Impact of high-fat diet consumption during prolonged period of pregnancy on placenta structures and umbilical vascular growth in goats

This study aimed to verify the impact of high-fat diet consumption for a prolonged period on oxidative stress, fetal growth, umbilical vascular system, and placental structures in pregnant goats. Twenty-two pregnant goats were grouped into the control diet (n= 11) and fat diet (n = 11). Flaxseed meal was added to the fat diet, replacing the corn grain of concentrate, from gestational day 100 to delivery date. Diets were isonitrogenous and isoenergetic, differing in fat content (2.8% vs. 6.3% dry matter). The fat group showed higher feed intake and total plasma lipid levels than the control group (P < 0.001). No difference was found in placentome, and umbilical vascular development. Fat diet-fed goats exhibited a lower systolic peak in the umbilical artery. At delivery, placental traits were similar with the exception of the cotyledon width (P = 0.0075), which was smaller in the fat group and cotyledon surface (P = 0.0047) for multiple pregnancy of fat diet. Cotyledonary epithelium showed more intense staining of lipid droplets and a greater area for lipofuscin staining in the fat group compared to control group (P < 0.001). The mean live weight of the kids was lower in the fat group in the first week after delivery than in control group. Thus, in goats, the continuous administration of a high-fat diet during pregnancy does not appear to modify the fetal-maternal vascular structures but has an impact on a part of the placental structure; therefore, its use must be carefully evaluated.(AU)

Glycerin supplementation strategies for three or seven days affects oxidative stress, follicle dynamics and ovulatory response in Morada Nova sheep

The Ziwuling black goat is an indigenously in China, their offspring are frequently affected by congenital cryptorchidism. The extracellular matrix (ECM) contains cytokines and growth factors that regulate the development of the testis, and component changes often result in pathological changes. Cryptorchidism is closely related to structural changes in ECM. In this study, the histochemical staining, immunohistochemical, immunofluorescence and Western blot combined with semi-quantitative analysis was used to describe the distribution of the important ECM components Collagen type IV (Col IV), laminin (LN)and heparan sulfate proteoglycans (HSPG) in the normal and cryptorchid testes of Ziwuling black goats. Results showed that: The histochemical staining showed that the dysplasia of seminiferous tubules and decreased number of Sertoli cells in cryptorchidism, as well as sparse collagen fiber. Meanwhile, the distribution of reticular fibers is relatively rich. Furthermore, the PAS and AB staining in the interstitial vessels and lamina propria of seminiferous tubules is weak. The immunohistochemical and immunofluorescence revealed that Col IV, LN was strongly expressed in Leydig, Sertoli cells of normal testes and moderately positive in the spermatogonia and spermatids, but HSPG was not expressed in the spermatogonia. However, cryptorchidism, the expression of Col IV, LN and HPSG in Leydig, Sertoli cells significantly decreased, as well as the expression of Col IV and LN in capillary endothelial cells, but HSPG was moderately expressed in spermatogonia. Based on these data, the underdevelopment of spermatogenic epithelium, decreased synthesis function of collagen fibers and Leydig cells develop usually in the cryptorchidism were shown to be closely related to the abnormal metabolism of Col IV and LN. The positive expressed of HSPG in the spermatogonia of cryptorchid testes is related to the compensatory development of spermatogonia.(AU)

Effectiveness of near-infrared spectroscopy as a non-invasive tool to discriminate spectral profiles of in vitro cultured oocytes from goats

Abstract Here, we aimed to discriminate between the spectral profiles of spent culture media after oocyte in vitro maturation (IVM) and culture (IVC) from goats of different ages subjected to repeated hormonal treatments. The profiles were discriminated using near infrared (NIR) spectroscopy combined with multivariate methods. A total of 19 goats (young = 10; old = 9) were subjected to serial hormonal stimulation (HS) with gonadotropins. Cumulus oophorus complexes (COCs) were collected using laparoscopic ovum pick-up (LOPU) and subjected to IVM and parthenogenetic activation. The initial embryos were subjected to IVC. Spent culture media were collected after oocyte IVM and on day 2 of IVC and analyzed using NIR spectroscopy. NIR spectral data were interpreted through chemometric methods, such as principle component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The results of PCA analysis clearly showed a separation in the spectral profiles between the experimental groups (HS sessions; young and old animals) both after IVM and IVC. Overall, the main absorption bands were attributed to the C-H group second overtone, first overtone of O-H and N-H, and C-H combinations and may serve as molecular markers. On the other hand, the spectral data obtained using PLS-DA models provided a better classification of the groups. The results showed the possibility of discriminating young and old groups as well as the three HS sessions with high specificity, sensitivity, and accuracy using NIR spectra. Thus, the culture medium analysis using NIR spectroscopy combined with multivariate methods indicated the dissimilarities between the groups and provided an insight into the in vitro development of goat oocytes. This technique serves as an efficient, objective, rapid, and non-invasive method to discriminate spectral profiles.

