Expression of CD44 in sheep oocytes and preimplantation embryos.

The CD44 family belongs to a larger group of hyaluronic acid-binding proteins and plays important roles in oocyte maturation, fertilization and preimplantational embryo development. We analyzed the CD44 receptor in sheep oocytes and embryos. Immature oocytes (N = 66) were obtained from a local abattoir; mature oocytes (N = 35) and embryos (N = 41) were obtained by laparotomy from adult hair ewes submitted to ovarian stimulation treatment. The CD44 mRNA was detected by hemi-nested PCR, after reverse transcription, while proteins were located by indirect immunofluorescence, using anti-human CD44 monoclonal antibody. Human lymphocytes and immature bovine oocytes were used as positive and negative controls, respectively. Assessment of the oocyte nuclear stages as well as classification of the embryonic development stage were made with Hoechst 33342 staining. Indirect immunofluorescence detected CD44 expression on the surface of mature oocytes and embryos; immature oocytes did not take up the stain. These findings were supported by the RT-PCR data, which showed no mRNA templates for CD44, even after two consecutive amplifications, in material from immature oocytes and cumulus cells. The CD44 amplicons were detected after a second hemi-nested PCR in mature oocytes and embryos. The finding of CD44 in mature oocytes and preimplantational embryos could reflect the expression profile of hyaluronic acid during terminal folliculogenesis and preimplantational embryo development in sheep.

Autoclaved, previously used intravaginal progesterone devices induces estrus and ovulation in anestrous Toggenburg goats.

Intravaginal progesterone devices are used worldwide for estrus induction in goats. Reused devices are able to induce estrus; however, this can be a health risk within a flock. The objective was to compare new and previously used (and autoclaved) progesterone-releasing intravaginal devices for induction of estrus and ovulation in seasonally anestrous Toggenburg goats. Anestrous goats (n=42) received new intravaginal devices containing 0.3g progesterone (CONTROL), or similar devices previously used for either 6 (USED6) or 12d (USED12) and subsequently autoclaved. All goats received 5mg dinoprost at device insertion and 200 IU eCG 5d later, and all devices were removed after 6d. After device removal, estrus was monitored and females displaying signs of estrus were mated by fertile bucks. Transrectal ovarian ultrasonography was performed after device removal until detection of ovulation. Blood samples were collected for determination of plasma progesterone concentration at different times. There was no difference (P>0.05) among groups CONTROL, USED6 or USED12 for: estrus response (87, 100 or 100%, respectively); duration of estrus (32.3±2.3, 25.2±3.4 or 27.3±4.1h); ovulation rate (100, 88 or 100%); number of ovulations (1.5±0.2, 1.9±0.3 or 1.7±0.3); and pregnancy rate (60, 58 or 67%). Plasma progesterone (P4) concentrations were greater (P<0.05) in CONTROL than in USED6-treated and USED12-treated goats (7.2±1.2, 4.7±0.7 and 4.3±0.6 ng/mL, respectively) at 6h after device insertion; these differences were maintained until 4d after device insertion (3.4±0.4, 2.3±0.2, and 2.5±0.2 ng/mL). Overall, plasma progesterone concentrations were greater (P<0.05) in nulliparous than in lactating goats (3.1±0.8 compared to 2.4±0.6 ng/mL, respectively). In conclusion, autoclaved, previously used intravaginal progesterone-releasing devices resulted in significant lesser plasma progesterone concentrations than new devices, but were similarly effective in inducing estrus and ovulation in anestrous goats.

Reproductive parameters of dairy goats submitted to estrus synchronization with prostaglandin F2α associated or not to hCG at estrous onset / Parâmetros ovarianos de cabras leiteira submetidas a sincronização do estro com protaglandina F2α associada ou não ao hCG no início do estro

