Advanced onset of puberty after metformin therapy in swine with thrifty genotype.

The prevention and treatment of obesity in children is based on adequate nutrition and exercise plus antihyperglycaemic drugs. Currently, the incidence of childhood obesity is aggravated in ethnicities with thrifty genotype, but there is no available information on the effects of metformin therapy. The relative effects of lifestyle and metformin on patterns of growth, fattening, metabolic status and attainment of puberty were assessed in females of an obese swine model (Iberian gilts), allocated to three experimental groups (group A, obesogenic diet and scarce exercise; group DE, adequate diet and opportunity for exercise; and group DEM, adequate diet and opportunity for exercise plus metformin). Group A evidenced high weight, corpulence and adiposity, high plasma triglycerides and impairments of glucose regulation predisposing to insulin resistance. These features were favourably modulated by adequate lifestyle (group DE), and these effects were strengthened by metformin treatment (group DEM), which induced an improvement in body development by favouring muscle deposition. However, contrary to expectations, metformin advanced the onset of puberty. Metformin treatments would have positive effects on growth patterns, adiposity and metabolic features of young females from ethnicities with thrifty genotype or developing leptin resistance, but a negative effect by advancing the attainment of puberty. This study provides a warning regarding the use of metformin, without further studies, in girls from these ethnicities.

Fertility of a high-altitude sheep model is compromised by deficiencies in both preovulatory follicle development and plasma LH availability.

At high altitude, hypoxia and/or oxidative stress may compromise fertility. This study tested the relative effect of short- or long-term exposure to high-altitude hypobaric hypoxia and oxidative stress in sheep on preovulatory follicle dynamics and gonadotrophin secretion. Thus, growth dynamics, stereidogenic function and competence to ovulate of preovulatory follicles, as well as FSH and LH availability throughout the entire oestrous cycle, were compared among sheep native from low and high altitude, and sheep newcomers to high altitude. The results indicates that short-term exposure in sheep newcomers to high altitude has a deleterious effect on both the ovarian function (affecting preovulatory follicular development) and the pituitary function (diminishing plasma LH availability). On the other hand, there were no detected differences in the preovulatory follicular development in sheep adapted to high altitude for generations and, conversely, LH secretion was increased, which suggests an adaptive mechanism. The treatment with antioxidant agents during a relative short period for the time of folliculogenesis (approximately 1 month and a half) changed substantially the development of preovulatory follicles in short-term exposed sheep to similar patterns than in sheep native and living to both high and low altitude. These results highlight the role of oxidative stress in the detriment of the reproductive function in individuals recently exposed to high-altitude hypoxic environment.

Reproductive response of fat-tailed Barbarine ewes subjected to short-term nutritional treatments including spineless cactus (Opuntia ficus-indica f. inermis) cladodes.

Reproductive outputs in fat-tailed Barbarine sheep in central Tunisia are often low because of feed shortage and the low nutritive value of diets. Supplementation with conventional concentrates is economically unsuitable in central Tunisia, so more cost-effective and sustainable alternative feeding strategies need to be developed. We tested effects of short-term nutritional treatments including cactus cladodes during the induction of 'male effect' on fertility and prolificacy parameters (follicular growth, ovulatory response and early embryo losses). One hundred and twenty ewes were distributed in 4 equal groups balanced for live weight grazed natural pastures and were supplemented for 21 days, starting day 10 after introduction of rams, with cactus cladodes (CA), cactus cladodes and soybean meal (CAS), concentrate (CC) or only soybean meal (S). Nutritional treatment did not affect live weight in this experiment. Ewes receiving cactus had higher number of large pre-ovulatory follicles (≥6 mm; 1.08 ± 0.05), between days 14 and 19 after introduction of rams, than females in the CC and S ewes (0.64 ± 0.06; p < 0.05). However, there were no differences in the onset of oestrous behaviour in response to 'male effect' or in the number of corpora lutea. Average ovulation rates were 1.42 ± 0.16 for CC, 1.47 ± 0.13 for CAS, 1.31 ± 0.15 for CA and 1.31 ± 0.13 for S groups respectively. Finally, reproductive wastages at day 35 after mating were not different between groups being 0.33 ± 0.19 for CC, 0.60 ± 0.17 for CAS, 0.43 ± 0.16 for CA and 0.31 ± 0.15 for S groups respectively. It is concluded that Barbarine ewes fed nutritional treatments including cactus performed similarly to those receiving diets including conventional concentrate feeds.

