler ultrasonography for early pregnancy diagnosis. Postpartum Bos taurus beef cows (n = 375) from four locations were enrolled in a completely randomized design, where cows were randomly assigned to 1 of the 2 treatments: 1) Natural Service (NS), cows were exposed to a 7-d CO-Synch + CIDR protocol followed by fixed-time artificial insemination (day 0). On day 15, cows were exposed to natural service for the remainder of the study; or 2) Early Resynchronization (ER), cows were exposed to a 7-d CO-Synch + CIDR protocol followed by fixed-time artificial insemination (day 0). On day 15, a controlled internal drug release (CIDR) device was inserted. On day 20, CIDR was removed, and pregnancy was estimated based on luteal blood perfusion using color Doppler ultrasonography. Non-pregnant cows received a 25-mg injection of prostaglandin F2α at CIDR removal and were fixed-timed-AI concurrently with a 100-μg injection of GnRH 60 to 66 h later. Pregnancy diagnosis and fetal aging were performed on day 53 using conventional ultrasonography. Pregnancy rate to the first AI was greater (P = 0.03) for NS compared with ER (70.25% vs 59.29%, respectively). Pregnancy rates for the second service were also greater (P = 0.01) for NS compared with ER (65.67 vs 42.93%, respectively). Cumulative pregnancy rates to AI did not differ between treatments (P = 0.38); however, cumulative pregnancy rates (first and second service) were greater (P < 0.01) for NS compared with ER (90 vs 77.73%, respectively). In conclusion, the proposed early resynchronization strategy using color Doppler ultrasonography resulted in fewer pregnancies compared with natural service as the second service strategy. Moreover, the proposed ER decreased pregnancy rates to the first insemination, indicating a detrimental effect of ER on pregnancy establishment.

A single dose of FSH or hCG during a split-time AI program did not enhance follicular growth or pregnancy per artificial insemination in beef heifers

ABSTRACT The objective of this study was to evaluate the effect of a low dose of hCG or FSH on follicle growth and pregnancy per artificial insemination (AI) in crossbreed beef heifers subjected to a split-time AI program. All heifers (n = 386) were subjected to a 7-d Co-synch protocol and an intravaginal controlled internal drug release (CIDR) device (GnRH and CIDR insertion; 7 days later PGF2α, CIDR removal, and application of a tail head estrus detection patch; 36 hours later AI of heifers detected in estrus, 24 hours later timed AI of remaining heifers). At CIDR removal, heifers were randomly assigned to receive either 150 IU of hCG, 20 mg of FSH, or remained as untreated controls. The diameter of the preovulatory follicle and the pregnancy per AI did not differ among treatments. A smaller (P = 0.035) proportion of heifers in the hCG group were detected in estrus compared with control. Additionally, the incidence of premature ovulation was greater (P < 0.01) among heifers treated with hCG compared with control. In conclusion, using a low dosage of hCG or FSH at the time of device removal did not affect pre-ovulatory follicle growth or the proportion of pregnant heifers.

55 Investigating the Potential Benefits of an Immunostimulant (Mycobacterium Cell Wall Fraction) Treatment on Increased Pregnancy Success of Embryo Transfer Recipients

Abstract Modulating the vaginal and uterine microbiome through the use of immunostimulants has the potential to improve fertility in cattle. The object of this study was to evaluate the potential benefits of an immunostimulant, mycobacterium cell wall fraction (MCWF), to increase pregnancy success of embryo transfer (ET) recipients. The three main objectives of this study were: determining if injecting ET recipient cows with MCWF increased proportion of cows with a corpus luteum (CL) on ET day, identifying differences in CL volume and progesterone concentration at the time of ET, and determining if ET recipients that received MCWF have greater pregnancy rate. A total of 926 beef cows at eight different locations were enrolled in this study. Primiparous and multiparous cattle were enrolled in a 7-day CO-Synch + CIDR estrous synchronization protocol. During this protocol, cows received an injection of GnRH (100-μg) at CIDR (controlled internal drug release) insertion [d -9], a prostaglandin F2α injection (25 mg), and CIDR removal [d-2] followed by another injection of GnRH (100 μg; d 0) within 60-66 hours after CIDR removal. At each location at (d 0), cows were sorted based on BCS and days post-partum and randomly assigned to one of two treatments: 1) MCWF- a single intramuscular injection (5 mL) of MCWF at (d 0; Amplimune, NovaVive Inc.; n = 452) or 2) Control- a single intramuscular injection (5 mL) of saline (n = 474). Seven days after (d 7), cows were examined by ultrasonography to determine if a CL was present. Recipient cows that had a CL received ET with a frozen or fresh in vivo or in vitro produced embryo in the ipsilateral uterine horn to the CL, with embryo type and grade balanced between treatments. Blood samples were collected in one location (n = 98) for analysis of plasma concentration of progesterone on (d 7) ET day. Pregnancy diagnosis was then performed by ultrasonography between 30 and 90 days of gestation. Data were analyzed using the GLIMMIX and MIXED procedures of SAS. Cow BCS and days postpartum were similar (P &amp;gt; 0.10) between treatments (5.5 ± 0.6 and 103 ± 14 days, respectively). Recipient cow utilization rate was similar between treatments (P = 0.49; 81.2 ± 0.42%). At the time of ET MCWF had larger CL than Control cows (5,768.6 ± 133.7 and 5,408.0 ± 128.5 mm3, respectively, P = 0.03). and increased progesterone concentration (2.68 ± 0.17 and 2.18 ± 0.14 ng/mL, respectively, P = 0.01). However, pregnancy rate to ET was similar (P = 0.922) between treatments (60.9 and 61.3 ± 0.05% for Control and MCWF, respectively). In conclusion, treatment of ET recipient cows with MCWF increased CL volume and concentration of progesterone at the time of ET but failed to generate greater pregnancy rates.

