Effect of one or two fixed glutamate doses on follicular development, ovarian-intraovarian blood flow, ovulatory rate, and corpus luteum quality in goats with a low body condition score.

This study aimed to investigate the effect of a short glutamate supply on the ovarian response in goats with low body condition scores. Twenty-one goats had their estrus and follicular waves synchronized using three injections of prostaglandin analog at seven-day intervals. Goats were allocated to groups receiving 10 mg/kg LW (iv) of glutamate administered in a single dose (group LBCG1, n = 7) or in two doses five days apart (group LBCG2, n = 7). The control group (LBC; n = 7) received saline solution. Glutamate treatment did not affect glucose, cholesterol, or glutathione peroxidase levels, body weight, or adipose deposits. During the experimental period, the LBCG2 group showed a higher (P < 0.05) number of follicles (> 3 mm) and an increase in follicle diameter (P < 0.05). Glutamate supply improved (P < 0.05) the intraovarian Doppler blood area size in the LBCG groups, and the second dose in LBCG2 also induced a higher (P < 0.05) systolic and diastolic peak of the ovary artery. After ovulation induction, LBCG2 exhibited a high (P < 0.05) volume of the corpus luteum and vascularized area. We concluded that the supply of two doses of glutamate five days apart was efficient in ovarian stimulation in goats with a low body condition.

Use of monosodium-glutamate as a novel dietary supplement strategy for ovarian stimulation in goats.

This study aimed to investigate the reproductive effects of adding monosodium glutamate (MSG) to the diet of goats. Eleven adult goats received synchronized estrus and follicular waves using three prostaglandin analog injections every seven days. Goats allocated to individual pens received 1 g/kg BW of MSG in their diet for 23 days (MOGLU group, n = 6), whereas the control group (n = 5) maintained the base diet. The supplemented animals showed an increase in dry matter intake (P < 0.0001) and a reduction in heart rate (P < 0.05), respiratory rate, and ruminal movement (P < 0.001). Surface and rectal temperatures were higher in the MOGLU group, (P < 0.0001) with a significant increase in the afternoon. There was an increase (P < 0.05) in the frequency of behaviors related to rumination, defecation, and urination in the MOGLU group, and a reduction in behaviors associated with stress (P < 0.05). No differences were observed in the plasma levels of proteins, albumin, urea, cholesterol, or triglycerides. Glucose levels were lower (P < 0.05) in the MOGLU group, which also showed increased glutathione peroxide levels during the induction of ovulation. Supplemented animals recorded a larger number (P < 0.05) of follicles throughout the experimental period and higher intraovarian blood perfusion (P < 0.05) during ovulation induction. We conclude that MSG exerts a positive effect on the reproductive response in goats and therefore represents an effective nutritional supplement.

Use of green microalgae Chlorella as a nutritional supplement to support oocyte and embryo production in goats.

This study aimed to evaluate the use of green microalgae as a nutritional supplement for oocyte and embryo production in goats. Two experiments were performed on adult goats to obtain oocytes (EVO; n = 14) and in vivo embryos (IVD; n = 14). In both, the donors were divided into control (n = 7) and Chlorella (n = 7) groups. All goats received a base diet, and donors were orally supplemented with Chlorella pyrenoidosa (CH) in the Chlorella groups. For EVO, donors received 10 g CH for 14 days, and for IVD, 20 g CH was given for six days before embryo recovery. In EVO and IVD, food intake in the CH group was comparatively low, and it showed relatively high subcutaneous adipose deposition. In addition, the CH group exhibited an increase in triglyceride, cholesterol, and plasma glucose levels. In IVD, a significant increase in peripheral glutathione peroxidase levels was noticed. In EVO, the CH group showed relatively large follicular size and an increase in intrafollicular levels of triglycerides, glucose, and glutathione peroxidase. No differences were observed in the oocyte collected, and CH oocytes showed a low intensity of MitoTracker fluorescence (MT). In IVD, the CH group had a high proportion of transferable embryos, and these structures exhibited high fluorescence intensities for MT and H2DCFDA probes. We concluded that under these conditions, CH did not enhance the quality of the recovered oocytes. However, a daily dose of 20 g CH improved the quality of embryos and stimulated their mitochondrial functionality.

