Conceptus-mediated endometrial vascular changes during early pregnancy in mares: an anatomic, histomorphometric, and vascular endothelial growth factor receptor system immunolocalization and gene expression study

This work examined how the conceptus modulates endometrial tissue remodeling and vascular development prior to implantation in mares. A macroscopic uterine examination was completed at day 21 of pregnancy. In situ morphology revealed that the endometrium involved in encroachment is restricted to the dorsal endometrium immediately overlying the yolk sac. The amount of stromal area occupied by blood vessels and the number of endometrial glands were increased during early pregnancy. Endometrial histomorphometry as well as the endometrial mRNA abundance and immunolocalization of VEGF, VEGFR1, VEGFR2, and Ki-67 was completed at days 14 and 21 of pregnancy, at day 10 of the estrous cycle, and during estrus. No obvious differences in VEGF and VEGFR1 protein localization were detected between pregnant and cycling mares but differential staining pattern for VEGFR2 and Ki-67 was observed. VEGFR2 localized to luminal and glandular epithelium of pregnant mares, while luminal epithelium was negative in cycling mares. Ki-67 staining was weak during the luteal phase but exhibited prominent luminal epithelium staining during estrus. In pregnant mares, all endometrial layers were Ki-67 positive. Quantitative RT-PCR revealed a greater abundance of VEGF mRNA during pregnancy. VEGFR2 transcript abundance was greatest in pregnant mares on day 21. This study supports the concept that the conceptus plays an active role in directing vasculogenesis within the uterus and thereby establishing hemotrophic nutrition that supports pregnancy after implantation. Reproduction (2011) 142 593-603

Use of oocyte transfer in a commercial breeding program for mares with reproductive abnormalities

In some mares with lesions of the reproductive tract, embryo collection and survival rates are low or collection of embryos is not feasible. For these mares, oocyte transfer has been proposed as a method to induce pregnancies. In this report, a method for oocyte transfer in mares and results of oocyte transfer performed over 2 breeding seasons, using mares with long histories of subfertility and various reproductive lesions, are described.Human chorionic gonadotropin or an implant containing a gonadotropin-releasing hormone analog was used to initiate follicular and oocyte maturation. Oocytes were collected by means of transvaginal ultrasound-guided follicular aspiration. Following follicular aspiration, cumulus oocyte complexes were evaluated for cumulus expansion and signs of atresia; immature oocytes were cultured in vitro to allow maturation. The recipient's ovary and uterine tube (oviduct) were exposed through a flank laparotomy with the horse standing, and the oocyte was slowly deposited within the oviduct. Oocyte transfer was attempted in 38 mares between 9 and 30 years old during 2 successive breeding seasons. All mares had a history of reproductive failure while in breeding and embryo transfer programs. Twenty pregnancies were induced. Fourteen of the pregnant mares delivered live foals. Results suggest that oocyte transfer can be a successful method for inducing pregnancy in subfertile mares in a commercial setting..

Early embryonic death in mares: clinical and hormonal aspects

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hematological values and total protein of Brasileiro de Hipismo and Breton mares during pregnancy

Estudaram-se as variáveis eritrométricas, leucométricas e a concentração de proteína total (PT) de éguas gestantes das raças Brasileiro de Hipismo (BH) e Bretão. Utilizaram-se 175 éguas sadias, com 3 a 13 anos de idade. Dentre as éguas utilizadas, 89 foram da raça BH, divididas em 14 não-prenhes e 75 prenhes, e 86 éguas da raça Bretão divididas em 11 não-prenhes e 75 prenhes. As éguas foram subdivididas em quatro grupos por raça: grupo controle (éguas não-prenhes, n=14 para BH e n=11 para Bretão); grupo I (primeiro período de gestação, 25-110 dias, n=25 para BH e Bretão), grupo II (segundo período de gestação, 111-210 dias, n=25 para BH e Bretão); e grupo III (terceiro período de gestação, 211-340 dias, n=25 para BH e Bretão). Os valores médios de cada variável eritrométrica, leucométrica e de proteína total foram obtidos para cada raça e, posteriormente, realizou-se comparação entre as mesmas. O número de hemácias (He), volume globular (VG) e teor de hemoglobina (Hb) aumentaram no segundo período de gestação e diminuíram no último período tanto para a raça BH, como para a raça Bretão. Além disso, essas mesmas variáveis apresentaram diferença estatística significativa tanto nas éguas não-prenhas como nas prenhes durante os três períodos gestacionais. Os valores médios obtidos de leucócitos e neutrófilos segmentados aumentaram a partir do segundo período de gestação para as duas raças estudadas.

