Genotyping of rams based on melatonin receptor 1A gene polymorphisms: a tool in sire selection?

Context Several polymorphisms in the melatonin receptor 1A gene (MTNR1A ) have been related to reproductive performance in ovine. Aims To investigate the effect of the Rsa I and Mnl I polymorphisms on ram seminal quality. Methods Eighteen Rasa Aragonesa rams were genotyped for the Rsa I (C/C, C/T, T/T) and Mnl I (G/G, G/A, A/A) allelic variants of the MTNR1A gene. Individual ejaculates were analysed once a month throughout the whole year. Sperm motility, morphology, membrane integrity, levels of reactive oxygen species (ROS), phosphatidylserine (PS) inversion, DNA fragmentation and capacitation status were assessed. The effect of the season and polymorphisms on seminal quality was evaluated by mixed ANOVA. Key results Both polymorphisms had an effect on membrane integrity and viable spermatozoa with low levels of ROS and without PS translocation, and Rsa I also on motile and DNA-intact spermatozoa. An interaction between both polymorphisms was found, pointing to a negative effect on seminal quality of carrying the T or A allele in homozygosity. Differences were higher in the reproductive than in the non-reproductive season. Conclusions Mutations substituting C by T and G by A at Rsa I and Mnl I polymorphic sites, respectively, in the MTNR1A gene in rams could decrease the seminal quality. Implications Genotyping of rams based on melatonin receptor 1A could be a powerful tool in sire selection.

The Effect of By-Pass Linseed Oil Supplementation on the Maternal Antioxidant System during the Embryo-Maternal Recognition Period in Ewes.

This study analyzed the effects of dietary supplementation with by-pass linseed oil (LO; rich in α-linolenic acid) on maternal antioxidant systems at Days 14 and 16 of pregnancy in Sarda ewes. This trial used sixteen dry ewes. Eight ewes (CT group) were fed with a control diet without LO, and eight ewes (LO group) were fed with a diet supplemented with LO (10.8 g of α-linolenic acid/ewe/day). Both diets had similar crude protein and energy levels. The experiment included 10 days of an adaptation period and 31 days of a supplementation period. This supplementation period was divided into Period -2 (from Day -15 to -8), Period -1 (from Day -7 to -1; before synchronized mating period/Day 0), Period +1 (from Day +1 to + 7 after mating), and Period +2 (from Day +8 to +15 after mating). Estrous synchronization was induced in all the ewes using an intravaginal sponge (45 mg fluorgestone acetate) for 14 days and equine chorionic gonadotropin (350 UI/ewe) at the end of the treatment. On Days 14 (CT, N = 4; LO, N = 4) and 16 (CT, N = 4; LO, N = 4) after mating, the ewes were slaughtered. Samples of plasma, uterine, and luteal tissues were collected. Thiols, total antioxidant activity (TEAC), superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content were measured. On Day 16, thiol and TEAC in luteal tissues were higher in the LO group when compared with the control one (p < 0.05). Moreover, TEAC was higher for the LO group in uterine tissues on Days 14 and 16 (p < 0.05). SOD activity was higher in the LO group in luteal and uterine tissues on Day 14 and Day 16, respectively (p < 0.001). On Day 16, uterine MDA content was lower for the LO group (p < 0.001). No differences were found between groups at the plasmatic level. However, the by-pass LO supplementation enhanced the analyzed antioxidant parameters in luteal and uterine tissues. In conclusion, these results demonstrate that by-pass LO supplementation exerted a positive effect on antioxidative defenses on maternal structures during the embryo-maternal recognition period in ewes. Thus, this could contribute to improving the maternal environment during the embryo-maternal recognition period in mammals.

Anatomy of hypothalamic and diencephalic nuclei involved in seasonal fertility regulation in ewes.

