Search

Recent studies document the LH-releasing pathway of nerve growth factor (NGF) in male camelids and that the LH response to seminal NGF is associated with elevated plasma testosterone concentration. Results provide rationale for the hypothesis that NGF in semen is associated with male fertility. In Experiment 1, the association between the amount of NGF in the ejaculate and characteristics of the male reproductive system was examined in alpacas. The concentration of NGF was measured by radioimmunoassay in semen samples collected from male alpacas (n = 47) and correlated with sperm morphology and motility, and measurements of the male reproductive anatomy. Most ejaculates had NGF concentrations that, based on previous studies, triggered ovulation in female camelids, however, we only found a positive correlation between NGF concentration with sperm concentration, thread formation and total NGF, and a negative correlation with pH. In Experiment 2, a retrospective analysis was carried out to determine if breeding performance during the previous season was related to recent concentrations of seminal NGF in male alpacas (n = 22). Birth rates tended to be correlated with sperm concentration and total amount of NGF in the ejaculate (P = 0.09). Experiment 3 was a prospective study to determine the relationship between seminal NGF (n = 8 male alpacas) and ovulation and pregnancy rates in a breeding trial. No association was detected between seminal NGF concentration and ovulation rate, pregnancy rate, or LH response in the female. We conclude that among the breeding males used in our study, the abundance of seminal NGF was correlated with sperm concentration and thread formation, however, it was not predictive of male fertility in alpacas. Examination of males not previously selected as breeding stock may be expected to include a broader range of seminal NGF and provide a more comprehensive understanding of the relationship between seminal NGF and male fertility., Competing Interests: Declaration of conflict of interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The ovulation-inducing effect of seminal plasma was first suggested in Bactrian camels over 30 years ago, initiating a long search to identify the 'ovulation-inducing factor' (OIF) present in camelids semen. During the last decade, primarily in llamas and alpacas, this molecule has been intensively studied characterizing its biological and chemical properties and ultimately identifying it as β-Nerve Growth Factor (β-NGF). The high concentration of OIF/β-NGF in seminal plasma of llamas and alpacas, and the striking effects of seminal fluid on ovarian function strongly support the notion of an endocrine mode of action. Also, have challenged the dogma of mating induced ovulation in camelid species, questioning the classical definition of reflex ovulators, which at the light of new evidence should be revised and updated. On the other hand, the presence of OIF/β-NGF and its ovulatory effect in camelids confirm the notion that seminal plasma is not only a transport and survival medium for sperm but also, a signaling agent targeting female tissues after insemination, generating relevant physiological and reproductive consequences. The presence of this molecule, conserved among induced as well as spontaneous ovulating species, clearly suggests that the potential impacts of this reproductive feature extend beyond the camelid species and may have broad implications in mammalian fertility. The aim of the present review is to provide a brief summary of all research efforts undertaken to isolate and identify the ovulation inducing factor present in the seminal plasma of camelids. Also to give an update of the current understanding of the mechanism of action of seminal β-NGF, at central and ovarian level; finally suggesting possible brain targets for this molecule., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The aim of this study was to evaluated changes in endometrial vascularization area (EVA) between the left and right uterine horn: a) during the ovarian follicular growth in intact llamas, and b) after exogenous estradiol administration of estradiol benzoate in ovariectomized (OVX) llamas. In experiment 1 follicle wave emergence was synchronized (n = 5 llamas) by follicle ablation (Day 0). Females were examined every other day from Day 1 to Day 27, using B mode ultrasonography to evaluate dominant follicle growth profile. Also, EVA was evaluated in each horn using Color-Power Doppler. Blood samples were taken every other day from Day 1 to Day 27 to measure estradiol (E2) plasma concentration by RIA. In experiment 2 OVX llamas (n = 4 llamas/group) were given a single im administration of: a) 1 mg of estradiol benzoate (EB) or b) 1 mL of saline. Females were subjected to ultrasound examinations every 48 h from Day -4 until treatment (Day 0), every 12 h from Day 0 to Day 4, and again every 48 h from Day 5 to Day 11. Evaluation of EVA in both uterine horns was performed as described for experiment 1. Blood sampling for the measurement of E2 was carried out at the same time points indicated for the ultrasound examinations. Serial data were analyzed by one way ANOVA for repeated measures using the MIXED Procedure in SAS. Also, Pearson's correlation was used to determine the relationship between variables. In intact llamas there was an effect of day on the dominant follicle growing profile (P < 0.01) and estradiol plasma concentration (P < 0.05). Dominant follicle diameter positively correlated (r = 0.4; P < 0.017) with estradiol plasma concentration. Also, EVA of right and left uterine horn did not differ (P = 0.89) during the evaluation period; however, it was affected by time (P < 0.05). In ovariectomized llamas estradiol concentration was significantly (P < 0.001) affected by treatment, time and their interaction. Accordingly, treatment with EB (P < 0.0001), time (P < 0.05) and their interaction (P < 0.01) affected EVA of both uterine horns; however, this variable did not differ between horns (P = 0.98). In conclusion, circulating concentrations of estradiol determined an increase in uterine vascularization, during the phase of follicular growth in intact llamas and after the exogenous administration of EB to ovariectomized females; however, no differential effect in endometrial vascularization area between right and left uterine horn was observed., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The synthesis of anti-Müllerian Hormone (AMH) by the Sertoli cells in males is crucial for sexual differentiation. There are no studies on AMH in Camelids. The objectives of this research were to 1) compare AMH serum concentrations in prepubertal and adult male alpacas and 2) determine the effect of castration on these concentrations in adult males to provide a validation of a commercial AMH test in alpacas. Serum samples were obtained from 15 prepubertal animals (5 for each age groups of 6, 7 and 8 months) and from 5 adult males (age 5-9 years), pre- and 24 h post-castration. AMH was determined with a quantitative ELISA according to the manufacture's instructions. There was not significant difference (P < 0.05) in AMH level (pg/ml) between pre-pubertal (549.9 ± 120.8, 789.4 ± 172.3, 597.5 ± 177.3 for ages 4, 7, and 8 months, respectively) and adult alpacas (938.7 ± 175.9). In adult males, AMH concentration decreased significantly following castration (P < 0.05) (938.7 ± 383.5 pg/ml) pre-castration, and 222.1 ± 116.5 pg/ml) after castration). There was a positive correlation between testosterone levels and AMH. In conclusion, the quantitative assay used is a reliable test to determine AMH in alpacas. The AMH level in prepubertal and adult alpacas appear to not differ, contrarily from other mammals, this requires further investigation. The decrease in serum AMH concentrations after castration suggests that measurement of this hormone can be used to diagnose bilateral cryptorchid or hemicastrated unilateral cryptorchid animals in this species., (Published by Elsevier Inc.)

