Importance of preovulatory estradiol on uterine receptivity and luteal function

Animals that exhibited estrus had greater pregnancy success compared to animals that did not exhibit estrus before fixed-time AI (FTAI). Estradiol is synthesized in bovine ovarian follicles under gonadotropin regulation and can directly and indirectly regulate the uterine receptivity and luteal function. Estradiol concentrations at FTAI impacted oviductal gene expression and has been reported to play an important role in establishing the timing of uterine receptivity. These changes have been reported to impact uterine pH and sperm transport to the site of fertilization. After fertilization, preovulatory estradiol has been reported to improve embryo survival likely by mediating changes in uterine blood flow, endometrial thickness and changes in histotroph. Cows with greater estradiol concentrations at the time of GnRH-induced ovulation also had a larger dominant follicle size and greater circulating progesterone concentrations on day 7. Therefore, it is impossible to accurately determine the individual benefit of greater estradiol concentrations prior to ovulation and greater progesterone concentrations following ovulation to pregnancy establishment, as these two measurements are confounded. Research has indicated an importance in the occurrence and timing of increasing preovulatory concentrations of estradiol, but increasing estradiol concentrations by supplementation may not be sufficient to increase fertility. Increased production of estradiol by the preovulatory follicle may be required to enhance fertility through the regulation of sperm transport, fertilization, oviductal secretions, the uterine environment, and embryo survival.(AU)

Early maturation of corpus luteum in rabbits - effect of sildenafil citrate on luteolytic capacity in the early luteal period

Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

Uso da ultrassonografia Doppler colorido na reprodução de pequenos ruminantes / Use of color Doppler ultrasonography in the reproduction of small ruminants

A ultrassonografia (US) Doppler colorido fornece uma ferramenta não invasiva valiosa para detectar e monitorar mudanças dinâmicas na rede vascular e fluxo sanguíneo em vários órgãos e tecidos reprodutivos. Em apoio as biotécnicas da reprodução animal assistida, a US Doppler colorido tem mostrada alta eficiência no monitoramento funcional das estruturas ovarianas. A previsão de respostas ovarianas e produções embrionária em ovinos pela identificação de sinais Doppler na parede folicular já se mostrou eficiente. A aplicação da US Doppler colorido para a identificação da funcionalidade do tecido luteal é ainda maior, desde acompanhamentos fisiológicos e diagnósticos de disfunções luteais até ampla aplicação em conjunto as diferentes biotécnicas reprodutivas. Destaca-se em ovelhas e cabras, a aplicação comercial da US Doppler colorido para o diagnóstico de gestação precoce, de disfunções luteais, de determinação de respostas ovarianas em fêmeas doadoras e receptoras de embriões, para identificar efeitos luteotrófico de estratégias hormonais, e ainda para amparar as estratégias de ressincronização de estro.(AU)

Color Doppler ultrasonography (US) provides a valuable non-invasive tool for detecting and monitoring dynamic changes in the vascular network and blood flow in various reproductive organs and tissues. In support of assisted animal reproduction biotechniques, color Doppler US has shown high efficiency in the functional monitoring of ovarian structures. The prediction of ovarian responses and embryonic production in sheep by identifying Doppler signals in the follicular wall has already proved to be efficient. The application of color Doppler US for identifying the functionality of the luteal tissue is even greater, from physiological monitoring and diagnosis of luteal dysfunctions to wide application together with different reproductive biotechniques. It stands out in sheep and goats, the commercial application of color Doppler US for the diagnosis of early pregnancy, luteal dysfunctions, determination of ovarian responses in embryo donor and recipient females, to identify luteotrophic effects of hormonal strategies, and even to support estrus resynchronization strategies.(AU)

Relationships between antral follicle counts and ovarian morphology of Bos indicus of different ages / Relações entre a contagem de folículos antrais e a morfologia ovariana de Bos indicus de diferentes idades

This study aimed to evaluate the relationship between the antral follicle count (AFC) and the ovarian morphology of Bos indicus in different age groups: 12-23 months (G1), 24-35 months (G2), 36-47 months (G3), 48-59 months (G4), and >60 months (G5). Ovaries were collected individually and sent to the laboratory, where we measured AFC, the diameter and weight of the ovaries, dominant follicle (DF, >8mm), corpus luteum (CL), and small follicles (SF, <8mm). AFC were classified as high, intermediate-high, intermediate-low, and low. A group of ovaries was subjected to follicular aspiration to evaluate the morphological quality of the recovered cumulus oocyte complexes (COCs). Mean AFC did not vary between ages. The morphological characteristic that was most closely correlated with AFC was the weight of the small follicles. There was no relationship between AFC and the weight and diameter of the DF and CL. Quality of the COCs was superior in ovaries in which the CL was present, but it did not vary between the AFC classes. We conclude that AFC can be performed on a single ovary, regardless of the presence, diameter, and weight of the CL and DF in zebu between 12 and 60 months old.

