José Alfonso Abecia

Pharmaceutical Control of Reproduction in Sheep and Goats

Small ruminant species such as sheep and goats are short-day breeders, which is a crucial factor affecting the offer of lambs and kids throughout the year. An appropriate management of reproduction allows ewes and does to breed in the spring to increase the supply of product to the marketplace on a year-round basis. Pharmaceutical control of reproduction is possible, usually through administration of hormones or analogues related to the natural estrous cycle, such as progesterone, prostaglandins, and/or melatonin.

Effect of undernutrition on uterine progesterone and oestrogen receptors and on endocrine profiles during the ovine oestrous cycle

In the present study, it was investigated whether undernutrition affected the binding capacity, immunoreactivity and mRNA expression for uterine oestrogen and progesterone receptors (ER and PR, respectively) in sheep, as well as whether the responses were associated with changes in plasma concentrations of progesterone (P4), oestradiol (E2), glucose, fatty acids, insulin, leptin and insulin-like growth factor (IGF)-I during the oestrous cycle. Twenty ewes were fed either 1.5 (C) or 0.5 (L) times their maintenance requirements and were killed on Day 5 or 14 of the cycle (Day 0 = oestrus). Compared with Group C, Group L had higher concentrations of non-esterified fatty acids and lower concentrations of insulin, leptin and IGF-I. Group L also had higher plasma concentrations of P4 during the final days of the luteal phase. At oestrus in both treatment groups, there were peaks in the concentrations of glucose, insulin and IGF-I. For ER and PR, transcript expression, binding capacity and immunoreactivity were higher on Day 5 than on Day 14 of the cycle. The binding capacities for ER and PR were lower in Group L than in Group C on Day 5. Group C showed more immunoreactive staining for ER than did Group L in two of five cell types, whereas no effect of treatment was observed for PR immunoreactivity. There was more PR mRNA in the uterine horn contralateral to the corpus luteum in Group C than in Group L ewes. We conclude that undernutrition impairs steroid receptor expression and binding capacity. This may alter the uterine environment and help explain the reductions in embryo survival.

Seasonal variations of melatonin in ram seminal plasma are correlated to those of testosterone and antioxidant enzymes

BackgroundSome breeds of sheep are highly seasonal in terms of reproductive capability, and these changes are regulated by photoperiod and melatonin secretion. These changes affect the reproductive performance of rams, impairing semen quality and modifying hormonal profiles. Also, the antioxidant defence systems seem to be modulated by melatonin secretion, and shows seasonal variations. The aim of this study was to investigate the presence of melatonin and testosterone in ram seminal plasma and their variations between the breeding and non-breeding seasons. In addition, we analyzed the possible correlations between these hormones and the antioxidant enzyme defence system activity.MethodsSeminal plasma from nine Rasa Aragonesa rams were collected for one year, and their levels of melatonin, testosterone, superoxide dismutase (SOD), glutathione reductase (GRD), glutathione peroxidase (GPX) and catalase (CAT) were measured.ResultsAll samples presented measurable quantities of hormones and antioxidant enzymes. Both hormones showed monthly variations, with a decrease after the winter solstice and a rise after the summer solstice that reached the maximum levels in October-November, and a marked seasonal variation (P < 0.01) with higher levels in the breeding season. The yearly pattern of GRD and catalase was close to that of melatonin, and GRD showed a significant seasonal variation (P < 0.01) with a higher activity during the breeding season. Linear regression analysis between the studied hormones and antioxidant enzymes showed a significant correlation between melatonin and testosterone, GRD, SOD and catalase.ConclusionsThese results show the presence of melatonin and testosterone in ram seminal plasma, and that both hormones have seasonal variations, and support the idea that seasonal variations of fertility in the ram involve interplay between melatonin and the antioxidant defence system.

Early pregnancy alters the metabolic responses to restricted nutrition in sheep

This study investigated whether a 27-day period of nutrition at half-maintenance during early pregnancy (up to Day 14) could alter maternal endocrine responses. Forty-six ewes were fed all or half of their maintenance requirements and slaughtered on Day 14 of the oestrous cycle or pregnancy. We used real time RT-PCR to study gene expression of growth hormone receptor (GHR) and leptin in adipose tissue and GHR, GHR1A and of the insulin-like growth factor I (IGF-I) in the liver. Blood profiles of metabolites and metabolic hormones were also determined. Throughout the experiment, underfed animals presented lower body weight and body condition, greater plasma concentrations of non-esterified fatty acids (NEFA), and lower plasma concentrations of leptin, compared to adequately fed animals. Undernutrition affected the patterns of gene expression in adipose and hepatic tissues, and the responses differed between pregnant and non-pregnant ewes. In adequately fed ewes, pregnancy up-regulated leptin mRNA expression in adipose tissue, a response that was impaired in underfed ewes. The hepatic expression of IGF-I mRNA was increased by pregnancy in underfed animals while no effect was observed in adequately fed ewes. It remains to be determined whether the changes in the endocrine milieu are paralleled by modifications in uterine gene expression that could alter the environment of the embryo during early pregnancy.

