Cervical relaxation for non-surgical uterus access in Santa Inês ewes

ABSTRACT The present study was composed by two experiments aiming to develop a cervical dilation technique for non-surgical access to Santa Inês ewe’s uterus. In Experiment 1, thirty ewes underwent four epidural treatments. The three experimental treatments used 2.0mg/kg ketamine. The group receiving this drug alone was denominated KG, whereas other group had ketamine associated with 0.1mg/kg morphine (KM) and KX a third group had ketamine associated with 0.05mg/kg xylazine (KX). Control treatment was 1mL/7.5kg saline solution epidurally (CON). Cervical dilation was evaluated in both experiments by attempting to pass a metal rod through the cervix. In Experiment 2, three different hormonal protocols for cervical dilation were tested in thirty ewes. Epidural anesthesia with 2.0mg/kg ketamine was the control treatment (KG) and was combined with hormonal treatments: Misoprostol (MI); Oxytocin + Estradiol (OE); Misoprostol + Oxytocin + Estradiol (MOE). In Experiment 1 transposition rate was not different among groups. In Experiment 2, OE presented the highest rate (90%) while MOE presented 86.2%, MI 68.9% and CON 62.1%. The study developed a pharmacological protocol that increased cervical transposition making the non-surgical access to the uterus feasible in Santa Inês ewes

Freezing goat embryos at different developmental stages and quality using ethylene glycol and a slow cooling rate

ABSTRACT The efficiency of an alternative freezing protocol for goat embryos of different morphology and quality was tested. Fifty-eight embryos on Day 6-7 stage were transferred as fresh or after freeze-thawing (n=29/group). For freezing, embryos were placed into 1.5M ethylene-glycol solution for 10min. During this time, they were loaded in the central part of 0.25mL straw, separated by air bubble from columns containing PBS/BSA 0.4% plus 20% BFS. Straws were then frozen using a freezing machine from 20ºC to -6ºC at a cooling rate of 3ºC/min, stabilization for 15min (seeding after 5min), from -6 C to -32ºC at 0.6 C/min,and plunged into liquid nitrogen. Frozen embryos were thawed for 30s at 37ºC in a water bath. Embryos subjected to fresh transfer were maintained in holding medium (37ºC). Fresh and frozen-thawed embryos were transferred at day 7 post-estrus to 30 recipients. Kidding and kid born rates were similar (P> 0.05), respectively, for recipients receiving fresh (66.7% or 10/15; 55.2% or 16/29) or frozen-thawed (60% or 9/15; 51.7% or 15/29) embryos. The cryopreservation of goat embryos using slow-freezing protocol and 1.5MEG resulted in similar efficiency rates of fresh embryos

Cervical penetration rates and efficiency of non-surgical embryo recovery in estrous-synchronized Santa Inês ewes after administration of estradiol ester (benzoate or cypionate) in combination with d-cloprostenol and oxytocin

The effects of estradiol esters, D -cloprostenol and oxytocin on induction of cervical dilation prior to non-surgical embryo recovery in Santa Inês ewes (Days 6–7 estrous cycle) were assessed in this study. In Trial 1, transcervical embryo flushing was performed in estrous-induced ewes ad- ministered 37.5 μg of D -cloprostenol i.m. 10 h before and 50 IU of oxytocin i.v. 20 min before uterine flushing with (EB-PGF-OT; n = 13) or without (PGF-OT; n = 11) 1 mg of estradiol benzoate i.m. administered concurrently with D -cloprostenol injection. In Trial 2, the estrous- synchronized animals were treated with 1 mg of estradiol benzoate (EB-PGF-OT; n = 12) or es- tradiol cypionate (EC-PGF-OT; n = 12) i.m. along with 37.5 μg of D -cloprostenol i.m. 16 h before and 50 IU of oxytocin i.v. 20 min before uterine flushing. In Trial 1, uterine flushing could be accomplished in 38% of ewes in the EB-PGF-OT and 27% those in the PGF-OT (P > 0.05) group. Flushing fluid recovery averaged 90% and there were 1.0 ± 1.1 embryos/ewe collected with mean duration of the flushing procedure being ̃36 min. In Trial 2, uterine flushing was accom- plished in 78% of ewes in the EB-PGF-OT and 44% of those in the EC-PGF-OT group (P > 0.05) with mean flushing fluid recovery rate being 88% and time elapsing to complete flushing being ̃33 min. Within the subsets of animals treated with EB, the percentages of successful transcer- vical penetrations were 38% compared with 78% in Trials 1 and 2, respectively (i.e., with EB administered 10 h compared with 16 h before uterine flushing: P < 0.05). The interval from EB administration to the beginning of transcervical penetration can affect the efficacy of embryo recovery procedures utilizing a combined EB/d-cloprostenol/oxytocin pre-treatment