O presente estudo teve como objetivo avaliar a relação entre a contagem de folículos antrais (CFA) e a morfologia ovariana de Bos indicus em diferentes faixas etárias: 12-23 meses (G1), 24-35 meses (G2), 36-47 meses (G3), 48-59 meses (G4) e >60 meses (G5). Os ovários foram coletados individualmente e enviados ao laboratório, onde suas características morfológicas foram medidas: CFA, diâmetro e peso dos ovários, do folículo dominante (FD, ≥8 mm), do corpo lúteo (CL) e dos folículos pequenos (SF, <8 mm). A CFA foi classificada como alta, intermediária-alta, intermediária-baixa e baixa. Um grupo de ovários foi submetido à aspiração folicular para avaliar a qualidade morfológica dos complexos cumulus oócitos (COCs) recuperados. A CFA média não variou entre as idades. A característica morfológica que se correlacionou mais intimamente com a CFA foi o peso dos folículos pequenos. Não houve relação entre a CFA e o peso e o diâmetro do FD e do CL. A qualidade dos COCs foi superior em ovários em que o CL estava presente, mas não variou entre as classes de CFA. Concluiu-se que a CFA pode ser realizada em um único ovário, independentemente da presença, do diâmetro e do peso do CL e do FD em zebuínos entre 12 e 60 meses de vida.

Reproductive performance of fixed-time artificial insemination in heifers supplemented with a homeopathic complex / Desempenho reprodutivo à inseminação artificial em tempo fixo em novilhas suplementadas com complexo homeopático

For this study, ½ blood Nelore × Aberdeen Angus heifers (n = 40; 14 months) were divided into two groups: control (n=20; 310 kg) and Pró-Cio (n=20; 304 kg). For Pró-Cio group, 20g of homeopathic product (Pró-cio®, Real H, Campo Grande, Brazil) was supplied together with corn eighteen days prior (D-18) to the beginning of the fixed-time artificial insemination protocol (FTAI). On D0, all females received an intravaginal P4 device and 2 mg BE. On D5, the AFC was assessed by ultrasound and blood was collected for AMH dosage. On D8, P4 device was removed; 300 IU eCG, 0.530 mg of cloprostenol sodium and 0.5 mg of EC were administered; an estrus-identifying adhesive was fixed at the tail insertion and the diameter of the dominant follicle (DF) was measured. On D10, all heifers were inseminated, estrus manifestation was assessed and the preovulatory follicle (POF) was measured. On D20, the CL area was measured and blood flow was assessed using Doppler ultrasound. Data were analyzed by Student, Mann-Whitney or Friedman test. The rates of heat expression, ovulation, and blood flow score with Doppler were analyzed using Fisher's exact test and pregnancy rate by binary logistic regression model (P ≤ 0.05). There was no difference in AFC or AMH dosage (P > 0.05). Pró-Cio group had greater diameters of the DF (9.94 ± 0.42 mm) and POF (11.61 ± 0.56 mm) than the controls (DF: 7.72 ± 0.34 mm and POF: 9.91 ± 0.37 mm), as well a larger CL area (3.26 ± 0.26 versus 2.35 ± 0.16 cm2) and a higher mean CL vascularization score (3.06 versus 2.26; P < 0.05). There was no significant difference (P > 0.05) between estrus manifestation or conception rate for control and Pró-Cio groups. Heifers supplemented with the homeopathic product showed larger follicular diameters at the end of the FTAI protocol and CL with better blood flow scores compared to the control group.

Para esse estudo, novilhas ½ sangue Nelore x Aberdeen Angus (n = 40; 14 meses) foram divididas em dois grupos: Controle (n = 20; 310 kg) e Pró-cio (n = 20; 304 kg). Para o grupo Pró-Cio, 20 g/animal de produto homeopático (Pró-cio®, Real H, Campo Grande, Brazil) foi fornecido juntamente com o milho dezoito dias antes (D-18) de inicar o protocolo de inseminaçãoa artificial em tempo-fixo (IATF). No D0, todas as fêmeas receberam um dispositivo intravaginal de P4 e 2 mg BE. No D5, avaliou-se a contagem de folículos antrais (CFA) por ultrassonografia e realizou-se coleta de sangue para dosagem de AMH. No D8, procedeu-se a retirada do dispositivo de P4; aplicação de 300 UI de eCG, 0,530 mg de cloprostenol sódico e 0,5 mg de CE; um adesivo identificador de estro foi fixado na inserção da cauda e foi feita a mensuração do diâmetro do folículo dominante (FD). No D10, todas as novilhas foram inseminadas, foi realizada a avaliação da manifestação de estro e mensuração do folículo pré-ovulatório (FPO). No D20, realizou-se mensuração da área e avaliação do fluxo sanguíneo do CL por ultrassonografia Doppler. Os dados foram analisados pelos testes de Student, Mann-Whitney ou Friedman. As taxas de expressão de estro, ovulação e escore de fluxo sanguíneo com Doppler foram analisadas usando o teste exato de Fisher e taxa de prenhez pelo modelo de regressão logística binária (p ≤ 0,05). Não houve diferença na CFA nem na dosagem de AMH (p > 0,05). O grupo PróCio apresentou maior diâmetro do FD (9,94 ± 0,42 mm) e FPO (11,61 ± 0,56 mm) em relação ao controle (FD: 7,72 ± 0,34 mm e FPO: 9,91 ± 0,37 mm), além de maior área do CL (3,26 ± 0,26 versus 2,35 ± 0,16 cm2; p < 0,05) e maior escore de vascularização médio do CL (3,06 versus 2,26; p < 0,05). Não houve diferença (p > 0,05) na taxa de manifestação de estro e nem na taxa de concepção entre os grupos Controle e Pró-Cio, respectivamente. Novilhas suplementadas com o produto homeopático apresentaram maiores diâmetros foliculares ao final do protocolo de IATF e CL com melhores escores de fluxo sanguíneo em relação ao grupo controle.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs

Abstract This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs

This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.(AU)

Comparison of cattle embryo collection between Ringer's lactate solution and Dulbecco's phosphatebuffered saline