Short-term beta-carotene-supplementation positively affects ovarian activity and serum insulin concentrations in a goat model.

In early October 2010, adult goats (no.=22, 3.5 yr old, 7/8 Sannen-Alpine, 26° N, 103° W, at 1117 m), were randomly assigned to: i) beta-carotene group (BC) [no.=10; live weight (LW)=45.9±1.97 kg, body condition score (BCS) =3.04±0.08; orally supplemented with 50 mg of BC per goat per day]; ii) control group (CONT) (no.=12; LW=46.2±2.04 kg, BCS=3.0±0.08). Animals received a basal diet of alfalfa hay, corn silage, and corn grain, having free access to water, shade, and mineral salts. During the second half of October, estrus was synchronized by using intravaginal sponges. Thereafter, by mid-follicular phase, an intensive blood sampling (6 h × 60 min) was performed to evaluate serum insulin concentrations (INS) by radioimmunoassay. By the end of the luteal phase, an ultrasonographic scanning was performed to evaluate total ovarian activity (TOA) [TOA=total follicles (TF) + total corpus luteum (TCL)]. The whole experimental period consisted of 34 days pre- and 17 days post-ovulation, for a total of 52 days. Average LW and BCS did not differ (p>0.05) during the experimental period. Nonetheless, increases in TF no. (5.0 vs 3.4±0.6 units; p=0.05), TCL no. (3.4 vs 2.8±0.2 units; p=0.05), TOA (8.1 vs 6.2±0.6 units; p=0.05) and INS (4.6 vs 3.9±0.4 ng ml-1; p=0.05) favored to the BC-supplemented group. A positive correlation between LW (r(2)=0.42; p=0.04) and BCS (r(2)=0.47; p=0.02) with respect to ovulation rate, was detected. BC-supplementation increased ovarian activity in the female goat while positively affected the release pattern of insulin, suggesting a potential role of BC as a central and/or pancreas-activating molecule in adult goats; such results may hold not only physiologic but also clinical significance.

In vitro release of ovarian progesterone is decreased during the oestrous cycle and pregnancy of swine with obesity/leptin resistance.

Previous studies indicate that reproductive prolificacy of obese swine breeds is markedly influenced by embryo losses in early pregnancy. In such period, adequate secretion of progesterone (P4) by the ovary is essential for pregnancy success. This study analyses the luteal functionality during the oestrous cycle and early pregnancy of Iberian sows and Large White x Landrace females, in terms of P4 secretion after in vitro culture of luteal tissue stimulated or not with luteinizing hormone (LH). The secretion of progesterone (expressed in ng/mg of luteal tissue or ng/mgLT) of the corpora lutea of obese Iberian swine was always hampered when compared to lean genotypes, either during early oestrous cycle (110.7 ± 37.8 vs 259.7 ± 10.2 ng/mgLT; p < 0.0001), late oestrous cycle (49.0 ± 3.5 vs 75.92 ± 7.14 ng/mgLT; p < 0.0001) or early pregnancy (38.4 ± 2.1 vs 70.7 ± 5.3 ng/mgLT; p < 0.0001). The differences in basal P4 secretion remained after stimulation with LH. Finally, P4 secretion during early pregnancy of Iberian sows decreased with age and, hence, with obesity features (46.6 ± 4.2 vs 65.5 ± 4.8 ng/mgLT; p < 0.001). In conclusion, the results of the present study provide convincing evidence of a reduced luteal function during oestrous cycle and early pregnancy of sows with obesity/leptin resistance like Iberian sows, which may contribute to the low reproductive efficiency reported in this breed.