Oviduct fluid during IVF moderately modulates polyspermy in in vitro-produced goat embryos during the non-breeding season

International audienceThe present study determined i) the presence of proteins (oviduct-specific glycoprotein, OVGP1; heat shock protein-70A, HSPA1A; heat shock protein-A8, HSPA8; annexin A1, ANXA1; annexin A5, ANXA5; and myosin-9, MYH9) known to be involved in early reproduction in the oviduct fluid (OF) of anestrous goats; and ii) the functional effect of during IVF on polyspermy modulation and embryonic development. In vitro-matured oocytes were co-cultured with spermatozoa (1.0, 2.0, or 4.0 x 106 cells/mL) for 18 h in SOF medium supplemented with 5 mg/mL of heparin, 4 mg/mL gentamicin, and 10% estrus sheep serum (CTRL1, CTRL2, and CTRL4 groups) or the same medium plus 10% OF (OF1, OF2, and OF4 groups) obtained from anestrus goats. The analysis of OF by western blotting confirmed the presence of the six proteins tested for. The increase in sperm concentration had no effect (P > 0.05) on the penetration rate in any group; however, monospermy rate decreased as sperm concentration was increased in both OF and CTRL. Regardless of the concentration used, when data were pooled, OF supplementation improved (P 0.057) to enhance IVF efficiency. Additionally, IVF efficiency was higher (P 0.05) by the sperm concentration and OF treatment, and the average values were cleavage (72 +/- 2.6%), blastocyst (37 +/- 3.0%), blastocyst in relation to the cleaved (51 +/- 4.8%), hatched (62 +/- 1.2%), and number of cells per blastocyst (174 +/- 1.8%). In conclusion, the six proteins analyzed are present in the OF of anestrous goats, and the supplementation of this OF during IVF may modulate the polyspermy incidence and enhance IVF efficiency, especially when 1x10(6) sperm per mL is used

Use of human intravaginal tampon embedded with natural progesterone induces synchronous estrus in Santa Inês ewes

RESUMO Este estudo avaliou a eficiência de três protocolos de indução de estro síncronizado em ovelhas da raça Santa Inês. Vinte e quatro ovelhas adultas foram equitativamente distribuídas em três grupos, de acordo com ordem de parto, peso corporal (kg) e escore da condição corporal. As ovelhas receberam implante vaginal de progesterona natural por seis dias mais 37,5μg de d-cloprostenol laterovulvar e 300UI de eCG i.m., 24 horas antes da remoção do dispositivo. Ovelhas controle receberam CIDR330mg de progesterona, e as demais receberam dispositivo absorvente intravaginal humano, tamanho mini, embebido com 200 (OB200) ou 400mg (OB400) de progesterona. Coletas de sangue foram feitas nos momentos D0 (antes da inserção dos dispositivos), D0+6h e diariamente, até um dia após retirada do dispositivo (D7). A progesterona (ng/mL) foi semelhante (P>0,05) em todos os tratamentos ao longo do período experimental, exceto no dia da remoção do dispositivo, quando as ovelhas controle (2,5±0,3) tiveram progesterona superior (P0,05) e o intervalo para o estro (46,3±3,9a, 26,4±4,5b e 31,2±5,8a,b) foi diferente (P0,05). Dispositivos humanos e fonte de progesterona podem ser usados para induzir o estro sincronizado em ovelhas Santa Inês