In this study, we describe in detail the anatomy of nuclei involved in seasonal fertility regulation (SFR) in ewes. For this purpose, the intergeniculate leaflet of the visual thalamus, the caudal hypothalamic arcuate nucleus, and suprachiasmatic, paraventricular and supraoptic nuclei of the rostral hypothalamus were morphometrically and qualitatively analyzed in Nissl-stained serial sections, in the three anatomical planes. In addition, data were collected on calcium-binding proteins and cell phenotypes after immunostaining alternate serial sections for calretinin, parvalbumin and calbindin. For a complete neuroanatomical study, glial architecture was assessed by immunostaining and analyzing alternate sections for glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (IBA1). The results showed a strong microglial and astroglia reaction around the hypothalamic nuclei of interest and around the whole 3rd ventricle of the ewe brain. Moreover, we correlated cytoarchitectonic coordinates of panoramic serial sections with their macroscopic localization and extension in midline sagittal-sectioned whole brain to provide guidelines for microdissecting nuclei involved in SFR.

Use of Acheta domesticus meal as a full soybean substitute in the feeding of slow-growing chicks.

Sustainable poultry meat production involves the use of slow-growing chick strains and the utilization of new protein sources as an alternative to the current monopoly of soybean meal. In this scenario, a study was conducted to assess the effect of replacing soybean meal with domestic cricket (Acheta domesticus) meal on the developing cycle of slow-growing chicks. To this end, a total of 128 one-day-old male chicks (Colorield) were randomly assigned into 16 experimental units, each consisting of 8 chicks, which in turn were grouped into 2 groups fed isoproteic and isoenergetic diets in which the protein source differed: the control group (C) fed soybean meal and the Acheta group (AD) fed Acheta domesticus insect meal as the main protein source. Chicks were slaughtered at 95 d of age. Three different diets (F1, F2 and F3) were used for each experimental group according to the nutritional needs of the birds during their growth. The F1 diet (1-29 d) resulted in higher feed and water intake and higher body weight gain for group C, but a lower feed conversion rate. On the contrary, during F2 (29-60 d) no differences in productive performances were observed between the 2 groups, except for a higher water intake for group C. Finally, during the period corresponding to diet F3 (60-95 d) there were only differences in feed intake, which was higher for the AD group. In conclusion, the substitution of soybean meal for Acheta domesticus meal caused a decrease in feed intake during the first month and consequently a lower body weight. During the first 4 weeks of life of the chicks, a partial replacement of soybean meal may be recommended, since high inclusions of A. domesticus meal in this period seem to be detrimental in young chicks. Given the absence of relevant differences in productive performances between both groups, it could be concluded that the use of A. domesticus cricket meal can be a potential protein alternative to soybean meal.

Effect of Strategic Supplementation of Dietary By-Pass Linseed Oil on Fertility and Milk Quality in Sarda Ewes.

The aim of the present study was to assess whether the strategic supplementation of bypass LO can enhance reproductive indexes­fertility, lambing rate, and prolificacy­in dairy Sarda ewes at the end of lactation. To assess whether LO supplementation leads to the adsorptions of PUFAs and their subsequent utilization by the body tissues, milk composition and fatty acid content were analyzed. Forty-eight ewes were assigned to the following groups: the control group (CT; N = 24), fed with a control diet without LO; and the treatment group (LO; N = 24), fed with a diet supplemented with LO (10.8 g/ewe/day). Both diets had similar crude protein and energy levels and were offered for 38 days (−21 to +17 days after artificial insemination). The trial included an adaptation period (7 days) followed by a regular supplementation (31 days) period. Estrus synchronization was induced in all the ewes using an intravaginal sponge and equine chorionic gonadotropin. Fifty-five hours after pessaries withdrawal, all ewes were inseminated using the cervical route and fresh semen. Cholesterol (p < 0.01), high-density lipoprotein (p < 0.001), and triglyceride (p < 0.05) levels in plasma were higher in the LO group. Plasmatic levels of non-esterified fatty acids were lower in the LO group after the end of the supplementation period (p < 0.05). Milk unsaturated fatty acids (UFAs), monounsaturated fatty acids (MUFAs), total polyunsaturated fatty acids (PUFAs), PUFAs omega 3 (PUFAs-ω3) and 6 (PUFAs-ω6), and trans fatty acids were higher in the LO group (p < 0.001), while saturated fatty acids (SFAs) were higher in the CT group during the supplementation period (p < 0.001). Three days after the end of the supplementation period, the content of milk UFAs (p < 0.05), PUFAs (p < 0.001), MUFAs, and PUFAs-ω6 (p < 0.01) were still higher in the LO group. whereas SFA was higher in the CT group (p < 0.01). There was no difference between groups in terms of ovulation rate, progesterone levels in plasma, fertility rate, prolificacy, and total reproductive wastage. However, the total area of luteal tissue was higher in the LO group (p < 0.01). Results obtained demonstrated that LO supplementation exerts a positive role in corpus luteum size at the onset of the peri-implantation period in Sarda dairy ewes. Additionally, the results obtained in the present study showed that the use of dietary bypass LO affects lipid metabolites in plasma and milk fatty acid profiles, demonstrating the ALA uptake by body tissues.