Gender-specific early postnatal catch-up growth after intrauterine growth retardation by food restriction in swine with obesity/leptin resistance.

The effects of undernutrition during pregnancy on prenatal and postnatal development of the offspring were evaluated in sows with obesity/leptin resistance. Females were fed, from day 35 of pregnancy onwards, a diet fulfilling either 100% (group control, n=10) or 50% of the nutritional requirements (group underfed, n=10). In the control group, maternal body weight increased during pregnancy (P<0.05) while it decreased or remained steady in the underfed group. At days 75 and 100 of gestation, plasma triglycerides were lower but urea levels were higher in restricted than in control sows (P<0.05 for both). Assessment of the offspring indicated that the trunk diameter was always smaller in the restricted group (P<0.01 at day 50, P<0.005 at days 75 and 100 and P<0.0001 at birth) while head measurements were similar through pregnancy, although smaller in the restricted than in the control group at birth (P<0.05). Newborns from restricted sows were also lighter than offspring from control females (P<0.01) and had higher incidence of growth retardation (P<0.01). Afterwards, during lactation, early postnatal growth in restricted piglets was modulated by gender. At weaning, males from restricted sows were still lighter than their control counterparts (P<0.05), while females from control and underfed sows were similar. Thus, the current study indicates a gender-related differential effect in the growth patterns of the piglets, with females from restricted sows evidencing catch-up growth to neutralise prenatal retardation and reaching similar development than control counterparts.

Glucogenic supply increases oocyte developmental competence in sheep.

The present study aimed to determine the influence of a glucogenic supply on oocyte developmental competence. Oestrous cycles were synchronised in 22 Sarda ewes by the insertion (Day 0) of one intravaginal progestagen-impregnated sponge that was removed after 6 days. After removal, the ewes were randomly allocated into two experimental groups (treated and control ewes) and, from Day 7 to Day 11, treated ewes received oral administration of a glucogenic mixture, whereas control animals received water. Follicular development was stimulated by FSH administration from Days 8 to 10. Glucose metabolism was assessed from Days 7 to 11, whilst follicle and corpus luteum growth dynamics and functionality were evaluated between Days 6 and 11. At Day 11 ovaries were collected and processed for in vitro embryo production. Glucogenic treatment increased both the plasma levels of glucose, progesterone, oestradiol and the number of 2-3-mm follicles (P < 0.05). Higher fertilisation and blastocyst rates (P < 0.05) were obtained after IVM of oocytes recovered from treated ewes compared with control ones. In conclusion, glucogenic treatment modifies follicle and corpus luteum functionality and improves oocyte quality, as evaluated by in vitro developmental kinetics and blastocyst output.

Reproductive, endocrine and metabolic feto-maternal features and placental gene expression in a swine breed with obesity/leptin resistance.

The current study was conducted in a swine breed (Iberian pig) with a genotype that predisposed the pig to obesity. The aim of the study was to determine the morphological, metabolomic and endocrine features of early conceptuses and to elucidate how placental gene expression (related to placentation, angiogenesis and fetal nutrition), maternal hormones and the metabolome affect the fetal environment and fetal growth. Conceptus viability and growth were found to be related to maternal endocrine (plasma progesterone levels) and metabolic features (plasma levels of leptin, cholesterol, HDL-c, LDL-c and triglycerides). These features were related to the placental expression of the vascular endothelial growth factor A (VEGFA) and leptin (LEP) genes, the placental efficiency and, thus, the nutrition and the metabolism of the fetus (availability of glucose, triglycerides and cholesterol, as HDL-c). Viability of conceptuses in females with evidence of dyslipidemia (low plasma levels of total cholesterol due to low HDL-c concentration but high levels of triglycerides) was diminished. The availability of nutrients and metabolic substrates to the conceptus was also affected in females with higher fat deposition and evidence of dyslipidemia. In conclusion, the conceptus viability and growth appear to be strongly related to maternal metabolic features and, thus, affected in females with alterations in lipid metabolism.