The effect of exogenous melatonin during the non-reproductive season on the seminal plasma hormonal profile and the antioxidant defence system of Rasa Aragonesa rams.

The aim of this study was to analyze the effect of melatonin implants, during the non-reproductive season, on the content of melatonin, testosterone and 17-β estradiol levels, and the antioxidant enzymes glutathione peroxidase, glutathione reductase, superoxide dismutase and catalase of seminal plasma in Rasa Aragonesa rams. Five rams were implanted with melatonin, and four others were used as a control group. Seminal plasma was separated from ejaculates collected one week before melatonin treatment until 21 weeks after melatonin placement, and the hormonal levels and the antioxidant enzyme activity were determined. Exogenous melatonin treatment significantly (P<0.05) increased the levels of endogenous melatonin in seminal plasma immediately, and this effect lasted for 14 weeks. Testosterone and 17-β estradiol levels significantly (P<0.05) increased four and eight weeks after melatonin treatment, respectively. As regards the antioxidant enzymes, melatonin treatment significantly increased (P<0.05) glutathione peroxidase and glutathione reductase activity only, and had no effect on superoxide dismutase and catalase. Therefore, melatonin treatment during the non-breeding season modifies the seminal plasma hormonal profile and some antioxidant enzyme activity in Rasa Aragonesa rams.

Effect of undernutrition on the uterine environment during maternal recognition of pregnancy in sheep

The effects of pregnancy and undernutrition on endometrial gene expression were investigated in ewes fed all or half their maintenance requirements and killed on Day 14 of pregnancy or of the oestrous cycle. The endometrial expression of progesterone, oestrogen, oxytocin and interferon receptors (PR, ERalpha, OXTR and IFNAR, respectively), cyclo-oxygenase (COX)-2, proliferating cell nuclear antigen (PCNA), insulin-like growth factors (IGF)-I and -II, and IGF-1 receptor (IGF-1R) was studied by immunohistochemistry or real-time reverse transcription-polymerase chain reaction. The luminal epithelium of cyclic control ewes was devoid of PR staining and had relatively high levels of ERalpha, OXTR, COX-2 and IFNAR2. The presence of a conceptus decreased the in vitro uterine secretion of prostaglandin (PG) F(2alpha) and the expression of IFNAR2 in most cell types, and increased the gene expression of IGF-I and IGF-II. Undernutrition tended to increase ERalpha protein and gene, but decreased in vitro uterine secretion of PGE(2) and the gene expression of IFNAR2 in cyclic ewes. There was no effect of undernutrition on pregnancy rates or the number of conceptuses recovered. Consistent with this, undernutrition of pregnant ewes did not have any effect on uterine gene expression. Moreover, in cases where changes were observed in cyclic ewes, these changes were negated when a conceptus was present.

Identification and immunolocalisation of melatonin MT1 and MT2 receptors in Rasa Aragonesa ram spermatozoa

The reproductive seasonality of sheep suggests that melatonin receptors may be present in ram spermatozoa. The present study confirms the presence of melatonin MT(1) and MT(2) receptors. The MT(1) receptor was detected using immunocytochemistry, with four sperm subpopulations identified based on the following labelling patterns: (1) one small subpopulation with labelling over the entire head and tail; (2) one of two main subpopulations that exhibited reactivity at the equatorial, post-acrosomal, neck and tail regions; (3) another main subpopulation with equatorial and tail labelling only; and (4) a subpopulation in which staining was detected only in the tail. Immunocytochemistry revealed the presence of the melatonin MT(2) receptor, with intense staining on the acrosome, post-acrosomal region and neck and tail regions of all cells, but not in the equatorial region. Western blot identification of ram protein extracts revealed a 39-kDa band compatible with both MT(1) and MT(2) receptors, a 75-kDa band compatible with MT(1)/MT(2) heterodimerisation, a 32-kDa band compatible with MT(1) receptor activation and a double band of 45-55 kDa that is compatible with MT(2) receptor homodimerisation or heterodimerisation with other G-proteins. In conclusion, we provide evidence of the presence of MT(1) and MT(2) receptors in ram spermatozoa, although the biochemical pathway triggered by these receptors and their function in terms of fertility remains to be elucidated.

