EFFECTS OF A SYSTEMIC HIGH UREA CONCENTRATION ON THE ENDOMETRIAL AND EMBRYONIC TRANSCRIPTOMES OF THE MARE

Pregnancy loss remains a major source of economic cost to the equine industry. Frequently, the exact causes of pregnancy loss remain unknown. It has been shown, in other species, that increased dietary protein leading to elevated blood urea nitrogen concentrations (BUN) can be a factor in decreased survival of the early embryo. Our studies provided novel information regarding the effects of elevated BUN on endometrium and embryos from mares as well as insights on changes in their gene expression. Our first objective was to develop an experimental model to elevate BUN during diestrus using intravenous urea infusion. We analyzed the effects of an acute elevation in BUN on uterine and vaginal pH along with changes in the endometrial transcriptome of mares with RNA sequencing. There was a significant increase in BUN and a decrease in uterine pH in the urea group compared to the control group. A total of 193 genes were differentially expressed (DEG) between the urea and control groups. The DEG were predicted to be related to cell pH, ion homeostasis, changes in epithelial tissue, fatty acid metabolism, and solute carriers. Our second objective was to evaluate the effects of elevated BUN in the endometrium of mares using a chronic oral urea administration to elevate BUN in mares. Uterine and vaginal pH were evaluated and RNA sequencing of the endometrium was again performed. There was an increase in BUN in the urea-fed mares, but no significant change in uterine or vaginal pH between the groups. A total of 60 DEG were characterized, with prediction of transcriptomic changes in the endometrium of mares related to cell death (necrosis) and cellular movement (invasion of cells). Our third objective was to determine the effects of a high BUN on the transcriptome of day-14 embryos. There was a positive correlation between plasma BUN and blastocoele fluid urea nitrogen concentration. Changes in embryo transcriptome were related to survival of organism, angiogenesis, adhesion, and quantity of cells. Our final objective was to evaluate the correlation between BUN and follicular fluid urea nitrogen and evaluate the survival of embryos collected from donor mares with high BUN concentrations. Urea nitrogen concentration was positively correlated between the plasma and follicular fluid of mares. Additionally, there was a higher pregnancy rate when embryos were collected from mares with lower BUN. Overall, these results further elucidate the mechanisms through which urea affects endometrial and embryonic transcriptome of mares with high BUN, serving to identify effects of a high BUN in the reproductive tract of mares that might lead to decreased fertility

Study of reproductive toxicity of Combretum leprosum Mart and Eicher in female Wistar rats

Most plants culturally used in Brazil for medicine do not have pre-clinical studies of reproductive toxicity, therefore risks of using such products on the reproductive system are unknown. The aim of the study was to evaluate possible reproductive toxicity of ethanolic extract of Combretum leprosum Mart and Eicher (EECL) in female Wistar rats. The animals, weighing between 180 to 250 g, were maintained in controlled environmental conditions of temperature, humidity, light/dark cycle of 12/12 h, water ad libitum and fed with commercial diet for rats. To verify the estrogenic activity of EECL, four groups of ovariectomized rats were used: saline + corn oil; saline + estradiol; EECL (500 mg/kg) + corn oil; EECL (500 mg/kg) + estradiol. To study the reproductive toxicity during the fecundation and implantation phases of the embryos and also during the organogenesis phase, two groups were used for each experiment, saline and EECL (500 mg/Kg). No estrogenic or anti-estrogenic activities were observed in the EECL. The ingestion of EECL did not cause modification in the number of implantation sites, which indicates a lack of toxicity during this phase. The EECL administered orally in the dose of 500 mg/kg did not produce adverse effects on the reproductive system of the female rats.Keywords: Estrogenic activity, organogenesis, teratogensAfrican Journal of Biotechnology Vol. 12(16), pp. 2105-210

Efeito da hCG ou deslorelina sobre a hemodinâmica folicular e perfil endogeno de LH em éguas cíclicas

