110 ASSISTED CONCEPTION IN THE FISHING CAT BY LAPAROSCOPIC INTRATUBAL INSEMINATION

Fishing cats (Prionailurus viverrinus) are small (6–15 kg) spotted cats from dispersed areas of Southeast Asia found mostly in wetland habitats. They are classified by the International Union for Conservation of Nature (IUCN) as endangered, with a decreasing population, due to habitat loss and degradation. Few studies have been done on applying assisted breeding techniques to the species, although the birth of a live kitten after IVF/embryo transfer (ET) has been reported (2006 Theriogenology 66, 1518–1524). Here, we describe the birth of a live fishing cat kitten using the technique of laparoscopic intratubal AI. A ten-year-old female who had served previously as an oocyte donor (5×) following gonadotropin treatment was administered a total of 5 IU of porcine FSH (Sioux Biochemical, Sioux City, Iowa) over 4 days (1×/day) followed by 10 IU of porcine LH on the fifth day. At approximately 28 h after LH treatment, the ovaries/oviduct were accessed by a laparoscopic technique comparable to that used for oviducal embryo transfer (ibid.). To deposit semen into the left oviduct, a 16-guage thin-wall trocar/needle was inserted into the abdominal cavity on the right side, approximately 1 cm lateral to the midline and approximately 2 to 3 cm below the umbilicus. A 14-cm open-end tom cat catheter was inserted into the 16-guage cannula (blunt) and the catheter tip was positioned underneath the fimbria overlaying the ovary. Then, a 50-mm length of 30-guage polytetrafluoroethylene (PTFE) thin-wall tubing containing approximately 30 μL of freshly collected semen was threaded through the catheter and the sample was expelled with positive pressure from a threaded-plunger 1-mL syringe. The left ovary contained 7 to 8 preovulatory (3–4 mm) follicles, 4 of which were manually ruptured immediately after deposition of semen with a 22-guage needle inserted through the 16-guage cannula. Then, with the 16-guage trocar/cannula in the same position (on the right side), the tip was redirected towards the right ovary and approximately 30 μL of semen was deposited underneath the fimbria as described above. The right ovary presented with 5 to 6 preovulatory (3–4 mm) follicles, 2 of which were punctured with the 22-guage needle after insemination. No ovulations were present on either ovary. The semen used for insemination was a fresh sample collected by electroejaculation from a 9-year-old male. The raw sperm concentration was 220 million mL–1, with 70% motility. The number of motile sperm deposited per oviduct was estimated to be approximately 4.6 million. The female was anesthetized 51 days later and radiography was done to determine her pregnancy status. A single fetus was present, so she was moved from an outdoor pen into a large indoor holding pen to allow for video-monitoring during the remainder of gestation. On Day 70, early signs of labour were observed and an elective Caesarean section was done approximately 4 h later. A live, healthy male kitten weighing 204 g was delivered. One year later, gonadotropin treatment/AI were repeated on the same pair. At approximately 30 h post-LH treatment, preovulatory follicles were present, but fewer than the previous treatment (5–6 total). Two fresh ovulation sites were seen on the left ovary. Pregnancy was not established. A reason for the failure was not apparent, unless it was age related.