The objectives of the study were to determine the effect of seminal plasma β-NGF on Corpus Luteum morphology and function and level of mRNA expression of steroidogenic enzymes. Llamas were assigned (n = 12/per group) to receive an intramuscular dose of: (a) 1 ml phosphate buffered saline (PBS), (b) 5 μg gonadorelin acetate (GnRH), or (c) 1.0 mg of purified llama spβ-NGF. Ovaries were examined by transrectal B-mode ultrasonography from treatment to ovulation (Day 0 = treatment). B mode/Power Doppler ultrasonography and blood samples collection were performed at Days 4, 8 and 10 (n = 3 llamas per treatment group/per time point) to determine CL diameter, vascularization and plasma progesterone concentration respectively. Plasma progesterone concentration was analyzed in all llamas at Day 0. Then females were submitted to ovariectomy at Days 4, 8 and 10 (n = 3 llamas/treatment/time), CL was removed to determine vascular area, proportion of luteal cells and CYP11A1/P450scc and STAR expression by RT-PCR. Ovulation was similar between llamas treated with GnRH or spβ-NGF and CL diameter did not differ between GnRH or spβ-NGF groups by Day 4, 8 or 10. Vascularization area of the CL was higher (P < 0.01) in llamas from the spβ-NGF than GnRH-treated group by Day 4 and 8. Plasma progesterone concentration was higher (P < 0.05) in llamas from the spβ-NGF compared to females of GnRH group by Day 4 and 8. The proportion of small and large luteal cells did not differ between GnRH or spβ-NGF groups by Day 8. CYP11A1/P450scc was upregulated 3 folds at day 4 and 10 by spβ-NGF compared to GnRH. STAR transcription was 3 folds higher at day 4 in females treated with spβ-NGF. In conclusion, the luteotrophic effect of spβ-NGF could be related to an increase of vascularization and up regulation of CYP11A1/P450scc and STAR transcripts enhancing progesterone secretion., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Fibroblast growth factor 21 (FGF21) is known as a metabolic regulator to regulate the metabolism of glucose and lipids. However, the underlying mechanism of FGF21 on melanin synthesis remains unknown. Therefore, the current study investigates the effect of FGF21 on melanogenesis in alpaca melanocytes. We transfected the FGF21 into alpaca melanocytes, then detected the melanin contents, protein and mRNA levels of pigmentation-related genes in order to determine the melanogenesis-regulating pathway of FGF21. The results showed that FGF21 overexpression suppressed melanogenesis and decreased the expression of the major target genes termed microphthalmia-associated transcription factor (MITF) and its downstream genes, including tyrosinase (TYR) and tyrosinase-related protein 2 (TRP2). However FGF21 increased the expression of phospho-extracellular signal-regulated kinase (p-Erk1/2). In contrast, FGF21-siRNA, a small interference RNA mediating FGF21 silencing, abolished the inhibition of melanogenesis. Altogether, FGF21 may decrease melanogenesis in alpaca melanocytes via ERK activation and subsequent MITF downregulation, which is then followed by the suppression of melanogenic enzymes and melanin production., (Copyright © 2017. Published by Elsevier Inc.)

The aim of this study was to estimate the genetic parameters for preweaning traits and their relationship with reproductive, productive and morphological traits in alpacas. The data were collected from 2001 to 2015 in the Pacomarca experimental farm. The data set contained data from 4330 females and 3788 males corresponding to 6396 and 1722 animals for Huacaya and Suri variants, respectively. The number of records for Huacaya and Suri variants were 5494 and 1461 for birth weight (BW), 5429 and 1431 for birth withers height (BH), 3320 and 896 for both weaning weight (WW) and average daily gain (DG) from birth to weaning, 3317 and 896 for weaning withers height (WH), and 5514 and 1474 for survival to weaning. The reproductive traits analyzed were age at first calving and calving interval. The fiber traits were fiber diameter (FD), standard deviation of FD (SD), comfort factor and coefficient of variation of FD and the morphological traits studied were density, crimp in Huacaya and lock structure in Suri, head, coverage and balance. Regarding preweaning traits, model of analysis included additive, maternal and residual random effects for all traits, with sex, coat color, number of calving, month-year and contemporary group as systematic effects, and age at weaning as linear covariate for WW and WH. The most relevant direct heritabilities for Huacaya and Suri were 0.50 and 0.34 for WW, 0.36 and 0.66 for WH, 0.45 and 0.20 for DG, respectively. Maternal heritabilities were 0.25 and 0.38 for BW, 0.18 and 0.32 for BH, 0.29 and 0.39 for WW, 0.19 and 0.26 for WH, 0.27 and 0.36 for DG, respectively. Direct genetic correlations within preweaning traits were high and favorable and lower between direct and maternal genetic effects. The genetic correlations of preweaning traits with fiber traits were moderate and unfavorable. With morphological traits they were high and positive for Suri but not for Huacaya and favorable for direct genetic effect but unfavorable for maternal genetic effect with reproductive traits. If the selection objective was meat production, the selection would have to be based on the direct genetic effect for WW but not on the maternal genetic effect that has been shown to have less relevance. Other weaning traits such as WH or DG would be indirectly selected.

