Research was conducted to define the basic reproductive physiology of killer whales (Orcinus orca) and to use this knowledge to facilitate the development of artificial insemination procedures. The specific objectives were 1) to determine the excretory dynamics of urinary LH and ovarian steroid metabolites during the estrous cycle; 2) to evaluate the effect of an exogenously administered, synthetic progesterone analog on reproductive hormone excretion; 3) to validate the use of transabdominal ultrasound for ovarian evaluation and timing of ovulation; 4) to examine the quality of semen after liquid storage and cryopreservation; and 5) to develop an intrauterine insemination technique. Based on urinary endocrine monitoring of 41 follicular phases and 26 complete cycles from five females, estrous cycles were 41 days long and comprised a 17-day follicular phase and a 21-day luteal phase. A consistent temporal relationship was observed between peak estrogen conjugates and the LH surge, the latter of which occurred approximately 0.5 days later. Two animals placed on oral altrenogest (three separate occasions for 30, 17, and 31 days, respectively) excreted peak urinary estrogen concentrations 25 days after withdrawal that were followed by sustained elevations in urinary pregnanediol-3alpha-glucuronide excretion. Mean preovulatory follicle diameter was 3.9 cm (n = 6), and ovulation occurred 38 h (n = 5) after the peak of the LH surge. Based on visual estimates of motility, liquid-stored semen maintained 92% of its raw ejaculate sperm motility index (total progressive motility x kinetic rating [0-5 scale, where 0 = no movement and 5 = rapid progressive movement]) when held at 4 degrees C for 3 days postcollection. Semen cryopreserved using a medium freezing rate demonstrated good postthaw total motility (50%), progressive motility (94%), and kinetic rating (3.5). Insemination during eight estrous cycles resulted in three pregnancies (38%), two from liquid-stored and one from cryopreserved semen. Two calves were delivered after gestation lengths of 552 and 554 days, respectively. These data demonstrate the potential of noninvasive endocrine monitoring combined with serial ultrasonography to improve our understanding of the reproductive biology of cetaceans. This fundamental knowledge was essential for ensuring the first successful conceptions, resulting in live offspring, using artificial insemination in any cetacean species.

To increase the basic understanding of killer whale (Orcinus orca) reproductive physiology necessary for the development of artificial breeding programs, we utilized radioimmunoassays (RIA) to detect urinary immunoreactive steroid metabolites (pregnanediol-3~-glucuronide [PdG] and estrone-conjugates [EC]) and gonadotropins (luteinizing hormone [LH] and follicle-stimulating hormone [FSH]) in urine samples from six female killer whales. Urine samples were collected from the whales by voluntary presentation behavior over a 2-to 4-year period. All urinary hormone values were corrected for intersample urine concentration variations by indexing with creatinine. Daily urine samples from four whales were collected during two conceptions and 18 complete estrous cycles. LH, FSH, EC, and PdG immunoreactive levels were determined and combined with observed copulatory activity in five cycles, including two conceptive cycles from two whales. Mean luteal phase lengths ranged from 9.7 to 19.2 days. Mean follicular phase lengths ranged from 6.5 to 16.8 days. Mean estrous cycle lengths based on the first detectable PdG levels were 41.6 2 6.72 S.E.M. days. After PdG nadir, immunoreactive FSH levels showed a bimodal pattern with the first peak being greater in size, and both preceding a follicular phase EC increase. LH levels > the 95% confidence interval of the mean were considered significant. Combined LH immunoreactive values from whales 2 and 6 during two and three estrous cycles, respectively, had significant LH peak concentrations on day minus 2. These significant LH peaks were assumed to represent the preovulatory LH surge. Eight copulations during two conceptive cycles were observed between whales 2 and 6 and a breeding male. Six of these copulations (3 with each female whale) occurred within 72 hours of the beginning or the end of the presumptive preovulatory LH surge. Estrous activity was seen throughout the year for the herd. However, individuals had varying periods of anestrus that could not be linked to environmental, social, or nutritional influences. The whales that were reproductively successful had anestrus intervals that were usually influenced by gestation, postparturient period, or lactation. The information obtained during this research enhances the foundation for future artificial reproductive management techniques. 0 1993 Wiley-Liss, Inc.

Diseases of the lens commonly affect captive pinnipeds. Access to UV-protective shade, early identification and medical management of ocular diseases, and prevention of fighting can limit the frequency or severity of lens-related disease in this population. An extended life span may result from captivity, but this also allows development of pathological changes associated with aging, including cataracts.

Sea World has maintained killer whales (Orcinus orca) since 1965. The total killer whale inventory has included 39 whales (25 females, 14 males); 28 were wild-caught and 11 captive-born, including one second-generation calf. As of September, 1993, there were 19 whales in the breeding program. Ten of these whales (53%) were captive-born, either at Sea World or other facilities in North America. The live wild-caught whales ranged in estimated age from 12-27 years (X * sd = 17.6 k 4.2 years). The captive-born whales ranged in age from <1 to 8 years. In the Sea World breeding program (through September, 1993), there have been nine live births and one stillbirth, with eight calves part of the current inventory. Births occurred from July to February. Calving intervals ranged from 32-58 months. Female age at birth of first calves ranged from 8 years to an estimated 17 years (% C sd = 12.7 5 3.0 years). Gestation, based on conception estimates from serum progesterone analysis, averaged 17 months (X 2 sd = 517 i 20 days), but successful pregnancies with viable calves occurred from 15-18 months (468-539 days). Females, in the presence and absence of males, were polyestrus with periods of cycling interspersed with individually variable noncycling (presumed anestrous) periods ranging from 3-1 6 months. Mean serum progesterone levels (ese) were as follows: noncycling periods = 121 -+ 20 pg/ml; peak elevations during nonconceptive ovulatory (estrous) cycles = 3,962 & 2,280 pglml; first pregnancies = 14,592 5 3,854 pg/ml; second pregnancies = 8,389 It 395 pg/ml; and third pregnancy = 8,180 t 4,556.