Effect of short-term supplementation with rumen-protected fat during the late luteal phase on reproduction and metabolism of ewes.

This study was designed to study the effect of short-term supplementation with rumen-protected fat during the late luteal phase on reproduction and metabolism of sheep during breeding season. Seventy-six ewes (Rahmani, Barki and Awassi × Barki) were allocated to two groups considering genotype: the control ewes (C-group) received a maintenance diet, and the fat-supplemented ewes (F-group) received the maintenance diet plus 50 g/head/day of rumen- protected fat (Megalac) for 9 days during which oestrus was synchronized. The latter had been accomplished using double intramuscular injection of prostaglandin F2α (PGF2α ) 11 days apart. Ovarian activity, serum concentration of cholesterol, glucose, insulin and reproductive performance variables were recorded. Data were analysed considering treatment (group) and genotype. Supplementation had positive effects on the overall mean serum concentrations of cholesterol (p < 0.05), glucose (p < 0.05) on day 6 of nutritional treatment and insulin (p = 0.07) on day 8. Fat supplementation did not affect the total number of follicles, follicle populations and ovulation rate. However, fat-supplemented Rahmani ewes tended to have higher ovulation rate compared with other breeds (treatment × breed interaction, p = 0.06). Treatment also did not affect the mean concentration of serum estradiol or progesterone. Supplemented ewes had higher conception (p = 0.06) and lambing rates (p < 0.05) compared with control. In conclusion, short-term supplementation with rumen-protected fat as a source of energy around breeding time improved metabolism, conception and lambing rates of ewes without effects on steroidogenic capacity and ovarian activity being apparent.

Blood metabolites and hormone-based programmed breeding treatments in anovular lactating dairy cows.

The objectives were to (1) compare blood metabolites and reproductive outcomes in lactating dairy cows not inseminated before (early) and after (late) 100 d in milk (DIM) because of prolonged anovulation or anestrus; and (2) evaluate reproductive responses of cows ≤100 DIM to GnRH+PGF(2α) treatments after a fixed-time artificial insemination (AI; Ovsynch) or after induced estrus (Select Synch). In blood samples collected before initiating hormone-based breeding programs, anovular cows ≤ 100 DIM had the greatest serum total protein and globulin concentrations and the lowest tri-iodothyronine concentrations. Anovular and ovular cows >100 DIM had the greatest serum urea concentrations. Ovaries in cows (n=40) >100 DIM were examined by transrectal ultrasonography, and those without a detectable corpus luteum (CL; anovular) were given GnRH and then PGF(2α) 7 d later (Select Synch), whereas cows with a CL (ovular) were given 2 PGF(2α) injections 12 d apart. Cows were inseminated at observed estrus after the second or only PGF(2α) injection. More ovular (79%; 15/19) than anovular cows (24%; 5/21) were detected in estrus. No differences were detected between ovular and anovular cows in DIM at first AI, conception rate to first AI, cumulative pregnancy rates, number of services per conception, or days open. Cows (n=93) ≤100 DIM were assigned randomly to 3 treatments: (1) control (n=20) AI at estrus; (2) GnRH and then PGF(2α) on 7d (Select Synch; n=42) and monitored for signs of estrus for 5d and AI accordingly; or (3) 2 GnRH injections 9 d apart with PGF(2α) given 48 h before second GnRH injection and AI at 16h after the second GnRH injection (Ovsynch; n=31). Among cows ≤100 DIM, controls had more days to first service (149±16 d) than Select Synch cows (117±7 d). Ovsynch cows had the fewest days to first service (84±10 d) compared with control (149±16 d) and Select Synch (117±7 d) cows. Conception rates in control (25%) and Select Synch (26.2%) cows did not differ from those in Ovsynch cows (29%). Ovsynch cows had greater cumulative pregnancy rates and fewer days open than control (161±20 vs. 258±29 d), but did not differ from Select Synch (233±19d). Timed AI produced comparable fertility and superior cumulative pregnancy rates, fewer days to first service, and fewer days open than AI at observed estrus in cows inseminated ≤100 DIM.

Effect of season, month of parturition and lactation on estrus behavior and ovarian activity in Barki x Rahmani crossbred ewes under subtropical conditions.