The objective of this study was to evaluate the efficiency of two doses of PGF associated or not to hCG on the associated reproductive parameters in dairy goats. A total of 29 goats received two doses of 30µg d-cloprostenol latero-vulvar at a 10 day intervals (Day 1 and Day 10). The does were allocated according to body weight and body condition score into two treatments, to receive hCG (250IU) or saline at estrus onset. After the second dose of PGF, estrus was monitored and ultrasound exams were performed twice daily. All does were inseminated 16h after estrus onset. Blood collection was performed every day for progesterone assay. The use of hCG at estrus onset did not affect any studied parameter and therefore the data were pooled. Estrous response rate was similar (P>0.05) after the first (75.9%, 22/29) and the second dose of PGF (79.3%, 23/29). The interval between the administration of PGF and estrus onset was greater (P<0.05) after Day 1 (75.8±53.9h) than Day 10 (47.7±10.1 h). Estrus duration was superior (P < 0.05) after Day 1 (35.4±15.9h) to Day 10 (26.8±15.0h). Ovulation rate was 79.3% (23/29) after the second dose of PGF. No differences (P>0.05) between both experimental groups were detected in the following parameters, averaging: the interval from the second dose administration to the ovulation (86.6±11.4h), interval from estrus to ovulation (39.9±12.3 h), diameter of largest follicle (7.2±1.4) and number of ovulations (1.8±0.6). At Day 1, 52.4% (11/21) of does presented progesterone concentrations <1ng/mL. At Day 10, 100% of the animals presented concentrations >1ng/mL. The results of the present study indicate that estrus can be efficiently synchronized in dairy goats with the use of two doses of PGF at a 10 day interval. Further research should be done evaluating hCG use in different doses or moments of administration.

O objetivo deste estudo foi avaliar a eficiência do uso de duas doses de PGF associadas ou não à administração de hCG no início do estro sobre os parâmetros reprodutivos de cabras leiteiras. Um total de 29 cabras receberam duas doses de 30 µg d-cloprostenol pela via latero-vulvar com 10 dias de intervalo (Dia 1 e Dia 10). As cabras foram alocadas para receberem o hCG (250 IU) ou salina i.m. no momento em que o estro foi detectado. Depois da realização da segunda dose de PGF, o estro foi monitorado e exames ultrassonográficos foram realizados duas vezes ao dia. Todas as fêmeas foram inseminadas 16 h após o inicio do estro. Amostras de sangue foram coletadas diariamente para determinação das concentrações plasmáticas de progesterona. O uso do hCG no momento do início do estro não afetou os parâmetros estudados e, portanto, os dados serão apresentados agrupados. A taxa de manifestação de estro foi similar (P > 0,05) na primeira (75,9% - 22/29) ou na segunda dose de PGF (79,3% - 23/29). O intervalo entre a administração de PGF e o início do estro foi maior (P < 0,05) no Dia 1 (75,8±53,9 h) que no Dia 10 (47,7±10,1 h). Duração do estro também diferiu (P < 0,05) 35,4±15,9 (Dia 1) vs 26,8±15, 0 h (Dia 10). A taxa de ovulação foi 79,3% (23/29) após a segunda dose PGF. Não foi encontrada diferença (P>0,05) entre os grupos experimentais quanto aos parâmetros reprodutivos: intervalo entre a aplicação da segunda dose e a ovulação (86,6±11,4h), intervalo do estro a ocorrência da ovulação (39,9±12,3h), diâmetro do maior folículo (7,2±1,4) e número de ovulações (1,8±0,6). No Dia 1, 52,4% (11/21) apresentavam concentrações de progesterona < 1 ng/mL. No Dia 10, 100% dos animais apresentavam concentrações >1ng/mL. O presente estudo permite concluir que o estro pode ser eficientemente sincronizado em cabras leiteiras com duas doses de PGF intervaladas em 10 dias. Novas pesquisas devem se realizadas para avaliar diferentes doses e momentos de utilização do hCG.

Reproductive parameters of Santa Inês ewes submitted to short-term treatment with re-used progesterone devices / Características reprodutivas de ovelhas Santa Inês que receberam dispositivos de progesterona reutilizados

The aim of this study was to verify the efficacy of reusing intravaginal progesterone (P4) devices on the reproductive parameters in Santa Inês ewes. Females received intravaginal P4 devices for their first, second or third use for five days plus 300 IU eCG IM and 5mg dinoprost laterovulvar 24h before device removal. Blood was collected at different moments. Transrectal ultrasonography was performed from device removal to ovulation. Part of the ewes were submitted to artificial insemination by laparoscopy (IAL - n=55) with fresh semen, whereas the rest were bred by fertile rams (n=41). On the initial 18 h, ewes that received devices for the first time showed higher P4 concentrations (5.1±1.8 vs 3.5±1.4 vs 2.4±1.1 - P<0.05). However, after the first 48h no difference was observed among all treatments and P4 supraluteal concentrations were detected in all ewes upon device removal. Estrous response, interval from device removal to estrus, rate of ovulating animals, number of ovulations, time from device removal to ovulation and average conception rates after IAL or natural mating were similar among all 3 groups. Intravaginal progesterone devices can be used up to three times without altering reproductive parameters in Santa Inês ewes.

