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Abstract Background Omega-3 PUFA or methionine (Met) supply during gestation alters offspring physiology. However, the effect of both nutrients on fetal development has not been explored. Our objective was to determine the effects of supplementation of these two nutrients during late gestation on fetal growth, DNA methylation, and mRNA expression of genes associated with the inflammatory response, and DNA methylation. Ewes (n = 5/treatment) were fed from day 100 to 145 of gestation one of the following treatments: 1) basal diet (NS) without fatty acids (FS) or methionine (MS) supplementation; 2) FS (10 g/kg Ca salts, source omega-3 PUFA); 3) MS (1 g/kg rumen protected methionine); and 4) FS and MS (FS-MS). On day 145, ewes were euthanized, and data from dams and fetus was recorded. Placenta (cotyledon), fetal liver, and blood samples were collected. Results A treatments interaction on fetal liver weight, ewe body weight and body condition score (BCS) was observed; FS-MS were heavier (P

Energy balance is regulated by ghrelin which is a neuroendocrine modulator. Ghrelin is expressed in repro- ductive organs. However, the role of ghrelin during in vitro maturation (IVM) and bovine preimplantational development is limited. The purpose of this study was to measure the expression of ghrelin (GHRL) and its receptor growth hormone secretagogue receptor 1A (GHS-R1A) mRNA, and determine cumulus oocyte complex (COC) viability after IVM with 0, 20, 40 and 60 pM of ghrelin. Also, pronuclear formation was recorded after in vitro fertilization (IVF). GHRL and GHS-R1A mRNA expression in oocyte and cumu- lus cells (CCs) was assessed using reverse transcription-polymerase chain reaction (PCR). Oocyte and CC viability were analyzed with the fluorescein diacetate fluorochrome-trypan blue technique. Pronuclear formation was determined 18 hours after IVF with Hoechst 33342. The results demonstrated that ghrelin mRNA is present in oocyte and CCs before and after 24 hours IVM with all treatments. Ghrelin receptor, GHS-R1A, was only detected in oocytes and CCs after 24 hours IVM with 20, 40 and 60 pM of ghrelin. Oocyte viability was not significantly different (P=0.77) among treatments. However, CC viability was significantly lower (P=0.04) when COCs were matured with ghrelin (77.65, 72.10, 66.32 and 46.86% for 0, 20, 40, and 60 pM of ghrelin, respectively). The chance of two pronuclei forming were higher (P=0.03) when ghrelin was not be added to the IVM medium. We found that ghrelin negatively impacts CC viability and pronuclear formation.

Electronic devices have been used to describe chemical compounds in the food industry. However, there are different models and manufacturers of these devices; thus, there has been little consistency in the type of compounds and methods used for identification. This work aimed to determine the applicability of electronic nose (e-nose) Cyroanose 320 to describe the differentiation of volatile organic compounds (VOCs) in fresh Mexican cheese (F-MC) formulated with milk from two different dairy cattle breeds. The VOCs were described using a device manufactured by Sensigent and Solid-Phase Micro-extraction (SPME) coupled to GC-MS as a complementary method. The multivariate principal components analysis (PCA) and the partial least squares discriminant analysis (PLS-DA) were used to describe the relationships of VOCs to electronic nose data, sensory data, and response levels. In addition, variable importance in projection (VIP) was performed to characterize the e-nose signals to the VOCs. The e-nose distinguishes F-MC prepared with milk from two dairy breeds. Sensor number 31 correlated with carboxylic acids most in F-MC from Jersey milk. The HS-SPME/GC-MS identified eighteen VOCs in F-MC made with Holstein milk, while only eleven VOCs were identified for F-MC made with Jersey milk. The more significant peaks in both chromatogram analyses were Propanoic acid, 2-methyl-, 1-(1,1-dimethylethyl)-2-methyl-1,3-propanediyl ester in cheese made from Holstein milk and Propanoic acid, 2-methyl-, 3-hydroxy-2,4,4-trimethylpentyl ester in Jersey milk cheese. Both compounds are considered essential carboxylic acids in the dairy industry. Thus, sensor 31 in the electronic nose Cyranose 320 increased its response by essential carboxylic acids identified by HS-SPME/GC-MS as a complementary method. The e-nose Cyranose 320 is potentially helpful for evaluating fresh Mexican cheese authentication independent of cows’ milk samples from different breeds.

