Bessie Urquieta

Biological activity of the seminal plasma of alpacas: stimulus for the production of LH by pituitary cells.

South American camelids are induced ovulators and require a stimulus to trigger the LH surge responsible for the ovulation. Seminal plasma (SP) of fertile alpacas (Lama pacos) was tested using a bioassay of pituitary cells to study the effect of seminal plasma on LH release. Plates containing rat pituitary cells (2 x 10(5) cells/90-95% viability) were cultured adding: (A) whole SP (WSP) treated with charcoal-dextran, or 1:2 or 1:4 proportions diluted in culture medium (DMEM/HEPES + antibiotics), or (B) 1:2 SP + anti-GnRH rabbit serum (inhibitory potency 10(-5) M), or (C) 1:2 SP + anti-GnRH + 100 nM synthetic GnRH (buserelin acetate) or (D) 100 nM, 50 nM, 10 nM, and 1 nM synthetic GnRH. Concentration (ng/ml) of LH secreted (Sec) and contained (Con) was analyzed using RIA 125I and the percentage of Sec and Con in each experiment was determined. The results of LH Sec for the cells treated with 50, 10, and 1 nM GnRH were 39, 13, and 1.5%, respectively (r2 = 98.41%, r = 0.9920) but cells treated with 100 nM GnRH secreted 10% of LH. With WSP, 1:2, or 1:4 SP the LH Sec was of 44.5% (3.25 ng/ml), 27% (1.9 ng/ml), and 18% (1.2 ng/ml), respectively. The exposure of cells to 1:2 SP + anti-GnRH, or to 1:2 SP + anti-GnRH/100 nM GnRH produced 31% (2.20 ng/ml) and 30% (1.8 ng/ml) of LH Sec, respectively. These results suggest that the SP of alpacas could have some factor(s) different from GnRH that would contribute to the mechanisms of LH secretion and to the induced ovulation in the female alpaca.

Effects of antioxidant vitamins on newborn and placental traits in gestations at high altitude: comparative study in high and low altitude native sheep

The present study evaluated the hypothesis that the effects of hypoxia on sheep pregnancies at high altitude (HA) are mediated by oxidative stress and that antioxidant vitamins may prevent these effects. Both HA native and newcomer ewes were maintained at an altitude of 3,589 m during mating and pregnancy. Control low altitude (LA) native ewes were maintained at sea level. Half of each group received daily oral supplements of vitamins C (500 mg) and E (350 IU) during mating and gestation. Near term, maternal plasma vitamin levels and oxidative stress biomarkers were measured. At delivery, lambs were weighed and measured, and placentas were recovered for macroscopic and microscopic evaluation. Vitamin concentrations in supplemented ewes were two- or threefold greater than in non-supplemented ewes. Plasma carbonyls and malondialdehyde in non-supplemented ewes were consistent with a state of oxidative stress, which was prevented by vitamin supplementation. Vitamin supplementation increased lamb birthweight and cotyledon number in both HA native and newcomer ewes, although placental weight and cotyledon surface were diminished. Placentas from vitamin-supplemented HA ewes were similar to those from ewes at sea level, making these placental traits (weight, number and diameter of cotyledons) similar to those from ewes at sea level. Vitamin supplementation had no effect on LA pregnancies. In conclusion, supplementation with vitamins C and E during pregnancy at HA prevents oxidative stress, improving pregnancy outcomes.

Alpaca semen characteristics previous to a mating period

Increasing the knowledge of the semen characteristics in the alpaca will contribute to understanding one of the many factors that affect the poor fertility rate in this species. Ten adult male alpacas, 2.6-10 years of age, average weight 64.7 +/- 4.7 kg were used. The animals were distributed randomly into two groups of five each and submitted alternatively to two semen collections, using an artificial vagina and sexually receptive females. For the first semen collection the animals had a sexual rest period of about 90 and 45 days before the second. Duration of semen collection, color and volume of ejaculate were recorded, and sperm concentration and morphology (light microscopy) were evaluated. Descriptive statistical analyses were used for each variable, considering all samples obtained (n= 19). An analysis of variance for animal groups and opportunity of collection were used for quantitative variables. Most frequent color was opalescent white (84.2%). There were no statistical differences among male groups or between semen collections. The average values and standard deviations for the quantitative variables were: 12.3 +/- 7.2 min for semen collection time, 1.8 +/- 0.8 ml for ejaculate volume, (17.6 +/- 26.1) x 10(6) sperm/ml for sperm concentration and 34.0 +/- 52.2 x 10(6) for total number of sperm per ejaculate. The percentage of normal spermatozoa was 51.0 +/- 12.4%. From the total abnormalities, that of mid piece segment (14.4%) was the most frequent. These results indicate that male alpaca have poor semen quality, when compared with other domestic species. Nevertheless, for the evaluation of male alpaca as breeders it would be necessary to create a protocol for the selection of them, where phenotypic, behavioral and seminogram aspects are considered. The values reported herein define the characteristics of the alpaca semen that could be considered as the initial base of the seminal analysis to select male alpacas before mating.