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.

Effect of short-term nutritional supplementation of green microalgae on some reproductive indicators of Anglo-Nubian crossbred goats.

Background and Aim: Despite the wide spectrum of uses, one of the chief drawbacks to expanding microalgae as a food supplement in livestock is the lack of a regimen protocol with established dosage and time length of supplementation. Therefore, this study aimed to investigate the effect of short-term supplementation with increasing doses of microalgae on ovarian response in goats reared in northeast Brazil. Materials and Methods: Twenty-eight goats had their follicular waves synchronized using three injections of a prostaglandin analog at 7-day intervals. Goats were allocated to groups that received daily oral Chlorella supplementation for 7 days, respectively: 5 g, GMA5 group (n = 7), 10 g (GMA10; n = 7), and 20 g (GMA20; n = 7). The control group (GMA 0; n = 7) received a drench of water. Results: The groups showed a quadratic increase (p = 0.0156) in kidney fat thickness but there was a significant reduction in dry matter intake in the GMA20 group. The GMA20 group showed higher glucose levels and glutathione peroxidase (p < 0.05). There was a decrease in plasma cholesterol (p < 0.05) in the 10 and 20 g treatments. The number of total follicles increased quadratically. Follicles <3 mm increased linearly (p = 0.0113) for microalgal supply. The GMA10 and GMA20 groups had the highest values (p < 0.05) among the treatments. After inducing ovulation, there was a significant increase in follicles >3 mm in the GMA10 group, which also showed a greater (p < 0.05) area of intraovarian blood perfusion and pulsatility index of the ovarian artery. Conclusion: We conclude that for 7 days of supplementation, the administration of 10 g of microalgae appears to be the most efficient dosage for stimulating the ovarian response in tropical goats.

Dyslipidemia induced by lipid diet in late gestation donor impact on growth kinetics and in vitro potential differentiation of umbilical cord Wharton's Jelly mesenchymal stem cells in goats.

Mesenchymal stem cells (MSC) from the umbilical cord (UC) have several attractive properties for clinical use. This study aimed to verify the impact of a lipid-rich diet during late gestation of donor goats on the growth and differentiation of MSCs from UC. From the 100th day of pregnancy to delivery, 22 goats were grouped based on their diet into the donor-lipid (DLD; n = 11) and donor-baseline (DBD; n = 11) diet groups. Diets were isonitrogenous and isoenergetic, differing in fat content (2.8% vs. 6.3% on a dry matter basis). Wharton's jelly (WJ) fragments were cultured. After primary culture, samples of WJ-MSCs were characterized by the expression of CD90, CD73, CD34, CD45, CD105, and Fas genes, mitochondrial activity using MitoTracker (MT) fluorescence probe, and growth kinetics. Population doubling time (PDT) was also determined. WJ-MSCs were differentiated into chondrocytes, adipocytes and osteocytes, and the mineralized area and adipocytes were determined. The lipid diet significantly increased triglyceride and cholesterol levels during pregnancy. The DLD group showed sub-expression of the CD90 gene, a high MT intensity, and a low proliferation rate at the end of the subculture. The mean PDT was 83.9 ± 1.3 h. Mineralized area and lipid droplet stain intensity from osteogenic and adipogenic differentiations, respectively, were greater in DLD. We conclude that in donor goats, dietary dyslipidemia during late pregnancy affects the ability of UC-derived MSCs to express their developmental potential in vitro, thus limiting their possible use for therapeutic purposes.

Effect of peri-conception high-fat diets on maternal ovarian function, foetal and placentome growth, and vascular umbilical development in goats.