The correlations between some meteorological parameters and fertility data were evaluated in Barki x Rahmani crossbred ewes using the records of five consecutive years (2003-2007). Additionally, estrus detection and ultrasonic evaluation were applied on eighteen mature dry ewes during breeding and non-breeding seasons. The effect of lactation was evaluated by monitoring estrus behavior in ninety four lactating ewes from 40 to 120 d after parturition. Moreover, ultrasonography was used to identify ovarian activity in six cyclic and six acyclic non-lactating ewes. Results revealed that relative estrus occurrence and fertile mating were positively correlated (P <0.05) with high temperature and long photoperiod (conditions of summer season), and were negatively correlated (P < 0.01) with rainfall (condition of winter season). During breeding season, estrus rate, serum progesterone concentration, and diameter of largest follicle were significantly (P < 0.05) higher than those observed during the non-breeding season. Furthermore, month of parturition had a significant effect (P < 0.05) on estrus rate of lactating ewes where ewes that lambed in August, September, and October recorded higher estrus rate than those lambed in November and December. However, the lactational strength did not exert any deleterious effect on the reproductive performance of lactating ewes. In conclusion, in Egypt under subtropical conditions, Barki x Rahmani crossbred ewes exerted optimum estrus behavior and fertile mating during summer season. The reduction in estrus activity during lactation was due to the seasonal effect rather than lactational stress.

Resynchronizing estrus with progesterone or progesterone plus estrogen in cows of unknown pregnancy status.

Two experiments were conducted to test 2 progesterone (P4)-based treatments that were applied to lactating dairy cattle of unknown pregnancy status to resynchronize estrus of nonpregnant cows. In experiment 1, cows were assigned randomly before a timed AI (TAI) to 1) treatment with a CIDR (controlled internal drug-releasing intravaginal insert containing P4) for 7 d starting on d 13 after TAI (CIDR; n = 300) or 2) no P4 treatment (control; n = 330). Compared with controls, P4 increased the synchrony of those detected in estrus, but failed to increase the overall return rates of non-pregnant cows during the 6 d after CIDR removal (27% vs. 31%; d 20 to 26 after TAI) and did not alter synchronized conception rates (32% vs. 20%) of those inseminated. Use of P4 did not compromise pregnancies resulting from TAI compared with controls (38% vs. 42%), but increased embryo survival between d 29 and 57 after TAI (65.5% vs. 44.3%). In experiment 2, on d 13 after TAI, 196 cows were treated with a CIDR insert for 7 d. Controls received no further treatment. Remaining cows were treated with 1 of 3 estrogen regimens: 1 mg of estradiol benzoate (EB), 0.5 mg of estradiol cypionate (ECP), or 1 mg of ECP on both d 13 and 21. Only 60% of nonpregnant, estrogen-treated cows were detected in estrus between d 20 and 26, and rates of return and conception did not differ among treatments. Estrogen on d 13 did not consistently turn over the dominant follicle when given at CIDR insertion but did increase concentrations of estradiol and reduced luteal function when administered on d 13 and 21 (24 h after CIDR removal). Treatments had no negative effects on milk yield, dry matter intake, or established pregnancies. Use of P4 alone had little effect on overall rates of return to estrus or conception at the first eligible estrus in experiment 1. Combining estrogen with P4 in experiment 2 had no detrimental effects on established pregnancies or subsequent conception and failed to improve return rates beyond P4 alone.

Pregnancy in dairy cows after synchronized ovulation regimens with or without presynchronization and progesterone.

Two experiments examined pregnancy after synchronized ovulation (Ovsynch) with or without progesterone (P4) administered via controlled internal drug release (CIDR) intravaginal inserts. In experiment 1, 262 lactating cows in one herd were in 3 treatments: Ovsynch (n = 91), Ovsynch + CIDR (n = 91), and control (n = 80). The Ovsynch protocol included injections of GnRH 7 d before and 48 h after an injection of PGF20. Timed artificial insemination (TAI; 57 to 77 d postpartum) was 16 to 20 h after the second GnRH injection. Cows in the Ovsynch + CIDR group also received a CIDR (1.9 g of P4) insert for 7 d starting at first GnRH injection. Control cows received A-I when estrus was detected using an electronic estrus detection system. Based on serum P4, 44.1% of cows were cyclic before Ovsynch. Pregnancy rates at 29 d (59.3 vs. 36.3%) and 57 d (45.1 vs. 19.8%) after TAI and embryo survival (75.9 vs. 54.5%) from 29 to 57 d were greater for Ovsynch + CIDR than for Ovsynch alone. In experiment 2, 630 cows in 2 herds received TAI at 59 to 79 d postpartum after 6 treatments. Estrous cycles were either presynchronized (2 injections of PGF2alpha 14 d apart; n = 318) or not presynchronized (n = 312). Within those groups, Ovsynch was initiated 12 d after second presynchronization PGF2alpha, and used alone (n = 318) or with CIDR inserts for 7 d (1.38 g of P4/insert, n = 124 or 1.9 g of P4/insert, n = 188). Before Ovsynch, 80% of cows were cyclic. Presynchronization increased pregnancy (46.8 vs. 37.5%) at 29 d after TAI, but CIDR inserts had no effect on pregnancy in experiment 2. Overall embryonic survival between 29 and 57 d in experiment 2 was 57.7%. Use of CIDR inserts with Ovsynch improved conception and embryo survival in experiment 1 but not in experiment 2, in part due to differing proportions of cyclic cows at the outset. Presynchronization before Ovsynch enhanced pregnancy rate.