Endometrial expression of IFNAR-1 and oxytocin receptor (OTR) is not improved by prostaglandin analogues when compared to progestagens in ewes.

The objective of this study was to investigate differences on the endometrial immunoexpression of type I IFN receptor subunit 1 (IFNAR1) and oxytocin receptor (OTR) during the time of maternal recognition of pregnancy in sheep, when oestrus is synchronized with either prostaglandin analogues (group PG) or conventional progestagens (group P). Plasma progesterone was measured from day 0 to 21 post-coitus (pc) (day 0 = day of oestrus). Immunohistochemistry was performed in samples of uterine horns from pregnant sheep on days 9pc, 13pc, 15pc, 17pc and 21pc to locate IFNAR1 and OTR expression in different endometrial compartments. Mean levels of plasma progesterone were different between treatments, obtaining higher levels in the PG group than in the P group (p < 0.05). Comparing days of pregnancy, IFNAR1 protein expression was different in the luminal epithelium (LE) (p < 0.05), while OTR was different in the LE and in the superficial glandular epithelium (SG) (p < 0.05). Temporal variation on the expression of both proteins from day 9pc to 21pc has been evidenced. IFNAR1 and OTR expression did not show significant differences between treatments. However, the response observed in the endometrium was highly inconsistent when prostaglandin analogues were used. Therefore, the protocol based on prostaglandin analogues still needs to be optimized before being considered as a better alternative to progestagens for oestrous synchronization in sheep.

Diet-induced swine model with obesity/leptin resistance for the study of metabolic syndrome and type 2 diabetes.

The objective of the present study was to determine the suitability of a swine breed with leptin resistance and predisposition to obesity (the Iberian pig) as model for studies on metabolic syndrome and type 2 diabetes. Thus, six Iberian sows had ad libitum access to food enriched with saturated fat (SFAD group; food consumption was estimated to be 4.5 kg/animal/day) whilst four females acted as controls and were fed with 2 kg/animal/day of a commercial maintenance diet. After three months of differential feeding, SFAD animals developed central obesity, dyslipidemia, insulin resistance and impaired glucose tolerance, and elevated blood pressure; the five parameters associated with the metabolic syndrome. Thus, the current study characterizes the Iberian pig as a robust, amenable, and reliable translational model for studies on nutrition-associated diseases.

The cactus effect: an alternative to the lupin effect for increasing ovulation rate in sheep reared in semi-arid regions?

The present study evaluated the effects of supplementation with cactus cladodes on follicular dynamics and ovulatory response of sheep reared in semi-arid areas. A total of 76 ewes were distributed into two equal groups supplemented with either concentrated feed or cactus cladodes. After 30 days of supplementation, no differences were found between feeding regimens on the final live weight (LW; 41.5 ± 0.6 and 42.1 ± 0.7 kg in the Concentrate and Cactus groups respectively) and body condition score (BCS; 1.8 ± 0.3 and 1.8 ± 0.4 for Concentrate and Cactus group respectively). Moreover, no differences were found between the initial and the final values of both LW and BCS; thus, there were no effects of supplementation on any of both parameters. Analysis of follicular population showed that, during the follicular phase induced by ram effect, the number of follicles reaching ovulatory size increased in both groups. However, the number was always higher in Cactus ewes and, at oestrus, Cactus ewes had 1.6 ± 0.2 and Concentrate sheep had 1.2 ± 0.2 large follicles (p < 0.05). Thereafter, ovulation rate was affected by duration of supplementation; being higher in sheep fed with cactus for 6-10 days (1.7 ± 0.1) than in ewes supplied with cactus for more than 11 days (1.3 ± 0.1; p < 0.05), in sheep fed with concentrate for 6-10 days (1.2 ± 0.1; p < 0.01) and even than in individuals subjected to classical flushing with concentrate (1.3 ± 0.1; p < 0.05).