Haematological Responses following Oestrus synchronisation and superovulation protocols in West African Dwarf Goats

Experiments were performed to evaluate the haematological (erythrocyte and leucocyte parameters) response of West African Dwarf goats following oestrus synchronisation and superovulation protocol. Twenty-four clinically healthy female goats aged 2.34 ± 0.4 years and average body weight of 13.83 ± 0.7 kg was used for the experiment. The goats were divided into two groups. Group I goats was synchronized using Controlled Internal Drug Release (CIDR) for 18 days with the first day of insertion as day 0. On day 16 following CIDR insertion into the vagina, a single dose of 500 IU equine chorionic gonadotropin (eCG) was administered to each doe intramuscularly while group II served as the control was subjected to the same housing and environmental conditions but was not synchronized. superovulation was induced with porcine follicle stimulating hormone (pFSH) which was injected intramuscularly at a rate of 5mg/kg in reducing doses twice a day at 06.00 h and 18.00 h for 3 days starting from 48hr i.e., 2 days before CIDR removal (day 17). A blood sample of 5 mL was collected by jugular venipuncture of each of the goats before the protocol (Day 0) and at the end of the protocol (Day 18). All erythrocyte parameters evaluated showed a significantly higher value in group II (control) compared to group I post-oestrus synchronisation while total leucocyte count, neutrophil count and neutrophil/lymphocyte ratio values obtained after the protocol were also higher in group I (P &lt; 0.05) compared to group II. It is therefore concluded that oestrus synchronisation and superovulation may elicit stress in West African Dwarf goats and an appropriate antioxidant should be part of the protocol.

Effect of progesterone infused controlled internal drug releasing (CIDR) device and timing of gonadotropin stimulation using P.G. 600 on reproductive success in ewes bred out of season.

Objectives of this study were to determine effects of exogenous progesterone (via controlled internal drug releasing devices; CIDR) in combination with exogenous gonadotropins (PMSG/hCG) use either at CIDR removal or 1 d before CIDR removal to induce estrus and cyclicity and subsequently enhance the proportion of ewes lambing, lambing rate, prolificacy, and days to lambing in ewes bred out of season. Multiparous ewes (n = 414) were randomly assigned to one of four treatments: untreated (U, n = 122), 7 d CIDR (C, n = 97), 7 d CIDR plus P.G. 600 (240 IU pregnant mare serum gonadotropin [PMSG] and 120 IU human chorionic gonadotropin [hCG]) at CIDR removal (CPG0, n = 97), and 7 d CIDR plus P.G. 600 (240 IU PMSG and 120 IU hCG) 1 d before CIDR removal (CPG-1, n = 98). Rams (n = 15) were joined with ewes immediately after CIDR removal and remained with ewes for a 21 d breeding period. Lambing data were summarized for the first 10 days of the lambing season and overall. Categorical data were analyzed using the GLIMMIX procedure of SAS whereas non-categorical data were analyzed using the mixed procedure. Proportion of ewes lambing in the first 10 d was greater (P < 0.05) for CPG0 and CPG-1 ewes compared with C ewes, which was greater (P < 0.0001) compared with U ewes. Overall proportion of ewes lambing was greater (P ≤ 0.0001) in all treatments utilizing CIDR compared with U ewes, but no differences (P ≥ 0.84) due to P.G. 600 were detected compared with C. Lambing rate in the first 10 d was greater (P < 0.05) for CPG-1 than C, with CPGO being intermediate, and all CIDR-treated ewes being greater than U (P < 0.0001). Overall lambing rate increased (P ≤ 0.0001) in all treatments utilizing CIDR compared with U ewes, but no differences (P ≥ 0.76) due to P.G. 600 were detected compared with C. Prolificacy was similar among all treatments both for the first 10 d of the lambing season (P = 0.86) and overall (P = 0.80). Day of lambing in the lambing season was reduced (P ≤ 0.03) for CPG0 and CPG-1 compared with CIDR-treated ewes, which was reduced (P < 0.0001) compared with U ewes (days 10.6, 9.0, 13.4, and 24.4 ± 0.9 for CPG0, CPG-1, CON, and U, respectively). Though timing of P.G. 600 did not influence results, the combination of CIDR and P.G. 600 enhanced the proportion of lambs born earlier in the lambing season, and incorporating a CIDR with or without P.G. 600 enhanced the overall proportion of ewes lambing and lambing rate in out-of-season breeding scenarios.