Transplantation of porcine umbilical cord matrix cells into the rat brain.

Immune rejection of transplanted material is a potential complication of organ donation. In response to tissue transplantation, immune rejection has two components: a host defense directed against the grafted tissue and an immune response from the grafted tissue against the host (graft vs host disease). To treat immune rejection, transplant recipients are typically put on immunosuppression therapy. Complications may arise from immune suppression or from secondary effects of immunosuppression drugs. Our preliminary work indicated that stem cells may be xenotransplanted without immunosuppression therapy. Here, we investigated the survival of pig stem cells derived from umbilical cord mucous connective tissue (UCM) after transplantation into rats. Our data demonstrate that UCM cells survive at least 6 weeks without immune suppression of the host animals after transplantation into either the brain or the periphery. In the first experiment, UCM cells were transplanted into the rat brain and recovered in that tissue 2-6 weeks posttransplantation. At 4 weeks posttransplantation, the UCM cells engrafted into the brain along the injection tract. The cells were small and roughly spherical. The transplanted cells were positively immunostained using a pig-specific antibody for neuronal filament 70 (NF70). In contrast, 6 weeks posttransplantation, about 10% of the UCM cells that were recovered had migrated away from the injection site into the region just ventral to the corpus callosum; these cells also stained positively for NF70. In our second experiment, UCM cells that were engineered to constitutively express enhanced green fluorescent protein (eGFP) were transplanted. These cells were recovered 2-4 weeks after brain transplantation. Engrafted cells expressing eGFP and positively staining for NF70 were recovered. This finding indicates a potential for gene therapy. In the third experiment, to determine whether depositing the graft into the brain protected UCM cells from immune detection/clearance, UCM cells were injected into the tail vein and/or the semitendinosis muscle in a group of animals. UCM cells were recovered from the muscle or within the kidney 3 weeks posttransplantation. In control experiments, rat brains were injected with PKH 26-labeled UCM cells that had been lysed by repeated sonic disruption. One and 2 weeks following injection, no PKH 26-labeled neurons or glia were observed. Taken together, these data indicate that UCM cells can survive xenotransplantation and that a subset of the UCM cells respond to local signals to differentiate along a neural lineage.

Supplemental norgestomet, progesterone, or melengestrol acetate increases pregnancy rates in suckled beef cows after timed inseminations.

In Exp. 1, 187 lactating beef cows were treated with injections of GnRH 7 d before and 48 h after prostaglandin F2alpha (PGF2alpha; Cosynch) or with Cosynch plus a 7-d treatment with an intravaginal progesterone (P4)-releasing insert (CIDR-B; Cosynch + CIDR). In Exp. 2, 183 lactating beef cows were treated with the Cosynch protocol or with Cosynch plus a 7-d treatment with norgestomet (Cosynch + NORG). In Exp. 1 and 2, blood samples for later P4 analyses were collected on d -17, -7 (first GnRH injection), 0 (PGF2alpha injection), and at timed artificial insemination (TAI; 48 h after PGF2alpha). In Exp. 3, 609 lactating beef cows were treated with the Cosynch + CIDR protocol or were fed 0.5 mg of melengestrol acetate (MGA) per day for 14 d before initiating the Cosynch protocol 12 d after the 14th d of MGA feeding (MGA + Cosynch). Blood samples were collected as in Exp. 1 and 2, plus additional samples on d -33 and -19 before PGF2alpha. In Exp. 4, 360 lactating beef cows were treated with a Cosynch + CIDR protocol, with TAI occurring at either 48 or 60 h after PGF2alpha, while receiving either GnRH or saline to form four treatments. Blood samples were collected as in Exp. 1 and 2. In Exp. 1, addition of P4 reduced the ability of the first GnRH injection to induce ovulation in anestrous cows with low P4 before PGF2alpha but improved (P = 0.06) pregnancy rates (61 vs 66%). In Exp. 2, the addition of NORG mimicked P4 by likewise increasing (P < 0.01) pregnancy rates (31 vs 51%) beyond those after Cosynch. In Exp. 3, the Cosynch + CIDR protocol increased (P < 0.001) pregnancy rates from 46 to 55% compared to the MGA + Cosynch protocol. In Exp. 4, administration of GnRH at TAI improved (P < 0.05) pregnancy outcomes (50 vs 42%), whereas timing of TAI had limited effects. We conclude that a progestin treatment concurrent with the Cosynch protocol improved pregnancy outcomes in all experiments, but pretreatment of cows with MGA was not as effective as the CIDR insert or NORG implants in this Cosynch-TAI model. Most of the improvement in pregnancy rates was associated with the increase in pregnancy rates of anestrous cows, regardless of whether ovulation was successfully induced in response to GnRH 7 d before PGF2alpha. Injection of GnRH at TAI following the Cosynch + CIDR protocol increased pregnancy rates in cycling cows with high P4 before the PGF2alpha injection and in anestrous cows with low P4 before PGF2alpha injection.