Exposure of rams in sexual rest to sexually activated males in spring increases plasma LH and testosterone concentrations.

Eight stimulating rams, and twelve stimulated rams, were used to determine whether a similar endocrine response to the introduction of sexually active males in spring in a flock of ewes is observed in a flock of rams. The stimulating rams (n = 4) were induced into a sexually active state by exposure to 2 months of long days (16 h light/d) (15 December-15 February). At the end of the long-day period, rams were returned to the natural photoperiod. Control-stimulating rams (n = 4) were kept under the natural photoperiod. On April 20, stimulated rams were divided into 2 groups, and joined with activated (ACT; n = 6) or control stimulating rams (C; n = 6). On the day of ram introduction, stimulated rams were blood sampled for 8 h at 20-min intervals, from 4 h before to 4 h after ram introduction, and next day from 24 to 28 h after ram introduction, and analyzed for plasma LH concentrations, and 10, 20 and 30 days after ram introduction to measure plasma testosterone levels. Mean (±SEM) plasma LH concentrations (ng/ml) of stimulated rams were similar during the 4 h before stimulating-ram introduction (ACT: 0.59 ± 0.03; C: 0.53 ± 0.04; P > 0.05). The introduction of the photoperiod-treated stimulating rams increased LH concentrations of stimulated rams during the 4 h after their introduction (1.14 ± 0.37) compared with the C group (0.51 ± 0.03; P < 0.05), especially during the first hour (ACT: 0.93 ± 0.16; C: 0.49 ± 0.03; P < 0.05), and during the blood sampling period 24-28 h after ram introduction (0.75 ± 0.07 vs. 0.58 ± 0.04; P < 0.05). Before the introduction of stimulating rams, the LH pulse frequencies and amplitudes did not differ between groups; however, LH pulsatility was higher at 4 h (0.58 ± 0.11 pulses/h; P < 0.05), and had trend to be higher 24 h (0.50 ± 0.06) (P = 0.10) after the introduction of the photoperiod-treated stimulating rams compared with the control-stimulating rams (0.29 ± 0.08 and 0.29 ± 0.10, respectively). As for LH pulses, there was an effect of group (P < 0.05) on LH amplitude, which presented a trend to be higher in ACT rams 4 h after ram introduction (1.68 ± 0.30; P < 0.10) and higher 24 h (1.07 ± 0.08; P < 0.05) after ram introduction, compared with LH amplitudes of C rams (0.71 ± 0.06 and 0.82 ± 0.07, respectively). Plasma testosterone concentrations of rams exposed to photoperiod-treated activated rams were higher than those of rams exposed to control-stimulating rams, at 4 h, 20 and 30 days after ram introduction (P < 0.05). In conclusion, sexually active rams in spring are able to stimulate LH and testosterone secretion of other rams in sexual rest, a phenomenon we called "ram-to-ram effect".

Effects of Implanting Exogenous Melatonin 40 Days before Lambing on Milk and Colostrum Quality.