Ovulation induction after treatment with hCG and GnRH analogs, such as deslorelin, occur through distinct mechanism. Meanwhile, it is possible that these treatments entail changes in follicular vascular perfusion during the pre-ovulatory period. Thus, color Doppler ultrasound presents a potential to be used as a tool for real-time assessment of follicles ovulatory potential in mares. The main purposes of this paper were to evaluate follicular vascular perfusion (FVP) after the use of ovulation induction hormones and the relationship with the plasmatic profile of LH. Additionally, evaluate if FVP can be used to indicate proximity to ovulation. Estrous cycles of mares were randomly divided in three experimental groups: GnRH group, hCG group and control group (n=10 mares/group). Treatments were done in the presence of a follicle with ≥35mm diameter associated with uterine edema. The moment immediately before treatment was considered H0. Blood collection and Doppler ultrasound exam with power-flow mode of the dominant follicle were done to evaluate, respectively, plasmatic concentration of LH and FVP. Parameters were evaluated every hour between H0-H12 and every six hours from H12 until H30. Additionally, GnRH and hCG groups were evaluated every hour from H30 until ovulation detection or until H48. In the control group, B-mode ultrasonography was completed every six hours after H30 and when detection of characteristics indicative of imminent ovulation Doppler ultrasonography was done every hour until ovulation detection. Control group presented a longer interval until ovulation (P < 0,0001). Group effect (P < 0.05) was detected for FVP. Animals from the GnRH group presented higher mean FVP (P < 0.05) when compared to the hCG and control groups since H0 until the moment of ovulation. No moment effect and interaction group:moment was found between experimental groups. Mean LH concentration between H1 and ...A indução da ovulação após o tratamento com hCG e análogos de GnRH, como a deslorelina, ocorrem por mecanismos distintos. Por sua vez, é possível que estes tratamentos acarretem alterações na perfusão vascular folicular durante o período pré-ovulatório. Desta forma, a ultrassonografia colorida Doppler apresenta potencial para ser utilizada como ferramenta na avaliação em tempo real do potencial ovulatório de folículos em éguas. Os objetivos principais deste trabalho foram avaliar as alterações na perfusão vascular folicular (PVF) após o uso de hormônios indutores da ovulação e sua relação temporal com o perfil plasmático de LH. Além de avaliar se a PVF pode ser utilizada para indicar a proximidade da ovulação. Ciclos estrais de éguas foram aleatoriamente divididos em três grupos experimentais: grupo GnRH, grupo hCG e grupo controle (n=10 éguas/grupo). Tratamentos foram realizados na presença de folículo com diâmetro ≥35mm associado à edema uterino. O momento imediatamente antes do tratamento foi considerado H0. Colheita de sangue e exame ultrassonográfico Doppler função power-flow do folículo dominante foram realizados para avaliar, respectivamente, a concentração plasmática de LH e a perfusão vascular folicular (PVF). Os parâmetros foram avaliados de hora em hora entre os intervalos H0-H12 e a cada seis horas da H12 até H30. Adicionalmente, os grupos GnRH e hCG foram acompanhados a cada hora a partir da H30 até a detecção da ovulação ou até H48. Já no grupo controle, foi realizada ultrassonografia modo-B a cada seis horas após H30 e ao se detectar características indicativas da proximidade de ovulação a US Doppler foi realizada de hora em hora até detecção da ovulação. O grupo controle apresentou um intervalo mais longo até à ovulação (P < 0,0001). Efeito de grupo (P < 0.05) foi detectado para PVF. Animais do grupo GnRH apresentaram maior PVF (P < 0.05) média quando ...Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Postnatal Dynamics of Circulating Steroid Hormones in Mule and Equine Neonates.

It is necessary to study hormonal patterns from mules to recognize alterations and neonatal maladaptation. Our objective was to evaluate concentrations of hormones in mule (n = 6) and equine foals (n = 6). Blood was collected at T0, 1, 6 and 12 h after birth. Hormone concentrations were evaluated using liquid chromatography tandem mass spectrometry. Effects of time, group and interactions and regression analysis were evaluated (p &lt; 0.05). There was a cubic and quadratic decline in mule and equine foals, respectively, for 3&beta;,20&alpha;-dihydroxy-DHP. Mule foals were born with lower circulating 3&beta;,20&alpha;-dihydroxy-DHP concentrations, which might be related to progestogen concentrations in mares with a hybrid placenta. Corticosterone and cortisol concentrations remained unchanged for the first hour post-foaling then declined in mule and equine foals (p &lt; 0.0001). Dehydroepiandrosterone was the main androgen present. There was a decrease in dihydrotestosterone at 12 h (p = 0.002). Differences in the temporal patterns of secretion within each steroid class, pregnanes, corticoids, and androgens, suggest they were derived from different tissue sources, presumptively the placenta, adrenals and gonads of the fetus/neonate, respectively. Mule and horse foals were born without evidence of testosterone secretion. For the first time, steroid hormone levels were measured in neonatal mules, and this will provide insight into neonatal physiology that differs from equine and allow us to gain an understanding of mules that have rarely been studied. Further studies are needed to elucidate the effects of hybrid pregnancies in the steroid endocrinology of neonates

Postnatal Dynamics of Circulating Steroid Hormones in Mule and Equine Neonates

It is necessary to study hormonal patterns from mules to recognize alterations and neonatal maladaptation. Our objective was to evaluate concentrations of hormones in mule (n = 6) and equine foals (n = 6). Blood was collected at T0, 1, 6 and 12 h after birth. Hormone concentrations were evaluated using liquid chromatography tandem mass spectrometry. Effects of time, group and interactions and regression analysis were evaluated (p &lt; 0.05). There was a cubic and quadratic decline in mule and equine foals, respectively, for 3β,20α-dihydroxy-DHP. Mule foals were born with lower circulating 3β,20α-dihydroxy-DHP concentrations, which might be related to progestogen concentrations in mares with a hybrid placenta. Corticosterone and cortisol concentrations remained unchanged for the first hour post-foaling then declined in mule and equine foals (p &lt; 0.0001). Dehydroepiandrosterone was the main androgen present. There was a decrease in dihydrotestosterone at 12 h (p = 0.002). Differences in the temporal patterns of secretion within each steroid class, pregnanes, corticoids, and androgens, suggest they were derived from different tissue sources, presumptively the placenta, adrenals and gonads of the fetus/neonate, respectively. Mule and horse foals were born without evidence of testosterone secretion. For the first time, steroid hormone levels were measured in neonatal mules, and this will provide insight into neonatal physiology that differs from equine and allow us to gain an understanding of mules that have rarely been studied. Further studies are needed to elucidate the effects of hybrid pregnancies in the steroid endocrinology of neonates

