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1
Kershaw-Young, C. M., X. Druart, J. Vaughan, and W. M. C. Maxwell. "β-Nerve growth factor is a major component of alpaca seminal plasma and induces ovulation in female alpacas." Reproduction, Fertility and Development 24, no. 8 (2012): 1093. http://dx.doi.org/10.1071/rd12039.
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Ovulation in camelids is induced by an unidentified protein in the seminal plasma of the male termed ‘ovulation-inducing factor’. This protein has been reported to be a 14-kDa protein under reducing conditions, which, when purified from seminal plasma, induces ovulation in llamas. The identification of this protein and investigation of its potential to induce ovulation in camelids may aid the development of protocols for the induction of ovulation. In the present study, alpaca seminal plasma proteins were separated using one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis and the most abundant protein of 14 kDa was identified as β-nerve growth factor (β-NGF) by liquid chromatography mass spectrometry. Female alpacas (n = 5 per group) were given intramuscular injections of: (1) 1 mL of 0.9% saline; (2) 4 µg buserelin, a gonadotrophin-releasing hormone agonist; (3) 2 mL alpaca seminal plasma; or (4) 1 mg human β-NGF. Ovulation was detected by transrectal ultrasonography 8 days after treatment and confirmed by plasma progesterone concentrations. Ovulation occurred in 0%, 80%, 80% and 80% of animals treated with saline, buserelin, seminal plasma and β-NGF, respectively. Treatment type did not affect the diameter of the corpus luteum, but plasma progesterone concentrations were lower in saline-treated animals than in the other treatment groups owing to the lack of a corpus luteum. The present study is the first to identify the ovulation-inducing factor protein in alpacas. β-NGF successfully induces ovulation in alpacas and this finding may lead to new methods for the induction of ovulation in camelids.
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Stuart, C. C., J. L. Vaughan, C. M. Kershaw-Young, J. Wilkinson, R. Bathgate, and S. P. de Graaf. "Effects of varying doses of β-nerve growth factor on the timing of ovulation, plasma progesterone concentration and corpus luteum size in female alpacas (Vicugna pacos)." Reproduction, Fertility and Development 27, no. 8 (2015): 1181. http://dx.doi.org/10.1071/rd14037.
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Ovulation in camelids is induced by the seminal plasma protein ovulation-inducing factor (OIF), recently identified as β-nerve growth factor (β-NGF). The present study measured the total protein concentration in alpaca seminal plasma using a bicinchoninic acid (BCA) protein quantification assay and found it to be 22.2 ± 2.0 mg mL–1. To measure the effects of varying doses of β-NGF on the incidence and timing of ovulation, corpus luteum (CL) size and plasma progesterone concentration, 24 female alpacas were synchronised and treated with either: (1) 1 mL 0.9% saline (n = 5); (2) 4 µg buserelin (n = 5); (3) 1 mg β-NGF protein (n = 5); (4) 0.1 mg β-NGF (n = 5); or (5) 0.01 mg β-NGF (n = 4). Females were examined by transrectal ultrasonography at 1–2-h intervals between 20 and 45 h after treatment or until ovulation occurred, as well as on Day 8 to observe the size of the CL, at which time blood was collected to measure plasma progesterone concentrations. Ovulation was detected in 0/5, 5/5, 5/5, 3/5 and 0/4 female alpacas treated with saline, buserelin, 1, 0.1 and 0.01 mg β-NGF, respectively. Mean ovulation interval (P = 0.76), CL diameter (P = 0.96) and plasma progesterone concentration (P = 0.96) did not differ between treatments. Mean ovulation interval overall was 26.2 ± 1.0 h. In conclusion, buserelin and 1 mg β-NGF are equally effective at inducing ovulation in female alpacas, but at doses ≤0.1 mg, β-NGF is not a reliable method for the induction of ovulation.
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Cervantes, M. P., T. Orban, and G. P. Adams. "208 OVARIAN FOLLICULAR DYNAMICS IN SOUTH AMERICAN CAMELIDS: EFFECT OF PLANE OF NUTRITION AND SPECIES." Reproduction, Fertility and Development 22, no. 1 (2010): 262. http://dx.doi.org/10.1071/rdv22n1ab208.
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Controversy exists regarding characteristics of follicular waves in llamas and alpacas. Lactational status has been shown to influence follicular dynamics, but the effects of species and nutrition have not been critically examined. A 2 × 2 experimental design was used to determine the effects of species (llama v. alpaca) and nutritional status (high-plane v. low-plane) on ovarian follicular wave dynamics. Adult female llamas (n = 16) and alpacas (n = 19), ≥ 3 years old, were assigned randomly to either a high or low plane of nutrition. Nutritional planes were defined by the grazing condition of the native pasture. The respective nutritional conditions were imposed 2 weeks before the start of the observational period. Body condition was estimated at the start of the observational period using a subjective scoring system (1 = very thin, 10 = very fat) and ovarian dynamics were monitored daily by transrectal ultrasonography for 38 days. Data were analyzed by two-way ANOVA and are expressed as mean ± SEM. Body condition scores were not different among groups (6.9 ± 0.35 and 6.6 ± 0.19 for llamas on high and low planes of nutrition, respectively, and 7.2 ± 0.25 and 6.8 ± 0.18 for alpacas on high and low planes of nutrition, respectively). The growing phase of the dominant follicle tended (P = 0.1) to be longer in llamas than in alpacas (9.8 ± 0.47 v. 8.8 ± 0.45 days) and in animals on a high plane of nutrition than in animals on a low plane (9.6 ± 0.50 v. 8.6 ± 0.42 days). Accordingly, the maximum diameter of the dominant follicle tended to be larger in llamas than in alpacas (10.1 ± 0.37 v. 9.1 ± 0.30 mm; P = 0.06) and in animals on a high plane of nutrition than in animals on a low plane (9.9 ± 0.39 v. 9.1 ± 0.27 mm; P = 0.14). The interwave interval was similar between llamas and alpacas (16.5 ± 0.66 v. 15.6 ± 0.42 days; P = 0.29), but was longer (P < 0.01) in animals on a high plane of nutrition than in animals on a low plane (16.9 ± 0.54 v. 15.0 ± 0.44 days); there was no interaction between main effects (P = 0.31). The total lifespan (duration of detection) of the dominant follicle was similar in both llamas and alpacas (22.9 ± 0.75 v. 21.9 ± 0.73 days; P = 0.38) and in animals on a high plane of nutrition than in animals on a low plane (22.7 ± 0.78 v. 22.0 ± 0.70 days; P = 0.53). There was no interaction between main effects (P = 0.21). All females (n = 35/35, 100%) had a follicle ≥ 7 mm (ovulatory size) from Days 7 to 12 after wave emergence. In conclusion, a low plane of nutrition had a suppressive effect on dominant follicle growth, resulting in a shortened interwave interval in llamas and alpacas. The interwave interval was not significantly longer in llamas than in alpacas despite a tendency for a longer growing phase and a larger dominant follicle. Research supported by Mitchell Group’s Mallkini Alpaca Breeding and Genetic Centre and the Natural Sciences and Engineering Research Council of Canada.
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Vivanco, H. W., E. Huaman, S. Leon, A. Gallegos, M. Asparrin, E. Alvarado, and G. Gamarra. "200 EVALUATION OF SUPEROVULATORY REGIMES FOR IN VIVO EMBRYO PRODUCTION IN ALPACAS (LAMA PACOS)." Reproduction, Fertility and Development 22, no. 1 (2010): 258. http://dx.doi.org/10.1071/rdv22n1ab200.
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The objective of the study was to evaluate 4 superovulatory regimes in terms of the quantity of transferable embryos recovered. A total of 48 female alpacas, 3 to 5 years of age and located at Malkini Alpacas Farm (4100 m elevation), were distributed into 4 treatments. In treatment 1, 13 female alpacas received on Day 0 an intravaginal device containing 0.78 mg of progesterone (Cue Mate®, Bioniche Animal Health, Belleville, Ontario, Canada) followed immediately by an i.m injection of estradiol (1 mg of estradiol benzoate) and an i.m. injection of PGF2α (Veyx®, 0.25 mg of cloprostenol). The intravaginal device was removed on Day 7, performing at removal time an i.m. injection of estradiol. From Days 8 to 16, the alpacas received an i.m injection twice per day and 12 hours apart of pFSH (FolltropinV®, Bioniche Animal Health) in decreasing doses totaling 420 mg of pFSH; on Day 16,300 IU of eCGi.m. (Pregnecol®, Bioniche Animal Health) was injected. In treatment 2, 13 alpacas received on Day 0 an intravaginal device of progesterone followed by an i.m. injection of PGF2; from Days 5 to 9, alpacas received injections twice per day of decreasing doses of pFSH (porcine FHS) totaling 320 mg; on Day 7, the intravaginal device was removed and 500 IU i.m. of eCG was injected. In treatment 3,13 alpacas received on Day 0 an intravaginal device of progesterone followed immediately by an i.m injection of GnRH (Conceptal®, 0.0042 mg of acetate of busereline); pFSH was injected i.m. from Days 5 to 9 in decreasing doses twice per day, totaling 440 mg; the intravaginal device was removed on Day 7. In treatment 4, 9 female alpacas received on Day 0 an i.m. injection of GnRH after verifying the presence of a preovulatory follicle (>8.0 mm diameter). On Day 2, the alpacas received 1000 IU i.m. of eCG followed on Day 7 by an i.m. injection of PGF2. In all cases, the donor alpacas were evaluated by ultrasonography. The matings for treatments 1, 2, and 3 were performed twice per donor alpaca at 12-hour intervals between Days 5 and 8 of the initiation of the pFSH treatments, whereas in treatment 4 the matings were made the following day after the application of the PGF2. In treatment 1, the donor alpacas received at time of first mating an i.m injection of 3.75 mg of LH (Lutropin®, Bioniche Animal Health); in treatments 2, 3, and 4, the donors received an i.m. injection of GnRH. In all treatments, embryo collection was performed by nonsurgical method 6.5 days after first mating. There were significant differences between treatments (P < 0.05) in the mean number of CL, with treatment 4 being the highest (4.7 ± 2.63, 4.1 ± 3.05, 1.8 ± 1.8, and 6.0 ± 3.16 for treatments 1 to 4, respectively). The total number of blastocysts recovered per treatment was 7, 16, 2, and 18 for treatments 1 to 4, respectively. The superovulatory strategy followed for treatment 4 showed to be the one resulting in the highest number of transferable embryos. Further comparative evaluations between FSH and eCG treatments are recommended. Research was partially funded by the contributions of Bioniche Animal Health.
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Vivanco, W., E. Huaman, S. Leon, T. Nunez, A. Gregoire, D. Ponce, E. Alvarado, and M. Asparrin. "159 COMPARISON BETWEEN EQUINE CHORIONIC GONADOTROPIN AND PORCINE FOLLICLE STIMULATING HORMONE FOR IN VIVO PRODUCTION OF EMBRYOS IN ALPACAS (VICUGNA PACOS) SHOWING NATURAL LUTEAL PHASE AFTER INDUCTION OF OVULATION." Reproduction, Fertility and Development 23, no. 1 (2011): 182. http://dx.doi.org/10.1071/rdv23n1ab159.
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Alpacas are animals with induced ovulation, andthey show high individual variation in the symptoms, duration, and regularity of oestrus or period of female receptivity to males; their follicular phase does not end in ovulation and subsequent luteal phase unless an external stimulation such as copulation or exogenous application of an ovulation inducing hormone is applied. The objective of the present study was to compare the use of eCG v. porcine (p)FSH as superovulatory hormones for the in vivo production of embryos in alpacas that were selected as being receptive to the male and were treated with an ovulation-inducing hormone to generate a luteal phase. Twenty adult (3 to 5 years old) female alpacas, located at Mallkini, Puno, Peru (at 4100 m elevation), were used for the trial. A group of females was exposed to males to test for breeding receptivity; 20 alpacas were receptive, adopting copulatory position. Each of the selected females received 3.75 mg of LH IM (Lutropin®, Bioniche Animal Health, Belleville, ON, Canada). Day 0 was then considered the date of LH injection. The 20 alpacas were then distributed into 2 treatments: Treatment 1 (T1 = 10 alpacas) received on Day 2, 1000 IU of eCG IM (Pregnecol®, Bioniche Animal Health) and on Day 7, a dose of PGF2α IM (0.263 mg of cloprostenol; Ciclar®, Andeanvet-Zoovet, Lima, Peru). Treatment 2 alpacas (T2 = 10 alpacas) received from Day 2 and up to Day 5, at 12-h interval, decreasing doses of pFSH IM (100 mg; Folltropin V®, Bioniche Animal Health) for 4 days, and on Day 7, a dose of PGF2α IM (0.263 mg of cloprostenol; Ciclar®, Andeanvet-Zoovet). All alpacas from T1 and T2 were mated twice with fertile males, the first mating at 24 h after the injection of PGF2α and the second at 12 h after the first mating. All females received a dose of GnRH IM (0.0084 mg of buserelin; Buserelina®, Andeanvet-Zoovet) at time of first mating. The embryos in both treatments were collected 6.5 days after the first mating by nonsurgical transcervical embryo flushing. There were no significant differences in the mean number of blastocysts collected per treatment (P > 0.05), being 3.0 ± 2.87 blastocyst for T1 and 1.6 ± 2.67 for T2. The number of blastocysts per treatment was 30 and 16 for T1 and T2, respectively. The results show that superovulatory treatment with eCG is more effective for the production of viable blastocysts than treatment with pFSH in alpacas treated for superovulation during the luteal phase. This work was partially funded by Bioniche Animal Health.