Why are long bones curved? It has long been considered a paradox that many long bones supporting mammalian bodies are curved, since this curvature results in the bone undergoing greater bending, with higher strains and so greater fracture risk under load. This study develops a theoretical model wherein the curvature is a response to bending strains imposed by the requirements of locomotion. In particular the radioulna of obligate quadrupeds is a lever operated by the triceps muscle, and the bending strains induced by the triceps muscle counter the bending resulting from longitudinal loads acting on the curved bone. Indeed the theoretical model reverses this logic and suggests that the curvature is itself a response to the predictable bending strains induced by the triceps muscle. This, in turn, results in anatomical arrangements of bone, muscle and tendon that create a simple physiological mechanism whereby the bone can resist the bending due to the action of triceps in supporting and moving the body. The model is illustrated by contrasting the behaviour of a finite element model of a llama radioulna to that of a straightened version of the same bone. The results show that longitudinal and flexor muscle forces produce bending strains that effectively counter strains due to the pull of the triceps muscle in the curved but not in the straightened model. It is concluded that the curvature of these and other curved bones adds resilience to the skeleton by acting as pre-stressed beams or strainable pre-buckled struts. It is also proposed that the cranial bending strains that result from triceps, acting on the lever that is the radioulna, can explain the development of the curvature of such bones., (Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.)

The objective of the study was to compare the pituitary and ovarian responses after intramuscular, intravenous, or intrauterine administration of β-nerve growth factor (β-NGF) of seminal plasma origin (SP-NGF) in llamas. In experiment 1, mature female llamas with a growing follicle of 7 mm or greater were assigned randomly to four groups (n = 7/group) and given 2 mg of purified SP-NGF in a volume of 2 mL by (1) intramuscular administration, (2) intravenous administration, and (3) intrauterine infusion, or (4) intrauterine infusion of 2 mL of PBS (negative control). Because ovulations were not detected after intrauterine infusion in experiment 1, a second experiment was done to determine if a higher dose of SP-NGF given by intrauterine infusion, similar to a natural dose during copulation, will elicit an ovulatory response. In experiment 2, llamas with a growing follicle of 7 mm or greater were assigned randomly to three groups (n = 6/per group) given an intrauterine infusion of (1) 4 mL of raw seminal plasma, (2) 4 mL of PBS containing 20 mg of purified llama SP-NGF, or 3) 4 mL of PBS (negative control). In both experiments, the ovaries were examined daily by transrectal ultrasonography using a B-mode scanner and power Doppler mode to detect ovulation and to monitor CL growth, regression, and vascularization. Blood samples were collected to determine plasma LH and progesterone concentrations. In experiment 1, only llamas treated by intramuscular or intravenous administration of SP-NGF ovulated (7 of 7 and 6 of 7, respectively). Plasma LH concentration did not differ between the intramuscular and intravenous SP-NGF-treated groups, nor did CL diameter, CL vascularization, or plasma progesterone concentration profiles. In experiment 2, the ovulation rate was 100% for llamas treated by intrauterine infusion of raw seminal plasma or llama SP-NFG, whereas no ovulations were detected in females treated with PBS. Plasma LH concentrations did not differ between groups that ovulated, nor did CL diameter, CL vascularization, or plasma progesterone concentration profiles. We conclude that β-NGF from llama seminal plasma origin elicits a preovulatory LH surge, followed by ovulation and the development of a functional CL, regardless of the route of administration. However, the dose required to elicit pituitary and ovarian responses is higher when administered by intrauterine infusion than by intramuscular or intravenous routes., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Twin births are rare in alpacas despite the high incidence of double ovulation and are undesirable because they contribute to early and late pregnancy loss, abortion, and birth of nonviable neonates. The objective of the present study was to determine the incidence and outcome of twin pregnancy in double-ovulating alpacas by retrospective and prospective analysis. Data from double-ovulating females (N = 41) presented for pregnancy diagnosis were analyzed to determine pregnancy status at three stages after mating (14-16, 25-30, and 45-53 days). In a prospective study, adult reproductively sound alpacas (n = 21) were examined by ultrasonography to determine the incidence of multiple ovulations. A subset of those alpacas (n = 10) were euthanized either at 9 days (n = 5) or 14 days (n = 5) after a single mating and administration of GnRH to determine presence and number of embryos. A total of 31 cycles were included in the study to determine the incidence of multiple ovulations. In the retrospective study, twin pregnancies were identified between 25 and 30 days in 47.1% of double-ovulating females. There were more twins from bilateral ovulations (62.5%) than from unilateral ovulations (37.5%). Twin pregnancies were either reduced to a singleton (62.5%) or lost completely (37.5%). One set of twins remained viable until Day 52. In the prospective study, double ovulation occurred in 18.8% of the cycles. Two embryos were collected from two of the four double-ovulating females. In conclusion, twin conception is not rare in alpacas. A high rate of spontaneous reduction of twin pregnancies occurs before Day 45. However, a significant proportion of twin pregnancy may result in loss of both embryos. Determination of the number of ovulations (CL) at the time of early pregnancy diagnosis (14-16 days) should be an integral part of any pregnancy evaluation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