In sheep, polyunsaturated fatty acid (PUFA) supplementations in late gestation increases the growth of offspring; however, there is a lack of evidence on the effect of PUFA supplementation during early gestation. Thus, the objective of this study was to evaluate the effect of dietary supplementation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in early gestation pregnant ewes on fatty acid concentration of fetal liver (FL) and fetal central nervous system (FCNS), and relative abundance of the mRNA for genes associated with transport and metabolism of fatty acids in FL and placenta. A total of 12 ewes, block for stage of gestation were fed a diet containing 1.6% (dry matter basis) monounsaturated fatty acids (MUFA) or EPA+DHA during the first 45 days of gestation. A cesarean section was conducted on day 45 of gestation to collect placenta (caruncle and cotyledon), FL, and FCNS. Relative abundance of mRNA in FL and FCNS and fatty acid concentration were analyzed using a 2x2 factorial arrangement of treatments considering fatty acid supplementation and tissue as the main factors. Concentrations of C18:1 isomers increase (P < 0.05) in FL and FCNS with MUFA supplementation; the FL and FCNS had a greater concentration of C20:3(n-6), C20:3(n-3), C22:1, C22:5 and C22:6 (P < 0.05) with EPA+DHA supplementation. In FL, the relative abundance of LPL mRNA was greater (P = 0.02) as a result of MUFA supplementation. In placenta, there was a FA x tissue interaction for relative abundance of DNMT3b and FFAR-4 mRNA (P < 0.05). Fetus from MUFA-supplemented dams had a greater relative abundance of FABP-4 mRNA (P < 0.05). Results indicate supplementation with EPA+DHA during early gestation increases the total EPA and DHA in FL. For the placenta, EPA+DHA supplementation led to an increase in the relative abundance of lipid mRNA for transport genes.

A negative energy balance (NEB) is detrimental to reproduction in animals. A suggested link between NEB and reproductive failure is the gastrointestinal hormone ghrelin, because of the association between ghrelin and the hypothalamo-pituitary-gonadal axis. The [D-Lys3]-Growth Hormone Releasing Peptide-6 ([D-Lys3]-GHRP-6) is a ghrelin antagonist that acts on ghrelin receptors (GHS-R1). The objective of this study was to evaluate the effect of [D-Lys3]-GHRP-6 on reproduction variables in feed restricted ewes. Two experiments were conducted. Experiment I was conducted for 30 days; and Experiment II for 13 days. In both experiments the ewes (n = 18) were randomly assigned to: Control (CO): fed to meet maintenance requirements; Feed restriction (FR): 80% of maintenance restriction; or Ghrelin antagonist (GA): feed restricted and daily subcutaneous of 7.5μg/kg of [D-Lys3]-GHRP-6. Plasma was collected to measure hormones and metabolite concentration. In Experiment II, the hypothalamus and ovaries were collected on day 13. In both Experiments, sheep allocated to the FR and GA treatments decreased their body weight compared with sheep in the CO group (P < 0.06); progesterone however, did not differ between treatments (P > 0.10). Experiment I: Plasma ghrelin concentration was greater (P < 0.01) in FR and GA compared with CO ewes. Plasma non-esterified fatty acids concentration was greater (P < 0.01) in GA and FR than CO. Experiment II: Kisspeptin1-Receptor (Kiss1-R) mRNA expression was greater in FR (P < 0.01) and tended to be greater in GA (P = 0.10) compared with CO ewes. The neuro peptide-Y (NPY) mRNA expression was greater (P = 0.03) in FR than CO; and tended to be greater (P = 0.06) compared with GA ewes. Growth hormone releasing hormone (GhRH) mRNA expression was greater in GA (P = 0.04) and tended to be greater in FR (P = 0.07) compared with CO ewes. Feed restriction increased GhRH, NPY, and Kiss-R mRNA expression in hypothalamus without affecting reproductive variables.Ghrelin antagonist may prevent an increase inNPY expression in ewes.

Lambs born from dams supplemented with different sources of fatty acids (FA) during late gestation have a different growth rate and plasma glucose concentration. The main objectives of this experiment were to evaluate the effect of supplementing different sources of FA during late gestation on offspring plasma metabolite concentrations, growth, and on a glucose tolerance test (GTT) during the finishing phase. Fifty-four lambs (18 pens, 3 lambs/pen) were born from ewes supplemented during late gestation with one of three treatments: (1) no FA (NF); (2) a source of monounsaturated FA (PDS, 1.01% of Ca salts); or (3) a source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (EDS, 1.01% of Ca salts containing). At birth (day 0), supplementation ceased, and all ewes and lambs were placed in a common pen. On day 60, lambs were weaned, grouped by sex, blocked by body weight (BW), and placed on a common finishing diet for 54 days (FP). One lamb per pen was used for the GTT after the FP. There was a tendency for FA × Sex × Day interaction (p = 0.08) on lamb growth during the finishing period, with PDS females being heavier than PDS males, while EDS males were heavier than EDS females at day 60. There was a tendency for FA × Sex interaction (p = 0.06) for plasma insulin concentration for the GTT. Plasma insulin concentration of wethers increased as FA unsaturation degree increased during the GTT; the opposite happened with the plasma insulin concentration of female lambs. In conclusion, FA supplementation during late gestation tended to modified growth and insulin response to a GTT; these changes differed with the degree of FA unsaturation of the supplement and lamb sex.