Effect of eCG dose on ovarian haemodynamics, hormonal profiles and prolificacy rate when oestrus was induced during low-breeding season in Beetal goats.

The objectives of the experiment were to determine the effect of two doses of equine chorionic gonadotropin (eCG) in a standard synchronization protocol based on a short-term progesterone (P4 ) priming on ovarian structures and haemodynamics, concentrations of steroid hormones and prolificacy rate when oestrus was induced during low-breeding season (LBS) in Beetal dairy goats. We hypothesized that inclusion of eCG in a short-term P4 priming-based synchronization protocol would increase the blood perfusion to ovarian structures leading to enhance oestrous and ovulatory responses and prolificacy rate in goats. Forty-two multiparous acyclic goats were blocked by body condition and, within block, assigned randomly to receive saline as control (CON), low eCG (L-eCG; 300 IU) or high eCG (H-eCG; 600 IU) dose. Initially, a controlled internal drug release (CIDR) device was placed in the anterior vagina on d -8, followed by removal of CIDR on d -3, concurrent with the administration of PGF2α and eCG according to their respective treatments. Goats were monitored for oestrous response. B-mode and Doppler ultrasonography was performed with 12-h interval, starting from day -3 until natural breeding (day 0), and then on days 5, 10, 15 and 20 post-breeding to monitor follicular and luteal dynamics and blood flow, respectively. Blood was sampled at 0, 12, 24, 36 and 60 h after CIDR removal to quantify plasma concentrations of estradiol-17β (E2 ), whereas plasma concentrations of P4 were assayed at days 5, 10, 15 and 20 after breeding. Pregnancy and prolificacy rates were determined at day 30 and 150 after breeding, respectively. Data were analysed with mixed-effects models, and orthogonal contrasts were used to evaluate the effect of treatment [Con vs. (½L-eCG + ½ H-eCG)] and dose of eCG (L-eCG vs. H-eCG). Data are presented in sequence as CON, L-eCG, H-eCG (LSM ± SEM). The oestrous intensity score (152.9 vs. 182.7 vs. 186.5 ± 15.1; p =.02) was greater in eCG-treated goats as compared to CON. Administration of eCG reduced the intervals to standing oestrus (66.2 vs. 41.8 vs. 48.9 h ± 5.5; p =.05), breeding (70.2 vs. 44.4 vs. 45.4 h ± 4.5; p =.03) and ovulation (84.5 vs. 61.2 vs. 63.4 h ± 6.2; p =.05) compared with CON goats. The mean growth rate of pre-ovulatory follicle was greater (1.11 vs. 1.49 vs. 1.45 mm ± 0.08; p =.01) in eCG-treated goats resulting in an increased diameter of pre-ovulatory follicle (6.27 vs. 7.20 vs. 7.31 mm ± 0.07; p <.01) and corpora lutea (6.75 vs. 8.26 vs. 8.07 mm ± 0.42; p =.04) than CON. The mean follicular blood flow did not differ among treatments; however, the mean luteal blood flow was greater in L-eCG-treated goats (0.81 vs. 1.61 vs. 1.07 cm2 ± 0.12; p =.001). The mean concentrations of E2 (4.03 vs. 5.21 vs. 4.78 pg/ml ± 0.42; p =.04) and P4 (4.85 vs. 6.39 vs. 6.22 ng/ml ± 0.34; p =.04) were greater in eCG-treated goats. The twinning rate did not differ between treatments; nevertheless, prolificacy rate was greater (p =.04) in L-eCG-treated goats. Collectively, our data suggest that the administration of eCG improves the induction of oestrous and ovarian dynamics. Administration of L-eCG enhances prolificacy rate, therefore, a low dose of eCG might be practically beneficial to improve reproduction during LBS in acyclic Beetal dairy goats.

Comparison of the effect of Gonadotropin-Releasing Hormone and Human Chorionic Go3nadotropin on Reproductive Performance of Romanov Ewes During the Natural Breeding Season.