The effects of exogenous melatonin implanted before lambing on the quality of colostrum and milk yield were quantified in 715 ewes. Forty days before lambing, 246 ewes (1M) received a melatonin implant; another 137 ewes (2M) received two implants, and the remaining 332 ewes (C) did not receive an implant (control). Milk analysis was based on individual monthly milk samplings (June, July, and August) after lambing. A colostrum sample was collected from 303 ewes (118 1M; 73 2M; and 112 C), and IgG concentrations were measured. Ewes implanted with melatonin had higher (p < 0.01) daily milk yield (DMY) in the three samplings than the C ewes. On average, 1M ewes produced more milk (p < 0.05) than ewes in the other two groups, and 2M ewes produced significantly (p < 0.05) more milk than C ewes. In the first and third controls, ewes that received two melatonin implants had a lower (p < 0.05) SCC than C and 1M ewes, and in the second sampling, 1M and 2M ewes had a lower (p < 0.01) SCC than C ewes. Ewes that received melatonin implants had a higher (p < 0.01) IgG concentration (21.61 ± 1.03 mg/mL) than non-implanted ewes (16.99 ± 1.13 mg/mL); 2M ewes had the highest IgG levels. In conclusion, ewes that received a melatonin implant 40 d before lambing produced colostrum that had a higher IgG concentration than the colostrum from nonimplanted ewes, and produced more milk, which had a lower SCC. The effect on SCC was prolonged if the sheep received a second melatonin implant.

Comparison of four strategies of ram management in a semen collection centre.

Thirty Merino rams were used to determine the effect of four management systems of rams on semen quality during the anoestrous season. Animals were divided into four groups: Artificial-Photoperiod group (AP; n = 8), which were isolated from females and exposed to artificial long days (16 hr/d) from 1 Feb to 15 Mar; Natural-Photoperiod (NP; n = 8), isolated from females and exposed to the natural photoperiod throughout the experiment; Oestrous-Ewe group (EE; n = 7), housed in a pen adjacent to another pen that housed three ewes in oestrus, and Anestric-Ewe group (AE; n = 7), housed adjacent to another pen that housed three ovariectomized ewes. From 20 Mar to the end of May (10 weeks), semen samples were collected weekly, and blood samples were collected to determine plasma testosterone concentrations. Mean plasma testosterone concentrations, ejaculate volume and reaction time were not affected either by treatment or week. There was a significant effect (p < .01) of ram treatment on sperm concentration, and both TM y PM, and their interaction, were significantly affected by group and week (p < .001). Rams exposed to ewes in oestrus presented the largest sperm concentration (p < .05) compared with the other three groups, although they had the lowest total and progressive motilities (p < .01). In conclusion, management strategy in spring affects semen quality of rams, with the presence of ewes in oestrus being the best plan to increase sperm concentration.

Performance of Slow-Growing Chickens Fed with Tenebrio molitor Larval Meal as a Full Replacement for Soybean Meal.

Insect larval meal is an increasingly common protein source in poultry systems. In this study, the effect of replacing soybean meal with Tenebrio molitor larval meal on the performance of slow-growing chickens was assessed. A total of 128 one-day-old chickens (Colorield) were randomly divided into a control group (C) (n = 64), fed with soybean meal, and an experimental group (TM) (n = 64), fed with T. molitor larvae meal. The chicks were slaughtered after 95 days. Three different isoenergetic and isoproteic diets (F1, F2 and F3) were used for each group. The F1 diet resulted in higher body weight gain and higher feed and water intakes in group C, but a lower feed conversion ratio. Contrarily, diets F2 and F3 did not produce differences in the studied parameters between the two groups, except for body weight gain in the case of diet F2, which was highest in group C. Therefore, weight gain and feed and water intakes were significantly higher in group C, but there were no differences in feed conversion ratio or live weight. In conclusion, the total replacement of soybean meal with T. molitor larvae meal resulted in a reduction in feed intake and a consequent reduction in weight. During this period, partial rather than total substitution may be recommended.

The effect of being housed with adult female flock-mates on the age at puberty of autumn-born ewe-lambs.