To understand the role of ovulation-inducing factor (or nerve growth factor) (OIF [NGF]) in bovine seminal plasma, we (1) used an in vivo llama bioassay to test the hypothesis that bovine seminal plasma induces ovulation and CL development in llamas similar to that of llama seminal plasma when the dose of seminal plasma is adjusted to ovulation-inducing factor content (experiment 1) and (2) determined the effect of bovine seminal plasma on the interval to ovulation and luteal development in heifers (experiment 2). Within species, seminal plasma was pooled (n = 160 bulls, n = 4 llamas), and the volume of seminal plasma used for treatment was adjusted to a total dose of 250 μg of ovulation-inducing factor. In experiment 1, mature female llamas were assigned randomly to four groups and treated intramuscularly with either 10 mL of PBS (negative control, n = 5), 50-μg GnRH (positive control, n = 5), 6-mL of llama seminal plasma (n = 6), or 12 mL of bull seminal plasma (n = 6). Ovulation and CL development were monitored by transrectal ultrasonography. In experiment 2, beef heifers were given a luteolytic dose of prostaglandin followed by 25-mg porcine LH (pLH) 12 hours later to induce ovulation. Heifers were assigned randomly to three groups and given 12 mL bovine seminal plasma intramuscularly 12 hours after pLH treatment (n = 10), within 4 hours after ovulation (n = 9), or no treatment (control, n = 10). Ovulation was monitored by ultrasonography every 4 hours, and the CL development was monitored daily until the next ovulation. In experiment 1, ovulation was detected in 0/5, 4/5, 4/6, 4/6 llamas in the PBS, GnRH, llama seminal plasma, and bovine seminal plasma groups, respectively (P < 0.05). Luteal development was not different among groups. In experiment 2, the interval to ovulation was more synchronous (range: 4 vs. 22 hours; P < 0.0001) in heifers treated with seminal plasma before ovulation compared with the other groups. Luteal development was not different among groups; however, plasma progesterone concentrations tended to be greater in the postovulation treatment group compared with other groups. In summary, results confirmed the presence of bioactive ovulation-inducing factor in bull seminal plasma and supported the hypothesis that bovine and llama seminal plasma have similar ovulatory effects, using a llama bioassay. Treatment with bovine seminal plasma resulted in greater synchrony of ovulation in heifers pretreated with pLH. Plasma progesterone concentration tended to be higher in heifers given bovine seminal plasma within 4 hours after ovulation, suggesting that bovine ovulation-inducing factor is luteotrophic., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Nowadays, the fibre diameter (FD) is considered the main selection objective in alpaca populations all over the world. International Committee for Animal Recording recommendations define the FD and its CV as the first two traits to be considered in breeding programmes for this specie. In addition to these main criteria, other selection criteria of economic value used are comfort factor (CF) or standard deviation (s.d.); also other less important traits being used as selection objectives are these morphological traits: density (DE), crimp (CR) or lock structure (LS) for, respectively, Huacaya (HU) and Suri (SU) ecotypes, head (HE), coverage (CO) and balance (BA). The goal of this study was to establish how to implement a combined selection index starting from genetic parameters and to study the expected correlation between genetic trends by considering different alternative procedures of weighting all the involved traits, and the consequences of a wrongly proceeding way. Heritabilities and genetic and phenotypic correlations were estimated from the data set belonging to the PACOMARCA experimental farm for SU and HU. Two approaches were used to check the consequences of a set of subjective weights essayed. The coefficients of selection indexes were obtained for two sets of reference weights. In addition, equivalent weights were drawn if applied those reference values as coefficients of hypothetical selection indexes directly on phenotypes; relative expected genetic responses were computed in different cases. Results showed that almost in all cases for both ecotypes, the weight applied to CF should be surprisingly negative. Concerning genetic responses, only CO was compromised in some cases for the HU ecotype. The essayed methodology allowed explaining the differences between ecotypes in the genetic trends. The proposed methodology was shown to be effective to study the relative importance of the traits granted by the manager of a breeding scheme.

In this study, a database of fetal and maternal parameters was developed in pregnant alpacas from 120 days of gestation to term for evaluation of fetal well-being and development. Ten pregnancies were examined every 2 weeks from 120 to 300 days of gestation, then weekly until parturition. Five more pregnancies were evaluated weekly during the last 4 weeks prepartum. Maternal weight and heart rate were recorded. Indicators of fetal well-being (mean, minimum, and maximum fetal heart rate, fetal movement, and tone), development (size of the aorta, thorax, abdomen, stomach, kidney, head, eye orbit, radius, and umbilical vessels), and uterine environment (combined thickness of the uterus and placenta, depth and character of allantoic and amnionic fluids) were assessed using transabdominal ultrasound. Reference values and fetal growth curves were created. Weights of the cria, fetal membranes, and dam at parturition were associated with parameters of fetal growth. Mean (R(2) = 0.65), minimum (R(2) = 0.57), and maximum (R(2) = 0.64) fetal heart rate decreased from midgestation to term (P < 0.0001). Fetal activity also decreased as pregnancy advanced (P < 0.0001). Fetal presentation was anterior in all fetuses by 10 months. Increases in depth of allantoic (R(2) = 0.11) and amniotic (R(2) = 0.02) fluid, combined thickness of the uterus and placenta (R(2) = 0.31), and maternal weight (R(2) = 0.09) were significantly but poorly associated with gestation length. All parameters of fetal development were positively associated with gestational age (P < 0.0001). Size of the aorta (R(2) = 0.88), abdomen (R(2) = 0.87), kidney (R(2) = 0.83), and thorax (R(2) = 0.82) provided the best estimates of gestational age. Birth weight was associated with the size of the aorta (R(2) = 0.71), kidney (R(2) = 0.61), and radius (R(2) = 0.73) (P < 0.05). This information might be of value in the assessment of fetal well-being and development in alpacas., (Published by Elsevier Inc.)

Gonadotrophin releasing hormone (GnRH) is commonly used in llamas to induce ovulation; however, the consequence of reduced doses of GnRH on luteinizing hormone (LH) release, ovulatory response, and subsequent corpus luteum (CL) development and function have apparently not been investigated. Hence, we examined the effect of gradual reduction of gonadorelin acetate (GnRH) dosage on pituitary LH release, ovulatory response, CL development, and plasma progesterone concentrations in llamas. Non-pregnant, non-lactating adult llamas were examined once daily by transrectal ultrasonography, and those with a follicle ≥8 mm in diameter that had grown for three consecutive days were randomly assigned to receive 50 (GnRH50, n = 23), 25 (GnRH25, n = 29), 12.5 (GnRH12.5, n = 29), or 6.25 μg (GnRH6.25, n = 29) of GnRH, or 0.5 mL of PBS (Control group, n = 16) im. In a subset (7 or 8 animals/group), intense blood sampling was done to measure LH concentrations. All females were examined by ultrasonography every 12 h from treatment (Day 0) to Day 2 to determinate ovulation, and thereafter on alternate days until Day 16 to evaluate CL development (9-13 animals/group). Also, blood samples for progesterone determination were taken (9 or 10 animals/group) on alternate days from Days 0-16. Ovulatory response (%) was highest (P < 0.05) in the GnRH50 (82.6), intermediate in the GnRH25 (72.3) and GnRH12.5 (75.9) groups, and lowest in the GnRH6.25 group (48.3). No ovulations were detected in the Control group. Mean peak LH concentrations (ng/mL) were highest (P < 0.05) for GnRH50 (6.2), intermediate for GnRH25 (4.4) and GnRH12.5 (2.9), and lowest for GnRH6.25 (2.2) groups. In addition, based on regression analysis, llamas with an LH peak <4 ng/mL were less likely to ovulate. Llamas given 50 μg of GnRH released more (P < 0.05) pituitary LH and had an LH surge of longer duration than those given 25, 12.5, or 6.25 μg. However, in those that ovulated, neither GnRH treatment nor treatment by time interaction affected (P > 0.05) CL diameter or plasma progesterone concentrations. In summary, reducing the dose of GnRH gradually decreased the magnitude of the preovulatory LH surge and ovulatory response; however, subsequent CL development and plasma progesterone concentrations were not affected., (Copyright © 2012 Elsevier Inc. All rights reserved.)