The role of nutrition in the regulation of LH secretion during anestrus by the serotoninergic and dopaminergic systems in Mediterranean ewes treated with melatonin

The steroid-dependent inhibition of LH secretion involves dopaminergic and serotoninergic systems but it is unclear how the plane of nutrition affects this inhibition during anestrus in melatonin treated ewes. Melatonin implants (18 mg) were inserted (Day 0) into ovariectomized, estradiol treated adult Rasa Aragonesa ewes on a high (H; n = 8) or low energy diet (L; n = 6) which were applied in early anestrus (Day 29-57) and late anestrus (Day 90-104). Cyproheptadine (0.1 mg/ kg), a serotoninergic (SHT2) receptor antagonist, was administered in early and late anestrus (Day 50 and 107) followed by pimozide (0.08 mg/kg), a dopaminergic2 receptor antagonist (Day 57 and 114). The H ewes had significantly higher LH concentrations (P < 0.05) before cyproheptadine treatment in early anestrus. The H and L ewes responded in a similar way to the antagonists in both early and late anestrus, except for L ewes who had a higher LH pulse amplitude after pimozide treatment in both periods (P < 0.05). During early anestrus, cyproheptadine tended to increase (P = 0.06) LH pulse frequency in L ewes and LH concentrations in H ewes. The LH secretion also increased in L ewes after pimozide administration during early anestrus (P < 0.05 for mean LH concentrations and LH pulse frequency and amplitude). However, pimozide dramatically increased LH secretion during late anestrus (Day 114) irrespective of the plane of nutrition (P = 0.06-0.08 for LH pulse frequency and P < 0.05 for LH concentrations and pulse amplitude). In melatonin treated Mediterranean ewes, the plane of nutrition appeared to modify the effect of dopaminergic and serotoninergic systems on the steroid-dependent inhibition of LH secretion throughout anestrus.

How pigs influence indoor air properties in intensive farming: practical implications – a review

Abstract Indoor environmental conditions in intensive pig farms are influenced by both the outdoor air temperature and humidity, and the heat, moisture and gas exchanges between the animal and the air. As ventilation rate in pig facilities is usually estimated in temperature, moisture and even CO2 balances, estimation of heat losses or gains, and moisture and CO2 production from the animal is needed, but the contribution of other sources of the barn, such as slurry or wet surfaces have also to be taken into account. Some recent studies have been conducted to update total heat and moisture production at farm level, showing that current, historical standards of latent heat transfer are consistently lower than those reported recently at facility level, for both adult and growing animals. Also, CO2 production needs to be updated by including an estimation of its release from slurry. These new values will help with updating the standards for ventilation rate recommendations and design of the modern intensive pig buildings.

Effect of a single-reduced dose of equine follicular fluid on the timing of oestrus and reproductive performances in goats

To evaluate the use of equine follicular fluid (EFF) in a single-reduced dose on the synchronisation of oestrus in Boer×Nubian goats, the following experimental treatments were applied: Goats in group T1 (FGA, n=10) were synchronised in oestrus by intravaginal sponges impregnated with 45 mg FGA (Chronogest-MSD) for 12 days; group T2 (T2, FGA + eCG, n=16) goats were synchronised as in T1, plus 400 IU eCG, 48 h before sponge withdrawal; T3 goats (FGA + PGF2α, n=11) received the sponge as T1 plus an i.m. dose of PGF2α (1 ml, Prosolvin-MSD) 48 h before sponge withdrawal; T4 (FGA + EFF, n=19) animals received the sponge as T1 plus 2 ml EFF at sponge withdrawal and finally, group T5 (FGA + PGF2α + EFF, n=10) was like T3 plus 2 ml EFF at sponge withdrawal. Presentation: onset and duration of oestrus were recorded, and fertility, prolificacy and fecundity were calculated. All females showed oestrus. The onset of oestrus was different (P < 0.01), T4 and T5 goats showing a delay (50.5±4.9 and 48.2±2.0 h for T4 and T5 groups, respectively). Litter size (2.10±0.11 kids/kidding) and fecundity (1.90±0.23 kids/goat) were higher (P<0.01) in T5 goats compared to T1 and T4, with no differences with T2 and T3. The combination of EFF with FGA and PGF2α in goats produces a delayed onset of oestrus and improves prolificacy and fertility in comparison with the use of FGA and EFF.