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.

Promoter-specific expression of the imprinted IGF2 gene in bovine oocytes and pre-implantation embryos.

The bovine IGF2 locus is a genomic region with alternative transcripts controlled by five promoters (P0, P1, P2, P3 and P4). As transcriptional regulation can affect messenger RNA (mRNA) stability and translation, and thus, subsequent biological effects, this study evaluated the bovine IGF2 promoter-specific expression patterns in oocytes and pre-implantation embryos produced in vitro by our standard IVP procedures. Immature and matured oocytes, and pre-implantation embryos at the 1-, 2-, 4-, 8- and 16-cell, and at early morula, compact morula, blastocyst and expanded blastocyst stages were collected in three pools of five structures per stage, in four replicates. Total RNA was extracted and subjected to RT-qPCR, using four sets of IGF2 promoter-specific primers covering transcripts driven by promoters P0/P1, P2, P3 and P4, with fragments sequenced for confirmation. Expression of P2- and P4-derived transcripts showed an initial peak between immature (P4) or matured (P2/P4) oocytes and 2-cell embryos, gradually falling until embryo genome activation (EGA), rising again at compaction and cavitation. P0/P1-derived transcripts were identified after EGA, during compaction, whereas P3 activity was not detected at any stage. Our findings suggest that P0/P1 and P2 likely have secondary roles during early stages, whereas P3 may be more relevant later in development. P4 seems to be the main pathway for bovine IGF2 expression during oocyte maturation and embryo development and, therefore, the main target to influence IVP in modulation of embryo growth and in studies in developmental biology.

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.

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.

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.

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 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.

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.

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.

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.