Low pregnancy rate is an important issue in small ruminants. Superovulation protocols would serve as the possible techniques to increase the pregnancy rate in small ruminants due to the use of synthetic hormones. The present study aimed to assess the effect of gonadotropin-releasing hormone (GnRH) and Human chorionic gonadotropin (hCG) after artificial insemination (AI) on the pregnancy rate of Romanov ewes breed. To this end, estrus was synchronized in 140 ewes using a controlled internal drug-releasing (CIDR) device. Following CIDR removal, all ewes received 400 IU Pregnant Mare's Serum Gonadotropin (PMSG). The ewes were assigned to three groups (control, n=20; experimental groups, each of them was divided into three subgroups, n=20). The first group in each experiment was assumed as control. The second and third groups received three levels of GnRH, (400, 500, and 600 µg, n=20) and hCG (200, 300, and 400 IU) administered by the intramuscular (IM) route. As soon as the signs of estrus were observed, the ewes were isolated from the others and artificially inseminated with fresh sperm using the transcervical method. The results indicated that there were no significant differences between experimental groups in ovulation and pregnancy rates on days 33 and 90 post-AI (P>0.05). The highest twin birth rate (20%) was recorded in the hCG group (300 IU), as compared to GnRH and control groups (P<0.05). The effect of GnRH and hCG on the plasma concentration of progesterone was significant (P<0.01). The groups significantly differed in fecundity rate (Single birth, twin birth, and triple birth) (P<0.05), all of these parameters were significantly higher in the hCG 300 IU group. It can be concluded that the injection of 300 IU of hCG hormone after insemination improves pregnancy rate in ewes.

19 Evaluation of Fixed-time Artificial Insemination and Estrus Expression in Angus-cross Heifers with Female Sex-sorted Semen Using 6-day and 7-day CIDR Protocols at Multiple Time Intervals

Abstract The objective was to investigate the influence of progesterone exposure time and breeding time on fixed-time artificial insemination (FTAI) protocols using sex-sorted semen in Angus-cross heifers (n = 85; mean BW = 324 kg.). Heifers were assigned randomly into four treatment groups. Treatments 1 and 2 used a 6-day CO-synch + controlled internal drug release (CIDR) protocol with gonadotropin releasing hormone (GnRH) administration and CIDR insertion on day 0 and prostaglandin F2α (PGF) administration on day 6, upon simultaneous CIDR removal. Treatment 1 heifers underwent FTAI 72 h post CIDR removal, and Treatment 2 heifers underwent FTAI 80 h post CIDR removal. Treatments 3 and 4 used a 7-d CO-synch + CIDR protocol with GnRH administration and CIDR insertion on d 0 and PGF administration on d 7, upon simultaneous CIDR removal. Treatment 3 heifers underwent FTAI 54 h post CIDR removal, and Treatment 4 heifers underwent FTAI 62 h post CIDR removal. Estrus expression was determined using an estrus detection patch (Estrotect™, Rockway Inc., Spring Valley, WI). Estrus expression was classified as a patch with 50% or more of the color rubbed off and the absence of estrus expression as a patch with less than 50% of the color rubbed off. All heifers were inseminated using female sex-sorted semen from a single Angus sire (SexedULTRA 4M™, STgenetics, Navasota, TX). Pregnancy was determined by ultrasound 57 d following FTAI. Pregnancy rates and patch scores were analyzed using the GLIMMIX procedure of SAS 9.4 with treatment as a fixed effect. There was no effect of treatment on the first service AI rates (P = 0.91) or on estrus expression (P = 0.17) in heifers. In conclusion, the combinations of progesterone and breeding time in this study did not affect first service AI conception rates or estrus expression in beef cattle heifers.

24 The Effects of Vaginal pH on Fertility of Beef Cows and Heifers Enrolled in the 7-d Co-synch + Controlled Internal Drug Release (CIDR) Protocol

Abstract The objective of this study was to evaluate the effect of vaginal pH on fertility of beef cows and heifers enrolled in a fixed-time artificial insemination (TAI) protocol. Year 1, 40 multiparous cows; and year 2, 48 multiparous cows and 41 heifers with no signs of vaginitis were enrolled. Vaginal flushes were collected for all females on D0, D7 and D10 and immediately analyzed for pH with a pH meter. As evidence of vaginitis, CIDR score was determined at CIDR removal (D7) following a 1 to 4 scoring system: 1=no secretion; 2=clear secretion; 3=purulent secretion; 4=purulent secretion and blood. Pregnancy status was determined by transrectal ultrasonography approximately 40 days after TAI. No animals had CIDR scores 1, whereas 23.7% of animals had a score 2, 69.6% a score 3, and 6.6% a score of 4. There was no effect of CIDR score on pregnancy outcome (P = 0.697). However, vaginal pH on D7, differed according to CIDR score (P = 0.045), where score 2 had greater pH (7.32±0.04) than score 4 (7.09±0.08), and score 3 were intermediate (7.25±0.02). No effect (P = 0.307) of CIDR score on vaginal pH on D10 was observed. There were no differences (P &amp;gt; 0.01) between vaginal pH of pregnant and open animals on D0 (7.06±0.05); D7 (7.26±0.03); and D10 (7.21±0.04). Nonetheless, on D10 females with vaginal pH 7.1 &amp;lt; pH ≤ 7.34 had greater (P = 0.005) PR (n=56; 66.1%) than cows with pH &amp;gt; 7.35 (n=33; 45.5%), and pH &amp;lt; 7.09 (n = 39; 33.3%). We conclude that vaginitis caused by the CIDR changes vaginal pH on D7, which is not carried on to D10. In addition, there is indication of an optimal vaginal pH range at the day of TAI for maximum PR to the protocol.

Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus, pregnancy outcomes, and cost per pregnancy

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF2α - PRID; d 6, PGF2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d -6, GnRH + controlled internal drug release (CIDR); d -1, PGF2α - CIDR; d 0, PGF2α; d 2, GnRH + TAI]; (2) CIDR6 (d -6, GnRH + CIDR; d -1, PGF2α; d 0, PGF2α - CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.

Effects of prolonging the interval from progestin removal to prostaglandin F2α injection from 16 to 17 d in a long-term estrus synchronization protocol in beef heifers.

To determine effects of delaying the injection of prostaglandin F2α (PGF) and fixed-time artificial insemination (TAI) in the 14-d CIDR-PG protocol, 1,049 Angus heifers at six locations were enrolled in a completely randomized design. Within location heifers were randomly assigned to one of two treatment groups: 1) PG16 (n = 518), heifers received a controlled internal drug release (CIDR) insert on d 0 for 14 d, a 25-mg injection of PGF 16 d after CIDR removal (d 30), and a 100-µg injection of gonadotropin-releasing hormone concurrent with TAI 66 ± 2 h later; or 2) PG17 (n = 531), heifers were treated the same as PG16, however, PGF was administered 17 d after CIDR removal (d 31), and heifers were TAI 66 ± 2 h later. Estrus detection patches were applied to a subset (n = 482) of heifers at the time of PGF administration and were examined for activation at TAI. Dominant follicle diameter was determined via transrectal ultrasonography at PGF administration and TAI in a subset of heifers (n = 116). Transrectal ultrasonography was performed to determine pregnancy rates to TAI (PR/AI) between 30 and 45 d after TAI. Estrus expression prior to TAI differed by treatment where PG17 heifers had greater (P < 0.01) expression of estrus than PG16 heifers (57.8 ± 6.1% vs. 43.4 ± 6.1%, respectively). Nevertheless, dominant follicle diameters at PGF and at TAI were similar (P ≥ 0.59) between PG16 and PG17 heifers. In addition, PR/AI did not differ (P = 0.29) between PG16 and PG17 treatments (50.5 ± 3.2% vs. 45.7 ± 3.1%, respectively). Results of this experiment indicate that delaying the injection of PGF and TAI in the 14-d CIDR-PG protocol increased estrus expression prior to TAI yet did not improve fertility in beef heifers.

35 Vaginal pH changes due to the use of a controlled internal drug release (CIDR) and its effects on fertility of beef cows

Abstract The objective of this study was to evaluate changes in vaginal pH of beef cows enrolled in a 7-d CO-Synch+controlled internal drug release (CIDR) protocol and its effects on pregnancy. A total of 46 multiparous beef cows with no signs of vaginitis were enrolled. Individual vaginal flushes were collected from cows on D0, D7 and D10. Individual flushes were immediately analyzed for pH with a portable pH meter. As evidence of vaginitis, a CIDR score was determined at CIDR removal (D7), following a 1 to 4 scoring system: 1 = clean, no secretion; 2 = clean, clear secretion; 3 = purulent secretion; 4 = purulent secretion and blood. Pregnancy status was determined by rectal ultrasonography approximately 40 days after TAI. No cows had CIDR scores of 1 or 4, 19.6% of cows had a score of 2 and 80.4% a score of 3. Vaginal pH increased from D0 (6.88±0.007) to D7 (7.32±0.047), regardless of CIDR score (P &amp;lt; 0.001). Cows with a CIDR score 2 had greater pH on D7 (7.35±0.084) than on D0 (6.90± 0.14; P = 0.001), and pH on D10 was intermediate (7.14±0.14; P = 0.14). Cows with CIDR score 3 had reduced pH on D10 (7.09±0.07) when compared to D7 (7.29±0.04; P = 0.003), but greater pH on D10 than on D0 (6.87±0.07; P = 0.007). There was a pregnancy by day interaction for pH values (P = 0.046). Pregnant cows had greater vaginal pH in comparison with non-pregnant cows on D0 (6.99±0.081 and 6.68±0.09, respectively; P = 0.01) and D10 (7.26±0.08 and 6.89±0.09, respectively; P = 0.006), but not on D7 (7.34±0.05 and 7.27±0.06, respectively; P = 0.38). Non-pregnant cows had reduced vaginal pH on D10 in comparison to D7 (6.89±0.09 and 7.27±0.06, respectively; P &amp;lt; 0.001), whereas pregnant cows had similar vaginal pH on D10 and D7 (7.26±0.08 and 7.34±0.05, respectively; P = 0.35). We conclude that CIDR increases vaginal pH overtime and greater pH at TAI is beneficial to fertility.