SNP discovery and association study for growth, fatness and meat quality traits in Iberian crossbred pigs.

Iberian pigs and its crosses are produced to obtain high-quality meat products. The objective of this work was to evaluate a wide panel of DNA markers, selected by biological and functional criteria, for association with traits related to muscle growth, fatness, meat quality and metabolism. We used 18 crossbred Iberian pigs with divergent postnatal growth patterns for whole genome sequencing and SNP discovery, with over 13 million variants being detected. We selected 1023 missense SNPs located on annotated genes and showing different allele frequencies between pigs with makerdly different growth patterns. We complemented this panel with 192 candidate SNPs obtained from literature mining and from muscle RNAseq data. The selected markers were genotyped in 480 Iberian × Duroc pigs from a commercial population, in which phenotypes were obtained, and an association study was performed for the 1005 successfully genotyped SNPs showing segregation. The results confirmed the effects of several known SNPs in candidate genes (such as LEPR, ACACA, FTO, LIPE or SCD on fatness, growth and fatty acid composition) and also disclosed interesting effects of new SNPs in less known genes such as LRIG3, DENND1B, SOWAHB, EPHX1 or NFE2L2 affecting body weight, average daily gain and adiposity at different ages, or KRT10, NLE1, KCNH2 or AHNAK affecting fatness and FA composition. The results provide a valuable basis for future implementation of marker-assisted selection strategies in swine and contribute to a better understanding of the genetic architecture of relevant traits.

Preovulatory follicle dynamics and ovulatory efficiency in sows with thrifty genotype and leptin resistance due to leptin receptor gene polymorphisms (Iberian pig).

This study compares follicular function and ovulatory efficiency in 20 sows with obesity/leptin resistance genotype (Iberian pig) and 20 females of lean commercial crosses (Large White × Landrace; LW×L). Estrous cycle was synchronized with progestagens; ovulation was induced with eCG and hCG, in half of the females of each group, to determine its effect. In females of both breeds not treated with gonadotropins, the number of follicles larger than 4.9 mm and the estradiol secretion increased throughout the follicular phase (P<0.05); estradiol values were similar at estrus detection (22.5±1.2 vs. 26.5±0.6 pg/ml respectively, for Iberian and LW×L sows). Moreover, ovulation rate was higher in Iberian pigs (15.3±1.3 CLs) than in LW×L (10.2±1.3 CLs; P<0.05), with mean progesterone values being 18.1±0.7 ng/ml in Iberian and 16.8±0.6 ng/ml in LW×L pigs. Thus, the preovulatory follicular growth and the ovulatory efficiency seem not to be the main limiting factors for reproductive efficiency in Iberian swine. The gonadotropins induced a significant increase, when compared to untreated females (P<0.05), in the number of follicles larger than 4.9 mm growing throughout the follicular phase; however, estradiol values at estrus were lower (P<0.05) in both breeds (9.2±0.7 pg/ml in Iberian vs. 8.6±0.8 pg/ml in LW×L), when compared with the nontreated animals, which suggests defective follicular function after gonadotropin stimulation. There were also no differences between genotypes in ovulation rate (15.2±1.3 vs. 12.7±1.8) and progesterone secretion (21.2±0.8 ng/ml in Iberian and 20.9±0.7 ng/ml in LW×L sows) in the treated animals. In conclusion, the current findings indicate that preovulatory follicular growth and ovulatory efficiency are not main limiting factors for prolificacy in a pig model of leptin resistance and obesity.

Effects of season and superovulatory treatment on embryo yields in fine-wool Merinos maintained under field conditions.

