Laparoscopic Ovum Pick-up For In Vivo Oocyte Retrieval In Alpacas

Alpacas are increasingly incorporated into reproductive and biomedical research due to distinctive physiological and immunological characteristics that extend their relevance beyond agricultural applications. As camelids, alpacas exhibit induced ovulation, prolonged follicular dynamics, asymmetric uterine physiology, and a unique immune system characterized by heavy-chain-only antibodies. These features support their use in comparative and translational research, particularly in reproductive biology and genetic engineering. Laparoscopic ovum pick-up (LOPU) is a minimally invasive approach for oocyte recovery in species with complex reproductive anatomy, allowing direct ovarian visualization, precise follicular aspiration, and reduced tissue trauma compared with transvaginal techniques. The objective of this study was to develop and standardize a reproducible in vivo oocyte retrieval protocol in alpacas using LOPU. Eight adult female alpacas (3-8 years of age) were enrolled in this study and subjected to repeated LOPU sessions conducted once per month throughout the study period. Alpacas were evaluated by transrectal ultrasonography, and animals presenting ovarian follicles ≥7 mm received 50 µg gonadotropin-releasing hormone intramuscularly. Ovarian superstimulation was induced using a total dose of 200 mg follicle-stimulating hormone administered in a decreasing twice-daily regimen over four days. LOPU was performed 12-14 h after the final hormone administration under general anesthesia induced with ketamine, xylazine, and butorphanol and maintained with isoflurane. A three-port laparoscopic approach with carbon dioxide insufflation was used. Follicles were aspirated using a 20-gauge needle connected to a regulated vacuum system (25-30 mmHg; 13-15 mL/min). Oocytes were recovered in TCM-199 medium and evaluated under a stereomicroscope. Alpacas exhibited a mean of 14.88 ± 3.85 follicles, with 10.94 ± 3.62 follicles aspirated per session and 6.68 ± 2.21 oocytes recovered, corresponding to a recovery rate of 61.30 ± 8.21%. These findings suggest that LOPU may represent a viable approach for in vivo oocyte retrieval in alpacas, with potential applications in reproductive biotechnology workflows.