EFICIÊNCIA DOS DILUIDORES TRIS E BOTU-CRIO® SOBRE OS PARÂMETROS SEMINAIS DE GARANHÕES DAS RAÇAS QUARTO DE MILHA E MANGALARGA MARCHADOR

The limiting factor in the cryopreservation process of equine semen is related to the species, as they present a great variability in the ejaculate’s characteristics after thawing. The aim of this study was to evaluate sperm viability after thawing, from Quarter Horse and Mangalarga Marchador stallions using two extenders (Botu-crio and Tris) in cryopreservation. To this end, we analyzed the physical characteristics of fresh semen, the sperm membrane functionality by the hypoosmotic swelling test (HOST), total motility and vigor by the Thermoresistance Test (TRT), progressive motility by a computerized system CASA (Computer-Assisted Semen Anlyses) and acrosomal membrane integrity and functionality of the mitochondria by epifluorescence microscopy. After thawing, the extender Botu-crio® better preserved motility, vigor and integrity of the plasma membrane. There was no significant difference between breeds for the thermoresistance test after twaing. Quarter Horse showed higher percentage of bigger defects in the sperm pathology analysis

Characterization of Luteal Blood Flow and Secretion of Progesterone in Mares Treated With Human Chorionic Gonadotropin for Ovulation Induction or During Early Diestrus

© 2015 Elsevier Inc. Human chorionic gonadotropin (hCG) has been used to induce ovulation and as a luteotrophic agent in cattle. However, the effect of hCG therapy on the functional status of the equine corpus luteum (CL) is unclear. This study aimed to characterize the hemodynamic and secretory function of early CL of mares treated with different doses of hCG at distinct stages of the estrous cycle. Mares were assigned to nine experimental groups (n = 6 mares/group) according to dose of hCG and time of treatment. A single injection of one of three different doses of hCG (750, 1,500, or 2,500 IU) was performed in one of three distinct stages of the estrous cycle: preovulatory follicle ≥35mm, day of ovulation (D0), or 48hours after ovulation (D2). In addition, a control group treated with NaCl 0.9% was included in the study. The end points evaluated daily from D0 to D8 were area of the CL, luteal vascularity, number of colored pixels and total pixel intensity, and concentrations of plasma progesterone (P4). No effect (. P \u3e .1) of dose or time of treatment was observed for any end point, within each day. Luteal area did not differ throughout the days (. P \u3e .1), whereas Doppler parameters and concentrations of plasma P4 presented a progressive increase (. P \u3c .05) after ovulation in all groups. Secretory function and luteal hemodynamic were not affected (. P \u3e .1) by hCG dose and time of treatment. In conclusion, hCG therapy during estrus or early diestrus, at the doses tested, did not improve P4 secretion or luteal blood flow

Characterization of Luteal Blood Flow and Secretion of Progesterone in Mares Treated With Human Chorionic Gonadotropin for Ovulation Induction or During Early Diestrus

© 2015 Elsevier Inc. Human chorionic gonadotropin (hCG) has been used to induce ovulation and as a luteotrophic agent in cattle. However, the effect of hCG therapy on the functional status of the equine corpus luteum (CL) is unclear. This study aimed to characterize the hemodynamic and secretory function of early CL of mares treated with different doses of hCG at distinct stages of the estrous cycle. Mares were assigned to nine experimental groups (n = 6 mares/group) according to dose of hCG and time of treatment. A single injection of one of three different doses of hCG (750, 1,500, or 2,500 IU) was performed in one of three distinct stages of the estrous cycle: preovulatory follicle ≥35mm, day of ovulation (D0), or 48hours after ovulation (D2). In addition, a control group treated with NaCl 0.9% was included in the study. The end points evaluated daily from D0 to D8 were area of the CL, luteal vascularity, number of colored pixels and total pixel intensity, and concentrations of plasma progesterone (P4). No effect (. P \u3e .1) of dose or time of treatment was observed for any end point, within each day. Luteal area did not differ throughout the days (. P \u3e .1), whereas Doppler parameters and concentrations of plasma P4 presented a progressive increase (. P \u3c .05) after ovulation in all groups. Secretory function and luteal hemodynamic were not affected (. P \u3e .1) by hCG dose and time of treatment. In conclusion, hCG therapy during estrus or early diestrus, at the doses tested, did not improve P4 secretion or luteal blood flow