The objective of this study was to determine the efficiency of Ringer's lactate solution (RL) and RL + 1% fetal bovine serum (FBS) and compare them with the efficiency of Dulbecco's phosphate-buffered saline (D-PBS). Twenty-two Wagyu female cattle were subjected to superovulation and were randomly distributed to form three groups: group 1 ­ uterine flushing with RL (n = 8), group 2 ­ uterine flushing with RL + 1% FBS (n = 7), and group 3 ­ uterine flushing with D-PBS (n = 7, control group). Cows received a CIDR® device containing 1.9 g of progesterone at random stages of the estrous cycle (day 0). Progesterone withdrawal occurred on day 8 in the morning. For heifers, 160 mg of porcine follicle-stimulating hormone (FSH-P) was used and for cows, 200 mg. Prostaglandin F2α was also injected on the eighth day of FSH-P administration. On day 9, in the morning, hCG was administered. Females were superovulated and inseminated twice in a fixed time for embryo transfer. On the 16th day, females were subjected to uterine flushing for embryo collection. We collected 76 embryos from 22 females subjected to superovulation, of which 52 were transferable and 24 had degenerated. The total of embryos collected was 23, 16, and 23 for groups 1, 2, and 3, respectively. The embryo recovery rates per group were 13.86±4.23, 15.39±4.61 and 27.16±13.33%, in groups 1, 2, and 3, respectively. The means for the total structures collected per female were 2.88±0.85, 3.00±1.23, and 4.57±1.72 in groups flushed with RL, RL + 1% FBS, and D-PBS, respectively. We conclude that Ringer's lactate solution and Ringer's lactate solution + 1% of FBS and Dulbecco's phosphate-buffered saline showed no significant differences in terms of embryo quality or quantity, suggesting that Ringer's lactate solution is an alternative for collecting embryos in cattle.

Immunolocalization of angiotensin-(1-7), angiotensin II and angiotensin- converting enzyme 2 in goat ovary / Imunolocalização de angiotensina-(1-7), angiotensina II e enzima-conversora de angiotensina 2 em ovário de cabras

There is increasing evidence as to the participation of the ovarian renin-angiotensin system in important reproductive processes. The inhibition of the angiotensin-converting enzyme (ACE) caused an increase in the rate of ovulation and pregnancy in the artificial insemination protocol has fixed time (TFIA). This study aimed to investigate the presence and location of Ang II, Ang- (1-7) and ACE2 in goat ovaries and the possibility of the involvement of these peptides in previous results. Ten ovaries from goats were collected in a slaughterhouse, washed in buffered PBS, perfused with protease inhibitor solution and processed for immunohistochemistry protocol. The search for peptides was performed using the avidin–biotin–peroxidase method. A strong immunoreactivity for Ang II in theca cells of antral follicles and corpus luteum was observed. Antral follicles (theca cells), corpus luteum and oocyte cytoplasm in early antral follicles exhibited strong immunoreactivity for Ang-(1-7). There was strong immunoreactivity for ACE2 in the cytoplasm of luteal cells and theca cells of antral follicles. In this study, for the first time, the presence and location of Ang II, Ang-(1-7) and ACE2 are reported in goat ovary, suggesting that there is participation in follicular development, oocyte maturation and corpus luteum development.

Há evidências crescentes quanto à participação do sistema renina-angiotensina ovariano em processos reprodutivos importantes. A inibição da enzima conversora de angiotensina (ECA) ocasionou aumento na taxa de ovulação e gravidez no protocolo de inseminação artificial por tempo fixo (TFIA). Este estudo teve como objetivo investigar a presença e localização de Ang II, Ang-(1-7) e ECA2 em ovários de cabras e a possibilidade do envolvimento desses peptídeos em resultados anterio-res. Dez ovários de cabras foram coletados em abatedouro, lavados em PBS tamponado, perfundidos com solução inibidora de protease e processados para protocolo de imunohistoquímica. A busca por peptídeos foi realizada usando o método avidina-bio-tina-peroxidase. Foi observada uma forte imunorreatividade para Ang II em células da teca de folículos antrais e corpo lúteo. Os folículos antrais (células da teca), corpo lúteo e citoplasma do oócito nos folículos antrais iniciais exibiram forte imunor-reatividade para Ang-(1-7). Houve forte imunorreatividade para ECA2 no citoplasma das células luteais e células da teca dos folículos antrais. Neste estudo, pela primeira vez, a presença e localização de Ang II, Ang- (1-7) e ECA2 são relatadas em ovário caprino, sugerindo que há participação no desenvolvimento folicular, maturação oocitária e desenvolvimento do corpo lúteo.

Immunolocalization of angiotensin-(1-7), angiotensin II and angiotensin- converting enzyme 2 in goat ovary / Imunolocalização de angiotensina-(1-7), angiotensina II e enzima-conversora de angiotensina 2 em ovário de cabras

There is increasing evidence as to the participation of the ovarian renin-angiotensin system in important reproductive processes. The inhibition of the angiotensin-converting enzyme (ACE) caused an increase in the rate of ovulation and pregnancy in the artificial insemination protocol has fixed time (TFIA). This study aimed to investigate the presence and location of Ang II, Ang- (1-7) and ACE2 in goat ovaries and the possibility of the involvement of these peptides in previous results. Ten ovaries from goats were collected in a slaughterhouse, washed in buffered PBS, perfused with protease inhibitor solution and processed for immunohistochemistry protocol. The search for peptides was performed using the avidin–biotin–peroxidase method. A strong immunoreactivity for Ang II in theca cells of antral follicles and corpus luteum was observed. Antral follicles (theca cells), corpus luteum and oocyte cytoplasm in early antral follicles exhibited strong immunoreactivity for Ang-(1-7). There was strong immunoreactivity for ACE2 in the cytoplasm of luteal cells and theca cells of antral follicles. In this study, for the first time, the presence and location of Ang II, Ang-(1-7) and ACE2 are reported in goat ovary, suggesting that there is participation in follicular development, oocyte maturation and corpus luteum development.(AU)