Fertility in a high-altitude environment is compromised by luteal dysfunction: the relative roles of hypoxia and oxidative stress

BackgroundAt high altitudes, hypoxia, oxidative stress or both compromise sheep fertility. In the present work, we tested the relative effect of short- or long-term exposure to high altitude hypobaric hypoxia and oxidative stress on corpora luteal structure and function.MethodsThe growth dynamics of the corpora lutea during the estrous cycle were studied daily by ultrasonography in cycling sheep that were either native or naïve to high-altitude conditions and that were supplemented or not supplemented with antioxidant vitamins. Arterial and venous blood samples were simultaneously drawn for determination of gases and oxidative stress biomarkers and progesterone measurement. On day five after ovulation in the next cycle, the ovaries were removed for immunodetection of luteal HIF-1alpha and VEGF and IGF-I and to detect IGF-II gene expression.ResultsThe results showed that both short- and long-term exposure to high-altitude conditions decreased luteal growth and IGF-I and IGF-II gene expression but increased HIF-1 alpha and VEGF immunoexpression. The level of plasma progesterone was also increased at a high altitude, although an association with increased corpus luteum vascularization was only found in sheep native to a high-altitude location. Administration of antioxidant vitamins resulted in a limited effect, which was restricted to decreased expression of oxidative stress biomarkers and luteal HIF-1alpha and VEGF immunoexpression.ConclusionsExposure of the sheep to high-altitude hypobaric hypoxia for short or long time periods affects the development and function of the corpus luteum. Moreover, the observed association of oxidative stress with hypoxia and the absence of any significant effect of antioxidant vitamins on most anatomical and functional corpus luteum traits suggests that the effects of high altitude on this ovarian structure are mainly mediated by hypoxia. Thus, these findings may help explain the decrease in sheep fertility at a high altitude.

Steroidogenesis in sheep pregnancy with intrauterine growth retardation by high-altitude hypoxia: effects of maternal altitudinal status and antioxidant treatment.

Sheep pregnancy in high-altitude environments frequently involves hypoxia and oxidative stress and causes intrauterine growth retardation. The adverse effects of altitude on fetal growth can be prevented by the administration of antioxidant vitamins, but the mechanisms responsible are not well known. The maintenance of a viable pregnancy depends largely on adequate placental steroidogenesis, especially in the last two-thirds of pregnancy. Thus, in the present study we evaluated the effect of antioxidant vitamins (C and E) on plasma concentrations of progesterone and 17β-oestradiol during the last two-thirds of high-altitude pregnancies in ewes both native and naïve to the high-altitude environment. In addition, pregnancy outcomes were evaluated by determining the bodyweight of newborn lambs. Sex steroid patterns differed between ewes with and without vitamin supplementation. Concentrations of plasma progesterone and 17β-oestradiol were significantly higher in the supplemented groups from approximately 40 days before parturition until near term. Newborn weights were significantly lower in animals not adapted to the higher altitude, and vitamin supplementation prevented this decrease. In conclusion, the administration of antioxidant vitamins in the present study enhanced placental steroidogenesis, thus favouring fetal development in pregnancies developing at high altitudes.

Distribution of keratins, vimentin, and actin in the testis of two South American camelids: Vicuna (Vicugna vicugna) and llama (Lama glama). An immunohistochemical study

The purpose of the present study was to investigate the pattern of distribution of cytokeratins, vimentin and muscular actin in the testis of vicuna (Vicugna vicugna) and llama (Lama glama) two species of camelids native of the Andean high plateau of South America.