The potential of volatile organic compound analysis in cervicovaginal mucus to predict estrus and ovulation in estrus-synchronized heifers

Cervicovaginal mucus is a mixture of mucins, ions, salts, and water, the proportions of which change during the reproductive cycle. It is suspected that this mucus emits an important volatile signal indicative of the reproductive state of the female. The objective of this study was to identify volatile organic compounds (VOC) in bovine cervicovaginal mucus that are modulated during the estrous cycle and could potentially be used as biomarkers of estrus and ovulation. Cervicovaginal mucus was collected from crossbred beef heifers (n = 8), which were synchronized using an 8-d controlled internal drug release (CIDR) protocol and in which onset of estrus and time of ovulation were determined by visual observation and ultrasonography, respectively. Mucus samples were collected between 0 and 96 h after CIDR removal (estrus onset occurred at 49.1 ± 3.3 h after CIDR removal). A validation study was performed on an independent group of 15 heifers from which cervicovaginal mucus samples were collected every 8 h from 40 to 80 h after CIDR removal. The VOC in mucus were identified using gas chromatography-mass spectrometry and selected compounds were quantified using selected-ion flow-tube mass spectrometry. The presence of 47 VOC was detected in mucus samples by gas chromatography-mass spectrometry with those exhibiting highest abundance including 2-butanone, acetone, 2-pentanone, 4-methyl-2-pentanone, 1-(1-methylethoxy)-2-propanone, ethanol, 2-methyl-2-propanol, and 2-butanol. All VOC peaked between 24 to 47 h after the onset of estrus (ovulation occurred 26.6 ± 5.6 h after estrus onset). Two VOC, 2-pentanone and 4-methyl-2-pentanone, exhibited a significant increase at the onset of estrus, whereas concentration of 2-butanone increased significantly just after estrus onset, indicating that these VOC may be used as putative biomarkers of estrus. The results of our study may contribute to the development of a sensor device based on VOC to aid the detection of estrus and ovulation in cattle, with particular relevance for the dairy industry where the majority of females are bred by artificial insemination.

Effects of flaxseed-rich diet on reproductive performance in estrous-synchronized Baluchi ewes.

The purpose of this study was to compare the effects of using different levels of flaxseed in diets on the reproductive performance of estrous-synchronized Baluchi ewes (para 3). Diets contained either basal diet (control) or different levels of extruded flaxseed (2%, 5%, 7%, 10%, and 12%) and were fed from lambing to 60 days after lambing. The ewe estrus cycles were synchronized using controlled internal drug release (CIDR) for 14 days starting from day 16 of fat supplementation. The rams were introduced 24 h after CIDR removal. The ewes fed control diets had the highest mean dry matter intake (1,800&amp;plusmn;35 g) which was declined with the increase of flaxseed levels. The experimental diets exerted no effects of urea concentration in blood plasma. However, plasma glucose concentration was lower (p &amp;lt;0.05) in the ewes fed the control diet and 2% flaxseed, compared to those in other groups. Nonetheless, there was no difference among the ewes fed 5%, 7%, 10%, and 12% flaxseed in terms of plasma glucose concentrations (p &amp;lt;0.05). The ewes fed 2% flaxseed had the highest level of plasma triglyceride concentration among other groups. In addition, the control group had the lowest level of plasma total cholesterol and low-density lipoproteins concentration in comparison to other groups (p &amp;lt;0.05). However, plasma nonesterified fatty acids and &amp;beta;-hydroxybutyrate concentrations were similar among the groups (P&amp;gt;0.05). The mean interval between CIDR removal and the exhibition of estrus ranged from 30 to 40 h with the shortest interval being recorded in the ewes fed 12% flaxseed (p &amp;lt;0.05). The control group had the lowest number of follicles on estrus day among other groups (p &amp;lt;0.05). Furthermore, the ewes fed 10% and 12% flaxseed had the highest ovulation, pregnancy, and lambing rates, compared to other groups (p &amp;lt;0.05). In conclusion, the findings revealed that feeding the ewes with 10% and 12% flaxseed resulted in the improvement of reproductive performance.

Supplementation frequency and amount modulate postweaning growth and reproductive performance of Bos indicus-influenced beef heifers.