The aim of the study was to assess the effects of superovulatory treatment (multiple FSH-dose vs single-shot FSH treatment) and seasonality on embryo yields in fine-wool Merino ewes. Treatment based on multiple FSH-dose consisted of 200 mg of FSH (Folltropin(®)) administered in seven decreasing doses. Single-shot treatment consisted of a single dose of 70 mg of FSH + eCG. In ewes treated with multiple FSH doses, number of recovered embryos was higher (6.0 ± 0.5 vs 3.5 ± 1.0), while non-fertilization rate was lower (12.8 ± 3.9 vs 40.3 ± 9.5) during the breeding season when compared to the non-breeding season (p < 0.05); although similar values of recovered Grades 1-2 embryos were observed between seasons. During the breeding season, proportion of responding ewes (98.1 vs 57.1%), ovulation rate (13.9 ± 0.8 vs 3.2 ± 1.2), recovered structures (7.9 ± 0.6 vs 1.7 ± 0.7), total recovered embryos (6.0 ± 0.5 vs 1.2 ± 0.6) and good-quality embryos (5.1 ± 0.5 vs 0.9 ± 0.6) were higher for the multiple FSH-dose treatment than for the single-shot protocol. In a similar way, in the non-breeding season, ovulation rate (11.3 ± 1.8 vs 6.0 ± 1.1) and recovered structures (6.6 ± 1.2 vs 2.7 ± 0.6) were higher for the multiple FSH injections protocol than those for the single-shot treatment, resulting in higher recovered Grades 1-2 embryos (3.2 ± 0.9 vs 1.4 ± 0.5). Current results indicate that seasonal anestrus affected embryo yields when applying multiple FSH-dose superovulatory treatment in Merino ewes, by decreasing the number of recovered embryos although the number of recovered good-quality embryos was not affected. During both seasons, multiple FSH injections produced higher ovarian response and number of viable embryos than the single-shot treatment.

Plasma leptin, ghrelin and indexes of glucose and lipid metabolism in relation to the appearance of post-weaning oestrus in Mediterranean obese sows (Iberian pig).

Iberian pig is the most abundant Mediterranean swine. The lack of knowledge of the reproductive physiology of Mediterranean genotypes, with predisposition to obesity, led us to evaluate the influence of body condition and metabolic status at weaning on the resumption of follicular growth and the appearance of post-weaning oestrus. Females failing to display post-weaning oestrus showed a high decrease in backfat mass during lactation; backfat depth at weaning was therefore lower than in sows becoming in oestrus. Females not bearing oestrus behaviour showed lower plasma leptin levels and higher ghrelin concentrations at weaning. Moreover, these sows evidenced dyslipidemic profile (increased triglyceridemia and cholesterolemia) and mobilization of fat reserves. Hence, changes in metabolic regulation of Iberian pigs may originate large effects on the resumption of ovulatory activity after weaning.

Deleterious effects of progestagen treatment in VEGF expression in corpora lutea of pregnant ewes.

The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri-implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen-treated group showed smaller corpora lutea than cloprostenol-treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen-treated group, which even showed a decrease between days 11 and 13. In progestagen-treated sheep, VEGF expression in granulosa-derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.

Patterns of corpora lutea growth and progesterone secretion in sows with thrifty genotype and leptin resistance due to leptin receptor gene polymorphisms (Iberian pig).

The current study aimed to compare luteal function, as measured by corpora lutea dynamics and progesterone secretion, in 10 sows with obesity/leptin resistance genotype (Iberian pig) and 10 females of lean commercial crosses (Large White × Landrace). In all the animals, the oestrous cycle was synchronized with progestagens, and ovulation was induced by exogenous gonadotrophins. Thereafter, number and size of follicles and plasma oestradiol concentration were determined at oestrus detection, and number and size of corpora lutea and progesterone concentration were evaluated from Day 4 to 12 of the cycle. There were no differences between genotypes in follicle population and oestradiol concentration, and ovulation rate (15.2±1.3 in Iberian vs 12.7±1.8 in LWxL sows); however, there was a higher percentage of Iberian than control sows showing luteal cysts (66.7% vs 30%, respectively; p<0.05). In both breeds, both total luteal area and plasma progesterone concentration grew linearly from Day 4 to 8 (p<0.01) and remained more stable between Days 8 and 12, without significant differences between genotypes. In conclusion, current study supports that ovulatory processes and luteal functionality are not the main limiting factors for prolificacy in a pig model of leptin resistance and obesity.