Stage of cycle, incidence, and timing of ovulation, and pregnancy rates in dairy cattle after three timed breeding protocols.

In experiment 1, 705 cows were assigned to three treatments: 1) the Ovsynch protocol (a GnRH injection given 7 d before and another 48 h after one PGF2alpha injection); 2) PGF2alpha + Ovsynch (one PGF2alpha injection given 12 d (d -22) before initiating Ovsynch (d -10); and 3) 2xPG12 (two PGF2alpha injections 12 d apart; d -15 and -3, followed 48 h later by a GnRH injection. All cows were inseminated (d 0) 16 to 20 h after the GnRH injection on d -1. Cyclic status was estimated by serum progesterone. More cows were in early diestrus at d -10 for PGF2alpha + Ovsynch (36%) and 2 x PG12 (29%) versus Ovsynch (19%). Multiparous cows receiving PGF2alpha + Ovsynch had greater pregnancy rates via ultrasonography at d 28 after AI (42%) than contemporaries after Ovsynch (28%) or 2xPG12 (27%) but did not differ significantly at palpation 10 to 30 d later (28, 19, and 17%, respectively). Pregnancy of first-parity cows was similar across treatments at 28 d (41%) or at palpation (33%). Pregnancy rates for 128 anestrous cows were lower, regardless of treatment. Overall embryo survival from d 28 until palpation was 72% but was only 44% in 2xPG12 cows that were anestrus through d -10. Experiment 2 included the three treatments above plus controls (one GnRH injection 7 d before PGF2alpha and AI after estrus). Preovulatory follicles were 6 to 11% larger near estrus in controls than on d -1 in cows receiving GnRH. More controls ovulated by 32 h after onset of estrus than were treated cows by 32 h after GnRH, but percentages (79 to 94%) were similar by 40 h. In multiparous cows, PGF2alpha before Ovsynch increased pregnancy rates, whereas the 2xPG12 protocol produced similar pregnancy rates as Ovsynch across parities. Ovulation was effectively induced by 40 h after GnRH.

An alternative AI breeding protocol for dairy cows exposed to elevated ambient temperatures before or after calving or both.

Our objective was to determine if a timed artificial insemination (AI) protocol (Ovsynch) might produce greater pregnancy rates than AI after a synchronized, detected estrus during summer. Lactating Holstein cows (n = 425) were grouped into breeding clusters and then assigned randomly to each of two protocols for AI between 50 and 70 days in milk. All cows were treated with GnRH followed 7 d later by PGF2alpha. Ovsynch cows then were treated with a second injection of GnRH 48 h after PGF2alpha and inseminated 16 to 19 h later. Controls received no further treatment after PGF2alpha and were inseminated after detected estrus. Pregnancy was diagnosed once by transrectal ultrasonography (27 to 30 d after AI) and again by palpation (40 to 50 d). Based on concentrations of progesterone in blood collected before each hormonal injection, only 85.4% of 425 cows were considered to be cycling. Although conception rates were not different between protocols at d 27 to 30, AI submission rates and pregnancy rates were greater after Ovsynch (timed AI) than after detected estrus. A temperature-humidity index > or = 72 was associated with fewer controls detected in estrus with lower conception than for controls detected in estrus when index values were < 72, whereas the reverse was true for cows after the Ovsynch protocol. We concluded that a timed AI protocol increased pregnancy rates at d 27 to 30 because its success was independent of either expression or detection of estrus. However, because of poorer embryonic survival in Ovsynch cows during heat stress only (39.5 vs. 69.2% survival for Ovsynch and control, respectively), pregnancy rates were not different by d 40 to 50 after timed AI.