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Dipaz-Berrocal, D. J., G. Rojas, C. Mamani, J. R. Figueiredo, and E. Mellisho. "87 Population estimate and morphology of ovarian preantral follicles in fetal and adult alpacas (Vicugna pacos)." Reproduction, Fertility and Development 33, no. 2 (2021): 151. http://dx.doi.org/10.1071/rdv33n2ab87.
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Preantral follicles are the largest ovarian follicle population and represent an important source of potentially competent oocytes. During the lifespan of the female this large population becomes atretic during their growth. In alpacas, there are few studies that estimate the number of preantral follicles. Therefore, the objective of the present study was to compare the population and morphology of preantral follicles in the ovaries of fetal and adult alpacas. Ovaries from alpacas in fetal (fetus during the last third of gestation, n=5) and adult stage (3–4 years, n=5) were collected at a local slaughterhouse. The whole ovaries were individually fixed overnight at room temperature, and later dehydrated in alcohol, cleared with xylene, and embedded in paraffin. Tissue were sectioned at 7μm with a rotating microtome. Then, sections were processed and stained with periodic acid Schiff and haematoxylin. Preantral follicles were classified for their development stage as primordial, transitional, primary, or secondary, according to the layer number and form of granulosa cells. Estimation of the number of preantral follicles was made by counting all follicles in each histological section. Only follicles in which the oocyte nucleus was visible were counted. In addition, for each follicle category (n=30 per group), oocyte and follicle diameters were measured using Motic Images Plus 2.0 software. The population estimate and follicular diameter were compared using Kruskal–Wallis test with significance set at P ≤ 0.05 using SPSS v.2 2 software (IBM Corp.). A total of 2174 histologic sections were analysed. The results showed a higher (P=0.045) number of preantral follicles (80 516.1±3623.9) for fetal alpacas compared with adult alpacas (67 870.8±2267.4). Also, primordial follicles population (31 543.4±2690) and morphologically normal follicles (98.2%) were higher (P=0.04) in fetus compared with those in the adult stage (2244.7±355.37; 76.35%) respectively. On the contrary, the diameters of primordial, transitional, and primary follicles (45.34±3.76; 52.38±6.22; 59.79±5.22µm) from adult alpaca were greater (P=0.04) than those of fetal preantral follicles (33.305±7.2; 36.715±3; 77.985±15.8µm). In conclusion, the preantral follicle population declines dramatically in adult alpaca and animals of this age show an increased percentage of degenerate primordial follicles.
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Dipaz-Berrocal, D. J., G. Rojas, C. Mamani, J. R. Figueiredo, and E. Mellisho. "87 Population estimate and morphology of ovarian preantral follicles in fetal and adult alpacas (Vicugna pacos)." Reproduction, Fertility and Development 33, no. 2 (2021): 151. http://dx.doi.org/10.1071/rdv33n2ab87.
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Preantral follicles are the largest ovarian follicle population and represent an important source of potentially competent oocytes. During the lifespan of the female this large population becomes atretic during their growth. In alpacas, there are few studies that estimate the number of preantral follicles. Therefore, the objective of the present study was to compare the population and morphology of preantral follicles in the ovaries of fetal and adult alpacas. Ovaries from alpacas in fetal (fetus during the last third of gestation, n=5) and adult stage (3–4 years, n=5) were collected at a local slaughterhouse. The whole ovaries were individually fixed overnight at room temperature, and later dehydrated in alcohol, cleared with xylene, and embedded in paraffin. Tissue were sectioned at 7μm with a rotating microtome. Then, sections were processed and stained with periodic acid Schiff and haematoxylin. Preantral follicles were classified for their development stage as primordial, transitional, primary, or secondary, according to the layer number and form of granulosa cells. Estimation of the number of preantral follicles was made by counting all follicles in each histological section. Only follicles in which the oocyte nucleus was visible were counted. In addition, for each follicle category (n=30 per group), oocyte and follicle diameters were measured using Motic Images Plus 2.0 software. The population estimate and follicular diameter were compared using Kruskal–Wallis test with significance set at P ≤ 0.05 using SPSS v.2 2 software (IBM Corp.). A total of 2174 histologic sections were analysed. The results showed a higher (P=0.045) number of preantral follicles (80 516.1±3623.9) for fetal alpacas compared with adult alpacas (67 870.8±2267.4). Also, primordial follicles population (31 543.4±2690) and morphologically normal follicles (98.2%) were higher (P=0.04) in fetus compared with those in the adult stage (2244.7±355.37; 76.35%) respectively. On the contrary, the diameters of primordial, transitional, and primary follicles (45.34±3.76; 52.38±6.22; 59.79±5.22µm) from adult alpaca were greater (P=0.04) than those of fetal preantral follicles (33.305±7.2; 36.715±3; 77.985±15.8µm). In conclusion, the preantral follicle population declines dramatically in adult alpaca and animals of this age show an increased percentage of degenerate primordial follicles.
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Piripi, Susan, Jaime Hustace, Katelyn R. Carney, Jerry R. Heidel, and Christiane V. Löhr. "Pulmonary arteriovenous malformation in two adult alpacas (Vicugna pacos)." Journal of Veterinary Diagnostic Investigation 24, no. 1 (December 6, 2011): 198–201. http://dx.doi.org/10.1177/1040638711425938.
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Two cases of pulmonary vascular anomaly in unrelated adult alpacas ( Vicugna pacos) are described. In the first case, a 9-year-old intact male alpaca presented at Oregon State University Veterinary Teaching Hospital with bilateral epistaxis and died the subsequent day following severe hemorrhage from the mouth and nostrils. At necropsy, a tortuous vascular lesion was identified in the right cranial lung lobe, associated with hemorrhage into airways. In the second case, a 2-year-old female alpaca presented with postpartum anorexia, opisthotonus, and recumbency. In this second case, a similar vascular lesion was identified in the right cranial lung lobe but without associated hemorrhage. Histopathological examination of the lesion in both cases revealed numerous dilated, irregular blood vessels with marked variation in wall thickness within vessels, surrounded by foci of extramedullary hematopoiesis. Diagnoses of locally extensive pulmonary vascular anomalies (arteriovenous malformations) were made.
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Huanca, W. F., F. Y. Hilari, J. C. Villanueva, M. Uchuari, and W. Huanca. "100 Use of Seminal Plasma, Human Chorionic Gonadotropin, and Follicular Ablation on the Interval to Follicular Wave Emergency and Dominant Follicle in Alpacas (Vicugna pacos)." Reproduction, Fertility and Development 30, no. 1 (2018): 189. http://dx.doi.org/10.1071/rdv30n1ab100.
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Alpacas, as other camelids, are inducer ovulators and FIO/NGF-β, a protein present in the seminal plasma (SP) is reported as the responsible of the ovulation (Kershaw-Young et al. 2012 Reprod. Fertil. Dev. 24, 1093-1097, 10.1071/RD12039). However, limited and controversial information exists regarding characteristics of follicular wave in alpacas post-induction of ovulation with SP or other stimulus. The experiment was designed to determine the effect of 3 external stimulations on the interval to follicular wave emergence and the interval to dominant follicle. Adult female alpacas between 5 and 6 years old were assigned to 1 of 3 treatments: (1) SP (n = 6): 1 mL of SP IM; (2) hCG (n = 5): 1000 IU of hCG (Pregnyl, Organon-Holland, Amsterdam, the Netherlands), via IM; or (3) follicular ablation (FA, n = 6): animals were induced by ultrasound-guided ablation of the dominant follicle ≥7 mm. Alpacas from treatments 1 and 2 were examined by ultrasonography (Aloka SSD 500, transducer 7.5 MHz; Aloka, Tokyo, Japan) at 1- to 2-h intervals between 22 and 30 h after treatment or until ovulation occurred, whichever occurred first. All animals were evaluated by ultrasonography every day from Day 2 to Day 7 post-treatment and after that on Days 9, 12, and 15 post-treatment. Data from one alpaca (FA group) was excluded because of problems in the timing of ablation. Therefore, the total number of alpaca used was 16 (SP = 6, hCG = 5, and FA = 5). Results of the effect in external stimulation were analysed using ANOVA. In conclusion, interval to the emergence of a new follicular wave on the detection of follicles ≥3 mm and interval to dominant follicle ≥7 mm differed for FA compared with hCG but not compared with SP treatment. Table 1.Follicular wave emergence (mean ± SEM) under 3 external stimulations: seminal plasma (SP), hCG, or follicular ablation (FA)
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Patino, Cristian, Eduardo Arroyo, Michela Ciccarelli, Jacobo Rodriguez, Alan Conley, and Ahmed Tibary. "Serum anti-Müllerian hormone concentrations in female alpacas: variations during the reproductive cycle and correlation with ovarian superstimulation response." Clinical Theriogenology 14, no. 2 (June 1, 2022): 91–97. http://dx.doi.org/10.58292/ct.v14.9139.
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Earlier, we validated an anti-Müllerian hormone (AMH) enzyme-linked immunosorbent assay kit for male alpacas. First, wecompared the validation data with another kit. There was a high correlation (R2 = 0.94) between these 2 kits. Second, we used thelatter kit to determine serum AMH concentrations during follicular and luteal phases of the reproductive cycle in female alpacas.There were no differences (p = 0.39) in serum AMH concentrations in alpacas (n = 11) between peak follicular and luteal phases(mean ± SEM, 1.33 ± 0.35 versus 1.18 ± 0.34 ng/ml, respectively). Third, we treated female alpacas (n = 13; 5 - 11 years) after 14-day treatment with decreasing doses of porcine follicle-stimulating hormone. There was no effect (p > 0.05) of day of treatmenton serum AMH concentrations. Number of follicles (7 - 10 mm; mean ± SD [as determined via transrectal ultrasonography]) atend of treatment (12.69 ± 5.25; range: 6 - 24) was positively correlated (R2 = 0.7; p < 0.01) with serum AMH concentrations. Toconclude, the kit tested is usable for female alpacas; serum AMH concentrations were not affected by the cycle stage nor by ovariansuperstimulation treatment. Furthermore, a significant correlation between serum AMH serum concentrations and response to superstimulationsuggested that estimation of serum AMH concentrations may be valuable in determining ovarian follicular reserve.
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Memon, Mushtaq A. "Camelids Orgling Sounds Help Female Alpacas to Ovulate." American Journal of Biomedical Science & Research 13, no. 2 (June 18, 2021): 205–6. http://dx.doi.org/10.34297/ajbsr.2021.13.001857.
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Vilca, Eliana, Miluska Navarrete, Gilberto Santillan, Alexander Chavez, and Francisco Santos. "MACROSCOPIC AND MICROSCOPIC DESCRIPTION OF ALPACA (Vicugna pacos) OVARIES DURING THE FETAL STAGE." SPERMOVA 12, no. 1 (July 31, 2022): 1–7. http://dx.doi.org/10.18548/aspe/0010.01.
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The aim of this work was to describe macroscopically and microscopically the ovaries of alpacas during the fetal stage. We worked with 18 female fetuses collected at the Huancavelica Municipal Slaughterhouse, from alpacas destined for human consumption. The process and analysis of the collected samples was performed at the Faculty of Veterinary Medicine - UNMSM, in Lima, Peru. Fetal age was calculated by measuring the biparietal diameter and divided into three groups: Group I (60 - 150 days), Group II (151 - 239 days) and Group III (240 - 335 days). The weight of the ovaries was 0.02 ± 0.01, 0.03 ± 0.01 and 0.03 ± 0.01 grams in the first, second and third group, respectively. The ovaries were paired, oval-shaped, with a smooth, cream surface, located in the sublumbar region at the level of the 6th and 7th lumbar vertebrae. The cortex and medulla were visible from the third month. Microscopically, in Group I, we observed oogonium, the distinction between cortex and medulla, unilaminar primary follicles, atresia and segmentation of the germ cells. In Group II, we observed preantral primary follicles. In Group III, we observed preovulatory follicles. We concluded that, from day 68 of the fetal stage of the alpaca, ovaries have macroscopic characteristics similar to those of an adult alpaca. Microscopically, they presented ovogonia, primordial follicles, unilaminar primary follicles, preantral follicles, preovulatory follicle, as well as the degeneration of the germ cells.