Effectiveness of near-infrared spectroscopy as a non-invasive tool to discriminate spectral profiles of in vitro cultured oocytes from goats

Here, we aimed to discriminate between the spectral profiles of spent culture media after oocyte in vitro maturation (IVM) and culture (IVC) from goats of different ages subjected to repeated hormonal treatments. The profiles were discriminated using near infrared (NIR) spectroscopy combined with multivariate methods. A total of 19 goats (young = 10; old = 9) were subjected to serial hormonal stimulation (HS) with gonadotropins. Cumulus oophorus complexes (COCs) were collected using laparoscopic ovum pick-up (LOPU) and subjected to IVM and parthenogenetic activation. The initial embryos were subjected to IVC. Spent culture media were collected after oocyte IVM and on day 2 of IVC and analyzed using NIR spectroscopy. NIR spectral data were interpreted through chemometric methods, such as principle component analysis (PCA) and partial least square discriminant analysis (PLS-DA). The results of PCA analysis clearly showed a separation in the spectral profiles between the experimental groups (HS sessions; young and old animals) both after IVM and IVC. Overall, the main absorption bands were attributed to the C-H group second overtone, first overtone of O-H and N-H, and C-H combinations and may serve as molecular markers. On the other hand, the spectral data obtained using PLS-DA models provided a better classification of the groups. The results showed the possibility of discriminating young and old groups as well as the three HS sessions with high specificity, sensitivity, and accuracy using NIR spectra. Thus, the culture medium analysis using NIR spectroscopy combined with multivariate methods indicated the dissimilarities between the groups and provided an insight into the in vitro development of goat oocytes. This technique serves as an efficient, objective, rapid, and non-invasive method to discriminate spectral profiles.(AU)

Impact of hypotensive action of angiotensin converting enzyme inhibitor on ovarian-intraovarian blood flow and follicles development in goats hormonally stimulated with repeated FSH-ECG treatment

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductiveprocesses, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed touse an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however,the presence of the main components of this system in reproductive tissues still need to be further investigated, since thephysiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate,an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats.Materials, Methods & Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n= 10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 ofenalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received thesame volume of saline solution. Estrus synchronization of all goats was made by intramuscular ad-ministration of PGF2αanalog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight h after devicewithdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetricsparameters of the ovarian artery and intraovarian blood flow, respec-tively, and B-mode real-time ultrasound scanner toevaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction ofsystolic and diastolic peak, without difference according to parity. In addition, in the third session of hor-monal stimulation,only the groups (nulliparous and multiparous) not treated with...(AU)

Impact of hypotensive action of angiotensin converting enzyme inhibitor on ovarian-intraovarian blood flow and follicles development in goats hormonally stimulated with repeated FSH-ECG treatment

Background: Recent evidence shows that the renin-angiotensin system (RAS) participates in important reproductiveprocesses, such as steroidogenesis, folliculogenesis, oocyte maturation and ovulation. Several studies have proposed touse an angiotensin-converting enzyme (ACE) as a RAS modulator, aiming to improve reproductive efficiency, however,the presence of the main components of this system in reproductive tissues still need to be further investigated, since thephysiological functions seem to be species-specific. The aim of this study was to assess the impact of enalapril-maleate,an ACE inhibitor, during repeated gonadotropins treatment on ovarian blood flow and follicular development in goats.Materials, Methods & Results: Twenty Anglo-Nubian cross-bred goats were equally grouped according to parity (n= 10/group): nulliparous and multiparous parity. In each group, five animals were randomly selected to receive 0.4 mg.kg-1 ofenalapril-maleate during 11 days of estrus synchronization and gonadotropins treatments. The other animals received thesame volume of saline solution. Estrus synchronization of all goats was made by intramuscular ad-ministration of PGF2αanalog, followed 48 h later by intravaginal insertion of a controlled internal drug release device. Forty-eight h after devicewithdrawal, a single dose of 60 mg of FSH plus 300 UI of eCG was administered and repeated every 4 days to complete 3treatments. Transrectal ultrasonography was performed using pulsed and color Doppler to evaluate Doppler velocimetricsparameters of the ovarian artery and intraovarian blood flow, respec-tively, and B-mode real-time ultrasound scanner toevaluate the follicular development. In the females treated with enalapril-maleate was observed a significant reduction ofsystolic and diastolic peak, without difference according to parity. In addition, in the third session of hor-monal stimulation,only the groups (nulliparous and multiparous) not treated with...