Twenty-five ewe-lambs born in November were used to measure the effect of the presence of cyclic or non-cyclic adult ewes on the age at first fertile estrus. Lambs were weaned at 45 days of age and kept in a single flock until 3 Mar (4 months old), when they were assigned to one of the following three groups: Group CE (n = 8), housed with six cyclic adult ewes; Group NCE (n = 8), housed with three non-cyclic ovariectomized adult ewes, and Group ISO (n = 9), housed isolated from adult ewes. On 2nd May (6 months old), two rams were introduced into each group, until 31st Oct (12 months old), when rams were removed. Adult ewes in group CE were separated from rams and ewe-lambs by a metal fence, allowing visual, olfactory, and nose-to-nose contact between animals. Average earliest fertile estrus was significantly (P < 0.01) earlier in NCE ewe-lambs (8th Aug) than it was in ISO (9th Sep) and CE (17th Sep) ewe-lambs, they lambed about 35 d earlier (1st Jan) than did the other two groups (2nd Feb and 10th Feb, respectively) (P < 0.01), and at an earlier age (14 mo, 15.2 mo, and 15.3 mo, respectively; P < 0.01). In conclusion, the presence of non-ovulatory adult ewes advanced puberty in autumn-born ewe-lambs of a Mediterranean genotype. The factor(s) that delayed age at puberty in isolated females and those housed with estrous adult ewes remains to be elucidated.

A High Cattle-Grazing Density Alters Circadian Rhythmicity of Temperature, Heart Rate, and Activity as Measured by Implantable Bio-Loggers.

Six cows managed under extensive grazing conditions were used to study the effect of moving the animals to a higher grazing density on the circadian rhythms of temperature (T), heart rate (HR), and activity (ACT), which were recorded by implantable bio-loggers. Cows were maintained at a density of 1.5 livestock units per hectare (LSUs/ha; low density, LD) until they were moved to a grazing area at 128 LSUs/ha (high density, HD). Animals were implanted subcutaneously with a T, HR, and ACT bio-logger, which was programmed to record data at 5-min intervals. For each animal, cosinor rhythmometry (the study of circadian rhythms by fitting a sine wave to a time series) was applied to the data recorded over 5 days in LD and HD. Mean Midline Estimating Statistic of Rhythm (MESOR; the average value around which the variable oscillates), amplitude (difference between the peak and the mean value of a wave), and acrophase (timing of peak activity) were calculated and evaluated statistically. Differences between mean day and nighttime values, and mean LD and HD values were calculated. Cows presented cosinor curves that fit a 24-h rhythm (p < 0.001) in T, HR, and ACT at both densities. MESOR (T: 37.98 vs. 38.02°C; HR: 69.12 vs. 65.91 bpm; ACT: 49.39 vs. 40.41 mg, for LD and HD, respectively) and amplitude (T: 0.28 vs. 0.28°C; HR: 4.12 vs. 3.14 bpm; ACT: 18.14 vs. 11.28 mg, respectively) did not differ significantly between the two densities; however, significant (p < 0.05) differences between densities occurred in the acrophase of the three variables; specifically, the T acrophase was 2 h later at HD (22:45 h) than LD (20:45 h), and HR (LD: 19:51; HD: 16:49 h) and ACT acrophases 3 and 2 h earlier at HD than LD (LD: 14:47; HD: 12:49 h), respectively. T and ACT differed significantly (p < 0.01) between daytime (mean ± SE; 37.92 ± 0.19°C, 40.39 ± 4.74 mg) and nighttime (38.14 ± 0.17°C, 29.93 ± 5.66 mg). In conclusion, our study suggests that a high animal grazing density might exacerbate the social competence for valuable resources for animals, resulting in shifting the circadian rhythmicity of temperature, heart rate, and activity of the cows, advancing or delaying their acrophases.

Performance Evaluation of Two Slow-Medium Growing Chicken Strains Maintained under Organic Production System during Different Seasons.