This study was designed to: 1) characterize the effect of ovulation-inducing factor (OIF) on pituitary LH secretion in ovariectomized (OVX) llamas; and 2) determine the effect of OIF on LH secretion in OVX llamas pretreated with estradiol-17β (E-17β) or estradiol benzoate (EB). In Experiment 1, intact and OVX llamas (n = 5 or 6 per group) were assigned to a two by two factorial design: 1) Intact llamas treated with 1 mL of phosphate buffered saline (PBS); 2) Intact llamas treated with 1 mg of purified OIF; 3) OVX llamas treated with 1 mL of PBS; or 4) OVX llamas treated with 1 mg of purified OIF. In Experiment 2, intact and OVX llamas (n = 5 or 6 per group) were randomly assigned to the following groups: 1) Intact llamas treated with 1 mg of purified OIF; 2) OVX llamas treated with 1.0 mL of PBS; 3) OVX llamas treated with 1.0 mg of purified OIF; 4) OVX llamas primed with E-17β, followed by 1.0 mg of purified OIF. Experiment 3 was similar as described for Experiment 2, except that priming was done with EB. In Experiment 1, animal category by treatment and animal category by treatment by time interactions tended (P = 0.08) to affect LH concentration. The effect of OIF on LH released was partly restored (P < 0.05), to the values observed for the intact OIF-treated females, when OVX llamas were primed with E-17β or BE (Experiments 2 and 3). We concluded that peripheral estradiol concentrations in llamas partially modulates the effect of OIF on pituitary LH secretion; however, other ovarian factor(s) could also participate in this modulatory action., (Copyright © 2012 Elsevier Inc. All rights reserved.)

In order to advance the development of assisted reproductive technologies in alpacas and other Camelids, the objective of this study was to explore the role of seminal plasma concentration on motility and functional integrity of alpaca sperm. Sixteen male alpacas > 3 y of age were used. In Experiment 1, epididymal sperm were incubated for 0 to 6 h in 0, 10, 25, 50, or 100% seminal plasma and motility was assessed. In Experiment 2, epididymal sperm were incubated in 0, 10, or 100% seminal plasma for 3 h and motility, acrosome integrity and DNA integrity were assessed. In Experiment 3, ejaculated sperm were incubated in 10, 25, 50, or 100% seminal plasma for 0 to 6 h and motility assessed. In Experiment 4, ejaculated sperm were incubated in 10 or 100% seminal plasma for 3 h and motility, acrosome integrity, DNA integrity, and viability were assessed. Epididymal and ejaculated sperm maintained motility longer when incubated in the presence of 10% seminal plasma compared to 0, 25, 50, or 100% seminal plasma (P < 0.001). The mean ± SEM percentage of epididymal sperm with intact acrosomes was less (P < 0.001) in samples incubated in 0% seminal plasma (39.4 ± 3.73) compared to 10% (75.3 ± 1.20) or 100% (77.4 ± 0.90) within 1 h after incubation. However, DNA integrity of ejaculated and epididymal sperm was not significantly affected by seminal plasma concentration. The mean viability of ejaculated sperm was reduced in the presence of 100 (12.7 ± 2.33) compared to 10% (36.2 ± 4.68) seminal plasma (P < 0.001) within 1 h of incubation. We concluded that alpaca semen should be diluted to a final concentration of 10% seminal plasma to prolong motility, preserve acrosome integrity, and maintain viability of sperm., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The objective was to evaluate the developmental competence of cumulus-oocyte complexes (COC) collected by follicular aspiration in llamas treated with FSH or eCG. Llamas were assigned randomly to two groups (n = 16 per group) and treated, at the time of ovarian follicular wave emergence, with either: 1) 25 mg of FSH im, twice daily for 4 d; or 2) 1000 IU of eCG as a single i.m. dose. The start of gonadotropin treatment was considered Day 0. Both groups were given 5 mg of Armour Standard LH im on Day 6, and COC were collected by follicle aspiration on Day 7. Expanded COC collected from FSH- (n = 157) and eCG-treated llamas (n = 151) were fertilized in vitro using epididymal sperm, and presumptive zygotes were in vitro cultured in SOF medium for 8 d. The FSH and eCG treatment groups did not differ with respect to: the number of follicles ≥7 mm (16.0 ± 2.7 vs 14.0 ± 1.9, respectively; P = 0.5); the number of COC collected (11.5 ± 1.9 vs 9.7 ± 1.2; P = 0.4); the number of expanded COC (9.8 ± 1.4 vs 9.4 ± 1.2; P = 0.8); or the percentage of presumptive zygotes which developed into 2 to 8 cell stage embryos (65.3 vs 63.1), morulas (46.2 vs 42.5), or blastocysts (23.1 vs 20.5; P > 0.05). In conclusion, FSH and eCG treatments were equally effective for recovery of a high number of expanded COC which were used directly for in vitro fertilization. Furthermore, rate of embryo development was not significantly affected by the gonadotropin treatment used., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Lowland mammals at high altitude constrict the pulmonary vessels, augmenting vascular resistance and developing pulmonary arterial hypertension. In contrast, highland mammals, like the llama, do not present pulmonary arterial hypertension. Using wire myography, we studied the sensitivity to norepinephrine (NE) and NO of small pulmonary arteries of fetal llamas and sheep at high altitudes. The sensitivity of the contractile responses to NE was decreased whereas the relaxation sensitivity to NO was augmented in the llama fetus compared to the sheep fetus. Altogether these data show that the fetal llama has a lower sensitivity to a vasoconstrictor (NE) and a higher sensitivity to a vasodilator (NO), than the fetal sheep, consistent with a lower pulmonary arterial pressure found in the neonatal llama in the Andean altiplano. Additionally, we investigated carbon monoxide (CO) in the pulmonary circulation in lowland and highland newborn sheep and llamas. Pulmonary arterial pressure was augmented in neonatal sheep but not in llamas. These sheep had reduced soluble guanylate cyclase and heme oxygenase expression and CO production than at lowland. In contrast, neonatal llamas increased markedly pulmonary CO production and HO expression at high altitude. Thus, enhanced pulmonary CO protects against pulmonary hypertension in the highland neonate. Further, we compared pulmonary vascular responses to acute hypoxia in the adult llama versus the adult sheep. The rise in pulmonary arterial pressure was more marked in the sheep than in the llama. The llama pulmonary dilator strategy may provide insights into new treatments for pulmonary arterial hypertension of the neonate and adult., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Nitric oxide (NO) and alpha-melanocyte-stimulating hormone (alpha-MSH) have been correlated with the synthesis of melanin. The NO-dependent signaling of cellular response to activate the hypothalamopituitary proopiomelanocortin system, thereby enhances the hypophysial secretion of alpha-MSH to stimulate alpha-MSH-receptor responsive cells. In this study we investigated whether an NO-induced pathway can enhance the ability of the melanocyte to respond to alpha-MSH on melanogenesis in alpaca skin melanocytes in vitro. It is important for us to know how to enhance the coat color of alpaca. We set up three groups for experiments using the third passage number of alpaca melanocytes: the control cultures were allowed a total of 5 days growth; the UV group cultures like the control group but the melanocytes were then irradiated everyday (once) with 312 mJ/cm(2) of UVB; the UV+L-NAME group is the same as group UV but has the addition of 300 microM L-NAME (every 6h). To determine the inhibited effect of NO produce, NO produces were measured. To determine the effect of the NO to the key protein and gene of alpha-MSH pathway on melanogenesis, the key gene and protein of the alpha-MSH pathway were measured by quantitative real-time PCR and Western immunoblotting. The results provide exciting new evidence that NO can enhance alpha-MSH pathway in alpaca skin melanocytes by elevated MC1R. And we suggest that the NO pathway may more rapidly cause the synthesis of melanin in alpaca skin under UV, which at that time elevates the expression of MC1R and stimulates the keratinocytes to secrete alpha-MSH to enhance the alpha-MSH pathway on melanogenesis. This process will be of considerable interest in future studies., ((c) 2010 Elsevier Inc. All rights reserved.)