Utilização de um sistema completamente portátil na produção in vitro de embriões bovinos / Use of a completely portable in vitro system in production of bovine embryos

A produção in vitro de embriões (PIVE) bovinos no Brasil é dependente de transporte de oócitos das fazendas para laboratórios, sendo útil um sistema de incubação que mantenha a competência ao desenvolvimento. Assim, comparou-se a dois sistemas de incubação (mini vs portátil) quanto à qualidade dos embriões produzidos. Oócitos bovinos foram maturados, fecundados e cultivados in vitro. Além disso, foi verificada a produção de espécies reativas de oxigênio (EROs). As taxas de clivagem e blastocisto foram similares (p>0,05) para ambos os sistemas. Os níveis de EROs foram significativamente maiores (p<0,05) em embriões de dia 2 produzidos na incubadora mini. Em conclusão, o sistema de incubação portátil mostrou ser uma alternativa viável para PIVE bovina.

The in vitro production of bovine embryos (PIVE) in Brazil, is dependent on oocyte transport from farms to laboratories, and an incubation system that maintains developmental competence is useful. Thus, it was compared to two incubation systems (mini vs portable) regarding the quality of the embryos produced. Bovine oocytes were matured, fertilized and cultured in vitro. In addition, the production of reactive oxygen species (ROS) was verified. The cleavage and blastocyst rates were similar (p>0.05) for both systems. ERO levels were significantly higher (p<0.05) in day 2 embryos produced in the mini incubator. In conclusion, the portable incubation system proved to be a viable alternative for bovine PIVE.

Efeito de diferentes sistemas de maturação e cultivo sobre a produção in vitro de embriões bovinos / Effect of different maturation and growth systems on in vitro production of bovine embryos

O objetivo desse estudo foi comparar dois diferentes sistemas de incubação (incubadora convencional – CONV e minibancada – MINI) sobre a produção in vitro de embriões (PIVE) de bovinos. Para tanto, complexos cumulusoócito (CCOs) foram submetidos a maturação in vitro e posterior fecundação e cultivo in vitro em ambos os sistemas. As estruturas foram avaliadas pós-maturação e pós-cultivo, para avaliação de maturação nuclear e quantidade de células/blastocisto, respectivamente. Não foram observadas diferenças estatísticas (p>0,05) nos diferentes parâmetros estudados. Concluímos que ambos os sistemas testados demonstraram ser eficientes para PIVE de bovinos.

The objective of this study was to compare two different incubation systems (conventional incubator - CONV and minibank - MINI) on the in vitro production of bovine embryos (PIVE). For this purpose, cumulus cytotoxic complexes (CCOs) were submitted to in vitro maturation and subsequent fertilization and in vitro culture in both systems. The structures were evaluated post-maturation and post-culture, for evaluation of nuclear maturation and amount of cells / blastocyst, respectively. No statistical differences (p>0.05) were observed in the different parameters studied. We conclude that both tested systems proved to be efficient for bovine PIVE.

