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A. M. Miller, M. E. Roelke, K. L. Goodrowe, J. G. Howard, and D. E. Wildt

Summary. Eight female pumas were treated i.m. with 1000 (N = 5) or 2000 (N = 3) i.u. PMSG followed 84 h later by 800 i.u. hCG. Eggs were recovered 24–26 h after hCG from ovarian follicles by using laparoscopy and transabdominal aspiration. Mature eggs were inseminated in vitro 4–6 h later whereas immature eggs were cultured for 24 h and then inseminated. Electroejaculates from 3 pumas were diluted with mKRB before insemination to evaluate the influence of sperm concentration on fertilization. Seven of 8 pumas responded with follicle development, and 140 eggs were recovered from 145 follicles (96·6%; 77 mature, 43 immature, 20 degenerate eggs; mean ± s.e.m., 20·0 ± 5·9 eggs/female). Overall fertilization rate was 43·5% (total eggs fertilized = 40) despite using inseminates containing 82·99% pleiomorphic spermatozoa. Of the 36 immature oocytes matured in vitro and inseminated, 12 were fertilized even though 50% of the inseminating spermatozoa contained an acrosomal defect. Fertilization rate of mature oocytes collected from follicles appeared unrelated (P > 0·05) to PMSG dose or number of spermatozoa/inseminate. This study demonstrates that a high proportion of follicular eggs can be recovered laparoscopically from adult pumas treated with PMSG and hCG. These gametes are capable of being fertilized in vitro (immediately or after maturation in vitro) even with low quality semen with a high incidence of sperm pleiomorphisms.

Keywords: in vitro fertilization; puma; oocyte maturation; teratospermia

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M. A. Barone, D. E. Wildt, A. P. Byers, M. E. Roelke, C. M. Glass, and J. G. Howard

Ovarian response to equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG), the effect of timing of anaesthesia relative to hCG injection and the use of laparoscopic intrauterine artificial insemination were examined in the puma (Fells concolor). In Expt 1, females were treated with 100 (n = 6) or 200 (n = 8) iu eCG (i.m.) followed 80 h later by 100 iu hCG (i.m.) and were then anaesthetized 40–43 h after hCG injection for ovarian assessment. Although there was no difference (P > 0.05) in the number of unovulated ovarian follicles, females treated with 200 iu eCG had more (P < 0.05) corpora lutea per female and more corpora lutea as a percentage of the total number of ovarian structures. In Expt 2, all females were treated with 200 iu eCG and 80 h later with 100 iu hCG, and then anaesthetized either 31–39 h (Group A; n = 8) or 41–50 h (Group B; n = 6) after hCG injection for ovarian assessment. All Group B pumas ovulated compared with only three (37.5%) Group A females (P< 0.05). Compared with Group A, Group B pumas had more corpora lutea per female, more corpora lutea as a percentage of the total number of ovarian structures, and fewer unovulated follicles (P < 0.05). One of nine post-ovulatory females laparoscopically inseminated in utero with 16 × 106 motile spermatozoa became pregnant and delivered a healthy cub. Administration of 200 iu eCG and 100 iu hCG followed by anaesthesia no earlier than 41 h after hCG treatment is most likely to result in ovulation in pumas, and laparoscopic artificial insemination can be used to produce pregnancy in this species.