The objective of the study was to compare the ovulatory response and embryo production in llamas (Lama glama) treated with a single dose of equine chorionic gonadotropin (eCG) alone or combined with intravaginal medroxyprogesterone acetate (MPA) at the time of follicular wave emergence. Llamas with a growing follicle >or=7 mm in diameter were assigned to one of the following groups: (1) Control (n=28): Nonstimulated llamas were mated and embryos were collected 7 d after mating. (2) eCG (n=32): Llamas were given 5mg luteinizing hormone (LH) (Day 0) to induce ovulation, 1000 IU eCG on Day 2, a luteolytic dose of prostaglandin F(2alpha) on Day 6, mating on Day 7, and embryo collection on Day 14. (3) eCG+MPA (n=34): Llamas were treated as those in the eCG group, but a sponge containing 60 mg MPA was placed intravaginally from Days 2 to 6. Llamas that did not respond to synchronization or superstimulation were excluded, leaving data from n=26, 26, and 27 in the control, eCG, and eCG+MPA groups, respectively, for statistical analysis. The mean (+/-SD) number of follicles>7 mm at the time of mating was greatest in the eCG group, intermediate in the eCG+MPA group, and lowest in the control group (16.6+/-5.3, 12.9+/-3.7, and 1.0+/-0.0, respectively, P<0.001). The number of corpora lutea was similar between eCG and eCG+MPA groups (10.1+/-2.9 and 8.6+/-3.7, respectively); both were higher (P<0.001) than in controls (0.9+/-0.3). The number of embryos did not differ significantly between the eCG and eCG+MPA groups (4.8+/-2.8 and 3.5+/-3.0, respectively), but both were higher (P<0.001) than in the controls (0.7+/-0.4). In conclusion, eCG, with or without MPA effectively induced a superovulatory response and multiple embryo production in llamas.

The role of nutrition, especially the role of energy and protein status, on reproductive performance of production animals has been well documented. Comparatively, there is a true paucity of literature regarding nutritional mediation of reproductive performance in llamas and alpacas. Following seasonal patterns of feed availability in South America, adverse effects of nutritional deprivation on reproductive performance are well recognized, suggesting similar nutrition-reproduction interrelationships. Camelids, with their unique metabolism, may have some peculiar interrelationships between reproduction and protein and phosphorus nutrition. This presentation will review basic issues of energy and protein nutrition relative to reproductive performance in llamas and alpacas, based primarily on hypotheses and extrapolation from other species. Opportunities for research on this topic will be discussed, including preliminary data from current research.