Há evidências crescentes quanto à participação do sistema renina-angiotensina ovariano em processos reprodutivos importantes. A inibição da enzima conversora de angiotensina (ECA) ocasionou aumento na taxa de ovulação e gravidez no protocolo de inseminação artificial por tempo fixo (TFIA). Este estudo teve como objetivo investigar a presença e localização de Ang II, Ang-(1-7) e ECA2 em ovários de cabras e a possibilidade do envolvimento desses peptídeos em resultados anterio-res. Dez ovários de cabras foram coletados em abatedouro, lavados em PBS tamponado, perfundidos com solução inibidora de protease e processados para protocolo de imunohistoquímica. A busca por peptídeos foi realizada usando o método avidina-bio-tina-peroxidase. Foi observada uma forte imunorreatividade para Ang II em células da teca de folículos antrais e corpo lúteo. Os folículos antrais (células da teca), corpo lúteo e citoplasma do oócito nos folículos antrais iniciais exibiram forte imunor-reatividade para Ang-(1-7). Houve forte imunorreatividade para ECA2 no citoplasma das células luteais e células da teca dos folículos antrais. Neste estudo, pela primeira vez, a presença e localização de Ang II, Ang- (1-7) e ECA2 são relatadas em ovário caprino, sugerindo que há participação no desenvolvimento folicular, maturação oocitária e desenvolvimento do corpo lúteo.(AU)

Influence of male on estrus behavior, estradiol and progesterone release and puberty onset on prepubertal Saanen goats / Influência do reprodutor sobre o comportamento de estro, liberação de estrógeno, progesterona e início da puberdade em cabras Saanen pré-puberes

The present study investigated the male effect on the estrus behaviors, estradiol and progesterone release in prepubertal Saanen goats. Twenty-nine female Saanen goats at 135 ± 10 days old with body weight of 22.8 ± 3.3 Kg were randomly assigned to three treatments: exposure to sexually active male (male treatment), exposure to androgenized females (androgenized female treatment), and prepubertal goats isolated from active male and androgenized females (control treatment). Sexual behaviors associated with estrus were recorded daily, and blood samples were taken weekly to determine estradiol and progesterone concentrations over 24 weeks. The experimental goats subjected to male or androgenized female had significantly higher frequency of estrus (mount acceptance) (P ≤ 0,02), progesterone (P ≤ 0,01), and estradiol (P ≤ 0,01) release than the control goats. Furthermore, goats exposed to a male showed estrus behavior two weeks earlier and maintained this estrus behavior for three weeks more than goats of both female and control treatments. Estrus was observed in 70 % of goats in male and female treatments during the breeding season versus 44 % of the control goats. Finally, significantly more goats subjected to male treatment (60 % of goats) showed progesterone concentrations higher than 1 ng mL-1 (which indicates the presence of a functional corpus luteum) compared to the female and control treatment (40 and 22 % of goats, respectively). These results shows that male treatment significantly increased the number of females showing estrus behavior, estradiol and progesterone release, and the number of animals with a functional corpus luteum, anticipating puberty for experimental goats, suggesting that the male effect could be used to anticipate the onset of puberty in goats.(AU)

O presente estudo investigou o efeito do macho sobre o comportamento do estro, liberação de estradiol e progesterona em cabritas Sannen pré-púberes. Vinte e nove cabritas com 135 ± 10 dias de idade e peso corporal de 22,8 ± 3,3 kg foram submetidas à três tratamentos: macho; fêmeas androgenizadas; controle (mantidas isoladas do efeito macho ou de fêmeas androgenizadas) . Os comportamentos sexuais foram registrados diariamente e as amostras de sangue foram colhidas semanalmente ao longo de 24 semanas. Os tratamentos macho e fêmea androgenizada aumentaram significativamente a ocorrência comportamental do estro (P ≤ 0,02), a concentração de progesterona (P ≤ 0,01) e estradiol (P ≤ 0,01) em comparação ao tratamento controle. As cabritas expostas ao efeito macho anteciparam o comportamento de estro em duas semanas, e o mantiveram por mais três semanas quando comparado às cabritas dos tratamentos fêmea androgenizada e controle. Apenas 44% das cabritas controle foram observadas em estro, sendo que 70% das cabritas submetidas aos tratamentos macho e fêmea androgenizada foram observadas em estro. Além disso, 60% das cabritas expostas ao efeito macho, 40 % das cabritas expostas ao efeito fêmea androgenizada e 22% das cabritas controle apresentaram concentrações de progesterona superiores a 1 ng mL-1, o que indica a presença de corpo lúteo funcional. De fato, o efeito macho aumentou significativamente o número de fêmeas em estro, a concentração de estradiol e progesterona, o número de fêmeas com corpo lúteo funcional, sugerindo que o efeito macho pode ser usado para antecipar o início da puberdade em cabritas Saanen.(AU)

Avaliação da eficiência de reutilização do CIDR® em protocolos de superovulação de ovelhas / Evaluation of efficiency of the reuse of CIDR® in sheep superovulation protocols