This 2-yr study evaluated the growth and puberty attainment of Bos indicus-influenced beef heifers offered 2 different postweaning concentrate supplementation amounts and delivery frequencies. On day 0 of each year, 64 Brangus crossbred heifers were stratified by initial body weight (BW) and age (mean = 244 ± 22 kg; 314 ± 17 d) and assigned into 1 of 16 bahiagrass pastures (4 heifers/pasture/yr). Treatments were randomly assigned to pastures in a 2 × 2 factorial design (4 pastures/treatment/yr) and consisted of concentrate dry matter (DM) supplementation at 1.25% or 1.75% of BW which were offered either daily (7×) or 3 times weekly (3×) for 168 d. On day 56 of each year, heifers were assigned to an estrus synchronization protocol consisting of intravaginal controlled internal drug release (CIDR) insertion on day 56, CIDR removal on day 70, i.m. injection of 25 mg of prostaglandin F2α (PGF2α) on day 86, and i.m. injection of 100 µg of gonadotropin-releasing hormone and timed-AI at 66 h after PGF2α injection (day 89). Heifers were exposed to Angus bulls from day 89 to 168 (1 bull/pasture). Pregnancy diagnosis was assessed on day 213 of each year. Supplementation amount × frequency effects were not detected (P ≥ 0.12) for any variable, except for plasma concentrations of glucose (P = 0.10) and urea nitrogen (PUN; P = 0.01). Herbage mass, herbage allowance, and nutritive value did not differ (P ≥ 0.12) among treatments. Increasing supplementation DM amount from 1.25% to 1.75% of BW increased (P ≤ 0.05) plasma concentrations of insulin-like growth factor 1 (IGF-1), overall average daily gain (ADG), final BW, percentage of pubertal heifers on day 89, pregnancy and calving percentages, and percentage of heifers calving within the first 21 d of the calving season. However, reducing the supplementation frequency from daily to 3× weekly, regardless of supplementation amount, did not impact overall pregnancy and calving percentages (P ≥ 0.42), but caused (P ≤ 0.05) fluctuations in plasma concentrations of insulin and IGF-1 and decreased (P ≤ 0.03) overall ADG, final BW, puberty attainment on days 56, 89, and 168, and percentage of heifers calving during the first 21 d of the calving season. Hence, increasing the supplement DM amount did not prevent the negative effects of reducing the frequency of supplementation (3× vs. 7× weekly) on growth and reproduction of replacement Bos indicus-influenced beef heifers.

Estrous response and pregnancy percentages following use of a progesterone-based, split-time estrous synchronization treatment regimens in beef heifers

Mean estrous response rate (%ERR) and pregnancy/AI percentages (%P/AI) were determined after imposing split-time AI (STAI) and fixed time AI (FTAI) following 14-d controlled internal drug release (CIDR)+PGF2α or 5-d Select Synch + CIDR regimens. In Experiment 1, 1152 heifers (five locations) were randomly assigned to 14- or 5-d and to 54 + 74- or 64 + 84-h STAI treatment combinations. Estrous detection patches were affixed at PGF2α administration (19 day after- and on day 5 at- CIDR removal for 14- and 5-d regimens, respectively), assessed at 54- or 64-h and again at 74- or 84-h after PGF2α. Heifers determined to be in estrus at respective times were inseminated and non-estrous heifers at 74- or 84-h were given GnRH and inseminated concomitantly. The %ERR between 54 + 74- and 64 + 84-h STAI combinations differed (73.2 % and 78.8 %, respectively; P < 0.05), but %P/AI did not. In Experiment 2, 2014 heifers (eight locations) were randomly assigned to 14- or 5-d regimens and were inseminated split-time (64+84-h combination, similar to Experiment 1) or at fixed time (72- or 56-h after PGF2α for 14- or 5-d regimens, respectively). There were differences (P < 0.01) between STAI and FTAI treatments for %ERR (81.3 % and 64.4 %) and %P/AI (61.2 % and 55.4 %). Estrous synchronization regimen by AI treatment interaction (P < 0.05) showed that the %ERR were 79.8 %, 82.6 %, 66.2 % and 62.8 % and the %P/AI were 58.9 %, 63.4 %, 56.5 % and 56.5 % (for 14-d/STAI, 5-d/STAI, 14-d/FTAI and 5-d/FTAI, respectively). In conclusion, the 5-d CIDR with 64+84-h STAI combination was the most effective because of the greater %P/AI when this regimen was imposed.

Greater concentrations of IGF-1 are associated with increasing pregnancy rate in melatonin implanted anestrous Barki ewes

The present study was conducted to determine if increases in IGF-1 concentration, associated with treatment of ewes with melatonin, has beneficial effects on pregnancy rates when there is induction of estrus in anestrous ewes. A total of 120 multiparous lactating ewes were assigned to three groups (n = 10/group). Ewes of Group 1 were treated with a melatonin implant for 42 days followed by insertion of a controlled internal drug release (CIDR) device for 14 days with administration of equine chorionic gonadotropin (eCG) at day of CIDR removal. The ewes of Group 2 were treated with a CIDR and eCG at the same times as ewes of Group 1 and ewes of Group 3 were assigned to an be untreated control group. Melatonin implantation resulted in an increase in IGF-1 concentrations and lesser estradiol (E2) and triiodothyronine (T3) concentrations. Ewes of Groups 1 and 2 had the greatest progesterone (P4) concentration compared ewes of Group 3. The E2:P4 ratio was less in ewes of Group 1 compared Group 3. Melatonin implantation of ewes resulted in a greater pregnancy rate compared to treatment with the CIDR and eCG which, in turn, had a greater rate than ewes of the control group. In conclusion, melatonin implantation modulates the hormonal milieu including P4, E2, T3 and IGF-1 in seasonally anestrous ewes. Increased IGF-1concentrations, as a result of melatonin treatment, are associated with a greater percentage pregnancy rate when there is treatment of anestrous ewes to induce onset of estrus.