Objective: To determine the minimum alveolar concentration (MAC) of desflurane in llamas and alpacas., Design: Prospective study. Animals Six healthy adult llamas and six healthy adult alpacas., Procedure: Anesthesia was induced with desflurane delivered with oxygen through a mask. An endotracheal tube was inserted, and a port for continuous measurement of end-tidal and inspired desflurane concentrations was placed between the endotracheal tube and the breathing circuit. After equilibration at an end-tidal-to-inspired desflurane concentration ratio >0.90 for 15 minutes, a 50-Hz, 80-mA electrical stimulus was applied to the antebrachium until a response was obtained (i.e. gross purposeful movement) or for up to 1 minute. The vaporizer setting was increased or decreased to effect a 10-20% change in end-tidal desflurane concentration, and equilibration and stimulus were repeated. The MAC was defined as the average of the lowest end-tidal desflurane concentration that prevented a positive response and the highest concentration that allowed a positive response., Results: Mean +/- SD MAC of desflurane was 7.99 +/- 0.58% in llamas and 7.83 +/- 0.51% in alpacas., Conclusions and Clinical Relevance: The MAC of desflurane in llamas and alpacas was in the range of that reported for other species.

We have recently reported the presence of an ovulation-inducing factor (OIF) in the seminal plasma of llamas and alpacas-species characterized as induced ovulators. The study was designed to test the hypothesis that the seminal plasma of bulls will induce ovulation in llamas, and to compare the ovulation-inducing effect of seminal plasma of conspecific versus hetero-specific males. The seminal plasma of alpacas, a closely related induced ovulator (Lama pacos), and cattle, a distantly related ruminant species (Bos taurus) considered to be spontaneous ovulators, were compared with that of the llama (Lama glama). Ovulation and maximum corpus luteum diameter were compared by ultrasonography among female llamas (n=19 per group) treated intramuscularly with 2 mL of phosphate buffered saline (PBS, negative control) and those treated with 2 mL of seminal plasma of bulls, alpacas, or llamas (conspecific control). The diameter of the preovulatory follicle did not differ among groups at the time of treatment. Bull seminal plasma induced ovulations in 26% (5/19) of llamas compared to 0% (0/19) in PBS group (P<0.001). The proportion of females that ovulated was lower (P<0.01) in bull seminal plasma group compared to the groups treated with alpaca or llama seminal plasma (100%). A corpus luteum was detected on Day 8 (Day 0=treatment) in all llamas in which ovulation was detected earlier (Day 2) by ultrasonography. The diameter of the CL did not differ among groups. Results document the presence of an ovulation-inducing factor in the seminal plasma of B. taurus. The interspecies effects of seminal plasma on ovulation and luteal development provide rationale for the hypothesis that OIF is conserved among both spontaneous and induced ovulating species.

Reproductive losses in camelids are due to infertility, pregnancy loss, udder diseases and neonatal mortality caused by a variety of infectious diseases. Uterine infection and abortion represent the major complaint in camelid veterinary practice. The major infectious organisms in endometritis and metritis are E. coli and Streptococcus equi subspecies zooepidemicus. Abortion rates due to infectious diseases vary from 10% to more than 70% in some areas. Leptospirosis, toxoplasmosis and chlamydiosis have been diagnosed as the major causes of abortion in llamas and alpacas. In camels, brucellosis and trypanosomiasis represent the major causes of infectious abortion in the Middle East and Africa. Mastitis is rare in South American camelids. The prevalence of subclinical udder infection in camels can reach very high proportions in dairy camels. Udder infections are primarily due to Streptococcus agalactiae and Staphylococcus aureus. Neonatal mortality is primarily due to diarrhea following failure of passive transfer and exposure to E. coli, rotavirus, coronavirus, Coccidia and Salmonella. This paper reviews the etio-pathogenesis of these causes of reproductive losses, as well as the major risk factors and strategies to prevent their occurrence.

Objective: To evaluate the anesthetic and cardiorespiratory effects of two doses of intramuscular xylazine/ketamine in llamas, and to determine if an intramuscular injection of tolazoline would shorten the anesthesia recovery time., Study Design: Prospective randomized study., Animals: Six castrated male llamas., Methods: Each llama received a low dose (LD) (0.4 mg kg(-1) xylazine and 4 mg kg(-1) ketamine) and high dose (HD) (0.8 mg kg(-1) xylazine and 8 mg kg(-1) ketamine). Time to sedation, duration of lateral recumbency and analgesia, pulse, respiratory rate, hemoglobin oxygen saturation, arterial blood pressure, blood gases, and the electrocardiogram were monitored and recorded during anesthesia. Three llamas in each treatment were randomized to receive intramuscular tolazoline (2 mg kg(-1)) after 30 minutes of lateral recumbency., Results: Onset of sedation, lateral recumbency, and analgesia was rapid with both treatments. The HD was able to provide at least 30 minutes of anesthesia in all six llamas. The LD provided only 30 minutes of anesthesia in two out of six llamas. Respiratory depression and hypoxemia were seen in the HD treatment during the first 10 minutes of lateral recumbency. Two llamas were severely hypoxemic during this period and were given nasal oxygen for five minutes. Heart rate decreased, but there were no significant changes in blood pressure. Tolazoline significantly shortened the duration of recumbency in the HD treatment., Conclusions: The HD provided more consistent clinical effects in llamas than did the LD. Intramuscular tolazoline shortens the duration of lateral recumbency in llamas anesthetized with this combination., Clinical Relevance: Both doses appear to be very effective in providing restraint in llamas. The LD may be used for procedures requiring a short period of anesthesia or restraint. The HD could be used when a longer duration of anesthesia is desired. Supplemental oxygen should be available if using the HD. Tolazoline (IM) shortened the recovery time with this combination in llamas.

The objective of the present study was to characterize ovarian activity in non-mated vicunas, relating ovarian structures (evaluated by transrectal ultrasonography, daily for 30 days) to changes in plasma concentrations of estradiol-17beta and progesterone. Ovarian follicular activity occurred in waves, characterized by the follicle emergence, growth and regression. The mean duration of follicular waves was 7.2+/-0.5 days (mean+/-S.E.M.), with a range of 4-11 days. The follicular growth phase averaged 3.0+/-0.2 days, the static phase 1.4+/-0.1, the regression phase 2.9+/-0.3 days, and the inter-wave interval was 4.2+/-0.3 days. The mean growth rate during the growing phase was 1.8+/-0.1mm/day, while the duration of the interval from 6mm to maximum diameter was 1.4+/-0.1 days. The mean maximum diameter of the dominant follicle was 8.4+/-0.3mm (range: 6.2-11.2) and mean diameter of the largest subordinate follicle was 5.4+/-0.1mm. There was an inverse relationship between the size of the largest follicle and the total number of follicles (r=-0.21, P=0.002). Follicle activity alternated between ovaries in 77% of the waves, with 40% of dominant follicles present in the left ovary and 60% in the right ovary. Plasma estradiol-17beta concentrations also had a wave-like pattern, varying between 12.0 and 62.8 pmol/l. Plasma progesterone concentrations remained below 5.0 nmol/l and there was no ultrasonographic evidence of ovulation during the study.