Sheep farming is an area of veterinary medicine of great importance at regional and national level, among the easily found breeds of economic relevance in northeastern Brazil are Morada Nova and Santa Inês. In addition, ultrasonography has been used systematically in animals superovulatory response to the sheep supervoulation protocols. In order to assess, based on the number of yellow bodies and number of viable embryos produced, the efficiency of the reuse of CIDR® in superovulation protocols in sheep using ultrasonography in color Doppler mode, a superovulatory protocol was performed in a total 40 sheep, with G-USED when 10 Morada Nova ewes and 10 Santa Inês ewes received reused CIDR® and G-NOVO when 10 Morada Nova ewes and 10 Santa Inês ewes received new CIDR®. The mean and standard error of yellow bodies was 10.35±1.33 for G-USED and 9.25±1.65 for G-NOVO, while the number of viable embryos was 6.50±2, 54 for G-USED and 8.25±1.49 for G-NEW. Thus, the reuse of CIDR® demonstrated efficiency in superovulation and did not interfere with the development of corpus luteum and the production of quality transferable embryos in sheep.

Low invasive estrous synchronization protocol for wild animals: an example with melengestrol acetate in brown brocket deer (Mazama gouazoubira)

Deer are sensitive to stressful stimuli by handling and their reproductive physiology could be altered by these procedures, making it necessary to develop less invasive protocols for ART. Melengestrol acetate (MGA), a synthetic progestin administered orally, appears as an alternative for estrous synchronization protocols (ESP), such as reported in cattle. Firstly, we compared two MGA doses (0.5 and 1.0 mg/day/animal), which would have suppression effect in estrous behavior (EB). Eight females were randomly and equally distributed in Group 1 (G1) and Group 2 (G2), which received 0.5 and 1.0 mg/day/animal respectively for 15 days (D1 to D15). Two cloprostenol (CP) applications were performed on D0 and D11. Estrus detection (ED) was performed every day. All females from G1 displayed estrus during treatment period, whereas all females from G2 displayed estrus after treatment, suggesting a suppressive effect of 1.0 mg in the EB. Once the suppressive MGA dose (1.0 mg) was defined, we used this dose for assessing ESP. The same eight females received 1.0 mg/animal for eight days (D-8 to D-1), followed by 0.25 mg of estradiol benzoate on D-8 and 265 g of CP on D0. Feces for fecal progesterone metabolites (FPM) measurement were collected from D0 until seven days after the last day of estrus. Seven females displayed estrus between 12 and 72 h after CP application, which was followed by a significant increase in FPM levels (except female MG6), suggesting the formation of corpus luteum. After ED, females were placed with a fertile male to assess the fertility of the protocol. Pregnancy was confirmed by ultrasound 30 days after mating in 3/6 individuals. Although the low effectiveness of MGA protocol, it should be considered as a promising alternative in deer ESP since this protocol has less stressful effect on the animal during reproductive management when compared to other ESP.

Avaliação da eficiência de reutilização do CIDR® em protocolos de superovulação de ovelhas / Evaluation of efficiency of the reuse of CIDR® in sheep superovulation protocols

Sheep farming is an area of veterinary medicine of great importance at regional and national level, among the easily found breeds of economic relevance in northeastern Brazil are Morada Nova and Santa Inês. In addition, ultrasonography has been used systematically in animals superovulatory response to the sheep supervoulation protocols. In order to assess, based on the number of yellow bodies and number of viable embryos produced, the efficiency of the reuse of CIDR® in superovulation protocols in sheep using ultrasonography in color Doppler mode, a superovulatory protocol was performed in a total 40 sheep, with G-USED when 10 Morada Nova ewes and 10 Santa Inês ewes received reused CIDR® and G-NOVO when 10 Morada Nova ewes and 10 Santa Inês ewes received new CIDR®. The mean and standard error of yellow bodies was 10.35±1.33 for G-USED and 9.25±1.65 for G-NOVO, while the number of viable embryos was 6.50±2, 54 for G-USED and 8.25±1.49 for G-NEW. Thus, the reuse of CIDR® demonstrated efficiency in superovulation and did not interfere with the development of corpus luteum and the production of quality transferable embryos in sheep.(AU)

Low invasive estrous synchronization protocol for wild animals: an example with melengestrol acetate in brown brocket deer (Mazama gouazoubira)

Deer are sensitive to stressful stimuli by handling and their reproductive physiology could be altered by these procedures, making it necessary to develop less invasive protocols for ART. Melengestrol acetate (MGA), a synthetic progestin administered orally, appears as an alternative for estrous synchronization protocols (ESP), such as reported in cattle. Firstly, we compared two MGA doses (0.5 and 1.0 mg/day/animal), which would have suppression effect in estrous behavior (EB). Eight females were randomly and equally distributed in Group 1 (G1) and Group 2 (G2), which received 0.5 and 1.0 mg/day/animal respectively for 15 days (D1 to D15). Two cloprostenol (CP) applications were performed on D0 and D11. Estrus detection (ED) was performed every day. All females from G1 displayed estrus during treatment period, whereas all females from G2 displayed estrus after treatment, suggesting a suppressive effect of 1.0 mg in the EB. Once the suppressive MGA dose (1.0 mg) was defined, we used this dose for assessing ESP. The same eight females received 1.0 mg/animal for eight days (D-8 to D-1), followed by 0.25 mg of estradiol benzoate on D-8 and 265 g of CP on D0. Feces for fecal progesterone metabolites (FPM) measurement were collected from D0 until seven days after the last day of estrus. Seven females displayed estrus between 12 and 72 h after CP application, which was followed by a significant increase in FPM levels (except female MG6), suggesting the formation of corpus luteum. After ED, females were placed with a fertile male to assess the fertility of the protocol. Pregnancy was confirmed by ultrasound 30 days after mating in 3/6 individuals. Although the low effectiveness of MGA protocol, it should be considered as a promising alternative in deer ESP since this protocol has less stressful effect on the animal during reproductive management when compared to other ESP.(AU)