Use of Propylene-Glycol as a Cosolvent for GnRH in Synchronization of Estrus and Ovulation in Sheep.

Simple SummaryThe foreseeable shortage of equine chorionic gonadotrophin (eCG) for estrus synchronization in sheep will have a huge impact on breeders using out-of-season breeding, artificial insemination, or embryo transfer because there is no other product with similar activity. Hence, there is a strong need for alternative protocols. The aim of the present work was to evaluate the reproductive response of sheep in breeding season to the administration of gonadotrophin-releasing hormone (GnRH) using propylene-glycol as a cosolvent, as well as the most adequate timing for such administration. From the results obtained, protocols based on a short-term treatment with a Controlled Internal Drug Release (CIDR) device and a single dose of GnRH in propylene-glycol at 36 h after CIDR removal may constitute an alternative to traditional protocols based on the administration of a single dose of eCG at CIDR removal.The foreseen shortage of eCG for estrus synchronization in sheep makes necessary the development of alternative protocols. The aim of the present work was to evaluate the reproductive response of sheep in breeding season to the administration of GnRH using propylene-glycol as a cosolvent and the subcutaneous route for slowing and extending the release of GnRH, as well as the most adequate timing for such administration. In the present study, protocols based on a short-term CIDR treatment and a single subcutaneous dose of GnRH in propylene-glycol at 36 h after CIDR removal induced a similar ovarian response to protocols based on administration of eCG at CIDR removal or intramuscular GnRH in distilled water at 56 h after. In such protocol, 80% of the animals developed estrus in a narrow timing (75% between 36 and 48 h after CIDR removal), and all of them also ovulated in a narrow window (87.5% between 72 and 76 h after CIDR removal, with 62.5% between 72 and 76 h) and showed a similar ovulation rate and plasma progesterone concentrations at the induced estrous cycle. Hence, administration of GnRH in propylene-glycol may constitute an alternative to traditional protocols based on the administration of eCG.

Consequences of twinning induction to Noemi ewes by a recombinant human follicle-stimulating hormone compared with pituitary-derived porcine follicle-stimulating hormone on follicular dynamics, maternal biochemical attributes, and neonatal traits.

Aim:The aim of this study was to investigate the effectiveness of using recombinant human follicle-stimulating hormone (FSH) compared with pituitary-derived porcine FSH given as one dose or multiple doses on the neonatal traits, follicular dynamics, and maternal blood biochemical constituents in Noemi ewes.Materials and Methods:A 3×2 factorial arrangement was designed utilizing 60 adults Noemi ewes to test the effects of using two sources of FSH (human vs. porcine) in addition to control, either given as a single total dose or six descending doses to provoke twinning. Six treatments (T) were tested (n=10 ewes/T). C1 and C6 served as control ewes given saline as one dose and six doses, respectively; H1 and H6 ewes were given human FSH as one and six doses; and P1 and P6 ewes were given porcine FSH similar to the above treatments. Saline and/or FSH administration were administered at days 8, 9, and 10 of the 10-day controlled internal drug release (CIDR) implant. At CIDR removal, fertile rams were used for natural mating. Blood samples for the assessment of serum metabolites were collected.Results:Twinning increased in FSH-treated ewes than control. However, giving FSH of either source as a single dose resulted in a higher incidence of stillbirths. Pregnancy rates were 30, 40, 50, 60, 70, and 80% in C1, C6, P1, P6, H1, and H6, respectively. Respective percent of ewes delivering twins/multiple birth was 0, 0, 80, 66.7, 71.4, and 87.5%. FSH of human source was more efficient for folliculogenesis than porcine FSH. Administration of FSH increased blood cholesterol, decreased high-density lipoprotein; however, low-density lipoprotein levels were not different than control. Moreover, an interaction (p<0.05) exists between source and type of FSH administration on blood glucose. Six doses of FSH elevated blood protein. Blood albumin decreased by porcine-FSH but not affected by human-FSH. Blood globulins were not different due to source of FSH, whereas giving FSH as six doses increased globulins than in single-dose protocol. Contrariwise, an interaction was found between source and type of FSH administration on elevating the activity of alanine aminotransferase and reducing the activity of aspartate aminotransferase.Conclusion:Administration of human FSH at 180 IU in six descending doses resulted in the best neonatal outcomes and maternal health in Noemi ewes.