A total of 160 1-day-old medium-growing male chicks (Gallus gallus domesticus) were raised for 120 days in a certified organic farming system. A total of two strains were studied (Coloryield, CY; RedBro, RB). Overall, two weather periods were considered based on the outdoor temperature, being S1 colder than S2. In total, 40 chicks per strain were assigned to each period (n = 80). Chickens were fed ad libitum with the same organic feeds. In the first month, chickens were kept indoors and, from day 30, they had access to the pasture. Slaughter live weight (LW), average daily gains, (ADG), the feed conversion ratio (FCR), and mortality rates did not differ between the two strains. LW was (p < 0.05) higher in the S1 and a trend (p = 0.084) was observed for ADG, which was higher in S1. No differences were found for feed intake, FCR, and mortality rates between weather periods. There were no differences for coefficient of variation (CV) between the strains studied, nevertheless, CV for LW in S2 was increased. Differences in the productive performance between these strains raised in organic production systems were slight. However, chickens raised in S1 had a better performance. It would be preferable to raise chickens in these weather conditions whenever possible.

Revisiting Cattle Temperament in Beef Cow-Calf Systems: Insights from Farmers' Perceptions about an Autochthonous Breed.

Understanding temperament is an important part of cattle production since undesirable temperament may cause serious problems associated with aggression, maternal care, and human safety. However, little is known about how farmers define or assess temperament, especially in autochthonous cattle breeds. The aim of this study was to explore perceptions of farmers about the temperament of the Pyrenean cattle breed with special attention to beef cow-calf systems in Spain. The methodology used to obtain the information was focus group discussions (FGD). Farmers defined temperament as a behavioural response to challenging situations imposed by human handling. Specific terms used were related to active or passive reactions to fear (e.g., "strong", "aggressive", "nervous", "fearful"). The speed of response to stimuli was also important. Female temperament was thought to become more docile with age while bull temperament was more variable. Maternal aggressiveness was highlighted as a potential human safety problem, but also desirable in an extensively bred animal who may need to defend calves against predators. Anatomical characteristics were seen as unreliable predictors of temperament, while behavioural indicators were more widely used, such as "alertness", which was a general trait of the breed, and "gaze", which, when associated with an alert expression, suggests a potential threat. Sensory acuity, such as sight and smell, were thought to be related with temperament in some FGDs but there was no overall agreement as to whether different behavioural responses were due to differences in sensory acuity. The results from the study could be useful during training programs or in the development of new genetic selection schemes and evaluation protocols involving cattle temperament.

The presence of an embryo affects day 14 uterine transcriptome depending on the nutritional status in sheep. b. Immune system and uterine remodeling.

Transcriptomics and bioinformatics were used to investigate the potential interactions of undernutrition and the presence of the conceptus at the time of maternal recognition of pregnancy on uterine immune system and remodeling. Adult Rasa Aragonesa ewes were allocated to one of two planes of nutrition for 28 days: maintenance energy intake (control; 5 cyclic, 6 pregnant ewes) providing 7.8 MJ of metabolisable energy and 0.5 maintenance intake (undernourished; 6 cyclic, 7 pregnant ewes) providing 3.9 MJ of metabolisable energy per ewe. Uterine gene expression was measured using Agilent 15 K Sheep Microarray chip on day 14 of estrus or pregnancy. Functional bioinformatics analyses were performed using PANTHER (Protein ANalysis THrough Evolutionary Relationships) Classification System. Pregnancy affected the expression of 18 genes in both control and undernourished ewes, underscoring the relevance for embryo-maternal interactions. Immune system evidenced by classical interferon stimulated genes were activated in control and -in a lesser extent-in undernourished pregnant vs cyclic ewes. Genes involved in uterine remodeling such as protein metabolism were also upregulated with the presence of an embryo in control and undernourished ewes. However, relevant genes for the adaptation of the uterus to the embryo were differentially expressed between pregnant vs cyclic ewes both in control and undernourished groups. Undernutrition alone led to an overall weak activation of immune system pathways both in cyclic and pregnant ewes. Data revealed that cellular and immune adaptations of the uterus to pregnancy are dependent on the nutritional status.

Vasectomy and Photoperiodic Regimen Modify the Protein Profile, Hormonal Content and Antioxidant Enzymes Activity of Ram Seminal Plasma.