Decreasing pH of mammary gland secretions is associated with parturition and is correlated with electrolyte concentrations in prefoaling mares

The objectives of this study were to determine pH of the mammary gland secretions and the corresponding electrolyte concentrations in prefoaling mares. Pregnant mares (seven primiparous and seven multiparous) were monitored daily from 310.320 days of gestation until parturition. Prefoaling mammary gland secretions were collected, and pH was immediately determined with a pH meter and pH strip test. An aliquot of prefoaling mammary secretions was frozen and stored until further analyses. After parturition, samples from day .4 to 0 (day of foaling) were thawed and electrolyte concentrations (ie, Ca2+, Mg2+, Na+, K+ and Cl-) were determined with an automated analyser. Data were analysed via a mixed model with the mare as a random effect. Correlations were determined between pH and electrolyte concentrations by the Pearson product-moment for each pair. There was significant reduction in pH of mammary secretions on the day of foaling (P<0.0001), and most mares (11/14) with a pH .7 foaled within 24 hours. There was high correlation between the two pH methods (r=0.93). Additionally, there were significant (P<0.05) increases in Ca2+ and K+ concentrations, and significant decreases in Na+ and Cl- concentrations from one day before to the day of foaling. The pH of mammary secretions was highly and significantly (P<0.001) correlated with Na + (r=0.87), Cl- (r=0.85), Ca2+ (r=-0.88); and K+(r=.0.80) concentrations, and moderately correlated with Mg 2+ (r=-0.58). Daily evening pH measure of the mammary gland secretions can predict foaling in most mares.

Effect of age and endometrial degenerative changes on uterine blood flow during early gestation in mares

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of transrectal and transabdominal ultrasound examination on salivary cortisol, heart rate, and heart rate variability in mares

Pregnancy diagnostics in equine reproduction are routinely performed using transrectal ultrasonography, although it is also possible to visualize the fetus by transabdominal ultrasound examinations from the 90th day of gestation onward. We hypothesized that ultrasound examinations may stress the mare and that the gestational stage status and lactation may influence the mare's stress reaction. To investigate the stress reaction, 25 thoroughbred mares of different age, pregnancy and lactational status underwent a transrectal examination. In pregnant mares, an additional transabdominal examination was performed. Salivary cortisol concentration, mean heart rate, and heart rate variability of mares were assessed to evaluate the reactions of hypothalamic–pituitary–adrenal (HPA) axis and of the autonomic nervous system. Significant differences were observed between lactating and nonlactating mares; with a lower responsiveness to stress in lactating mares. The transrectal ultrasound examination in nonlactating mares induced a significant increase in salivary cortisol (P < 0.05), and in the heart rate variability parameter, ratio of low to high frequencies (P < 0.05). This reflects an activation of the HPA axis and a shift to more sympathetic dominance. In contrast, a transabdominally performed pregnancy check did not induce an activation of the HPA axis over basal level but increased the mean heart rate and low to high frequency ratio. The results of this study indicate that checks of advanced pregnancies can be easily performed by transabdominal ultrasonography. With regard to animal welfare, this technique should be preferred during midgestation in nonlactating mares.

Auscultatory and electrocardiographic characteristics of Crioulo horses

In order to determine auscultatory and electrocardiographic characteristics of Crioulo horses, one hundred animals ranging between one and twenty-six years of age (21 stallions, nine geldings, 27 pregnant mares e 43 not pregnant mares) were evaluated. The cardiac auscultation was performed during the clinical examination of the cardiovascular system, evaluating frequency, rate, normal and abnormal heart sounds (heart murmurs). The electrocardiographic examination followed the bipolar base-apex derivative system with animals at rest, by using an ECG-PC TEB equipment. The cardiac frequency, heart rate, morphology, duration, wave and complex amplitudes and interval durations were determined. The results were submitted to ANOVA and Tukey tests with an error probability of 5%. The cardiac auscultation revealed presence of functional systolic and diastolic murmur (10.00%) and systolic murmur compatible with tricuspid regurgitation besides normal heart sounds S1 (100.0%), S2 (100.0%), S3 (19.0%) and S4 (34.0%). The cardiac frequency obtained the average of 43.64 bpm, observing significative differences in relation to sexual and age factors and training level. The sinus rhythm was the most frequent (57.00%), followed by sinus tachycardia (38.00%) and sinus arrhythmia (5.00%), being observed rhythm disturbances in 16% of tracings. The P and T waves were observed more frequently in their forms P bifida positive (95.00%) and biphasic T (91.00%), being variable at tracing. There were also observed Q waves in 12.00% of the tracings. Thus, it was concluded that the auscultatory characteristics of Crioulo horses are according to the described in the literature for the species and the sexual factor, category, age factor and training level can influence some electrocardiographic parameters.