Ovarian follicular dynamics and hormonal secretory profiles in guanacos (Lama guanicoe)

The objective of the present study was to describe ovarian activity in 11 adult non-mated guanacos, evaluated every second day for 40 days by transrectal ultrasonography and by plasma estradiol-17beta and progesterone concentrations. An inverse relationship (r = -0.29, P < 0.001) was observed between the diameter of the largest ovarian follicle and the total number of follicles indicating that follicular growth in guanacos occurs in waves. The mean duration of follicular wave was 15.1 +/- 4.2 days with a range from 9 to 26. The follicular growth phase was 7.0 +/- 2.4 days, the static phase 3.0 +/- 1.2 days, the regression phase 5.2 +/- 2.1 days and the inter-wave interval was 12.6 +/- 5.6 days. The maximum follicular diameter in each follicular wave was 10.2 +/- 2.1 mm with a range from 7.2 to 16.1 mm. Inter-wave intervals of longer duration were associated with a larger maximum follicle diameter (P < 0.001). Follicular activity alternated between ovaries in 93% of the waves with 48% of dominant follicles occurring in the left and 52% in the right ovary without differences (P > 0.05). Plasma estradiol-17beta concentrations showed a wave-like pattern, varying from 20.0 to 92.1 pmol/L. Plasma progesterone concentrations remained below 1 nmol/L without any ultrasonic evidence of ovulation during the study. These results in guanacos suggest a follicular wave pattern more similar to the llama (Lama glama) than previously described in other South American and Old World camelid species.

Plasma progesterone in alpaca (Lama pacos) during pregnancy, parturition and early postpartum

Plasma progesterone concentration during pregnancy and the early postpartum period was measured by radioimmunoassay in five alpacas (Lama pacos), of the Huacaya breed, whose pregnancy length had a mean of 344.8 +/- 4.4 days. Concentration of progesterone increased (P < 0.001) from low premating values (0.11 +/- 0.06 nmol/l) to greater values at 30 days of pregnancy (8.05 +/- 1.13 nmol/l) and remained high up to 2 months of pregnancy. A slight transitory decline was observed between 3 to 7 months. Concentration of plasma progesterone dropped markedly during the 72 h before parturition especially, at the day of parturition.

Endocrine changes during pregnancy, parturition and post-partum in guanacos (Lama guanicoe)

Plasma concentrations of progesterone (P4), estradiol-17beta (E2), estrone (E1) and estrone sulfate (E1S) were measured during gestation in eight guanacos kept in captivity. Gestational length was 346.1+/-9.8 days. P4 plasma concentrations increased after ovulation and remained elevated until parturition. However, during the last 4 weeks of gestation, a gradual decrease from 4.17x1.17(+/-1)nmol/L to 2.02x1.95(+/-1)nmol/L on day 5 before parturition was observed, followed by a more abrupt final decline to baseline concentrations which were reached on the day after parturition. Mean E2 plasma concentrations started to increase during the eighth month of gestation, and were significantly elevated up to maximum concentrations of 484.7x1.21(+/-1)pmol/L during the last 2 months of pregnancy. Concentrations returned to baseline during the last 2 days of gestation. An increase of E1S concentrations (p<0.01) was observed in the eleventh month of gestation. Mean E1S concentrations remained rather constant during the last 3 weeks of gestation between 4 to 8nmol/L until parturition, when a steep precipitous decline was observed. E1 concentrations were slightly elevated during the last 4 weeks of gestation, however, maximum concentrations did not exceed 1.5nmol/L. The results show distinct species specific features of gestational steroid hormone profiles in the guanaco in comparison to domestic South American camelids, such as a more pronounced gradual prepartal decrease of P4 concentrations prior to the final decline to baseline, and clearly lesser E1S concentrations during the last 4 weeks of gestation, which lack a continuous prepartal increase.

Histological Characteristics and Steroid Concentration of Ovarian Follicles at Different Stages of Development in Pregnant and Non-pregnant Dairy Cows

The aim of the study was to investigate the histological characteristics and steroid concentrations in follicular fluid of different populations of follicles at different stages of development, during pregnancy and the oestrous cycle in cows. Follicles from ovaries collected at a slaughterhouse were allocated into three size categories (small, 2–5.9 mm; medium, 6–13.9 mm; and large, 14–20 mm) in pregnant and non-pregnant cows. Slices were stained with HE and PAS for histological analysis. Follicular fluid was pooled according to size and pregnancy status and estradiol, testosterone and progesterone concentrations in follicular fluid were determined by RIA. Characteristics of healthy follicles did not differ, regardless of follicle size or pregnancy status. Total histological atresia was significantly higher in pregnant cows than in non-pregnant cows (p < 0.05). Estradiol increased and testosterone decreased significantly, while follicles increased in size, in both non-pregnant and pregnant cows (p < 0.05). Nonpregnant cows had the highest estradiol values in follicles of all sizes. Medium and large follicles from pregnant cows showed the lowest testosterone concentration (p < 0.05). Progesterone levels increased with follicle size only in non-pregnant animals. In large follicles, progesterone concentration was significantly higher in non-pregnant cows than in pregnant cows (p < 0.05). Considering steroid concentration and histological findings, most large follicles might be atretic during pregnancy in cattle.