This work aimed to determine the contribution of the testis and epididymis and the effect of the photoperiodic regimen on ram seminal plasma (SP). Semen was collected from 15 mature rams located in an equatorial (Colombian Creole and Romney Marsh, eight intact and two vasectomized) or a temperate climate (Rasa Aragonesa, three intact and two vasectomized). SP proteins were analyzed by Bradford, SDS-PAGE and difference gel electrophoresis (DIGE). Melatonin and testosterone concentrations were quantified by ELISA, and activity of glutathione peroxidase (GPx), glutathione reductase (GRD), and catalase by enzymatic assays. Vasectomy increased protein concentration and the intensity of high molecular weight bands (p < 0.001), with no differences between breeds. DIGE revealed the absence of six proteins in vasectomized rams: angiotensin-converting enzyme, lactotransferrin, phosphoglycerate kinase, sorbitol dehydrogenase, epididymal secretory glutathione peroxidase and epididymal secretory protein E1. Vasectomy also decreased melatonin concentrations in seasonal rams, and testosterone in all of them (p < 0.001), but did not affect antioxidant enzyme activity. Equatorial rams showed lower melatonin and testosterone concentration (p < 0.01) and catalase, but higher GPx activity (p < 0.05). In conclusion, vasectomy modifies the protein profile and hormonal content of ram seminal plasma, whereas the exposure to a constant photoperiod affects hormonal concentration and antioxidant enzymes activity.

Semen Quality of Rasa Aragonesa Rams Carrying the FecXR Allele of the BMP15 Gene.

The FecXR mutation is a variant of the ovine gene BMP15 in the Rasa Aragonesa breed. Information on the physiological importance of carrying the FecX polymorphism in rams is limited. The aim of this study was to compare semen quality, testicle characteristics, and fertility rate of rams that carry the FecXR allele. Rams (n = 15) were either FecXR allele carriers (n = 10) or non-carriers, wild type (++) (n = 5). FecXR rams exhibited higher mass motility (p < 0.05), proportion of rapid sperm (p < 0.05), and a lower proportion of slow sperm (p < 0.0001) than did ++ rams. The presence of the FecXR allele was not associated with mean scrotal circumference or testicular length and diameter, although season had a significant (p < 0.05) effect on these traits. Genotype (p < 0.05) and season (p < 0.01) had a significant effect on mean fertility rate, FecXR rams had a higher proportion of pregnant ewes than did ++ rams (p < 0.05). In conclusion, the FecXR allele produced high-quality semen throughout the year, and corresponded with an improvement in some sperm parameters, particularly, mass motility and the proportion of rapid sperm.

Exogenous melatonin and male foetuses improve the quality of sheep colostrum.

Colostrum is the first product secreted by the mammary gland to transfer immunity to the newborn, especially through immunoglobulins (Ig) G. Melatonin is an immunomodulatory factor and there is evidence that it has a direct effect on IgG production. To evaluate the effects of melatonin treatment during pregnancy, litter size and offspring sex on colostrum quality, sixty pregnant Rasa Aragonesa ewes were divided into three groups: one group received a melatonin implant at the third month of pregnancy (3M, n = 13), another group at the fourth month (4M, n = 18) and the remaining ewes were not implanted (Control, C, n = 29). Immediately after lambing, a sample of colostrum was collected and IgG, crude protein and fat content analysed. Timing of melatonin implantation (p < .001), and offspring sex (p < .01) had a significant effect on IgG concentration. Colostrum of treated ewes had a higher mean (±SEM) IgG concentration than that of the control ewes (55.54 ± 3.09 and 49.50 ± 4.36 mg/ml, respectively; p < .05), mainly because the concentration in the 4M group was significantly (p < .001) higher than it was in the other groups. The relationship between lamb sex and IgG (p < .01) and %CP (p < .05) was evident in singletons (ewes with a male lamb: 54.57 ± 5.37 mg IgG/ml, 15.42 ± 0.82%CP; ewes with a female lamb: 34.66 ± 4.30 mg/ml, 13.18 ± 0.73%CP). The presence of a female in the litter was associated with significantly (p < .01) lower colostrum IgG levels (litters with at least one female: 49.33 ± 3.42 mg/ml; litters with no females: 58.24 ± 4.00 mg/ml). Among 4M ewes, female foetuses had significantly (p < .01) lower IgG levels whether they carried singletons or multiple lambs. In conclusion, treatment with melatonin implants at the fourth month of pregnancy resulted in a higher colostrum quality based on IgG concentration.