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Simons, J. A., D. L. Waldron, and D. P. Hennessy. "Clinical biochemical reference ranges for female alpacas (Lama pacos)." Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 105, no. 3-4 (July 1993): 603–8. http://dx.doi.org/10.1016/0305-0491(93)90095-m.
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Huanca, W., K. Garcia, W. F. Huanca, A. Cordero, and J. Malaga. "108 Use of seminal plasma as ovulation inductor in alpacas (Vicugna pacos) embryo recipient and its effect on pregnancy rate." Reproduction, Fertility and Development 32, no. 2 (2020): 181. http://dx.doi.org/10.1071/rdv32n2ab108.
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Alpacas, like other camelids, are classified as induced ovulators because an external stimulus is required for the occurrence of ovulation. Recent studies have identified the β-nerve growth factor (β-NGF), a protein present in the seminal plasma (SP), as responsible for inducing ovulation in this species and having potent luteotropic function. We speculate that SP can be used in reproductive biotechnologies, such as embryo transfer (ET), to increase the number of genetically superior animals produced in breeding programs. The objective of this study was to evaluate the effect of inducing ovulation with SP or a gonadotrophin-releasing hormone (GnRH) analogue on pregnancy rate in recipients of an alpaca ET program. Semen from 5 adult male alpacas was collected with an artificial vagina and diluted 1:1 (v/v) with phosphate-buffered saline (PBS; Gibco-BRL). The diluted semen was centrifuged for 30min at 1200g, and the supernatant or SP was decanted and examined for absence of sperm. Then, the SP was centrifuged again for 20min at 1200×g. The SP was stored at −80°C until the use. Female alpacas (n=38; 6-8 years) with a body condition score of 2.5-3.5 (scale 1-5) were used for the experiment. Animals were evaluated daily by transrectal ultrasonography to determine the presence of a dominant follicle ≥7mm and randomly assigned to one of two groups: (1) GnRH (n=20), 0.04mg of Buserelin acetate IM, or (2) SP (n=18): 1.0mL of seminal plasma IM. Seven days after GnRH or SP treatment (Day 0) the recipients received a fresh embryo. Alpaca donors (n=18) were given GnRH (0.04mg IM) and treated 36h later with 700IU of equine chorionic gonadotropin (ECG). Donors were mated with fertile males 5 days after ECG (Day 0 of recipients), and embryos were recovered 7 days later. Embryos of similar quality were assigned to both groups and transferred nonsurgically to the uterine horn ipsilateral to the corpus luteum. Ultrasonography examinations were performed on Day 2 to confirm ovulation and Day 25 to determine pregnancy in all of the recipients. Data was analysed by chi-squared test. Ovulation rate was not different between groups (100% each). Pregnancy rate was 45% (9 out of 20) and 44% (8 of 18) in GnRH and SP groups, respectively (P=0.77). In conclusion, SP was effective to induce ovulation in alpacas and was able to produce a pregnancy rate similar to that of GnRH as an ovulation-inducing treatment. Thus, SP can be used as an alternative for ET programs in alpacas. Research was funded by the project Role of Seminal Plasma in Reproductive Physiology and Application of Biotechnologies in Camelids (149-2017-CIENCIACTIVA).
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Lutz, J., S. Johnson, K. Duprey, P. Taylor, H. Vivanco, M. Ponce-Salazar, M. Miguel, and C. Youngs. "7 Pregnancy from a vitrified-warmed alpaca pre-implantation embryo." Reproduction, Fertility and Development 32, no. 2 (2020): 128. http://dx.doi.org/10.1071/rdv32n2ab7.
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The alpaca (Vicugna pacos) is a ruminant livestock species in the South American camelid family. There are more than 9 million South American camelids globally that make important contributions to the livelihoods of rural farmers through conversion of low quality roughages to high quality food and fibre. Reproductive biotechnologies for alpacas are not well developed compared with those for other ruminant livestock species. In particular, embryo cryopreservation technologies are lacking. The objective of this study was to evaluate under field conditions a vitrification protocol originally developed for old world camels that we adapted for use in alpacas. Potential donors were evaluated for follicular development using a 7.5-MHz ultrasound probe. Hembras (sexually mature female alpacas) with ovarian follicles 7-10mm in diameter were behaviour tested to determine sexual receptivity, and receptive females were naturally mated to a proven herd sire. At the time of breeding, non-superovulated donors (n=4) received 30μg gonadorelin. Embryos were nonsurgically collected 7 days after breeding and handled at 20°C. Diameter of harvested embryos (n=4 quality grade 1 hatched expanded blastocysts) was measured using an eyepiece reticle. All recovered embryos were placed individually into 0.5-mL drops of vitrification solution (VS1: 1.4M glycerol) for 5min, 0.5-mL drops of VS2 (1.4 M glycerol + 3.6M ethylene glycol) for 5min, 0.05-mL drops of VS3 (3.4 M glycerol + 4.6M ethylene glycol) for 20s, and 0.05-mL drops of VS3 for 20s while loading into open-pulled straws (OPS). Each OPS was plunged directly into liquid nitrogen for storage for 29 days. At warming, each OPS was submerged into a 1-mL drop of warming solution 1 (WS1: 0.5M galactose) for 1min followed by 1min in WS2 (0.25 M galactose) for 5min before being incubated at 37°C in 5% CO2 in humidified air for 21h in 1mL of Syngro holding medium supplemented with 10% (vol/vol) alpaca serum. Embryos that grew during culture (n=2) were transferred individually into synchronous recipients, and embryos that did not appear to grow (n=2) were transferred together as a pair. Prior to embryo transfer, potential recipients were evaluated ultrasonographically as described previously. Hembras with ovarian follicles 7-10mm in diameter were behaviour tested, and sexually receptive females received 30μg gonadorelin 6 days before embryo transfer. Final selection of recipients (n=3) was based on presence of a corpus luteum and nonreceptive behaviour to a herd sire 24h before transfer. Pregnancy was detected ultrasonographically, and fetal heartbeat was detected 29 days post-transfer in one of the three recipients. Ultrasound at 177 days post-transfer revealed that the pregnancy, generated from a 400μm×375μm vitrified-warmed embryo that had grown in culture, was still ongoing. If this pregnancy results in the birth of a live cria (newborn alpaca), it would represent-to the best of our knowledge-the world's first cria born from a cryopreserved alpaca pre-implantation embryo. It would also demonstrate the potential utility of this protocol under field conditions.
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Tibary, Ahmed, Alexis Campbell, Jacobo S. Rodriguez, Agustin J. Ruiz, Cristian Patino, and Michela Ciccarelli. "Investigation of male and female infertility in llamas and alpacas." Reproduction, Fertility and Development 33, no. 2 (2021): 20. http://dx.doi.org/10.1071/rd20257.
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Llamas and alpacas are important production animals in South America, with increasing interest in other parts of the world. Poor reproductive efficiency combined with several unique anatomical and physiological reproductive features offer challenges in the diagnosis and treatment of infertility in camelids. This review presents an approach to the clinical investigation and common causes of infertility and subfertility in the male and female. The selection of males for breeding should be made based on complete evaluation to eliminate congenital and possibly hereditary disorders. Common disorders of the male reproductive system include testicular hypoplasia, testicular and epididymal cysts and testicular degeneration. Semen evaluation presents some challenges owing to the viscous nature of the ejaculate in these species. Females should be screened for congenital genital defects before breeding. Causes of subfertility in the female are dominated by ovarian and uterine disorders. A systematic clinical approach and the use of endometrial biopsy and advanced techniques, such as laparoscopy, allow early identification of these disorders. Further research is needed for continued understanding of the reproductive pathological processes in these species.
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Tibary, Ahmed, Alexis Campbell, Jacobo S. Rodriguez, Agustin J. Ruiz, Cristian Patino, and Michela Ciccarelli. "Investigation of male and female infertility in llamas and alpacas." Reproduction, Fertility and Development 33, no. 2 (2021): 20. http://dx.doi.org/10.1071/rd20257.
Full textAbstract:
Llamas and alpacas are important production animals in South America, with increasing interest in other parts of the world. Poor reproductive efficiency combined with several unique anatomical and physiological reproductive features offer challenges in the diagnosis and treatment of infertility in camelids. This review presents an approach to the clinical investigation and common causes of infertility and subfertility in the male and female. The selection of males for breeding should be made based on complete evaluation to eliminate congenital and possibly hereditary disorders. Common disorders of the male reproductive system include testicular hypoplasia, testicular and epididymal cysts and testicular degeneration. Semen evaluation presents some challenges owing to the viscous nature of the ejaculate in these species. Females should be screened for congenital genital defects before breeding. Causes of subfertility in the female are dominated by ovarian and uterine disorders. A systematic clinical approach and the use of endometrial biopsy and advanced techniques, such as laparoscopy, allow early identification of these disorders. Further research is needed for continued understanding of the reproductive pathological processes in these species.
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Diaz, A., W. Huanca, A. Ampuero, H. Huaman, J. Camacho, T. Huanca, D. Quispe, and H. Diaz. "113 EFFECT OF SOME FACTORS ON CONCEPTION RATE IN ALPACAS UNDER PERUVIAN HIGHLAND CONDITION." Reproduction, Fertility and Development 23, no. 1 (2011): 161. http://dx.doi.org/10.1071/rdv23n1ab113.
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Alpacas are a domestic species of South American camelids with a great importance to highland people because of their high-quality fibre production. However, their reproductive performance has been reported to be poor, with a birth rate of 50% under the Andean community’s conditions. Two experiments were designed to evaluate the effect of some factors on the pregnancy rate of alpacas at first service in lactating, 3- to 8-years old, without parity problems. The objective of the first experiment (n = 85) was to evaluate the effect of age (G1: 3, G2: 4 to 5, G3: ≥ 6 years old); mating time (G1: 15 min, G2: 16–24 min, G3: ≥ 25 min), and interval from parturition to mating (G1: 20 days, G2: ≥ 20 days) on first service conception rate. A second experiment (n = 174) evaluated the effect of month of calving (January, February, or March) on conception rate in females with a postpartum interval ≥20 days. Animals were mated with male of good fertility after a receptivity test and confirmation of presence of a dominant follicle ≥7 mm by ultrasonography. Pregnancy was determined by ultrasonography 25 days after mating. Proportional data were compared by Fisher’s exact test. In the first experiment, conception rates were 57.9, 66.7, and 47.2% in females of G1, G2, and G3, respectively (P ≤ 0.05); 50.0, 54.5, and 59.0% in females with mating time of 15, 16–24, or ≥ 25 min of mating; 48.0 and 59.4% in those with a postpartum interval <20 days and ≥20 days (P ≤ 0.05). In the second experiment, conception rates were 58.3, 70.7, and 82.1% in alpacas calving in January, February, and March, respectively (P ≤ 0.05). The results suggest that age of female, postpartum interval, and month of calving are factors that affect conception rates in alpacas. Therefore, a reproductive management system that includes these factors would improve the pregnancy rate in alpacas under highland Peruvian conditions. Consejo Superior de Investigacion – UNMSM.
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Arana, N. Z., H. C. Ygnacio, F. T. Zárate, E. A. Malca, and A. Gallegos-Cárdenas. "88 Plasma anti-Müllerian hormone as a marker for ovarian follicular population and oocyte quality in alpacas." Reproduction, Fertility and Development 33, no. 2 (2021): 151. http://dx.doi.org/10.1071/rdv33n2ab88.
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The limited success of embryonic transfer technique in alpacas is related to the high variability of superovulation protocols, the response to which depends on, among other factors, the number of antral follicles growing along a follicular wave. The concentration of anti-Müllerian hormone (AMH) is correlated with antral follicle count, making it a reliable biomarker for identifying the number of available follicles and allowing us to ensure the reproductive potential of females. Studies in different production animals exist, but, thus far, no published studies exist on the use of AMH as a biomarker in alpacas. Thus, its study is necessary in order to be able to choose the best donors for embryonic transfer, reducing response variability and unnecessary expenses in superovulation protocols. The objective of this study was to determine whether AMH is a reliable marker for the number of follicles and quality of oocytes in an induced follicular wave in alpacas. We used 52 female, nonpregnant Suri alpacas of reproductive age (3–7 years), with a body condition score between 2.5 and 3.5 (where 1 is extremely thin and 5 is obese), and with the presence of a preovulatory follicle (diameter ≥7mm) determined through transrectal ultrasound. Blood samples were collected from nonpregnant Suri alpacas 5 days post-induction of ovulation and divided into groups according to AMH concentration. AMH concentrations were then correlated with the number of follicles and percentage by sizes of follicles using the least significant difference test with adjusted means. To relate the quality of oocytes with the AMH concentration groups, a Chi-square test was used. The high-AMH group had a greater number of follicles than the low-AMH group (20.51±2.76 vs. 11.58±2.55; P=0.036). There were no significant differences between AMH groups and percentage of follicle sizes. However, the high-AMH group had a greater percentage of follicles 3- to 7-mm long (67.49%). Furthermore, the high-AMH group showed dependence on the quality of oocytes (P=0.02). These results indicate that the plasma concentration of AMH can be a reliable marker for follicle and oocyte quality in Suri alpacas. Future investigations are needed to ensure the optimal timing to collect blood samples, a kit individualized to the species, and improvement of the future donors for current superovulation protocols in alpacas.