Influence of leptin on in vitro maturation and steroidogenic secretion of cumulus-oocyte complexes through JAK2/STAT3 and MEK 1/2 pathways in the rabbit model.

Extreme body mass indexes may impair reproductive outcome in assisted reproductive technologies. Leptin reflects the amount of body fat and could act as a modulator of oocyte quality through activation of specific transcription factors. The aim of this work was to establish whether: 1) leptin influences meiotic and cytoplasmic oocyte maturation; 2) STAT3 and MAPK mediate the effects of leptin and 3) leptin modulates steroid secretion by cumulus-oocyte complexes (COC) during in vitro maturation (IVM). We confirmed immunolocalisation of leptin receptor in oocytes, cumulus/granulosa cells during the peri-ovulatory period. The confocal study showed that COC supplemented with 1, 10 and 100 ng/ml leptin had a significantly higher metaphase II (MII) percentage than those IVM without leptin (P<0.05) and a similar MII index compared to the group supplemented with 10% FCS. Leptin did not increase the percentage of cytoplasmically matured oocytes in terms of cortical granule migration rate, whereas a significantly higher index was found in the FCS group (P<0.001). Oestradiol concentrations in spent media were higher in the FCS group compared to other treatments (P<0.001). Leptin-stimulated nuclear oocyte maturation was significantly impaired when leptin-induced JAK2/STAT3 and MEK 1/2 activation was suppressed by the inhibitors (P<0.001). Steroid secretion of COC was not affected by leptin activation of JAK2/STAT3 or MEK 1/2 pathways. In conclusion, JAK2/STAT3 and MEK 1/2 pathways mediate the enhancement of nuclear oocyte maturation by leptin; however, neither cytoplasmic oocyte maturation nor steroidogenic response of COC were improved in the present rabbit model.

Ultrasonographic imaging in small ruminant reproduction.

CONTENTS: Ultrasonography is a simple, reliable and non-invasive imaging technique without secondary effects. Application of ultrasonography in veterinary practice, and specifically in small ruminants, has developed to become the most efficient diagnostic tool for managing reproduction. The objectives of current revision are to offer an overview, after a short reminder of equipment and techniques for imaging genitalia in small ruminants, of the uses and utility of ultrasonography for the evaluation of physiological and pathological conditions in males and females and for the application of assisted reproductive technologies.

Short-term undernutrition affects final development of ovulatory follicles in sheep synchronized for ovulation.

The objective of this study was to determine, in sheep, the effect of a short-term undernutrition on growth dynamics and competence of pre-ovulatory follicles. Synchronization of sexual cycles and induction of ovulation were performed, with progestagens and gonadotrophins, in 14 adult female sheep. Morphological characteristics and developmental competence of ovarian follicles to achieve ovulation were determined by imaging techniques (ultrasonography and laparoscopy) and blood sampling. All the animals ovulated and mean ovulation rates were similar between groups (2.0 ± 0.6 corpora lutea in control ewes and 2.2 ± 0.8 in undernourished sheep). However, nutritional restriction, even during a short period, was related to the presence of large follicles in static growing phase which, despite reaching ovulation, persisted static during the induced follicular phase and evidenced functional alterations as there was no inhibition of the development of subordinate follicles. Thus, this study suggests the existence of deleterious effects from short-term undernutrition on functionality of pre-ovulatory follicles, which can compromise fertility.