Ultrassonografia Dopper em ovelhas / Doppler ultrasonography in sheep

O uso da técnica de ultrassonografia em modo B, como uma ferramenta pouco invasiva na reprodução animal, passou por uma evolução significativa na pesquisa e na prática. Na exploração comercial de animais de produção seu uso é, praticamente, concentrado ao manejo reprodutivo. Embora a ultrassonografia convencional em Modo B tenha passado por avanços tecnológicos esta não possibilita o acesso às informações da função do órgão, demonstradas pela perfusão vascular. Em ovelhas, o estudo da hemodinâmica e funcionalidade do útero, ovário e suas estruturas podem ser úteis para avaliar a viabilidade de programas reprodutivos e, por conseguinte, exibirem impacto econômico diante do uso da modalidade colorida ou pulsada (ou espectral) da ultrassonografia doppler. A ultrassonografia doppler é baseada no efeito Doppler na qual as frequências dos feixes de ultrassom refletidos pelos eritrócitos em movimento variam conforme as células se movem com diferentes velocidades. No tocante a reprodução de ovelhas, uma série de estratégias tem sido apresentadas para uso desta tecnologia, dada a necessidade e intempéries apresentadas a campo. Suas principais aplicações baseiam-se na avaliação da gestação, hemodinâmica uterina, funcionalidade luteal e respostas à superovulação, dentre outras. O presente trabalho teve como objetivo apresentar os aspectos relacionados ao uso e aplicações da ultrassonografia doppler espectral e colorida na reprodução de ovelhas.

The use of B-mode ultrasound technique, as a little invasive tool in animal reproduction, has undergone a significant evolution in research and practice. In farm animals, its use is practically concentrated on reproductive management. Although conventional B-mode ultrasonography has undergone technological advances, it does not allow access to organ function information, as demonstrated by vascular perfusion. In sheep, the study of the hemodynamics and functionality of the uterus, ovary and its structures can be useful to assess reproductive programs viability and, therefore, exhibit economic impact when using the color or pulsed (or spectral) modalityof doppler ultrasound. Doppler ultrasonography is based on the Doppler effect in which the frequencies of the ultrasound beams reflected by the erythrocytes in motion vary as the cells move at different speeds. A lot of strategies have been presented using this technology, related to reproduction of sheep animals, in farm conditions. Applications are based on the evaluation of pregnancy, uterine hemodynamics, luteal functionality and responses to superovulation, among others. This work aimed to present aspects regarding to applications of spectral and color doppler ultrasound in sheep reproduction.

Pregnancy rates and luteal phase characteristics of bovine embryo recipients treated with flunixin meglumine / Taxas de prenhez e características da fase lútea de receptoras de embriões tratadas com flunixina meglumina em bovinos

The aim of this study was to evaluate the effects of flunixin meglumine administration on pregnancy rates and luteal phase characteristics in bovine embryo recipients at the moment of embryo transfer. In experiment 1, in vitro produced embryos were transferred to 184 females divided as control and treated group (recipients treated with 1.1mg/kg flunixin meglumine). In experiment 2, 22 females were divided as control group; group 2 (animals submitted to a reproductive tract manipulation similar to an embryo transfer on the 7th day after estrous); and group 3 (females submitted to a manipulation and treatment with 1.1mg/kg flunixin meglumine). In experiment 1 no difference was observed between control and treated groups (40.2% and 44.6%, respectively) for pregnancy rates. In experiment 2 no difference was observed on the length of luteal phase between groups, however, animals in group 2 presented lower plasma progesterone concentrations than the control group and group 3. Therefore, we concluded that although the administration of flunixin meglumine at the moment of embryo transfer inhibited the reduction plasma progesterone concentrations, it was not effective in increasing pregnancy rates of bovine recipients.(AU)

O objetivo deste estudo foi avaliar os efeitos da administração de flunixina meglumina sobre as taxas de prenhez e características da fase lútea da receptora no momento da transferência de embriões em bovinos. No experimento 1, embriões produzidos in vitro foram transferidos para 184 fêmeas, divididas em grupos controle e tratado (tratados com 1,1mg/kg de flunixina meglumina). No experimento 2, 22 fêmeas foram divididas em grupo controle (n=7); grupo 2 (n=8; animais submetidos à manipulação do trato reprodutivo semelhante à transferência de embriões no sétimo dia pós-cio); e grupo 3 (n=7; fêmeas submetidas à manipulação e ao tratamento com 1,1mg/kg de flunixina meglumina). No experimento 1, não foi observada diferença nos grupos controle e tratado (40,2% e 44,6%, respectivamente) para as taxas de prenhez. No experimento 2, não houve diferença na extensão da fase lútea entre os grupos, entretanto os animais do grupo 2 apresentaram concentrações plasmáticas de progesterona mais baixas que o grupo controle e o grupo 3. Portanto, conclui-se que a administração de flunixina meglumina no momento da transferência de embriões inibiu a redução das concentrações plasmáticas de progesterona, no entanto não foi eficaz para aumentar as taxas de prenhez de receptoras em bovinos.(AU)