Background Manipulating the dietary fatty acid (FA) content can alter FA profiles of reproductive tissues. Numerous researchers have evaluated the effect of fish oil (FO) sup- plementation on reproductive characteristics in domestic animals, but reliable informa- tion concerning dietary FO effects on semen quality and testosterone concentrations in dogs has not been reported. Therefore, this study evaluated the effects of dietary FO on semen quality and serum testosterone concentrations in dogs. Materials and Methods In this cross-over experimental study, 5 male dogs consumed either a control diet or the same diet supplemented with 54 mg FO/kg metabolic body weight (BW) for 120 days. After the 120-day wash-out period, control (C) dogs received FO and FO-fed dogs consumed the control diet. In the first period, 2 dogs were allocated to the FO group and 3 to the C group. In the second period, 3 dogs were allocated to the FO group and 2 to the C group. Semen samples collected on days 0, 60, 90 and 120 were evaluated by standard methods. Day 120 semen samples were analyzed for FA profiles. Blood samples were collected on days 0, 30, 60, 90 and 120 to measure serum testoster- one concentrations. Data were analyzed by analysis of variance with repeated measures using the Mixed Models procedure of SAS (version 9.0, SAS Institute Inc., Cary, NC, USA). Animals and period of time (first or second 120 days) were random variables; and treatment, time, and the treatment by time interaction were considered fixed effects. Results FO supplementation increased the percentage of motile sperm (P=0.02), total sperm count (P < 0.01), total sperm viability (P < 0.01), and total morphologically normal sperm (P < 0.01). Supplementation decreased the percentage of viable sperm (P=0.03) and serum testosterone concentration (P < 0.01). FO supplementation also increased the per- centage of arachidonic acid, eicosapentaenoic acid, (EPA) and total n-3 in semen samples (P≤0.05). Conclusion These results are consistent with the concept that long-term FO supplemen- tation influences semen quality and testosterone concentrations in dogs by altering semen FA profiles.

Herbal formulas during pregnancy have been used in developing countries. Despite that, the potential effects on the mother and offspring and whether those supplements elicit epigenetic modifications is still unknown. Therefore, our objectives were to determine the effects of supplemental herbal choline source (BCho) on the percentage of 5-hmC in whole blood from gestating ewes and their offspring, as well as determining the milk quality and growth of the offspring. Thirty-five gestating Rambouillet ewes were randomly assigned to five treatments: T1, supplementation of 4 g per day (gd−1) of BCho during the first third of gestation; T2, supplementation of 4 gd−1 of BCho during the second third of gestation; T3, supplementation of 4 gd−1 of BCho during the last third of gestation; T4, supplementation of 4 gd−1 of BCho throughout gestation; and T5, no BCho supplementation (control). For the 5-hmC DNA analysis, whole blood from ewes was sampled before pregnancy and at each third of gestation (50 days). Whole blood from lambs was sampled five weeks after birth. The evaluation of the nutritional programming effects was conducted through the percentages of 5-hmC in the lambs. Compared with other treatments, the whole blood from ewes supplemented during T1 and T4 had the greatest 5-hmC percentages (p < 0.05). However, only ewes fed BCho throughout gestation (T4) maintained the greatest percentages of 5-hmC (p < 0.05). The lamb growth performance indicated that the BCho maternal supplementation did not affect the nutritional programming. However, the lambs born from ewes supplemented during T2 had the greatest 5-hmC percentages (p < 0.05). Our data suggest that ewes supplemented during T4 with BCho increase and maintain the percentages of 5-hmC in whole blood, and the offspring born from ewes supplemented with BCho during T2 maintained the greatest percentages of 5-hmC 35 d after they were born.

The objective of the study was to evaluate the effect of time of gestation on fatty acid transporter mRNA expression in maternal and fetal bovine placenta. Placentas from twelve cows at different thirds of gestation (n=4 per third) were sampled at slaughter to measure FATP-1, FATP-4, FABP-1 mRNA concentration in maternal (caruncles) and fetal (cotyledons) side. Once the placenta was removed, 1cm2 was dissected and, divided into caruncles and cotyledons, stored in sterile tubes, dropped into liquid nitrogen and kept at -80° C until rtPCR analysis. Data were analyzed as a complete randomized design with a 3 x 2 factorial arrangement, using the mixed procedure (SAS 9.3) with repeated measurements on space. Time of gestation, side of the placenta and their interaction were fixed factors, whereas animal was a random factor. There was a time by treatment interaction (P < 0.01) on FATP-1 mRNA expression due to a greater mRNA expression in cotyledons on the first third of gestation as compared with the concentration in caruncles. On the second and third stages of gestation, the mRNA concentration in cotyledons decreased, reaching a similar concentration to that observed in caruncles. Fatty acid transport protein -4 and FABP-1 mRNA concentration were not different (P >0.1). We conclude that FATP-1 might play an important role in fatty acid transport during early fetal development.

The objective of the present study was to evaluate the effect of calcium propionate (CaPr) on in vitro ruminal fermentation using a factorial arrangement 2 x 2 evaluating CaPr (0 vs. 1%) and grain level (55 vs. 65%). There was a CaPr x Grain interaction in the volume of gas produced (V; p = 0.04). Addition of CaPr prolonged Lag time (1.4 vs. 1.04 h; P