Utilização de um sistema completamente portátil na produção in vitro de embriões bovinos / Use of a completely portable in vitro system in production of bovine embryos

A produção in vitro de embriões (PIVE) bovinos no Brasil é dependente de transporte de oócitos das fazendas para laboratórios, sendo útil um sistema de incubação que mantenha a competência ao desenvolvimento. Assim, comparou-se a dois sistemas de incubação (mini vs portátil) quanto à qualidade dos embriões produzidos. Oócitos bovinos foram maturados, fecundados e cultivados in vitro. Além disso, foi verificada a produção de espécies reativas de oxigênio (EROs). As taxas de clivagem e blastocisto foram similares (p>0,05) para ambos os sistemas. Os níveis de EROs foram significativamente maiores (p<0,05) em embriões de dia 2 produzidos na incubadora mini. Em conclusão, o sistema de incubação portátil mostrou ser uma alternativa viável para PIVE bovina.(AU)

The in vitro production of bovine embryos (PIVE) in Brazil, is dependent on oocyte transport from farms to laboratories, and an incubation system that maintains developmental competence is useful. Thus, it was compared to two incubation systems (mini vs portable) regarding the quality of the embryos produced. Bovine oocytes were matured, fertilized and cultured in vitro. In addition, the production of reactive oxygen species (ROS) was verified. The cleavage and blastocyst rates were similar (p>0.05) for both systems. ERO levels were significantly higher (p<0.05) in day 2 embryos produced in the mini incubator. In conclusion, the portable incubation system proved to be a viable alternative for bovine PIVE.(AU)

Efeito de diferentes sistemas de maturação e cultivo sobre a produção in vitro de embriões bovinos / Effect of different maturation and growth systems on in vitro production of bovine embryos

O objetivo desse estudo foi comparar dois diferentes sistemas de incubação (incubadora convencional CONV e minibancada MINI) sobre a produção in vitro de embriões (PIVE) de bovinos. Para tanto, complexos cumulusoócito (CCOs) foram submetidos a maturação in vitro e posterior fecundação e cultivo in vitro em ambos os sistemas. As estruturas foram avaliadas pós-maturação e pós-cultivo, para avaliação de maturação nuclear e quantidade de células/blastocisto, respectivamente. Não foram observadas diferenças estatísticas (p>0,05) nos diferentes parâmetros estudados. Concluímos que ambos os sistemas testados demonstraram ser eficientes para PIVE de bovinos.(AU)

The objective of this study was to compare two different incubation systems (conventional incubator - CONV and minibank - MINI) on the in vitro production of bovine embryos (PIVE). For this purpose, cumulus cytotoxic complexes (CCOs) were submitted to in vitro maturation and subsequent fertilization and in vitro culture in both systems. The structures were evaluated post-maturation and post-culture, for evaluation of nuclear maturation and amount of cells / blastocyst, respectively. No statistical differences (p>0.05) were observed in the different parameters studied. We conclude that both tested systems proved to be efficient for bovine PIVE.(AU)

Efeito de diferentes meios e concentrações espermáticas na fecundação in vitro curta sobre a produção de embriões bovinos / Effect of different mediums and sperm concentrations in the short-in vitro fertilization on the bovine embryo production

tThe present study investigated the effect of two different mediums (IVF-TALP and BO) and spermconcentrations (16  106 and 20  106), during the short-in vitro fertilization (IVF) on bovine embryoproduction. Thus, after in vitro maturation (IVM), oocytes were randomly distributed into three groups:traditional IVF (control), IVF-TALP and Brackett-Oliphant (BO). The groups IVF-TALP and BO weresubdivided according sperm concentration (16  106 and 20  106) used for short-IVF (6 h). For the controlgroup, the IVF was performed for 18 h. Concerning the cleavage rate, the BO-20 group showed the best results(77,1%). On the other hand, for the blastocyst yield, the highest results were obtained in control and BO-20groups: 20.8% and 24.6%, respectively. In conclusion, it seems that BO medium with 20  106 sptz/mL positivelyinfluence the bovine embryo in vitro production. However, further studies should be performed.(AU)

Avaliação do desenvolvimento in vitro de embriões bovinos após exposição à crotamina linear / Evaluation of bovine embryo in vitro development after exposure to linear crotamine

The present study investigated the effects of different concentrations of linear crotamine (CrL), used asagent for cell transfection, on in vitro development of bovine embryos. Embryos were exposed to 0; 5 and 12.5µM of CrL in Synthetic Oviductal Fluid (SOF) for 6 h. Vehicle solution was NaCl 150 mM and not exposedembryos (in vitro fertilized-IVF group) was included. In vitro developmental competence was evaluated bycleavage (day 2), blastocyst (day 7 and 8), and hatching (day 8) rates. No difference (P > 0.05) was observedamong CrL groups and control group (IVF) for all observed parameters. In conclusion, the use of the tested CrLconcentrations for 6 h during in vitro culture of bovine embryos have no deleterious effects on embryodevelopment.(AU)