The study was designed to compare the efficacy of treatments intended to induce follicular wave synchronization among llamas (Experiment 1), and to determine the effect of these treatments on pregnancy rates after fixed-time natural mating (Experiment 2). In Experiment 1, llamas were treated with: (1) saline (control, n=20); (2) estradiol and progesterone (E/P, n=20); (3) LH (LH, n=20); or (4) transvaginal ultrasound-guided follicle ablation (FA, n=20). The ovarian response was monitored daily by transrectal ultrasonography. The intervals from treatment to follicular wave emergence and to the day on which the new dominant follicle reached >/=7 mm, respectively, did not differ between the LH (2.1+/-0.3 days and 5.2+/-0.5 days, respectively) and FA groups (2.3+/-0.3 days and 5.0+/-0.5 days), but both were shorter (P<0.05) and less variable (P<0.01) than in the control group (5.5+/-1.0 days and 8.4+/-2.0 days), while the E/P group (4.5+/-0.8 days and 7.7+/-0.5 days) was intermediate. In Experiment 2, llamas at unknown stages of follicular development were assigned randomly to control, E/P, and LH groups (n=30 per group). A single, fixed-time natural mating was permitted 10-12 days after treatment. Ovulation rates did not differ among groups (control, 93%; E/P, 90%; LH, 90%; P=0.99), but the pregnancy rate was higher (P<0.05) for synchronized llamas (LH and E/P groups combined, 41/54) than for non-synchronized llamas (control group, 15/28). In conclusion, LH and FA treatments were most effective for inducing follicular wave synchronization, while E/P treatment was intermediate. Synchronization treatments did not influence ovulation rate subsequent to fixed-time natural mating, but a higher pregnancy rate in synchronized than non-synchronized llamas warrants critical evaluation of the effects of follicular status on the developmental competence of the contained oocyte.

Ultrasound-guided transvaginal follicle aspiration has been described as a noninvasive and repeatable procedure for oocyte collection in several species, but its use has not been described for any of the members of the family, Camelidae. A study was designed to determine the feasibility of an ultrasound-guided transvaginal approach for oocyte collection in llamas. Fifteen non-pregnant, adult female llamas (10 non-stimulated and 5 superstimulated) were examined by transrectal ultrasonography with a 7.5-MHz linear-array transducer to determine the number and diameter of follicles available for aspiration. After caudal epidural anesthesia was induced, the 7.5-MHz linear-array transducer was fastened to a long rigid handle and inserted intravaginally. The free hand was placed into the rectum to manipulate the ovaries, one at a time, in position against the vaginal wall over the face of the transducer. A 20-gauge, 55-cm-long, single-lumen needle was advanced through the vaginal fornix and into follicles > or = 3 mm in diameter. Follicular contents were aspirated using a regulated vacuum pump (flow rate = 33 mL/min; approximately 150 mm Hg) into a tube containing 3 mL of phosphate buffered saline and 0.2% BSA. Fluid was filtered (75 microm mesh), and oocytes were located and morphologically evaluated using a stereomicroscope. Overall, 134 follicles were aspirated, and 76 oocytes were collected (collection rate = 57%). Thirty-two oocytes (42%) were surrounded by multiple layers of compacted granulosa cells and had homogenous dark ooplasm; 13 oocytes (17%) were surrounded by the corona radiata layer only and had heavily granulated ooplasm; 9 oocytes (12%) were denuded and had homogenous dark ooplasm; and 22 oocytes (29%) were denuded and displayed signs of ooplasm degeneration. The ultrastructure of llama oocytes was similar to that of cattle except for conspicuous accumulation of large lipid droplets in the cytoplasm. Twenty-four hours after follicle aspiration, the ovaries were examined by transrectal ultrasonography and intrafollicular hematomas were detected in 3 llamas (9 of 48 follicles aspirated). Results demonstrate the potential utility of a transvaginal ultrasound-guided technique for oocyte collection and in vitro embryo production in llamas. Oocytes of llamas bear an ultrustructural resemblance to those of cattle, but are distinguished by a predominance of cytoplasmic lipid.

As part of a larger project investigating the development and heritability of choanal atresia glama), it was necessary to develop a protocol for aborting llamas at various stages of gestation. Twenty-seven animals between 4 and 7 mo of gestation were successfully aborted a total of 53 times following two 250 microg intramuscular injections of cloprostenol at 24 h intervals. Abortion was induced once in 10 animals and multiple times (range 2 to 5) in 17 animals. Twenty-four animals (45.2%) aborted 3 d following the first injection, with 20 animals (37.7%) aborting 4 d post prostaglandin administration. Other animals aborted at 2 d (n=6, 11.3%), 5 d (n=2, 3.8%), and 7 d (n=1, 1.9%) following drug administration. Forty-nine (92.5%) of the abortions occurred following a single series of injections, while 4 animals (7.5%) aborted following a second series of injections. No confirmed pregnant animals failed to abort following the second series of cloprostenol injections. Conception rates in animals rebred 2 to 4 wk following an abortion were comparable to those of untreated animals in the research herd. Unlike the severe hypertension and death that has been reported following dinoprost tromethamine administration in the llama, no adverse reactions were observed in this study following cloprostenol administration. The results demonstrate that llamas can be safely and effectively aborted up to 7 mo of gestation (normal full term gestation = 342 +/- 10 days) without adverse effects on subsequent fertility.