Ultrastructural observations on the placenta of the alpaca (Lama pacos).

The alpaca is one of the four South American species of the family Camelidae. Its placenta, like that of other camelids, is diffuse and epitheliochorial in type. The chorionic epithelium is thrown into unbranched villi or folds which are closely apposed to corresponding undulations of the uterine epithelium, and the fetal-maternal interface consists of an intricate interdigitation of fetal and maternal microvilli. In late gestation both chorionic and uterine epithelia are deeply indented by placental capillaries, so that the minimum intercapillary distance across the diffusion pathway may be as little as 2 microns. This distance appears to be less than that found in the epitheliochorial placenta of any other species of domestic ungulate in late gestation: it may be one of several adaptations to pregnancy at high altitude. Delivery of the fetal membranes occurs some 45 minutes after the birth of the fetus. The placenta is non-deciduate.

Plasma concentrations of 15-ketodihydro-PGF2 alpha, progesterone, oestrone sulphate, oestradiol-17 beta and cortisol during late gestation, parturition and the early post partum period in llamas and alpacas.

Plasma concentrations of 15-ketodihydroprostaglandin (PG) F2 alpha, progesterone, oestrone sulphate, oestradiol-17 beta and cortisol during late gestation, parturition and the early post-partum period were measured in six llamas and five alpacas. During the last 100 days of pregnancy, 15-ketodihydro-PGF2 alpha concentrations increased steadily until the day of parturition when a massive release was detected (P < 0.01) concomitant with a decrease in progesterone concentrations (P < 0.01). The highest PGF2 alpha metabolite concentrations (159 +/- 35 nmol l-1 and 92 +/- 29 nmol l-1 in llamas and alpacas respectively) were detected in the sample collected during the morning on the day of parturition. Basal concentrations were registered by day 3 after delivery. Plasma concentrations of oestrone sulphate started to increase 80 days before parturition and reached peak concentrations immediately before parturition (15 +/- 3 nmol l-1 in llamas and 18 +/- 5 nmol l-1 in alpacas). Oestrone sulphate concentrations dropped sharply (P < 0.01) on the day of parturition in llamas and one day later in alpacas, whereupon they remained relatively unchanged until at least 20 days postpartum. Oestradiol-17 beta concentrations were higher than 180 pmol l-1 during the last 45 days of pregnancy, began to decrease on the day of parturition and reached very low concentrations within the following two days. High oestradiol-17 beta concentrations were registered 7 days postpartum in all alpacas (P < 0.05) and within 10 days of parturition in five of six llamas (P < 0.01). No significant cortisol peaks were observed around parturition, but mean concentrations were increased in both species.

Hemorrhagic ovarian follicles in llamas.

The ovaries of 74 llamas were examined daily by transrectal ultrasonography for at least 30 d. Hemorrhagic follicles were observed in 13 (18%) llamas (incidence per anovulatory dominant follicle, 16%). The proportion of llamas in which a hemorrhagic follicle was detected was different among groups (nonmated, 8/25; mated to a vasectomized male, 4/21; mated to an intact male, nonpregnant, 1/10; mated to an intact male, pregnant, 0/18; P<0.05). A hemorrhagic follicle, observed grossly after ovariectomy, was large (13 mm) and fluctuant, with a thin translucent wall and dark red contents. No ovulatory stigma was detected, and after incising the wall, bloody fluid escaped and the follicle collapsed leaving only a small blood clot within the antrum. Ultrasonically, the formation of a hemorrhagic follicle was indicated by scattered free-floating echogenic spots within the follicular antrum which swirled upon ballottement of the ovary. The antral contents appeared to become organized (did not swirl when ballotted) after follicle growth ceased. Ultrasonic indications of antral hemorrhage were not observed in any follicles in which ovulation was later detected (0/45 ovulatory follicles). All of the hemorrhagic follicles (13/13) involved the dominant follicle of a wave during which no copulatory stimulus was applied. Hemorrhagic follicles were apparently anovulatory and were repeatable (P<0.05) within individuals. The interval from first detection to the first day of maximum diameter was longer (P<0.05) and maximum diameter was greater (P<0.0001) for hemorrhagic follicles than nonhemorrhagic follicles (16.4 versus 13.1 d and 22.1 versus 12.8 mm, respectively); however, the interwave interval was not affected by the presence of a hemorrhagic follicle. Luteinization of the hemorrhagic follicle was indicated (thickened wall) in two llamas by an elevated plasma progesterone concentration and/or by ultrasound. By their large size, hemorrhagic follicles may be interpreted as hemorrhagic follicular cysts; however, they were not associated with other ovarian irregularities or with infertility.

In situ observation of the ovaries of llamas and alpacas by use of a laparoscopic technique.

A laparoscopic technique was used to observe in situ the ovaries of llamas and alpacas. Use of this technique results in minimal surgical stress and allows repetitive observation of various reproductive stages in the same animal. Ovarian follicular activity before and after breeding, time of ovulation, corpus luteum formation and regression, early embryo death, failure of ovulation, and the early events of pregnancy can be assessed quickly and safely. For 1,291 laparoscopies performed, postsurgical disturbance was not observed, and animals were released from the clinic 1 hour after laparoscopy.