The objective of this study was to determine whether a high-fat diet (HFD) fed to goats for a brief period during peri-conception would optimize reproductive and foetal responses. Thirty-four Anglo-Nubian crossbred adult goats were allocated into three groups: control (n = 11), fed with a total mixed ration (TMR) based on chopped elephant grass and concentrate; HFBM (n = 11), given TMR supplemented with soybean oil on a 0.5% dry matter basis for 11 days starting nine days before mating (BM); and HFAM (n = 12), fed with soybean oil included in the TMR for 15 days after mating (AM). The TMR diets differed in their fat content (7.5% vs. 2.9%). All goats had oestrus synchronized for 14 days BM by intravaginal administration of 60 mg MPA sponge for 12 days. Forty-eight hours BM, the sponge was removed and 0.075 mg PGF2α was applied intramuscularly. After 36 h, 1 ml GnRH was administered intramuscularly, and goats were mated after sponge removal. The fat groups showed lower feed intake (p < .001) and higher cholesterol levels (p < .001) when HFD was administered. Doppler and B-mode ultrasound evaluations revealed a greater (p < .05) number of small (<3 mm, 10 ± 0.6 vs. 8 ± 0.5) and large (≥3 mm, 6 ± 0.4 vs. 5.0 ± 0.2) follicles and intraovarian blood area (p < .05) in the HFBM group during sponge removal (57.6%) and mating (24.2%) than those of the no-fat group. During AM, the fat-fed groups exhibited higher glutathione peroxidase levels (p < .05) and a reduction (p < .001) in corpus luteum size (19%) and vascularized Doppler area (41%). No difference (p > .05) between groups was found in foetal traits, placentome and umbilical vascular development, except for the embryonic vesicle where HFAM twin pregnancy showed a smaller size than the control (26.1 ± 3.5 cm vs. 33.7 ± 2.7 cm; p < .01). Thus, HFD applied during peri-conception of goats has no impact on later foetal development but improved the follicular growth when given before the mating. Thus, the use of HFD in periconception has no impact on foetal development but increases follicular growth before breeding time.

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

This study examined the effect of glycerin supply strategies in different short-term protocols on follicular dynamics and ovulatory rate in Morada Nova sheep. Eighteen Morada Nova ewes with body condition > 2.9 had their estrus and follicular waves synchronized using three injections of prostaglandin analogue at seven-day intervals. All animals received the same diet during 21 days, which consisted of a total mixed ration (TMR) based on chopped elephant grass and concentrate twice daily. In the control group (n=9), ewes were fed the TMR diet. In the other four groups, ewes received 150 mL of glycerol daily, supplied as an oral drench or mixed in the TMR during three or seven days prior to the application of the third PGF2 alfa analogue. These groups were named as follows: Drench3d (n=10), Drench7d (n=8), TMR3d (n=9) and TMR7d (n=9). Follicle dynamics were monitored by ultrasonography, and plasma glucose and glutathione peroxidase levels were measured at the third prostaglandin administration. Six days after the final PGF2 alfa analogue dose, ovulatory rate was measured by laparoscopy. Glucose was higher (P< 0.001) in the glycerin-treated groups than in control group (83.7 ± 1.7 vs. 68.4 ± 4.5 mg. dL-1; P < 0.001). Ewes in the TMR3d, Drench7d and TMR7d groups had a greater (P < 0.001) number of large follicles (≥ 3 < 5 mm), and the presence of follicles larger than 5 mm was observed. In the same groups, at the third PGF2 alfa analogue dose, a greater (P < 0.001) number of growing follicles (> 3 mm) and a larger size of the largest follicle (P < 0.001) were also recorded. Ovulation rate was 30% higher in the groups that received glycerin for seven days (1.6 ± 0.1 53 vs. 1.1 ± 0.1; P < 0.05), and they also exhibited a 38% reduction in glutathione peroxidase. Thus, the use of glycerin in Morada Nova sheep as a source of energy in short-term supplementation for increase ovulation rate is an efficient strategy when provided for seven days, either orally or in the feed.

Impact of donor nutritional balance on the growth and development of mesenchymal stem cells from caprine umbilical cord Wharton´s jelly.