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Arana, N. Z., H. C. Ygnacio, F. T. Zárate, E. A. Malca, and A. Gallegos-Cárdenas. "88 Plasma anti-Müllerian hormone as a marker for ovarian follicular population and oocyte quality in alpacas." Reproduction, Fertility and Development 33, no. 2 (2021): 151. http://dx.doi.org/10.1071/rdv33n2ab88.
Full textAbstract:
The limited success of embryonic transfer technique in alpacas is related to the high variability of superovulation protocols, the response to which depends on, among other factors, the number of antral follicles growing along a follicular wave. The concentration of anti-Müllerian hormone (AMH) is correlated with antral follicle count, making it a reliable biomarker for identifying the number of available follicles and allowing us to ensure the reproductive potential of females. Studies in different production animals exist, but, thus far, no published studies exist on the use of AMH as a biomarker in alpacas. Thus, its study is necessary in order to be able to choose the best donors for embryonic transfer, reducing response variability and unnecessary expenses in superovulation protocols. The objective of this study was to determine whether AMH is a reliable marker for the number of follicles and quality of oocytes in an induced follicular wave in alpacas. We used 52 female, nonpregnant Suri alpacas of reproductive age (3–7 years), with a body condition score between 2.5 and 3.5 (where 1 is extremely thin and 5 is obese), and with the presence of a preovulatory follicle (diameter ≥7mm) determined through transrectal ultrasound. Blood samples were collected from nonpregnant Suri alpacas 5 days post-induction of ovulation and divided into groups according to AMH concentration. AMH concentrations were then correlated with the number of follicles and percentage by sizes of follicles using the least significant difference test with adjusted means. To relate the quality of oocytes with the AMH concentration groups, a Chi-square test was used. The high-AMH group had a greater number of follicles than the low-AMH group (20.51±2.76 vs. 11.58±2.55; P=0.036). There were no significant differences between AMH groups and percentage of follicle sizes. However, the high-AMH group had a greater percentage of follicles 3- to 7-mm long (67.49%). Furthermore, the high-AMH group showed dependence on the quality of oocytes (P=0.02). These results indicate that the plasma concentration of AMH can be a reliable marker for follicle and oocyte quality in Suri alpacas. Future investigations are needed to ensure the optimal timing to collect blood samples, a kit individualized to the species, and improvement of the future donors for current superovulation protocols in alpacas.
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Pollard, J. C., R. P. Littlejohn, and I. C. Scott. "The effects of mating on the sexual receptivity of female alpacas." Animal Reproduction Science 34, no. 3-4 (January 1994): 289–97. http://dx.doi.org/10.1016/0378-4320(94)90024-8.
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Bravo, P. Walter, Daniel Diaz, Virgilio Alarcón, and Cesar Ordoñez. "Effect of the reproductive state of female alpacas on embryonic mortality rate." American Journal of Veterinary Research 71, no. 9 (September 2010): 1096–99. http://dx.doi.org/10.2460/ajvr.71.9.1096.
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McGregor, B. A., and K. L. Butler. "Sources of variation in fibre diameter attributes of Australian alpacas and implications for fleece evaluation and animal selection." Australian Journal of Agricultural Research 55, no. 4 (2004): 433. http://dx.doi.org/10.1071/ar03073.
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Sources of variation in fibre diameter attributes of Australian alpacas and implications for fleece evaluation and animal selection were investigated using data collected in the years 1994–97, from 6 properties in southern Australia. Data were analysed using REML (multiple regression analysis) to determine the effect on mean fibre diameter (MFD) and coefficient of variation of MFD (CV(FD)) of age, origin (property), sex (entire male, female), breed (Huacaya, Suri), liveweight, fibre colour, individual, and interactions of these effects. The mean (n = 100) age (range) was 4.2 years (0.1–11.9), liveweight 72.0 kg (12.0–134 kg), MFD 29.1 μm (17.7–46.6 μm), CV(FD) 24.33% (15.0–36.7%). A number of variables affected MFD and CV(FD). MFD increased to 7.5 years of age, and correlations between MFD at 1.5 and 2 years of age with the MFD at older ages were much higher than correlations at younger ages. Fibre diameter 'blowout' (increase with age) was positively correlated with the actual MFD at ages 2 years and older. There were important effects of farm, and these effects differed with year and shearing age. Suris were coarser than Huacayas with the effect reducing with increased liveweight; there was no effect of sex. Fleeces of light shade were 1 μm finer than dark fleeces. CV(FD) declined rapidly between birth and 2 years of age, reaching a minimum at about 4 years of age and then increasing; however, CV(FD) measurements on young animals were very poor predictors of CV(FD) at older ages, and the response of CV(FD) to age differed with farm and year. Suris had a higher CV(FD) than Huacayas on most properties, and MFD, liveweight, and sex did not affect CV(FD). Fleeces of dark shade had higher CV(FD) than fleeces of light shade in 2 of the years. It is concluded that there are large opportunities to improve the MFD and CV(FD) of alpaca fibre through selection and breeding. The potential benefit is greatest from reducing the MFD and CV(FD) of fibre from older alpacas, through reducing the between-animal variation in MFD and CV(FD). Sampling alpacas at ages <2 years is likely to substantially decrease selection efficiency for lifetime fibre diameter attributes.
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Vivanco-Mackie, H. W., M. D. Ponce-Salazar, M. Miguel-Gonzales, C. R. Youngs, C. Jara, and M. Asparrin. "112 Comparative study between slow freezing and vitrification on the survival rate of cryopreserved alpaca embryos post-transfer." Reproduction, Fertility and Development 31, no. 1 (2019): 182. http://dx.doi.org/10.1071/rdv31n1ab112.
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The objective of this study was to compare the effectiveness of cryopreserving in vivo-produced alpaca embryos by slow freezing v. vitrification. The embryos were produced from 9 female alpacas at Fundo Mallkini, Puno, Peru, located at 4300m elevation. The donor alpacas were synchronized by induction to ovulate with an injection of gonadotropin-releasing hormone (0.0084mg of buserelin acetate) and natural mating with vasectomized males to male receptive donors (day of ovulation induction was considered Day 0). On Day 2, the donors were injected 700IU of eCG. On Day 7, the donors received an injection of prostaglandin F2α (0.25mg of cloprostenol) and were mated on Day 8 by fertile males (2 matings 12h apart: 0600 and 1800h). The embryos were collected at 5.5 days after fertile mating and were graded as per IETS recommendations; most of the embryos were already expanded and hatched blastocysts. Embryos were washed and maintained in holding medium (1L PBS+1g Glucose+36mg sodium pyruvate+0.4% BSA+50mg kanamycin monosulfate) at 23°C for up to 1h and distributed into 2 groups for either slow freezing for direct transfer (n=14 embryos) or vitrification (n=10 embryos). Slow freezing consisted of transfer into freezing medium (9mL of 1.5M ethylene glycol+1mL of 1.0M sucrose prepared in holding media) at 23°C, placing in 0.25-mL straws and subjected to freezing at a rate of −0.5°C/ minute to −35°C and then plunging into LN. Vitrification followed a procedure described for camel embryos whereby embryos were exposed to solutions containing increasing amounts of glycerol and ethylene glycol for fixed periods and were then loaded into an open pull straw and plunged directly into LN for storage. The cryopreserved embryos were transferred into adult alpacas at the Community of Suitucancha, Junin, Peru (1500km from the farm where the embryos were collected and cryopreserved, 4200m elevation). Embryos in the slow-freezing group were thawed in warm water at 37°C for 30s and loaded directly into the embryo transfer gun for direct transfer into 7 alpaca recipients (2 embryos per recipient). Vitrified embryos were warmed by removing the open pull straw from the LN and transferring the embryos to 2 warming solutions at 37°C with decreasing levels of vitricants and containing 0.5M galactose with a final incubation at room temperature in holding media and then transferred into 5 alpaca recipients (2 embryos per recipient). The embryos were transferred into synchronized recipients by transcervical nonsurgical method. Pregnancy diagnosis was made by transrectal ultrasound examination at 45 days post-transfer. The pregnancy rates in the slow-freezing and vitrification groups, respectively, were 2/7 (29%) and 0/5 (0%); the difference was not significant (P>0.05) based on Fisher’s exact test. Twin pregnancies were not detected. We consider the result with slow freezing very promising, as in previous trials we had less than 18% pregnancies. More trials with larger number of embryos per cryopreservation method are being programmed.
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Vivanco-Mackie, H. W., M. D. Ponce Salazar, M. M. Gonzales, and M. A. Tapia. "108 COMPARATIVE EFFICIENCY OF GONADOTROPIN-RELEASING HORMONE AND LUTEINIZING HORMONE IN THE INDUCTION OF OVULATION IN SUPEROVULATED ALPACAS." Reproduction, Fertility and Development 26, no. 1 (2014): 168. http://dx.doi.org/10.1071/rdv26n1ab108.
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Alpacas are induced ovulators, responding to copulation and (or) exogenous application of ovulation-inducing hormones. The objective of this study was to determine the efficiency of the injection of a gonadotropin-releasing hormone (GnRH) agonist versus LH in the induction of ovulation and the presence and size of non-ovulated follicles at the time of embryo collection and its relationship to the yield of transferable embryos in superovulated alpacas. Twenty-one adult (3 to 7 years old) female alpacas under extensive grazing at 4300 m elevation in the Peruvian Andes that had been synchronized and treated for superovulation were induced to ovulate 6 days after the application of the superovulatory hormone (1000 IU of eCG, Folligon®, Intervet International BV, Boxmeer, the Netherlands) by mating with fertile males and injection immediately after copulation of either an IM dose of 0.0084 mg of buserelin acetate (Buserelina Zoovet®, Laboratorio Zoovet, Santa Fe, Argentina) to 10 alpacas (T1) or an IM dose of 5-mg Armour standard of LH (Lutropin®, Bioniche Animal Health, Belleville, ON, Canada) to 11 alpacas (T2). All alpacas had a second mating 12 h after the first mating and were subjected to ovarian inspection by ultrasonography and previous embryo collection by nonsurgical transcervical embryo flushing 6.5 days after the first mating. On average, the embryo recovery rate for T1 was 34.6% and there were 7.8 ± 3.99 corpora lutea (CL), 2.7 ± 4.08 collected embryos, and 3.6 ± 2.95 total, 0.5 ± 0.85 small (<6 mm), 1.8 ± 1.99 medium (≥6 but ≤14 mm), and 1.3 ± 2.11 large (≥15 mm) non-ovulated follicles. For T2, the embryo recovery rate was 59.4% and there were 6.73 ± 1.49 CL, 4.0 ± 2.57 collected embryos, and 0.64 ± 0.81 total, 0.0 ± 0.0 small, 0.36 ± 0.67 medium, and 0.27 ± 0.47 large non-ovulated follicles. The differences between treatments were nonsignificant (P > 0.05) for all the parameters; however, there was a clear tendency for a better recovery rate, more embryos collected, and lower number of non-ovulated follicles in T2. The Pearson correlation coefficient between the presence of large follicles in the ovaries at the time of embryo collection and the total number of embryos collected was negative (r = –0.26) and highly significant (P ≤ 0.001). The use of LH for ovulation induction of superovulated alpacas seems to be more advisable than the use of GnRH agonist; further comparisons with larger number of observations per treatment are recommended. This study was financed by the Peruvian Fund for Innovation, Science and Technology (FINCYT).
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Huanca, W., F. Hilari, M. Ticona, B. Lira, J. C. Villanueva, and W. F. Huanca. "96 Follicular Fluid and Serum Biochemical Composition in Alpacas (Vicugna pacos) with Different Nutritional Planes." Reproduction, Fertility and Development 30, no. 1 (2018): 187. http://dx.doi.org/10.1071/rdv30n1ab96.