Ultrassonografia Dopper em ovelhas / Doppler ultrasonography in sheep

O uso da técnica de ultrassonografia em modo B, como uma ferramenta pouco invasiva na reprodução animal, passou por uma evolução significativa na pesquisa e na prática. Na exploração comercial de animais de produção seu uso é, praticamente, concentrado ao manejo reprodutivo. Embora a ultrassonografia convencional em Modo B tenha passado por avanços tecnológicos esta não possibilita o acesso às informações da função do órgão, demonstradas pela perfusão vascular. Em ovelhas, o estudo da hemodinâmica e funcionalidade do útero, ovário e suas estruturas podem ser úteis para avaliar a viabilidade de programas reprodutivos e, por conseguinte, exibirem impacto econômico diante do uso da modalidade colorida ou pulsada (ou espectral) da ultrassonografia doppler. A ultrassonografia doppler é baseada no efeito Doppler na qual as frequências dos feixes de ultrassom refletidos pelos eritrócitos em movimento variam conforme as células se movem com diferentes velocidades. No tocante a reprodução de ovelhas, uma série de estratégias tem sido apresentadas para uso desta tecnologia, dada a necessidade e intempéries apresentadas a campo. Suas principais aplicações baseiam-se na avaliação da gestação, hemodinâmica uterina, funcionalidade luteal e respostas à superovulação, dentre outras. O presente trabalho teve como objetivo apresentar os aspectos relacionados ao uso e aplicações da ultrassonografia doppler espectral e colorida na reprodução de ovelhas.(AU)

The use of B-mode ultrasound technique, as a little invasive tool in animal reproduction, has undergone a significant evolution in research and practice. In farm animals, its use is practically concentrated on reproductive management. Although conventional B-mode ultrasonography has undergone technological advances, it does not allow access to organ function information, as demonstrated by vascular perfusion. In sheep, the study of the hemodynamics and functionality of the uterus, ovary and its structures can be useful to assess reproductive programs viability and, therefore, exhibit economic impact when using the color or pulsed (or spectral) modalityof doppler ultrasound. Doppler ultrasonography is based on the Doppler effect in which the frequencies of the ultrasound beams reflected by the erythrocytes in motion vary as the cells move at different speeds. A lot of strategies have been presented using this technology, related to reproduction of sheep animals, in farm conditions. Applications are based on the evaluation of pregnancy, uterine hemodynamics, luteal functionality and responses to superovulation, among others. This work aimed to present aspects regarding to applications of spectral and color doppler ultrasound in sheep reproduction.(AU)

Pregnancy rates and luteal phase characteristics of bovine embryo recipients treated with flunixin meglumine / Taxas de prenhez e características da fase lútea de receptoras de embriões tratadas com flunixina meglumina em bovinos

The aim of this study was to evaluate the effects of flunixin meglumine administration on pregnancy rates and luteal phase characteristics in bovine embryo recipients at the moment of embryo transfer. In experiment 1, in vitro produced embryos were transferred to 184 females divided as control and treated group (recipients treated with 1.1mg/kg flunixin meglumine). In experiment 2, 22 females were divided as control group; group 2 (animals submitted to a reproductive tract manipulation similar to an embryo transfer on the 7th day after estrous); and group 3 (females submitted to a manipulation and treatment with 1.1mg/kg flunixin meglumine). In experiment 1 no difference was observed between control and treated groups (40.2% and 44.6%, respectively) for pregnancy rates. In experiment 2 no difference was observed on the length of luteal phase between groups, however, animals in group 2 presented lower plasma progesterone concentrations than the control group and group 3. Therefore, we concluded that although the administration of flunixin meglumine at the moment of embryo transfer inhibited the reduction plasma progesterone concentrations, it was not effective in increasing pregnancy rates of bovine recipients.(AU)

O objetivo deste estudo foi avaliar os efeitos da administração de flunixina meglumina sobre as taxas de prenhez e características da fase lútea da receptora no momento da transferência de embriões em bovinos. No experimento 1, embriões produzidos in vitro foram transferidos para 184 fêmeas, divididas em grupos controle e tratado (tratados com 1,1mg/kg de flunixina meglumina). No experimento 2, 22 fêmeas foram divididas em grupo controle (n=7); grupo 2 (n=8; animais submetidos à manipulação do trato reprodutivo semelhante à transferência de embriões no sétimo dia pós-cio); e grupo 3 (n=7; fêmeas submetidas à manipulação e ao tratamento com 1,1mg/kg de flunixina meglumina). No experimento 1, não foi observada diferença nos grupos controle e tratado (40,2% e 44,6%, respectivamente) para as taxas de prenhez. No experimento 2, não houve diferença na extensão da fase lútea entre os grupos, entretanto os animais do grupo 2 apresentaram concentrações plasmáticas de progesterona mais baixas que o grupo controle e o grupo 3. Portanto, conclui-se que a administração de flunixina meglumina no momento da transferência de embriões inibiu a redução das concentrações plasmáticas de progesterona, no entanto não foi eficaz para aumentar as taxas de prenhez de receptoras em bovinos.(AU)

Reproductive biology, morphology, and morphometry of ovaries and oviducts of Trachemys scripta elegans in Brazilian Cerrado / Biologia reprodutiva, morfologia e morfometriade ovários e ovidutos de Trachemys scripta elegans no Cerrado Brasileiro