Mesenchymal stem cells (MSCs) from the umbilical cord (UC) have aroused considerable interest. However, little is known about the maternal effect on these cells. The aim of this study was to verify the impact of the nutritional status of donor goats on the growth and differentiation of MSCs from the UC. At parturition, 19 goats were grouped based on their low or high body mass index (low BMI, LBMI, n = 9; and high BMI, HBMI, n = 10). UCs were collected during delivery and Wharton's jelly (WJ) fragments cultured. WJ-MSCs were differentiated into osteocytes, adipocytes, chondrocytes, and the population doubling time (PDT) was determined. Samples of WJ-MSCs were also used to verify the expression of the CD90, CD73, CD34, CD45, and CD105 genes. Media used for WJ-MSC primary cultures were analyzed using near-infrared spectroscopy. The lag phase was 7.5 ± 0.6 days and the entire culture took 26.7 ± 1.3 days, with a cell proliferation rate of 8.500 cells/day. The mean PDT from subculture was 30.0 ± 0.7 h. The CD105 gene was sub-expressed in LBMI, and the spectra of the spent media from the second to fourth day of WJ-MSC primary culture were segregated into negative scores by multivariate analysis. We conclude that, in goats, the nutritional balance of the donor did not affect the in vitro growth of MSCs derived from the UC. However, the molecular profile observed in the low BMI group suggests that the use of MSCs for therapeutic purposes should be considered more carefully.

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.

Impact of short nutrient stimuli with different energy source on follicle dynamics and quality of oocyte from hormonally stimulated goats.

The aim of this study was to verify the effect of the energy source for a short-term diet supplementation on follicular dynamics, ovarian response and oocyte recovery in goats. Thirty Anglo Nubian crossbred does received a diet for 4 weeks to satisfy the nutritional requirements of breeding for adult non-dairy goats. Seven days prior to oocyte recovery (OR), a group of does (n = 10) was supplemented with ground full-fat linseed in the diet (Diet A), whereas a second group of does (n = 10) received crude glycerine in the diet (Diet B). The total mixed ration (TMR) diet was maintained as the Control Diet (n = 10). All animals were oestrous-synchronized by the use of a progesterone insert for 12 days prior to OR. Follicles were stimulated by using pFSH (five 40-mg/ml doses) during the supplementation time. At OR, follicles were counted and recovered oocytes were classified as viable or degenerated. Follicular dynamics was monitored by ultrasonography, and plasma glucose, cholesterol and triglyceride levels were measured during supplementation. Glucose was higher in Diet B and cholesterol in Diet A. Diet B had a lower proportion of small (<3 mm) and large follicles (≥3 mm; p = 0.01). The follicular growth rate was higher in Diet A (p < 0.01), with follicles emerging in the 5th day of supplementation. No differences were observed for follicles counted and oocytes recovered. Thus, the type of energy source supplemented for a short term was capable to alter the follicular dynamics, without affecting the proportion of morphologically viable oocytes upon recovery.

Milk production in Saanen goats treated with repeated low doses of intermediate-release insulin during early lactation / Produção de leite em cabras Saanen tratadas com repetidas doses baixas de insulina de liberação intermediária durante a lactação inicial

ABSTRACT: The effect of insulin administration on the productive responses of Saanen goats during early lactation was investigated. Ten of 20 adult females were subjected to subcutaneous administration of intermediate-acting insulin (0.14UI/kg body weight) at 2, 9, and 14 days postpartum. Milk yield was measured twice daily for 13 weeks and milk samples were collected to measure protein and fat contents. Plasma levels of progesterone, insulin, non-esterifies fatty acids, glucose and other metabolites were measured. Results showed a significantly increased effect of insulin treatment on the content of milk fat and protein; moreover, milk production in the first and second postpartum weeks were higher than control group. The peak of lactation in the insulin group was achieved one week earlier in comparison to the control group. In addition, the milk production rate showed lower persistency (milk yield 13 week/milk yield at peak) in the same group. During the first four weeks of postpartum, treated animals showed greater weight loss and higher non-esterified fatty acid concentration, whereas no effect was observed on the concentration of progesterone and other metabolites. The above results indicated that repeated administration of insulin in dairy goats during early lactation increase yield and qualitative components of milk, but has substantial consequences on animal productive rate and metabolic response.