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The importance of nutrition on composition of follicular fluid and oocyte quality is recognised in several species but limited information is available on alpacas and would explain the high rate of early embryo mortality. The aim of this study was to analyse the biochemical composition of serum and follicular fluid (FF) from alpacas (Vicugna pacos) with different nutritional planes. Twelve adult female alpacas between 6 and 8 years old were assigned randomly to either a high (n = 6) or low (n = 6) plane of nutrition. Nutritional planes were defined by the grazing condition of the natural pasture with (high) or without (low) supplementation with 200 g of concentrate/day (total digestible nutrients: 52%, protein: 16%, and metabolizable energy: 2.8 Mcal kg−1). The nutritional conditions were imposed 1 month before the start of the experiment.The concentrations of total glucose, cholesterol, triglycerides, and proteins were determined with a semi-automatic biochemical analyzer (SINOWA, China). When an ovulatory follicle ≥7 mm was detected by ultrasonography using a 5-MHz linear-array transducer (SSD-500, Aloka, Tokyo, Japan), a transvaginal transducer with an attached needle guide (UST-945BP-5, Aloka) was used to collect follicular fluid. Follicular puncture was performed using a disposable 19-gauge # 12 mm hypodermic needle connected to a 50-mL conical tube via a silicon tube. A caudal epidural anaesthesia was induced with 4 mL of 2% lidocaine. Blood samples were obtained by jugular venipuncture into serum vacutainers and kept at room temperature for 30 min. Follicular fluid and serum were centrifuged at 1500 × g for 20 min, decanted, and stored at –20°C until analysed. Results were analysed by Student’s t-test. The results are presented in Table 1. The results suggest differences in biochemical composition of glucose and cholesterol in follicular fluid, which could explain the oocyte quality and embryo survival in alpacas. Table 1.Biochemical composition of follicular fluid (FF) and serum from alpacas (Vicugna pacos) under 2 nutritional planes Research was supported by INNOVATE PERU Grant 405-PNICP-PIAP-2014.
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Hilasaca Mamani, Madeley Gladys, Jesús Martín Urviola Sánchez, Francisco Halley Rodríguez Huanca, and Víctor Raúl Leyva Vallejos. "Efecto de la duración de cópula en la respuesta ovulatoria y tasa de preñez en alpacas." Revista de Investigaciones Altoandinas - Journal of High Andean Research 23, no. 4 (October 31, 2021): 229–35. http://dx.doi.org/10.18271/ria.2021.296.
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An experiment was carried out at the Experimental Center La Raya-Universidad Nacional del Altiplano Puno, to determine the effect of copulation duration on ovulation and pregnancy in alpacas. 47 adult females were used (postpartum time ≥ 20 days and presence of a follicle ≥ 7 mm) distributed in three groups, according to copulation time: 20, 35 and 50 min, for G1 (n = 15), G2 (n = 16) and G3 (n = 16), respectively. For the mating, 6 reproducers of proven fertility were used, interrupting the copulation at the established time. Ovulation and pregnancy rates were evaluated on days 7 and 30 post service respectively (by ultrasound and receptivity of the female), the data were analyzed using X2 and simple correspondence. Both the ovulation rate (G1: 73.33%, G2: 81.25% and G3: 81.25%); and pregnancy (G1: 53.33%, G2: 81.25% and G3: 75%) were not different (P> 0.05). Pregnancy rates between G1 and G2 tended to be different (P <0.10), consistent with the simple correspondence analysis. Despite there being no differences in ovulation and pregnancy rates, there was a higher percentage trend in G2 and G3, suggesting that the longer duration of mating would have some positive effect on them, according to the simple correspondence analysis, and the trend of low significance (P <0.10) in favor of pregnancy in G2 compared to G1. In conclusion, there were no significant differences in the effect of copulation duration on ovulation; however, there was a 90% trend for the difference between mating duration of 35 minutes versus 20 minutes (in pregnancy).
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Huanca, W. F., J. M. Palomino, J. C. Villanueva, J. Malaga, and W. Huanca. "228 Effect of seminal plasma on the interval to application of equine chorionic gonadotrophin for the recovery of cumulus-oocyte complexes in alpacas (Vicugna pacos)." Reproduction, Fertility and Development 32, no. 2 (2020): 242. http://dx.doi.org/10.1071/rdv32n2ab228.
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Alpacas as other camelids are induced ovulators and require an external stimulus-mounting by the male-for ovulation. NGF-β, protein present in the seminal plasma (SP) is reported as being responsible for stimulating ovulation. However, limited information exists on the effect of ovulation inducers in the new follicular wave emergence with application on protocols of superstimulation. The aim of the study was to determine the effect of two ovulation induced techniques and two different times on the interval to application of equine chorionic gonadotrophin (eCG) for the ovarian superstimulation for the recovery of COCs by ovum pick- up (OPU). Alpacas were evaluated by transrectal ultrasonography with an Aloka SSD 500 ultrasound and transducer (7.5MHz) to determine the presence of a dominant follicle=7mm. A 2×2 experimental design was used with adult female alpacas, 6 to 8 years old, which were assigned to one of the treatments when a dominant follicle=7mm was present. Alpacas were assigned randomly 1 of 2 groups (Day 0) in which follicular ovulation was induced by seminal plasma 1mL IM (SP, n=13; group 1) with application of 650IU of eCG at 36h (n=6) or 48h (n=7) post-application of SP, or by GnRH with 0.008mg of Buserelin IM (GnRH, n=10; group 2) with application of 650IU eCG at 36h (n=5) or 48h (n=5) post-application of GnRH. On Day 7, COCs were counted and the OPU procedure was performed in every group. Data were analysed using ANOVA. Results are presented in Table 1. In conclusion, both inducers of ovulation and both times of application of eCG were effective for producing follicles of 7mm and COCs acceptable for recovery in Alpacas. Table 1.Number of follicles=7mm (top) and number of COCs recovered (mean±s.e.m.) Outcome and group 36 h 48 h Total Follicles GnRH 6.8±1.93 13.4±4.34 10.10±2.50a SP 5.5±2.23 5.29±1.15 5.38±1.25b Total 6.09±1.44 8.67±2.18 7.43±1.33 COCs GnRH 5±0.97 3±1.26 4.33±0.74a SP 1±0.41 0.33±0.22 0.67±0.25b Total 3.29±0.67x 1.4±0.6y 2.5±0.53 a,bValues within columns with different letters differ significantly (P=0.05). x,yValues within columns with different letters differ significantly (P=0.05). This research funded by CIENCIACTIVA-CONCYTEC as part of the project title “Role of seminal plasma in reproductive physiology and application of biotechnologies in camelids” (149-2017).
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VAUGHAN, JL, KL MACMILLAN, GA ANDERSON, and MJ D'OCCHIO. "Effects of mating behaviour and the ovarian follicular state of female alpacas on conception." Australian Veterinary Journal 81, no. 1-2 (January 2003): 86–90. http://dx.doi.org/10.1111/j.1751-0813.2003.tb11442.x.
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Huanca, W., A. Castro, N. Gomez, and A. Cordero. "111 BIOCHEMICAL COMPOSITION OF FOLLICULAR FLUID IN RELATION TO THE STIMULUS TO INDUCE OVULATION IN ALPACAS (VICUGNA PACOS)." Reproduction, Fertility and Development 29, no. 1 (2017): 164. http://dx.doi.org/10.1071/rdv29n1ab111.
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Alpacas, like other camelids, are induced ovulators. A study was designed to determine the effect of the ovulation-inducing stimulus on the biochemical composition of follicular fluid. Adult female alpacas (n = 18) were examined daily for 3 days by transrectal ultrasonography using a 5-MHz linear-array transducer (Aloka SSD-500, Tokyo, Japan). When the largest growing ovarian follicle was ≥7 mm, alpacas were given 1.0 mL of seminal plasma intramuscularly (SP, n = 9) or 40 µg of busereline acetate intramuscularly (GnRH, n = 9). A transvaginal transducer with an attached needle guide (Aloka UST-945BP-5) was used for collection of follicular fluid 22 h post-induction. Follicular contents were then centrifuged at 800 × g for 20 min to separate the fluid from the cells. The follicular fluid was collected and stored at –20°C until analysis with a semi-automatic biochemical analyzer (SINOWA, China). The results were glucose 49.17 and 47.95 (mg/dL; P > 0.05), total protein 1.85 and 1.15 (g/dL; P < 0.05), albumin 1.11 and 1.13 (g/dL; P > 0.05), triglycerides 3.94 and 3.16 (mg/dL; P > 0.05), cholesterol 39.01 and 42.5 (mg/dL; P > 0.05), phosphatase 32.68 and 21.36 (IU/L; P < 0.05), alanine aminotransferase 3.66 and 5.07 (IU/L; P > 0.05), and lactate dehydrogenase 42.17 and 27.27 (IU/L; P > 0.05) for SP or GnRH treatments, respectively. Results suggest the need to continue research to explain the effect of possible differences in total protein, cholesterol, and phosphatase on oocyte-expressed genes and follicular development. Research was supported by the project no. 405-PNICP-PIAP-2014-UNMSM.
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Byers, Stacey R., James F. Evermann, Daniel S. Bradway, Steven M. Parish, and George M. Barrington. "Evaluation of a commercial bovine viral diarrhea virus vaccine in nonpregnant female alpacas (Vicugna pacos)." Vaccine 28, no. 3 (January 2010): 591–93. http://dx.doi.org/10.1016/j.vaccine.2009.10.026.
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Gamarra, G., A. Gallegos, E. Alvarado, M. Asparrin, and W. Vivanco. "159 TECHNIQUES FOR OVUM PICK-UP IN GONADOTROPIN-TREATED ALPACAS." Reproduction, Fertility and Development 20, no. 1 (2008): 159. http://dx.doi.org/10.1071/rdv20n1ab159.
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The objective of the present study was to evaluate the quantity and quality of oocytes collected when using 2 methods for ovum pick-up and 2 different regimens for ovarian stimulation in live alpaca donors. Thirty-four non-pregnant female alpacas of 3 to 5 years of age maintained at 4100 m elevation in southern Peru were randomly distributed into 4 experimental groups. Groups 1 (n = 8) and 3 (n = 9) received an intravaginal device containing 0.78 mg of progesterone (Cue-Mate�, Bioniche Animal Health, Belleville, Ontario, Canada) plus an i.m. injection of 1 mg of estradiol benzoate on Day 0; the intravaginal device was removed on Day 7. Groups 2 (n = 7) and 4 (n = 10) received an i.m. injection of 3.1 mg of LH (Lutropin�, Bioniche Animal Health) on Day 0. Females received 700 IU of eCG (Pregnecol�, Bioniche Animal Health) i.m. on Day 7 (Groups 1 and 3) or Day 2 (Groups 2 and 4). In all groups, oocyte collection was done 2 days after the injection of eCG. Groups 1 and 2 were subjected to ventral laparotomy aspirating the oocytes from follicles >3 mm in diameter using a 10-mL hypodermic syringe containing 1 mL of aspiration media (Ringer's lactate solution plus 10% bovine serum) and connected to an 18 G � 1 inch aspiration needle. After collection, the follicular fluid was searched and the COC were graded. Groups 3 and 4 were subjected to ovum pick-up by transvaginal recovery using an ultrasound scanner (Parus 240�, Pie Medical, Maastricht, the Netherlands) equipped with a vaginal probe of 7.5 MHz (MEVA�, Pie Medical) and a 17G � 55 cm aspiration needle introduced through a needle guide. Follicles >3 mm in diameter were aspirated into 50-mL centrifuge tubes containing 5 mL of aspiration media with 75 IU mL–1 of heparin. The aspirated fluid was filtered and rinsed using an embryo filter (EmCon�, Immunosystems, Menomonie, WI), and COC were searched and graded under a microscope based on the intactness of the cumulus cell layers. Data were analyzed by ANOVA. There were no differences (P > 0.05) between groups in the mean number of follicles aspirated per donor (11.0, 13.8, 9.4, and 9.1 for Groups 1 to 4 respectively), and in the mean number of COC recovered per donor (7.6, 7.0, 6.0, and 6.1 respectively for Groups 1 to 4). The proportions of good quality COC were significantly (P < 0.01) different between surgical (81.0 and 79.5% for Groups 1 and 2) and transvaginal/ultrasound-guided (7.4% for Group 3) methods of collection; however, they were similar to the proportion in Group 4 (64.9%) retrievals. The results show that in the absence of an intravaginal device, a similar quantity and quality of alpaca oocytes can be collected when using a surgical approach or minimally invasive ultrasound-guided transvaginal follicular aspiration.
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Kubátová, A., T. Fedorova, I. Skálová, and L. Hyniová. "Non-invasive Pregnancy Diagnosis from Urine by the Cuboni Reaction and the Barium Chloride Test in Donkeys (Equus asinus) and Alpacas (Vicugna pacos)." Polish Journal of Veterinary Sciences 19, no. 3 (September 1, 2016): 477–84. http://dx.doi.org/10.1515/pjvs-2016-0060.
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Abstract The aim of the research was to evaluate two chemical tests for non-invasive pregnancy diagnosis from urine, the Cuboni reaction and the barium chloride test, in donkeys (Equus asinus) and alpacas (Vicugna pacos). The research was carried out from April 2013 to September 2014. Urine samples were collected on five private Czech farms from 18 jennies and 12 alpaca females. Urine was collected non-invasively into plastic cups fastened on a telescopic rod, at 6-9 week intervals. In total, 60 and 54 urine samples from alpacas and jennies, respectively, were collected. The Cuboni reaction was performed by the State Veterinary Institute Prague. The barium chloride test was done with 5 ml of urine mixed together with 5 ml of 1% barium chloride solution. Results of the Cuboni reaction were strongly influenced by the reproductive status of jennies; the test was 100% successful throughout the second half of pregnancy. However, no relationship was found between the real reproductive status of alpaca females and results of the Cuboni reaction. It was concluded that the barium chloride test is not suitable for pregnancy diagnosis either in donkeys, due to significant influence of season on the results, or in alpacas, because no relationship between results of the test and the reproductive status of alpaca females was found. In conclusion, the Cuboni reaction has potential to become a standard pregnancy diagnostic method in donkeys.