The embryo affects day 14 uterine transcriptome depending on nutritional status in sheep. a. Metabolic adaptation to pregnancy in nourished and undernourished ewes.

This study investigated the effects of undernutrition and the presence of the conceptus at the time of maternal recognition of pregnancy on the expression of uterine indicators of metabolism in ewes. Adult Rasa Aragonesa ewes were allocated to one of two planes of nutrition for 28 days: maintenance energy intake (control; 5 cyclic and 6 pregnant ewes) providing 7.8 MJ of metabolisable energy, and 0.5 maintenance intake (undernourished; 6 cyclic and 7 pregnant ewes) providing 3.9 MJ of metabolisable energy per ewe. RNA from intercaruncular uterine tissue was harvested at slaughter on Day 14 of estrous cycle or pregnancy, and hybridized to the Agilent 15K Sheep Microarray chip. Functional bioinformatics analyses were performed using PANTHER (Protein ANalysis THrough Evolutionary Relationships) Classification System. The presence of the embryo upregulated expression of genes encoding peptide and monocarboxylate transporters regardless of nutritional treatment, although the degree of gene expression was lower in undernourished ewes. Genes encoding enzymes involved in glycolysis were downregulated both in pregnant control and undernourished ewes, probably as a compensatory mechanism for the increased glucose transport to the uterus. Compared with control cyclic ewes, control pregnant ewes had greater expression of genes involved in oxidation of fatty acids, suggesting increased uterine energy demands. This was not observed in undernourished pregnant animals when compared to undernourished cyclic ewes; nevertheless, those animals had lower uterine expression of enzymes involved in fatty acid biosynthesis. The presence of the embryo upregulated genes involved in electron transport probably as a result of increased energy demands for pregnancy. Overall, the data indicate that depending on the nutritional status of ewe, pregnancy alters gene expression of metabolic pathways related to energy generation in the uterus. An impairment in nutrient transport and metabolism in the uterus of pregnant undernourished ewes may explain the greater embryo mortality associated with undernutrition.

Role of melatonin on embryo viability in sheep.

Melatonin is a natural hormone synthesised in the pineal gland, the activity of which is regulated by day-night perception and dictates seasonal rhythms in reproduction in ovine species. Exogenous melatonin, administered via subcutaneous implants, is used to prolong the breeding season of ewes and can increase the proportion of pregnant ewes (fertility rate) and litter size. The increased proportion of ewes that become pregnant and the number of lambs born per lambing among melatonin-treated sheep may be caused by increased embryo survival, through enhanced luteal function, reduced antiluteolytic mechanisms, or improved embryo quality. This review focuses on the effects of melatonin on embryo viability and summarises the processes by which this hormone affects the ovary, follicle, oocyte, corpus luteum and embryo. Moreover, the effects of melatonin on the mechanisms of invivo maternal recognition of pregnancy in sheep and the protective action that it appears to have on the invitro procedures that are used to obtain healthy embryos are reviewed.

Influences of nutrition and metabolism on reproduction of the female ruminant.

Beef cows and ewes grazing native pastures are exposed to cycles of undernutrition that reflect the seasonal variations of biomass production. In grazing dairy cows, the physiological undernutrition during early lactation due to increased demands for lactation and low dry matter intake is exacerbated by the need to get sufficient intake from pasture and the extra grazing energy costs. Undernutrition has profound impacts on reproduction by affecting multiple reproductive processes at different levels of the reproductive axis. The objective of this paper is to review the influence of undernutrition on reproductive events of the adult female ruminant, with emphasis on both grassland and mixed rain-fed grazing farming systems. The comparative endocrinology and reproductive biology among ewes, beef and dairy cows may provide a comprehensive knowledge of the metabolic and reproductive adaptation to feed restriction. Understanding the critical underlying physiological mechanisms by which nutrition affects reproduction is the base of focus feeding strategy to improve the reproductive performance of the female ruminant.