Avaliação do desenvolvimento in vitro de embriões bovinos após exposição à crotamina linear / Evaluation of bovine embryo in vitro development after exposure to linear crotamine

The present study investigated the effects of different concentrations of linear crotamine (CrL), used asagent for cell transfection, on in vitro development of bovine embryos. Embryos were exposed to 0; 5 and 12.5µM of CrL in Synthetic Oviductal Fluid (SOF) for 6 h. Vehicle solution was NaCl 150 mM and not exposedembryos (in vitro fertilized-IVF group) was included. In vitro developmental competence was evaluated bycleavage (day 2), blastocyst (day 7 and 8), and hatching (day 8) rates. No difference (P > 0.05) was observedamong CrL groups and control group (IVF) for all observed parameters. In conclusion, the use of the tested CrLconcentrations for 6 h during in vitro culture of bovine embryos have no deleterious effects on embryodevelopment.

Efeito de diferentes meios e concentrações espermáticas na fecundação in vitro curta sobre a produção de embriões bovinos / Effect of different mediums and sperm concentrations in the short-in vitro fertilization on the bovine embryo production

tThe present study investigated the effect of two different mediums (IVF-TALP and BO) and spermconcentrations (16  106 and 20  106), during the short-in vitro fertilization (IVF) on bovine embryoproduction. Thus, after in vitro maturation (IVM), oocytes were randomly distributed into three groups:traditional IVF (control), IVF-TALP and Brackett-Oliphant (BO). The groups IVF-TALP and BO weresubdivided according sperm concentration (16  106 and 20  106) used for short-IVF (6 h). For the controlgroup, the IVF was performed for 18 h. Concerning the cleavage rate, the BO-20 group showed the best results(77,1%). On the other hand, for the blastocyst yield, the highest results were obtained in control and BO-20groups: 20.8% and 24.6%, respectively. In conclusion, it seems that BO medium with 20  106 sptz/mL positivelyinfluence the bovine embryo in vitro production. However, further studies should be performed.

Avaliação da maturação nuclear de oócitos bovinos submetidos a três diferentes sistemas de incubação / Evaluation of nuclear maturation of bovine oocytes submitted to three different incubation systems

This study aimed to evaluate three different incubation systems in nuclear maturation of bovine oocytes.Follicles of 2 to 8 mm in diameter were aspirated for obtaining cumulus-oocyte complexes (COCs). Thereafter,COCs were subjected to in vitro maturation (IVM) in TCM-199 medium supplemented at 38.5 °C for 23 h indifferent incubation systems: bench incubator with high oxygen (20% O2 in air), bench incubator with lowoxygen (5% O2) and portable incubator with low oxygen. After IVM, cumulus cells were removed and oocyteswere stained with Hoechst 33342. The slides were analyzed by a fluorescence microscope and the oocytes wereclassified into: degenerated (DG), metaphase I (MI) and metaphase II (MII). No difference (P > 0.05) wasobserved among the rates of DG, MI and MII for the different incubation systems. Therefore, the use of portableincubation system is a viable alternative for in vitro maturation of bovine oocytes.(AU)

Avaliação da maturação nuclear de oócitos bovinos submetidos a três diferentes sistemas de incubação / Evaluation of nuclear maturation of bovine oocytes submitted to three different incubation systems

This study aimed to evaluate three different incubation systems in nuclear maturation of bovine oocytes.Follicles of 2 to 8 mm in diameter were aspirated for obtaining cumulus-oocyte complexes (COCs). Thereafter,COCs were subjected to in vitro maturation (IVM) in TCM-199 medium supplemented at 38.5 °C for 23 h indifferent incubation systems: bench incubator with high oxygen (20% O2 in air), bench incubator with lowoxygen (5% O2) and portable incubator with low oxygen. After IVM, cumulus cells were removed and oocyteswere stained with Hoechst 33342. The slides were analyzed by a fluorescence microscope and the oocytes wereclassified into: degenerated (DG), metaphase I (MI) and metaphase II (MII). No difference (P > 0.05) wasobserved among the rates of DG, MI and MII for the different incubation systems. Therefore, the use of portableincubation system is a viable alternative for in vitro maturation of bovine oocytes.