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Brennan, Patricia L. R., Maya Sterett, Mary DiBuono, Genesis Lara Granados, Kay Klo, Rebecca Marsden, Pearl Schleinig, Louise Tanner, and Stephen Purdy. "Intra-horn Penile Intromission in the Alpaca Vicugna pacos and Consequences to Genital Morphology." Integrative and Comparative Biology 61, no. 2 (May 10, 2021): 624–33. http://dx.doi.org/10.1093/icb/icab050.
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Synopsis Copulatory behavior and genital morphology interact to deliver sperm more effectively during mating, but the nature of this interaction has not been explored in depth in most vertebrates. Alpacas have unusually long copulations lasting 15–20 min, and a unique copulatory behavior, where the penis intromits all the way past the cervix, into the uterine horns. Here we describe the morphology of male and female genitalia and report unique morphological characteristics that may be associated with this unusual insemination mode. Vaginal shape is highly variable, and seemingly not associated with age or parity. The cranial vagina varies between bulbous and cylindrical, while the caudal vagina is typically narrower and always cylindrical. The cervix consists of a series of two to three spirals or rings, and it is often found in a relaxed state that may prevent damage caused by the cartilaginous penis tip as it pushes through the cervix to reach the uterine horns. The uterus and uterine horns have a complex shape with multiple constrictions. The cartilaginous penis tip has a sharp urethral process that may help to push against these constrictions. The diameter of the vaginal lumen is much greater than the diameter of the penis suggesting that there is little direct interaction between them, and that female vaginal shape is not under strong copulatory selection. In effect, the entire female reproductive tract of the female is interacting with the penis during copulation.
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Huanca, W., G. Marin, A. Cordero, M. Uchuari, and W. F. Huanca. "28 Evaluation invitro of two protocols of vitrification from alpaca (Vicugna pacos) embryos." Reproduction, Fertility and Development 33, no. 2 (2021): 121. http://dx.doi.org/10.1071/rdv33n2ab28.
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The reproductive efficiency of South American camelids as the alpaca is low, with a few number of animals having a good genetic characteristic. The transfer of cryopreserved embryos has great potential to disseminate valuable genetic, but the suitable protocol for such cryopreservation still needs to be developed. In this study, two protocols of vitrification of alpaca embryos were tested. Day 6.5 post-mating, embryos (n=66) were recovered from 14 female alpacas through a non-surgical technique and classified according to the characteristics of old world camelids reported by Skidmore et al. 2004 (Reprod. Fertil. Dev. 16, 605–609). Only quality 1 and 2 embryos were used for the study. They were placed together in 50-µL drops of holding medium for 30min and transferred to a 100-µL drop of equilibration solution 1, consisting of 7.5% (v/v) ethylene glycol (EG) + 0.25M sucrose. After 1min, embryos were transferred to equilibration solution 2, consisting of 15% (v/v) EG + 0.5M sucrose. After 2min, embryos were transferred into 2 consecutive drops of vitrification solutions A [SA: 30% (v/v) EG + 1M sucrose] for 20s each, then in 2 other drops of vitrification solution B [SB: 30% (v/v) EG + 3% glycerol + 1M sucrose] for 20s each. Thereafter, embryos were quickly loaded into open pulled straws (OPS) in a volume of 10µL and then plunged into liquid nitrogen. For warming, the OPS were held in air for 5s and subsequently thawed at 37°C for 50s. Straws were emptied into 1mL of prewarmed holding medium solution (HMS1) containing 1M sucrose for wash and the thawed blastocysts were transferred into a second 1mL of prewarmed HMS1. After 5min incubation at 37°C, the blastocysts were transferred into 1mL of warmed Holding medium solution 2 (HMS2) containing 0.5M sucrose maintained at room temperature (∼24°C) for evaluation. Data were analysed by the Chi-squared test. Post-thaw embryo expansion results were 81.3% and 58.8% for SA and SB (P<0.05), respectively. Post-thaw embryo quality (1 and 2) were found at 62.5% and 29.1% with SA and SB, respectively (P<0.05). In conclusion, the vitrification of alpaca embryos with the ethylene glycol:sucrose solution results in better post-thaw outcomes than the ethylene glycol:sucrose:glycerol. Further experiments with embryo transfer are needed. This research was funded by FONDECYT project no. 149-2017.
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Huanca, W., G. Marin, A. Cordero, M. Uchuari, and W. F. Huanca. "28 Evaluation invitro of two protocols of vitrification from alpaca (Vicugna pacos) embryos." Reproduction, Fertility and Development 33, no. 2 (2021): 121. http://dx.doi.org/10.1071/rdv33n2ab28.
Full textAbstract:
The reproductive efficiency of South American camelids as the alpaca is low, with a few number of animals having a good genetic characteristic. The transfer of cryopreserved embryos has great potential to disseminate valuable genetic, but the suitable protocol for such cryopreservation still needs to be developed. In this study, two protocols of vitrification of alpaca embryos were tested. Day 6.5 post-mating, embryos (n=66) were recovered from 14 female alpacas through a non-surgical technique and classified according to the characteristics of old world camelids reported by Skidmore et al. 2004 (Reprod. Fertil. Dev. 16, 605–609). Only quality 1 and 2 embryos were used for the study. They were placed together in 50-µL drops of holding medium for 30min and transferred to a 100-µL drop of equilibration solution 1, consisting of 7.5% (v/v) ethylene glycol (EG) + 0.25M sucrose. After 1min, embryos were transferred to equilibration solution 2, consisting of 15% (v/v) EG + 0.5M sucrose. After 2min, embryos were transferred into 2 consecutive drops of vitrification solutions A [SA: 30% (v/v) EG + 1M sucrose] for 20s each, then in 2 other drops of vitrification solution B [SB: 30% (v/v) EG + 3% glycerol + 1M sucrose] for 20s each. Thereafter, embryos were quickly loaded into open pulled straws (OPS) in a volume of 10µL and then plunged into liquid nitrogen. For warming, the OPS were held in air for 5s and subsequently thawed at 37°C for 50s. Straws were emptied into 1mL of prewarmed holding medium solution (HMS1) containing 1M sucrose for wash and the thawed blastocysts were transferred into a second 1mL of prewarmed HMS1. After 5min incubation at 37°C, the blastocysts were transferred into 1mL of warmed Holding medium solution 2 (HMS2) containing 0.5M sucrose maintained at room temperature (∼24°C) for evaluation. Data were analysed by the Chi-squared test. Post-thaw embryo expansion results were 81.3% and 58.8% for SA and SB (P<0.05), respectively. Post-thaw embryo quality (1 and 2) were found at 62.5% and 29.1% with SA and SB, respectively (P<0.05). In conclusion, the vitrification of alpaca embryos with the ethylene glycol:sucrose solution results in better post-thaw outcomes than the ethylene glycol:sucrose:glycerol. Further experiments with embryo transfer are needed. This research was funded by FONDECYT project no. 149-2017.
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Skidmore, Julian A., M. Billah, R. V. Short, and W. R. Allen. "Assisted reproductive techniques for hybridization of camelids." Reproduction, Fertility and Development 13, no. 8 (2001): 647. http://dx.doi.org/10.1071/rd01057.
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The camelid family comprises the Old World camelids (or dromedary and Bactrian camels) and the New World camelids (namely the llamas, alpacas, guanacos and vicunas). Although the species within each group can hybridize among themselves to produce fertile offspring, it is only recently that a hybrid between New and Old World camelids has been reported. To create this hybrid, semen was collected from male camels by artificial vagina (AV) and inseminated into female guanacos (n= 9) and llamas (n= 3) at the appropriate stage of their follicular wave cycle. Similarly, guanaco and llama semen was collected, also by AV, and inseminated into female camels (n= 42). Although several conceptions occurred, only one hybrid (camel sire×guanaco dam) continued to term and was born alive after 328 days of gestation, and another is pregnant at the time of writing (camel sire×llama dam). Further studies are presently being carried out using extraspecific embryo transfer to try and improve the success rate of live offspring being born. Female guanacos (n= 4) are treated with hormones to stimulate their ovaries to produce several follicles before being inseminated with camel semen. Of the 12 camel recipients that have to date received hybrid embryos (camel sire×guanaco dam), 10 conceived, but 9 of these subsequently aborted between 30 and 365 days and only one recipient was still pregnant at the time of writing.
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Cockcroft, PD, I. Mackie, J. Perry, C. Caraguel, K. Townsend, and MP Reichel. "Cross-sectional observational survey of serum biochemistry values in a population of 69 adult female alpacas (Vicugna pacos) in South Australia." Australian Veterinary Journal 94, no. 4 (March 28, 2016): 125–26. http://dx.doi.org/10.1111/avj.12421.
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Gamarra, G., A. Gallegos, M. Asparrin, and H. W. Vivanco-Mackie. "244 DEVELOPMENT OF SUPEROVULATORY STRATEGIES IN ALPACAS." Reproduction, Fertility and Development 19, no. 1 (2007): 238. http://dx.doi.org/10.1071/rdv19n1ab244.
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The objective of the present study was to determine the ovarian response of alpacas to different treatments for follicular development and superovulation. Twenty-nine mature, lactating alpacas, between 31 and 56 days postpartum, managed in the Peruvian highlands (altitude = 4100 m) were randomly distributed into 4 experimental groups. Groups 1 (n = 7) and 3 (n = 8) received a homemade intravaginal sponge containing 60 mg of medroxyprogesterone acetate (MAP; Sigma Chemical Co., St Louis, MO, USA) plus 2 mL of PGF2α (IM; Illiren�; Intervet International, Boxmeer, The Netherlands) on Day 0. Groups 2 (n = 7) and 4 (n = 7) received 2 mL of PGF2α (IM) on Day 0, but did not receive a MAP sponge. All groups received 6 injections (IM) of FSH (Folltropin V�; Bioniche Animal Health, Beltsville, Ontario, Canada) in decreasing dosages of 50, 50, 30, 30, 20, and 20 mg, respectively, every 12 h (at 0700 and 1900 h each day), plus 300 IU of eCG (IM; Folligon�; Intervet International) at the time of the last FSH treatment, with the aim of increasing LH levels. The FSH treatments started on Days 7, 5, 9, and 7 (from Day 0) in groups 1–4, respectively. MAP sponges were removed at the time of the last FSH treatment in groups 1 and 3. All alpacas were naturally mated twice at 12 and 24 h after the last FSH treatment. Alpacas in groups 1 and 2 received 3000 IU of hCG (IM; Corulon�; Intervet International) and alpacas of groups 3 and 4 received 2.5 mL of GnRH (IM; Conceptal�; Intervet International) immediately after the first mating. Seven days after the first mating, ovaries of all alpacas were examined by transvaginal ultrasonography. Ovarian response was estimated by determining the number of CL present on each ovary. The numbers of follicles that were at least 8 mm in diameter were also counted. Data were analyzed as a complete randomized design with 4 treatments. The average number of CL per alpaca was 1.3, 1.00, 1.00, and 0.9 for groups 1 to 4, respectively (P > 0.05). The average number of follicles that were at least 8 mm in diameter per alpaca was 9.4, 20.4, 0.9, and 3.9 for groups 1 to 4, respectively (P ≤ 0.05) with females in group 2 showing the highest response. We conclude that progestin treatment did not affect ovulatory response of lactating alpacas to exogenous gonadotropins. An effective ovarian stimulation strategy for achieving superovulation in alpacas remains to be developed.
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Van Steelandt, M. D., V. M. Tanco, M. H. Ratto, and G. P. Adams. "164 EFFECT OF OVULATION-INDUCING FACTOR (OIF) ON OVARIAN FUNCTION IN CATTLE." Reproduction, Fertility and Development 20, no. 1 (2008): 162. http://dx.doi.org/10.1071/rdv20n1ab164.