RESUMO: O objetivo do estudo foi avaliar o efeito da administração de insulina sobre a resposta produtiva de cabras Saanen durante a lactação inicial. Dez de vinte fêmeas adultas foram sujeitas à administração subcutânea de repetidas e baixas doses de insulina de liberação intermediária aos 2, 9 e 14 dias pós-parto. A produção de leite foi mensurada duas vezes ao dia, por 13 semanas, e amostras de leite foram coletadas para mensurar teores de proteína e gordura. Os níveis plasmáticos de progesterona, insulina, ácidos graxos não-esterificados (AGNE), glicose e outros metabólitos foram mensurados. Os resultados mostraram um efeito significativamente maior nos animais tratados com insulina sobre o teor de gordura e proteína no leite. Além disso, a produção de leite na primeira e segunda semana pós-parto foi maior no grupo tratado do que no grupo controle. O pico de lactação no grupo insulina foi alcançado uma semana antes em comparação ao grupo controle. Além disso, a taxa de produção de leite nos animais tratados mostrou uma menor persistência de produção de leite durante o período analisado. Durante as primeiras quatro semanas pós-parto, os animais tratados com insulina mostraram maior perda de peso e maior concentração de AGNE, enquanto não se observou nenhum efeito sobre a concentração de progesterona ou outros metabólitos. Os resultados acima indicam que repetidas doses de insulina em cabras leiteiras durante a lactação inicial aumenta o rendimento de produção e concentração de componentes qualitativos do leite, mas apresenta consequências consideráveis sobre taxa de produção animal e resposta metabólica.

Proteome of the periovulatory oviduct and uterus of goats as related to nutritional balance.

This study was conducted to evaluate the effects of different feeding levels on the proteome of oviduct and uterus tissues of hormonally stimulated goats during the periovulatory period. Forty goats were separated into four different diet groups: Diet 1.0 M (n = 11), Diet 1.3 M (n = 10), Diet 1.6 M (n = 9), Diet 1.9 M (n = 10), fed with 1.0, 1.3, 1.6 and 1.9 times live weight maintenance, respectively. After four weeks of treatment, six hormonally stimulated females per treatment group were randomly selected for collection of uterine and the oviduct tissue samples. Samples were collected after animals were slaughtered in a commercial unit. Feeding goats with 1.3 to 1.9 times more nutrients than a control group directly influenced the proteome of the oviduct and uterus, altering the expression of proteins that participate in biological processes such as apoptosis, antioxidant, and immunological activities. These events are crucial for fertilization and early embryonic survival. Expression of oviduct proteins such as Tubulin Beta 2B, Transferrin and Disulphide-isomerase A3 increased in the 1.9 M group in relation to the other feeding levels. Disulphide-isomerase A4 showed higher expression in the 1.0 M group compared to diets with higher energetic levels. As energy intake increased in the diets, there was higher expression of Alpha-1-antitrypsin and downregulation of Profilin-1 in the uterus of the goats. In conclusion, this study showed that specific proteins of the goat oviduct and uterus expressed during the periovulatory period are modified as the result of nutritional balance.

Developmental Outcome and Related Abnormalities in Goats: Comparison Between Somatic Cell Nuclear Transfer- and In Vivo-Derived Concepti During Pregnancy Through Term.

Cloning by somatic cell nuclear transfer (SCNT) is characterized by low efficiency and the occurrence of developmental abnormalities, which are rather poorly studied phenomena in goats. This study aimed at comparing overall SCNT efficiency in goats by using in vitro-matured (IVM) or in vivo-matured oocytes and fibroblast donor cells (mock transfected, transgenic, or wild type), also characterizing symptoms of the Abnormal Offspring Syndrome (AOS) in development, comparing results with pregnancies produced by artificial insemination (AI) and in vivo-derived (IVD) embryos. The SCNT group had lower pregnancy rate (18.3%, 11/60), total number of concepti (20.0%, 12/60), term births (3.3%, 2/60), and live births (1.7%, 1/60) than both the IVD (77.8%, 7/9; 155.5%, 14/9; 122.2%, 11/9; 88.8%, 8/9) and the AI (71.4%, 10/14; 121.4%, 17/14; 100%, 14/14; 78.5%, 11/14) groups, respectively (p < 0.05). No SCNT pregnancies reached term using IVM oocytes, but in vivo-matured oocytes resulted in two term transgenic cloned kids. The proportion fetal membrane (FM) weight/birth weight reflected an increase in FM size and cotyledonary enlargement in clones, for disproportionally bigger newborns in relation to cotyledonary numbers. Overall, goat cloning showed losses and abnormality patterns similar to the AOS in cloned cattle and sheep, which have not been previously well recognized in goats.