Although Trachemys scripta elegans is an exotic species popular as a pet in Brazil, studies on reproductive biology and capacity are non-existent in the Brazilian Cerrado. This study analyzed ovarian and oviduct characteristics and the egg production capacity of T. scripta elegans grown in this biome. The findings will associate with the size of the specimens and the sexual maturity, aiming at comparisons with native and exotic populations, as well as interspecific and contributing to the understanding of its impact on the invaded ecosystems and the establishment of eradication programs. Thus, 39 females had evaluated the body biometry and the morphology and morphometry of the ovaries and oviducts. G2 (N=20): with Class I (>5-10mm) follicles, with Class I and Class II (>10-fold) follicles, 25mm) and G3 (N=9) with Class I, Class II and Class III (>25mm) follicles. Analysis of variance, Scott-Knott's test, and Pearson's correlation analysis showed that there was no significant difference between the groups in body biometry; in the mean gonadosomatic index and gonadal morphometry, only the width of the oviducts in the right antimer and the mass and width in the left antimer were higher in G3, the only one that presented eggs. There was positive and harmonic development between body mass, carapace, and plastron, and gonadal growth occurred concomitantly with body growth, indicating a higher reproductive potential and a positive relationship between the size of the litter and the female litter. The gonadosomatic index proved to be an excellent reproductive indicator, and the ovarian evaluation was a better indicator of sexual maturity than the maximum carapace length. Ovaries were irregular structures, without delimitation between the cortical and medullary regions and filled with vitelogenic follicles of different diameters, atresic follicles, and corpora lutea, which reflected the ovarian complexity of the species and the presence of follicular hierarchy. In the scarce stroma, two germinative beds were observed per ovary and the presence of gaps very close to the follicles and associated with the blood vessels. Analysis of gonadal tissue revealed three types of oocytes according to cytoplasmic characteristics: homogeneous, vesicular or vesicular in the cortex with apparent granules. Oviducts were functional and separated, joining only in the final portion to form the cloaca and subdivided into infundibulum, tuba, isthmus, uterus, and vagina. The structure of the uterine tube was composed of serosa, muscular and mucous, which was full of glands. The presence of eggs in the oviducts indicated that the specimens can reproduce in the Brazilian Cerrado. This study provides necessary and relevant information on the reproductive biology and capacity of T. scripta elegans in the Brazilian Cerrado and can contribute to the understanding of its impact on the invaded ecosystems and the establishment of eradication programs. The extraction of females with capacity can reduce the annual reproductive yield of the species and decrease its effect on local biodiversity.(AU)

Embora Trachemys scripta elegans seja uma espécie exótica popular como animal de estimação no Brasil, estudos sobre biologia e capacidade reprodutivas são inexistentes no Cerrado brasileiro. Este estudo analisou características ovarianas e do oviduto e a capacidade de produção de ovos em T. scripta elegans criadas neste bioma, correlacionando estes achados ao tamanho dos espécimes e a maturidade sexual, visando comparações com populações nativas e exóticas, bem como interespecíficas e contribuir para a compreensão de seu impacto nos ecossistemas invadidos e com o estabelecimento de programas de erradicação. Assim, 39 fêmeas tiveram avaliadas a biometria corporal e a morfologia e morfometria dos ovários e ovidutos. De acordo com o tamanho dos folículos ovarianos as fêmeas foram separadas em G1 (N= 10): com folículos Classe I (>5-10 mm), G2 (N= 20): com folículos Classe I e Classe II (>10-25 mm) e G3 (N= 9) com folículos Classe I, Classe II e Classe III (>25 mm). À análise de variância, teste de Scott-Knott e à análise de correlação de Pearson verificou-se que não houve diferença significativa entre os grupos na biometria corporal; no índice gonadossomático médio e na morfometria gonadal, apenas a largura dos ovidutos no antímero direito e a massa e a largura no antímero esquerdo foram maiores no G3, o único que apresentou ovos. Houve desenvolvimento positivo e harmônico entre massa corporal, carapaça e plastrão e o crescimento gonadal ocorreu concomitante ao crescimento corporal, indicando maior potencial reprodutivo e relação positiva entre o tamanho da ninhada de ovos e o da fêmea. O índice gonadossomático mostrou-se um bom indicador reprodutivo e a avaliação ovariana um melhor indicador da maturidade sexual que o comprimento máximo da carapaça. Ovários foram estruturas irregulares, sem delimitação entre a região cortical e medular e repletos de folículos vitelogênicos de diferentes diâmetros, folículos atrésicos e corpos lúteos, que refletiram a complexidade ovariana da espécie e a presença de hierarquia folicular. No estroma escasso foram observados dois leitos germinativos por ovário e a presença de lacunas muito próximas aos folículos e associadas aos vasos sanguíneos. A análise do tecido gonadal revelou três tipos de oócitos de acordo com as características do citoplasma: homogêneo, vesicular ou vesicular no córtex com grânulos aparentes. Ovidutos eram funcionais e separados, unindo-se apenas na porção final para formar a cloaca e subdividiam-se em infundíbulo, tuba uterina, istmo, útero e vagina. A estrutura da tuba uterina era constituída de serosa, muscular e mucosa, a qual era repleta de glândulas. A presença de ovos nos ovidutos indicou que os espécimes podem se reproduzir no cerrado brasileiro. Este estudo fornece informações básicas e relevantes da biologia e capacidade reprodutivas de T. scripta elegans no Cerrado brasileiro e pode contribuir com a compreensão de seu impacto nos ecossistemas invadidos e com o estabelecimento de programas de erradicação, uma vez que a extração de fêmeas com capacidade reprodutiva pode contribuir com a diminuição do rendimento reprodutivo anual da espécie e diminuir seu efeito sobre a biodiversidade local.(AU)