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Systemic administration of ovulation-inducing factor (OIF), discovered recently in seminal plasma of llamas, alpacas (induced ovulators), and cattle (spontaneous ovulators), stimulated ovulation in >90% of female llamas and alpacas. The objective of the present study was to test the hypothesis that purified OIF from llama seminal plasma would induce ovulation in cattle. Peripubertal heifers, weighing 323 � 27 kg, were used to minimize the confounding effect of spontaneous ovulation. Heifers (n = 11/group) were treated intramuscularly with 1.0 mg/100 kg of purified OIF, 100 µg of GnRH (positive control), or 2.5 mL of phosphate-buffered saline (negative control). Ovarian dynamics were monitored daily by transrectal ultrasonography for 10 days post-treatment. Blood samples were collected at 0.5- to 1-h intervals for 8 h, beginning at the time of treatment. Ovulation occurred in 9/11 (82%) of GnRH-treated heifers and in 1/11 (9%) heifers in each of the OIF- and saline-treated groups (P < 0.05). A surge in plasma LH concentration was detected within 30 min of treatment in the GnRH group (2.2 � 0.1 ng mL–1; P < 0.05), but remained at the basal level in the OIF- and saline-treated groups (0.3 � 0.1 and 0.2 � 0.1 ng mL–1, respectively). The onset of regression of the dominant follicle present at the time of treatment was earlier (P < 0.05) in OIF- v. saline-treated heifers (3.1 � 0.6 days v. 6.0 � 0.7 days). The interval from treatment to follicular wave emergence was shorter (P < 0.05) in GnRH- and OIF-treated heifers than in those treated with saline (1.1 � 0.4 days, 1.5 � 0.3 days, and 3.1 � 0.3 days, respectively). A similar pattern was observed for emergence of the second follicular wave (5.1 � 0.7 days, 4.6 � 0.5 days, and 6.6 � 0.4 days, respectively). Purified OIF did not induce ovulation in heifers but hastened both the regression of the extant dominant follicle and follicular wave emergence. Results provide a rationale for the hypothesis that OIF from seminal plasma is involved in controlling follicular wave dynamics in spontaneously ovulating species (e.g., Bos taurus) through a suppressive effect on the dominant follicle. The mechanism of action on ovarian follicular wave dynamics, as well as species specificity, remains to be elucidated.
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Huanca, W., R. L. Condori, M. A. Chileno, J. Cainzos, J. J. Becerra, L. A. Quintela, and P. G. Herradon. "211 IN VIVO MATURATION AND IN VITRO FERTILIZATION OF ALPACA OOCYTES." Reproduction, Fertility and Development 23, no. 1 (2011): 204. http://dx.doi.org/10.1071/rdv23n1ab211.
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The objectives of the study were to evaluate the ovarian follicular response, cumulus–oocyte complex (COC) collection rate, fertilization, and culture of COC collected from alpacas after treatment with 2 different gonadotropins. Female alpacas were assigned to Group 1 (n = 8), 200 mg of FSH (Folltropin, Bioniche, Belleville, Ontario, Canada) divided b.i.d. for 3 days, plus a single IM dose of 1000 IU of hCG (Chorulon, Intervet, Salamanca, Spain) 24 h after the last FSH treatment; or Group 2 (n = 10), 750 IU of eCG (Folligon, Intervet) as a single dose, plus a single IM dose of 1000 IU of hCG on Day 3 after eCG treatment (Day 0 = start of the superstimulatory treatment). At 20 to 22 h post-hCG treatment, the ovaries were surgically exposed and COC were aspirated from follicles ≥6 mm and evaluated. The COC with a homogeneous cytoplasm and 2 or more layers of cumulus cells were transferred to plates with a 40-μL drop of TCM-199 maturation medium supplemented with 10% FCS (vol/vol) plus 0.5 μg mL–1 of FSH, 10 μg mL–1 of hCG, 0.2 mM sodium pyruvate, 50 μg mL–1 of gentamicin, and 1 μg mL–1 of oestradiol under mineral oil with 10 to 12 oocytes/drop and maturated 24 h at 39°C in an atmosphere of 5% CO2 and high humidity. After maturation, COC were removed and fertilized in vitro using epididymal sperm. Testes were collected from mature males from a slaughterhouse and transported to the laboratory. The caudal epididymide was isolated. A prick was made on the convoluted tubules with a sterile hypodermic needle and the fluid, rich in spermatozoa, was aspirated in syringes containing 2 mL of Tris-fructose egg yolk extender. Motile spermatozoa were obtained by centrifugation at 600 × g on a Percoll discontinuous gradient (45.0:22.5%) for 10 min. The supernatant was then removed by aspiration and the pellet was resuspended in TL-HEPES and centrifuged again at 300 × g for 5 min. The pellet was resuspended in TL-stock. Gametes were co-incubated for 18 h at 39°C with 5% CO2 and high humidity. Presumptive zygotes were cultured in KSOM medium supplemented with 1 mM glutamine, 0.3 mM sodium pyruvate, 50 μg mL–1 of gentamicin, EDTA, essential and nonessential amino acids, and BSA for 3 days and cultured in SOF medium for 7 days. Embryo development was evaluated at 72 h and 7 days. Data were subjected to ANOVA. The number of follicles ≥6 mm did not differ at the time of COC collection (19.3 ± 5.7 and 21.5 ± 7.3), and the number of COC collected was 16.7 ± 5.3 and 17.3 ± 6.6 for the FSH group and the eCG group, respectively. The cleavage rate was 45.2 and 42.1% for the FSH group and the eCG group, respectively, at 72 h of culture, and the blastocyst stage at Day 7 (22.2 v. 19.3) did not differ between treatments. In conclusion, the FSH and eCG treatments did not differ in the ovarian follicular response, COC collection rate, fertilization, and culture of COC. Both gonadotropins can be used in the IVF protocol for alpacas. Grant 064 FINCyT-PIBAP 2008 and Grant 032-2009 PROCYT–CONCYTEC.
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Bogle, O. A., M. H. Ratto, and G. P. Adams. "Evidence for the conservation of biological activity of ovulation-inducing factor in seminal plasma." REPRODUCTION 142, no. 2 (August 2011): 277–83. http://dx.doi.org/10.1530/rep-11-0042.
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An ovulation-inducing factor (OIF) in the seminal plasma of llamas and alpacas (induced ovulators) and cattle (spontaneous ovulators) suggests that OIF is a conserved constituent of seminal plasma among mammals. In this study, three experiments were designed to determine the biological effects of OIF in different species. In experiment 1, superstimulated prepubertal female CD-1 mice (n=36 per group) were given a single 0.1 ml i.p. dose of 1) phosphate-buffered saline (PBS), 2) 5 μg gonadotropin-releasing hormone (GNRH), 3) 5 IU hCG, or 4) llama seminal plasma. The proportion of mice that ovulated was similar among groups treated with GNRH, hCG, or seminal plasma, and all were higher than the saline-treated group (P<0.001). In experiment 2, female llamas (n=8 or 9 per group) were intramuscularly treated with 1) 2 ml PBS, 2) 1 ml diluted llama seminal plasma, 3) 3 ml equine seminal plasma, or 4) 3 ml porcine seminal plasma. Experiment 3 was the same as experiment 2 except that the dose of equine and porcine seminal plasma was increased to 8 and 10 ml respectively. All llamas that were treated with llama seminal plasma ovulated and none that were treated with saline ovulated (P<0.0001). The proportion of llamas that ovulated in response to equine and porcine seminal plasma was intermediate. We conclude that the mechanism for the biological response to OIF is present in prepubertal CD-1 mice and that OIF is present in equine and porcine seminal plasma.
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Volkery, J., J. Gottschalk, A. Sobiraj, T. Wittek, and A. Einspanier. "Progesterone, pregnanediol-3-glucuronide, relaxin and oestrone sulphate concentrations in saliva, milk and urine of female alpacas (Vicugna pacos) and their application in pregnancy diagnosis." Veterinary Record 171, no. 8 (August 2, 2012): 195. http://dx.doi.org/10.1136/vr.100393.
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Cuya, R., W. Huanca, G. Medina, R. Sanchez, and W. F. Huanca. "99 Effect of application of seminal plasma on Day 0, 5, or 7 postmating on pregnancy rate and embryonic survival in alpacas (Lama pacos)." Reproduction, Fertility and Development 31, no. 1 (2019): 175. http://dx.doi.org/10.1071/rdv31n1ab99.
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Alpacas, similar to other camelids, are classified as induced ovulators, as an external stimulus is required for the occurrence of ovulation. A protein identified as β-nerve growth factor, present in the seminal plasma (SP), has the capacity to induce ovulation and corpus luteum formation. Alpacas exhibit poor reproductive efficiency, with birth rates below 50% due in part to high embryo mortality before 35 days post-mating. A study was carried out to evaluate the effect of the application of SP on Day 0, 5, or 7 post-mating on pregnancy rate and embryo survival, defined as the difference in the numbers of pregnant females between Day 35 and 25. Nonpregnant adult alpaca females (n=124) between 5 to 6 years old were evaluated by transrectal ultrasonography to determine presence of a follicle ≥7mm, and then 2 days later to confirm permanence of the follicle. Alpacas were then bred by natural mating and assigned randomly to 1 of 4 treatments: 1mL of SP IM at mating; 1mL of SP IM Day 5 post-mating; 1mL of SP IM Day 7 post-mating; and control. Semen was collected from adults male and ejaculates were diluted 1:1 with PBS and then centrifuged for 30min at 3000 rpm. Supernatant was separated and a drop evaluated to determine absence of spermatozoa and SP-free sperm was stored at −20°C. Twenty adult males with optimal reproductive performance were used for mating with females assigned to the different treatments. Animals were evaluated by ultrasound with an Aloka SSD 500 (Aloka, Tokyo, Japan) and 5.0-MHz linear transducer on Day 25 and 35 to determine pregnancy rate and embryonic survival. Data were analysed by chi-square. Results are present in Table 1. The results differ from our initial hypothesis and a possible explanations may be that additional application of SP IM could saturate receptors and block the action of the seminal plasma present in the ejaculate of males. Table 1.Pregnancy rate and embryonic survival in alpacas with application of seminal plasma on Day 0, 5, or 7 Study was supported by project no. 405-PNICP-PIAP-UNMSM.
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Huanca, W. F., C. Mamani, and W. Huanca. "165 EFFECT OF INJECTION OF SEMINAL PLASMA ON OVULATION RATE, CORPUS LUTEUM DEVELOPMENT, AND SENSITIVITY TO PROSTAGLANDIN IN ALPACAS (VICUGNA PACOS)." Reproduction, Fertility and Development 27, no. 1 (2015): 173. http://dx.doi.org/10.1071/rdv27n1ab165.
Full textAbstract:
The seminal plasma (SP) of camelids contains a protein identified as β Nerve Growth Factor with capacity of induced ovulation and develop a corpus luteum. A study was designed to evaluate the effect of application of SP on the interval of time from injection of stimulus to the ovulation and corpus luteum (CL) size (Experiment 1, n = 24) and on the sensitivity of CL to the injection of prostaglandin (196 µg of tiaprost) (PG) at different periods from ovulation (Experiment 2, n = 86). Exp. 1: Adult female alpacas with presence of a follicle ≥7 mm were assigned to the application of 1 mL of SP via IM (T: n = 12) or application of 50 µg of acetate of busereline IM (T2: n = 12). Exp. 2: Alpacas with presence of a follicle ≥7 mm were induced to ovulation with 50 µg of acetate of busereline or 1 mL IM of SP. Animals were evaluated by ultrasound to confirm the ovulation and were assigned to the following treatment: T1 (n = 8): SP + PG Day 4; T2 (n = 8): buserelin acetate + PG Day 4; T3 (n = 8): SP + PG Day 5; T4 (n = 8): buserelin acetate + PG Day 5; T5 (n = 8): SP + PG Day 6; T6 (n = 8): buserelin acetate + PG Day 6; T7 (n = 8): SP + PG Day 7; T8 (n = 8): buserelin acetate + PG Day 7; T9 (n = 8): SP + PG Day 8; T10 (n = 8): buserelin acetate + PG Day 8 and T11 (n = 6) (Control): Application of 1 mL of saline solution. The animals were evaluated by ultrasound with an Aloka SSD500 (Aloka, Tokyo, Japan) and 7.5-MHz linear transducer each 2 h (Exp. 1) and each 12 h (Exp. 2) after of application of PG. In Exp. 1, the ovulation rate was 95.7% to T1 and T2 and an interval of time between injection of stimulus and ovulation was 27.4 ± 2.5 h and 26.8 ± 1.8 to T1 and T2, respectively. In Exp. 2, luteolysis was 0.0%, 0.0%, 25.0%, 0.0%, 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, and 0.0% for the treatments T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, and T11 respectively. The results suggest that no differences exist between in the ovulation rate and interval to the ovulation between the application of buserelin acetate or SP and that the CL was sensible at Day 5 to the prostaglandin respect SP and with similar response to the sensibility of CL from Day 6 to Day 8.
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Bogle, O. A., M. H. Ratto, and G. P. Adams. "221 PREPUBERTAL MOUSE BIOASSAY FOR OVULATION-INDUCING FACTOR IN SEMINAL PLASMA." Reproduction, Fertility and Development 20, no. 1 (2008): 190. http://dx.doi.org/10.1071/rdv20n1ab221.