Evaluation of intra-abdominal vasectomy in llamas and alpacas.

A method of intra-abdominal vasectomy (deferentectomy), using a laparoscopic technique in llamas and alpacas, was evaluated. Food was withheld from 14 animals for 24 hours before sedation and laparoscopy. The ductus deferens was located close to the bladder, and a 2- to 3-cm section of ductus deferens was resected with the aid of a pair of hook scissors. The procedure was completed in 5 to 8 minutes, and animals did not have any adverse effects. Forty-five days after vasectomy, animals were used for detecting sexually receptive females or inducing ovulation during reproductive physiologic studies. The procedure did not impair libido and had no effect on male sexual behavior for many years. The technique is simpler, safer, and faster than the traditional external approach.

Endocrine changes after mating in pregnant and non-pregnant llamas and alpacas.

Plasma concentrations of oestradiol-17 beta, progesterone, 15-keto-dihydro-PGF2 alpha and luteinizing hormone (LH) were monitored in llamas and alpacas after mating with an intact male. Concentrations of LH and PGF2 alpha metabolite were high immediately after copulation. Ovulation occurred in 92% of the animals. The first significant increases in progesterone were recorded on day 4 after mating. In non-pregnant animals the lifespan of the corpus luteum was estimated to be 8-9 days. Luteolysis occurred in association with the release of PGF2 alpha. In pregnant animals, a transient decrease in progesterone concentrations was observed between days 8 and 18 in both species. No significant changes in PGF2 alpha secretion were registered during this period. Oestradiol-17 beta concentrations were high on the day of mating, declined to low values on day 4, and started to increase again on day 8. Peak values after luteolysis in non-pregnant animals were significantly higher than those registered in pregnant ones. Furthermore, concentrations of oestradiol-17 beta were elevated for a longer period in non-pregnant than in pregnant animals. The results suggest that progesterone from the corpus luteum exerts a negative influence on follicular activity in pregnant animals by reducing oestradiol-17 beta secretion.

Chronology of early embryonic development and embryo uterine migration in alpacas.

The objectives were to: (1) describe the chronology of early embryonic development from ovulation to entry into the uterus; and (2) to determine the timing of embryo migration to the left uterine horn when ovulation occurred from the right ovary. The experiment was conducted in Peru. Females (n = 132) were randomly assigned to 15 experimental groups. All females were mated to an intact male, given 50 µg GnRH im (Cystorelin) and ovulation time determined by transrectal ultrasonography, conducted every 6 hours, starting 24 hours postmating. Animals were slaughtered at a specific intervals postovulation and reproductive tracts were recovered and subjected to oviductal and uterine flushing for females slaughtered between 1 and 6 days postovulation (dpo; Day 0 = ovulation) and uterine flushing for females slaughtered from 7 to 15 dpo for recovery of oocytes/embryos. Season of mating did not influence the interval from mating to ovulation (winter: 29 ± 6 hours vs. summer: 30 ± 6 hours; P = 0.49). Ovulation rates for females mated during winter and summer were 92% versus 100%, respectively (P = 0.05). Fertilization rates for winter and summer mated females were 72% and 82% (P = 0.29). Unfertilized ova were not retained in the uterine tube. All embryos collected were in the uterine tube ipsilateral to the side of ovulation between 1 and 5 dpo. Embryos reached the uterus on 6 dpo. Embryos began to elongate on 9 dpo; at this time, 83% of embryos derived from right-ovary ovulations were collected from the left uterine horn. Embryos occupied the entire uterine cavity by 10 dpo. In conclusion, we characterized early embryo development and location of embryo during its early developmental stages in alpaca. This was apparently the first report regarding chronology of embryo development and migration to the left horn in alpaca which merits further investigation regarding its role in maternal recognition of pregnancy.

Reproduction in female South American domestic camelids.

Alpacas and llamas are induced ovulators. They show marked reproductive seasonality in the Andean region, but under Northern Hemisphere conditions of feeding and management, they are non-seasonal breeders. Puberty is attained when they reach 50% of adult body weight. When they are not exposed to a male, females show successive waves of follicular maturation and atresia. Growth, maintenance and regression of a follicle each require an average of 4 and 6 days in alpacas and llamas, respectively. After sterile mating, progesterone concentrations in blood were increased from day 5, reached maximum concentrations on day 7-8, and declined rapidly at 9-10 days after mating. A fertile mating results in formation of a corpus luteum that remains functional throughout gestation. The duration of gestation is 340-346 days. Almost all fetuses were found to occupy the left uterine horn, even though ovulation occurs from both ovaries with equal frequency. Several methods of pregnancy diagnosis have been described. Mating is recommended within 15-20 days after parturition to obtain good fertility rates and one offspring per year. The factors that contribute to high rates of embryonic mortality are unknown. Reproductive technologies, such as AI, superovulation, embryo transfer and IVF, have not been used very extensively in these species but can be successfully applied.