Avaliou-se a eficácia da reutilização de dispositivos intravaginais de progesterona (P4) sobre características reprodutivas em ovelhas Santa Inês. As fêmeas receberam dispositivos intravaginais contendo P4 para o seu primeiro, segundo ou terceiro uso por cinco dias, associado a 300 UI eCG IM e 5mg dinoprost laterovulvar 24h antes da remoção dos dispositivos. Ultrassonografia transretal foi realizada da remoção dos dispositivos até a ovulação. Parte das ovelhas foi submetida à inseminação artificial laparoscópica (IAL- n=55) com sêmen a fresco, enquanto outra parte foi acasalada por machos férteis (n=41). Nas 18 h iniciais, as ovelhas que receberam dispositivos pela primeira vez apresentaram maior concentração de P4 (5.1±1.8 vs 3.5±1.4 vs 2.4±1.1 - P<0.05). Entretanto, após as primeiras 48h nenhuma diferença foi observada entre os tratamentos quanto as concentrações circulantes de progesterona. Foram encontradas concentrações de progesterona supraluteais em todas as ovelhas na retirada dos dispositivos. Estro, intervalo da retirada ao estro, taxa de animais que ovularam, número de ovulações, momento da retirada à ovulação e taxa média de concepção após IAL ou monta natural foram similares entre os tratamentos. Dispositivos intravaginais de progesterona podem ser usados até três vezes sem alterar as características reprodutivas de ovelhas Santa Inês.

Effects of prostaglandin administration 10 days apart on reproductive parameters of cyclic dairy nulliparous goats / Efeitos da aplicação de prostaglandinas intervaladas de 10 dias sobre características reprodutivas de cabras leiteiras nulíparas cíclicas

This study reported the effects of prostaglandin (PGF2a) administration 10 days apart on reproductive parameters of cyclic artificial inseminated (AI) nulliparous Alpine (n=9) and Saanen (n=9) goats. Animals received two doses of 22.5mg PGF2a 10 days apart. After 1st and 2nd PGF2a administrations, estrus was monitored at 12 h intervals, with a buck teaser. Plasma progesterone concentration (ng/mL) was determined from blood sampled on day 0 (1st PGF2a) and the following 5, 10 (2nd PGF2a), 15, 20, 25 and 30 days. After the onset of the second estrus, females were transrectally (5 MHz probe) scanned at 4 hour intervals until at least 8h after ovulation. Pregnancy was checked through transrectal ultrasound on days 20, 25, 30, 35 and 90 after insemination. All parameters studied did not differ between breeds (P>0.05). Estrous response and interval to estrus, respectively, after 1st (78.9% and 50.6±17.2h) and 2nd PGF2a (88.9% and 50.0±14.8h) administration did not differ (P>0.05). Overall animals ovulating (100.0%), interval to ovulation after 2nd PGF2a (64.5±19.5h) and after estrous onset (18.0±9.1h), ovulation rate (1.3±0.5), diameter of ovulatory follicle (8.1±1.1mm) were recorded. Embryo loss occurred before day 30 of pregnancy. Estrus can be efficiently synchronized in nulliparous Alpine and Saanen goats with two doses of prostaglandin 10 days apart.

Relataram-se os efeitos da aplicação de prostaglandina sobre características reprodutivas de cabras leiteiras nulíparas cíclicas. Cabras Alpinas (n=9) e Saanen (n=9) receberam duas doses de 22,5mg PGF2a com 10 dias de intervalo. A progesterona plasmática (ng/mL) foi determinada a partir de amostras de sangue coletadas nos dias 0 (primeira dose), 5, 10 (segunda dose), 15, 20, 25 e 30. Após início do segundo estro, as fêmeas foram monitoradas por ultrassonografia transretal a cada quatro horas até oito horas após a ovulação. A gestação foi verificada por ultrassonografia transretal nos dias 20, 25, 30, 35 e 90 após a segunda dose. As características estudadas foram semelhantes entre as raças (P>0,05). Animais em estro e o intervalo parto-estro de, respectivamente, 78,9% e 50,6±17,2h e 88,9% e 50,0±14,8h após a primeira e segunda administrações de prostaglandina, não diferiram (P>0,05). Todas as cabras ovularam e registraram-se valores do intervalo parto-ovulação após a segunda aplicação de prostaglandina de 64,5±19,5h e após início do estro de 18,0±9,1h, a taxa de ovulação de 1,3±0,5 e diâmetro do folículo ovulatório de 8,1±1,1mm. Perda embrionária ocorreu antes de 30 dias de gestação. O estro pode ser eficientemente sincronizado em cabras leiteiras núliparas com duas doses de prostaglandina intervaladas de 10 dias.