Full textAbstract:
A substance in the seminal plasma of llamas and alpacas has been discovered that induces ovulation and growth of the corpus luteum (CL) in the female of the same species. The ovarian effects of the ovulation-inducing factor (OIF) are associated with a surge release of LH into circulation. Ultrasonographic detection of ovulation and CL development is currently the only method available for testing the bioactive effects of OIF. The purpose of this study was to determine if a superstimulatory prepubertal mouse model could be developed as an in vivo bioassay for OIF. Prepubertal female CD1 mice (n = 144), 20 days of age and weighing 20–25 g, were housed at 24�C with lights on from 0500 to 1900 h and free access to food and water. An intramuscular dose of 5 IU of eCG (Novormon, Bioniche Animal Health, Belleville, ON, Canada) was given (Day 0) for ovarian superstimulation. On Day 2, mice were assigned randomly to 4 groups (n = 36 per group) and given a single 0.1 mL intraperitoneal dose of (1) 5 IU of hCG (Chorulon, Intervet Canada, Ltd., Whitby, ON, Canada), (2) 5 µg GnRH (gonadotropin-releasing hormone: Cystorelin, Merial, Ltd., Iselin, NJ, USA), (3) llama seminal plasma, or (4) phosphate-buffered saline (negative control). On Day 3, females were euthanized by an overdose of inhaled halothane. Oviducts were collected and oocytes were counted using trans-illumination stereomicroscopy. The proportion of mice that ovulated did not differ among groups treated with hCG, GnRH, and seminal plasma (31/36, 31/36, 28/36, respectively); however, the proportion of mice that ovulated in each treatment group was greater than that in the saline-treated group (9/36) (P < 0.001). The number of oocytes counted (mean � SEM) was also similar among groups treated with hCG (25.8 � 2.9), GnRH (27.4 � 2.7), and seminal plasma (19.2 � 2.8), all of which were greater (P < 0.01) than in the saline-treated group (6.2 � 2.1). We conclude that the superstimulated prepubertal CD1 mouse model is effective as an in vivo bioassay for OIF in seminal plasma. Whether the bioassay may be used for quantitative estimates of OIF activity will require dose-response trials using serial dilutions of seminal plasma.
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Perez Guerra, Uri, Manuel Perez Durand, Lourdes Limache Mamani, Vilma Condori Villegas, Rassiel Macedo Sucari, Eloy Condori Chuchi, Oscar Orós Butrón, Saul Espinoza Molina, and María Ignacia Carretero. "Evaluation of fertility and subfertility in adult alpacas and tuis using ultrasonography, endometrial cytology and bacterial isolation." SPERMOVA 10, no. 2 (December 31, 2020): 57–63. http://dx.doi.org/10.18548/aspe/0008.09.
Full textAbstract:
The objective of this study was to compare the uterine health between fertile, sub-fertile alpacas and tuis using transrectal ultrasonography, endometrial cytology and bacterial isolation. A total 10 tuis (young mature females without breeding with average age of 1.5 years) and 20 adult alpacas of the Suri breed were used. In turn, the adult females were divided into two groups of 10 animals each according to their reproductive history: fertile group (parturition every year) and sub-fertile group (1 to 2 years without pregnancy). In all females, the thickness of the cervix and uterine horns was determined by transrectal ultrasonography. On the other hand, endometrial cytology and bacterial isolation were performed from samples obtained by uterine flushing. A Kruskal-Wallis and a Chi-square tests were used to compare ultrasonography and cytology groups. A greater thickness of the cervix and both uterine horns (p˂0,05) was observed in the fertile alpacas with respect to the sub-fertile and tuis. The percentage of PMN in tuis and sub-fertile alpacas was < 2%, while in fertile alpacas the percentage of PMN were: 6 animals with < 2% PMN, 2 animals with 2-5% PMN and two other alpacas with > 5% PMN. The bacteria isolated were: Bacillus lechiniformis and Escherichia coli in the three groups studied, Staphylococcus saprophyticus and Bacillus cereus in tuis and fertile alpacas, Staphylococcus aureus in tuis and sub-fertile, Bacillus spp. and Micrococcus spp. in fertile and sub-fertile alpacas, Bacillus lactic acid, Staphylococcus epidermidis and Citrobacter spp. in fertile alpacas, Enterococcus spp., Bacillus subtilis and Klebsiella spp. in sub-fertile and Enterobacter spp. in tuis. The low percentage of PMN in endometrial cytology in sub-fertile alpacas would indicate the absence of endometritis at the time of the study. However, the lower thickness of the cervix and uterine horns observed in sub-fertile alpacas suggest that it would be necessary to perform uterine biopsies in order to evaluate if there is any association between the thickness of the uterine wall and the presence of degenerative and/or inflammatory changes observed on histopathological examination.
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Perez Guerra, Uri, Manuel Perez Durand, Lourdes Limache Mamani, Vilma Condori Villegas, Rassiel Macedo Sucari, Eloy Condori Chuchi, Oscar Orós Butrón, Saul Espinoza Molina, and María Ignacia Carretero. "Evaluation of fertility and subfertility in adult alpacas and tuis using ultrasonography, endometrial cytology and bacterial isolation." SPERMOVA 10, no. 2 (December 31, 2020): 57–63. http://dx.doi.org/10.18548/aspe/0008.09.
Full textAbstract:
The objective of this study was to compare the uterine health between fertile, sub-fertile alpacas and tuis using transrectal ultrasonography, endometrial cytology and bacterial isolation. A total 10 tuis (young mature females without breeding with average age of 1.5 years) and 20 adult alpacas of the Suri breed were used. In turn, the adult females were divided into two groups of 10 animals each according to their reproductive history: fertile group (parturition every year) and sub-fertile group (1 to 2 years without pregnancy). In all females, the thickness of the cervix and uterine horns was determined by transrectal ultrasonography. On the other hand, endometrial cytology and bacterial isolation were performed from samples obtained by uterine flushing. A Kruskal-Wallis and a Chi-square tests were used to compare ultrasonography and cytology groups. A greater thickness of the cervix and both uterine horns (p˂0,05) was observed in the fertile alpacas with respect to the sub-fertile and tuis. The percentage of PMN in tuis and sub-fertile alpacas was < 2%, while in fertile alpacas the percentage of PMN were: 6 animals with < 2% PMN, 2 animals with 2-5% PMN and two other alpacas with > 5% PMN. The bacteria isolated were: Bacillus lechiniformis and Escherichia coli in the three groups studied, Staphylococcus saprophyticus and Bacillus cereus in tuis and fertile alpacas, Staphylococcus aureus in tuis and sub-fertile, Bacillus spp. and Micrococcus spp. in fertile and sub-fertile alpacas, Bacillus lactic acid, Staphylococcus epidermidis and Citrobacter spp. in fertile alpacas, Enterococcus spp., Bacillus subtilis and Klebsiella spp. in sub-fertile and Enterobacter spp. in tuis. The low percentage of PMN in endometrial cytology in sub-fertile alpacas would indicate the absence of endometritis at the time of the study. However, the lower thickness of the cervix and uterine horns observed in sub-fertile alpacas suggest that it would be necessary to perform uterine biopsies in order to evaluate if there is any association between the thickness of the uterine wall and the presence of degenerative and/or inflammatory changes observed on histopathological examination.
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Tribulo, P., O. A. Bogle, and G. P. Adams. "128 BOVINE SEMINAL PLASMA INDUCES OVULATION IN LLAMAS." Reproduction, Fertility and Development 24, no. 1 (2012): 176. http://dx.doi.org/10.1071/rdv24n1ab128.
Full textAbstract:
Ovulation-inducing factor (OIF) is a protein present in the seminal plasma of several species, including llamas, alpacas, pigs, cattle, sheep, horses and rabbits. In an initial study (Ratto et al. 2006 Theriogenology 66, 1102–1106), bovine seminal plasma induced ovulations in 26% (5/19) of llamas compared with 0% (0/19) in the placebo group, but induced proportionately less than in llamas treated with alpaca or llama seminal plasma (100%). It is important to highlight that treatments were based on volume of seminal plasma; the actual dose of OIF was unknown. In a later study (Tanco et al. 2011 Biol. Reprod. doi:10.1095/biolreprod.111.091876), OIF from llama seminal plasma had a dose-dependent effect on ovulation rate, corpus luteum (CL) diameter and progesterone production in llamas. The present study was designed to test the hypothesis that bovine seminal plasma induces ovulation and CL development in llamas comparable with that of llama seminal plasma, based on total dose of OIF. Within species, seminal plasma was pooled from 1 to 4 ejaculates per male (n = 145 bulls, n = 4 llamas). The concentration of OIF in the pooled seminal plasma was measured by radioimmunoassay and the volume of seminal plasma used for treatment was adjusted to reach a total dose of 250 μg of OIF. Mature female llamas were assigned randomly to 4 groups and given a single intramuscular dose of 10 mL of PBS (negative control, n = 5), 50 μg of gonadotropin-releasing hormone (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. The incidence of ovulation was compared among groups by Fisher's exact test. Nonserial data (i.e. follicle size at treatment, maximum CL diameter, day of maximum CL diameter and first day of CL detection) were compared among groups by ANOVA. The diameter of the preovulatory follicle at treatment did not differ among groups (P = 0.10). The incidence of ovulation was 0/5, 4/5, 3/6 and 4/6 in the groups treated with PBS, GnRH, llama seminal plasma and bovine seminal plasma, respectively (P < 0.05). The incidence of ovulation did not differ among llamas treated with GnRH, llama seminal plasma, or bovine seminal plasma. Among the treatments that elicited ovulation, neither the maximum CL diameter nor the day of maximum CL diameter differed (P = 0.30 and P = 0.24, respectively). In addition, no difference was detected in the day of first detection of the CL (P = 0.25). Results document the bioactivity of OIF in the bovine seminal plasma of Bos taurus. These findings further support the notion that OIF is highly conserved among mammals and that seminal plasma exerts its effect in an OIF dose-related manner. This research was supported by the Natural Sciences and Engineering Research Council of Canada.
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Landeo, L., M. Zuñiga, T. R. Gastelu, and J. A. Ruiz. "19 Invitro embryonic development from oocytes collected by ovum pickup of superstimulated females and nonstimulated slaughterhouse ovaries of alpaca (Vicugna pacos)." Reproduction, Fertility and Development 33, no. 2 (2021): 117. http://dx.doi.org/10.1071/rdv33n2ab19.
Full textAbstract:
The objective of this study was to evaluate the embryonic development of alpaca oocytes collected by ovum pickup from superstimulated females (OPU, Group 1) and from slaughterhouse ovaries of 8 non-superstimulated females (SHO, Group 2) using a conventional aspiration technique (20G needle and a 3-mL syringe). A total of 8 nonpregnant alpacas, 3 to 4 years old, were superstimulated with a single dose of 200IU of equine chorionic gonadotrophin (eCG, Day=0). Three days later, alpacas were examined by transrectal ultrasonography with a 7.5-MHz linear-array transducer to determine the number and diameter of follicles available for aspiration. A total of 101 follicles were aspirated, recovering 67 oocytes (66.3%) by OPU using an endocavity transducer attached to a 21G needle adapted for alpacas. The follicular fluid was aspirated using a regulated vacuum pump (40 mmHg) into a tube containing 5mL of phosphate-buffered saline (PBS), 0.2% bovine serum albumin (BSA), and 10IUmL−1 heparin, at 37°C. In the SHO group we used 16 ovaries maintained at 28°C. The recovery of oocytes was carried out within 3h of ovary collection. We aspirated 155 follicles from SHO and recovered 117 oocytes (75.5%). After collection, all oocytes recovered were morphologically classified into categories (I and II) and cultured for 26h in an incubator (5% CO2 in air at 38.5°C), in TCM-199 supplemented with 0.2mmol sodium pyruvate, 50µgmL−1 gentamicin sulphate, 0.02IUmL−1 FSH, 1µgmL−1 oestradiol 17β, and 10% fetal calf serum (FCS). After maturation, oocytes were invitro fertilized with epididymal spermatozoa recovered from postmortem males and co-cultured for 18 to 20h. After this period, all cleaved oocytes were incubated (5% CO2 in air, 38.5°C) for 6 days in synthetic oviductal fluid-serum medium. Number and morphological quality of oocytes collected, invitro cleaved, and embryos ratea were registered and compared between groups. Statistical significance was determined using Kruskal–Wallis test. The mean and standard error were calculated from average of the percentages obtained in each repetition. Results indicated that the mean number of oocytes collected per ovary was higher (P<0.05) using SHO (7.8±2.4) than OPU (4.5±3.0). Also, the number of oocytes classified as category I, was higher in the SHO compared with OPU group (56% vs. 30% respectively; P<0.05); however, category II oocytes were the same (16% vs. 15%, respectively). There was no difference in early development (cleavage) rate between OPU (57±2.0) and SHO (49±1.5) groups. However, there was difference in the rate of development (P<0.05) between OPU and SHO groups to reach the morula stage (56±2.0 vs. 42±1.7, respectively) and early blastocyst stage (55±2.0 vs. 34±1.4, respectively). In conclusion, oocyte quality could be affected by hormonal stimulation or by the quality of follicles aspirated by OPU. In contrast, oocytes recovered from live animals by OPU have greater capability of embryonic development invitro than oocytes recovered from slaughterhouse ovaries.