Avaliação da taxa de fertilidade em éguas da raça puro sangue lusitano : efeito da idade da égua e do tipo de cobrição (cobrição natural vs inseminação artificial)

Ao longo dos meses de estágio acompanhei diversos procedimentos médicos veterinários, sendo que a maior parte deles diziam respeito à área da reprodução equina. A Raça Puro Sangue Lusitano (PSL) é a mais importante raça portuguesa autóctone de equinos. Contudo, não existem muitos estudos no que respeita à sua eficiência reprodutiva. Este trabalho pretende fazer um estudo sobre o efeito da idade da égua e do tipo de cobrição (natural e inseminação artificial - IA) na taxa de fertilidade de éguas PSL, através do cálculo das diferentes taxas de gestação. Neste estudo, verificou-se que a percentagem de éguas gestantes no final da época reprodutiva foi de 90%, encontrando-se dentro dos valores esperados (71% - 96%). Os dados foram recolhidos durante a época reprodutiva de 2011/2012 e permitiram-me elaborar um estudo retrospectivo para melhor perceber a prática do maneio reprodutivo equino e a influência de alguns parâmetros na taxa de fertilidade das éguas. Relativamente aos diferentes tipos de sémen, verificou-se que a taxa de fertilidade em éguas cobertas por cobrição natural (n=14) foi de 78,6% e por IA (n=36) foi de 94,4%, resultado que se situa dentro dos valores esperados. No que respeita à IA, a taxa de fertilidade com sémen fresco (n=16) foi de 93,8%, com sémen refrigerado (n=19) foi de 94,7% e com sémen congelado (n=1) foi de 100%. Os valores para sémen fresco e refrigerado encontram-se dentro do esperado, sendo que para sémen congelado este é muito superior aos valores reportados na bibliografia, visto ser o tipo de sémen com menores taxas de fertilidade. Este facto deve-se, provavelmente, à dimensão da amostra, e ao facto de o sémen congelado ser de elevada qualidade. Neste estudo verificamos também que não há relação estatística entre a idade das éguas e a taxa de gestação, contrariamente ao que é referido na bibliografia, o que se deverá provavelmente ao tamanho da amostra (n=50).

Características eletrocardiográficas em eqüinos clinicamente normais da raça Puro Sangue Inglês

Determinaram-se as características eletrocardiográficas de amplitude, duração, ritmo e eixo cardíaco no plano frontal de potros, sobreanos e éguas prenhes clinicamente normais, da raça Puro Sangue Inglês. Estudaram-se 50 potros e 50 sobreanos, machos e fêmeas, e 11 éguas prenhes. Concluiu-se que a freqüência cardíaca diminui com a idade e que nos potros e sobreanos as características apresentaram valores intermediários entre os valores padrão de neonatos e adultos. Nos sobreanos eles estão mais próximos dos de referência para adultos. Nas éguas em gestação a única diferença observada com o padrão foi o leve desvio para a direita do eixo cardíaco.

Uso do ácido mefenâmico em receptoras de embriões equinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito de dois períodos de tratamaneto com altrenogest sobre a manutenção da gestação em éguas receptoras acíclicas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Perfil bioquímico sérico de éguas gestantes e não gestantes das raças brasileiro de hipismo e bretão

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência das alterações degenerativas endometriais e da idade hemodinâmica do trato reprodutivo de éguas após a inseminação artificial e durante as fases iniciais do desenvolvimento embrionário

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Utiização de altrenogest injetável par o estabelecimento e manutençao da gestação em éguas receptoras de embriões acíclicas

Pós-graduação em Medicina Veterinária - FMVZ