Effects of lactational and reproductive status on ovarian follicular waves in llamas (Lama glama).

The effects of lactational status and reproductive status on patterns of follicle growth and regression were studied in 41 llamas. Animals were examined daily by transrectal ultrasonography for at least 30 days. The presence or absence of a corpus luteum and the diameter of the largest and second largest follicle in each ovary were recorded. Llamas were categorized as lactating (N = 16) or non-lactating (N = 25) and randomly allotted to the following groups (reproductive status): (1) unmated (anovulatory group, N = 14), (2) mated by a vasectomized male (ovulatory non-pregnant group, N = 12), (3) mated by an intact male and confirmed pregnant (pregnant group, N = 15). Ovulation occurred on the 2nd day after mating with a vasectomized or intact male in 26/27 (96%) ovulating llamas. Interval from mating to ovulation (2.0 +/- 0.1 days) and growth rate of the preovulatory follicle (0.8 +/- 0.2 mm/day) were not affected by lactational status or the type of mating (vasectomized vs intact male). Waves of follicular activity were indicated by periodic increases in the number of follicles detected and an associated emergence of a dominant follicle that grew to greater than or equal to 7 mm. There was an inverse relationship (r = -0.2; P = 0.002) between the number of follicles detected and the diameter of the largest follicle. Successive dominant follicles emerged at intervals of 19.8 +/- 0.7 days in unmated and vasectomy-mated llamas and 14.8 +/- 0.6 days in pregnant llamas (P = 0.001). Lactation was associated with an interwave interval that was shortened by 2.5 +/- 0.05 days averaged over all groups (P = 0.03). Maximum diameter of anovulatory dominant follicles ranged from 9 to 16 mm and was greater (P less than 0.05) for non-pregnant llamas (anovulatory group, 12.1 +/- 0.4 mm; ovulatory group, 11.5 +/- 0.2 mm) than for pregnant llamas (9.7 +/- 0.2 mm). In addition, lactation was associated with smaller (P less than 0.05) maximum diameter of dominant follicles averaged over all reproductive statuses (10.4 +/- 0.2 vs 11.7 +/- 0.3 mm). The corpus luteum was maintained for a mean of 10 days after ovulation in non-pregnant llamas and to the end of the observational period in pregnant llamas. The presence (ovulatory non-pregnant group) and persistence (pregnant group) of a corpus luteum was associated with a depression in the number of follicles detected and reduced prominence of dominant follicles (anovulatory group greater than ovulatory non-pregnant group greater than pregnant group).(ABSTRACT TRUNCATED AT 400 WORDS)

Niveles de Progesterona periférica en alpacas y llamas y su aplicación en el diagnóstico precoz de Gestación y otros usos clínicos / Progesterone profiles levels in alpaca and llama females and its aplication in early pregnancy diagnosis and another medical uses

Se estudiaron los niveles de progesterona periférica (P4)después del servicio fértil, en 12 alpacas y 12 llamas en celo y con cría al píe. Fueron servidas por una sola vez con machos fértiles y por un período de 20 minutos. Se tomó una muestra diaria de sangre yugular los días 1(día del servicio), 5, del 8 al 20, 25 y 30 post servicio. Se separó el plasma por centrifugación y se congeló a -20 grados centígrados hasta su análisis, por la técnica de radioinmunoensayo de Fase Sólida. Todas las alpacas y 10 llamas ovularon y desarrollaron cuerpo lúteo, que se observo al 3er día post-servicio, por laparoscopía. Los niveles de P4 el día 1 del estudio (celo y servicio) fueron 0.32 y 0.53 nmol/L en alpacas y llamas respectivamente. Seguidamente, los niveles basales de P4 se incrementaron hasta el día 8 en alpacas no preñadas (12.03 nmol/L) y hasta el día 9 en llamas no preñadas (14.10 nmol/L). Más adelante se observó una caída rápida de la P4 circulante, hasta niveles basales en los días 10 y 11 post servicio en alpacas y llamas respectivamente (p<0.001). Los niveles de P4 en alpacas preñadas se mantuvieron altos después del día 8 hasta el día 30 post servicio, en que se condujo el presente estudio, fluctuando entre 12,32 y 17,36 nmol/L. En llamas preñadas, las concentraciones de P4 siguen el mismo perfil que en alpacas preñadas, aunque con niveles más altos, fluctuando entre 17,51 y 24,66 nmol/L. En las dos llamas que presentaron falla de ovulación, los niveles de P4 se mantuvieron basales. En este estudio, el nivel mínimo de P4 para un diagnóstico positivo de gestación, entre 9 y 30 días post- servicio, fué de 4.0 nm/L (1.25 ng/ml) en ambas especies. El diagnóstico precoz de gestación mediante la determinación de la progesterona periférica, puede llevarse a cabo a partir del día 12 post-servicio. aquellas alpacas o llamas que mostraron bajos niveles de P4 después de la ovulación (día 8 o 9), fueron susceptibles a perder el embrión muy tempranamente, por insuficiencia luteal. Niveles basales de P4 el día 4to. post-servicio, indicarían falla de ovulación.