Enhancing captive Indian rhinoceros genetics via artificial insemination of cryopreserved sperm.

The objective of this study was to design an artificial insemination (AI) protocol using cryopreserved spermatozoa to obtain pregnancies in captive Indian rhinoceroses (Rhinoceros unicornis). Four methods developed varied by timing and approach, as follows; Method 1: females (n=2) were inseminated pre- and post-ovulation under general anesthesia, Method 2: females (n=2) were inseminated pre-ovulation without anesthetic via endoscopy, Method 3: females (n=1) were inseminated pre-ovulation without anesthetic via manual insertion of an insemination catheter, Method 4: females (n=2) were inseminated same as Method 3 with the addition of standing sedation. Semen deposition site varied as a result of changes in AI technology and experience. All females conceived following intrauterine AI using three methods. Four pregnancies (n=3 females) produced via Method 3 and 4 resulted in term births (n=2 male calves, n=2 female calves) at 481.8±12.8days post-AI. Unfortunately, two early pregnancy losses were documented in a fourth female conceiving via Method 2. Pregnancy rates were 0%, 22%, 17%, and 50% for Method 1-4, respectively. Method 3 and 4 rates improved to 29% and 67%, respectively when accounting for AI's conducted only on ovulatory estrous cycles. Spermatozoa (n=5 males) were cryopreserved 0.3-9.3 y prior to successful AI procedures. The lowest dose of frozen-thawed sperm resulting in conception was 500×10(6) motile sperm. Mean time from AI to ovulation in conceptive and non-conceptive cycles was 26±11.8h and 66±80.7h, respectively.

Urinalysis in three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis).

This study reports urinalysis values for three species of captive rhinoceros (Rhinoceros unicornis, Dicerorhinus sumatrensis, and Diceros bicornis) and evaluates individual and species differences. Repeated urinalysis was conducted on 11 individuals to establish normal reference ranges. Although no individual or species differences existed in urinary values for pH, all species differed in specific gravity. Rhinoceros urine demonstrated many physical and chemical properties similar to that of the horse, but reliability of this comparison was limited. Urinary pH in the rhinoceros was within range of that established for the horse and other large herbivores. However, all rhinoceros species exhibited urinary specific gravities below the lower limit of the normal equine reference range. Comparative urinalysis using an outside laboratory source confirmed the results of this study and illustrated the value of conducting in-house analysis. These results are the first data available on reference ranges for urine parameters in the greater one-horned, Sumatran, and African black rhinoceros and provide a useful diagnostic tool for the veterinary care of individuals in captivity.

Follicular, endocrine and behavioural dynamics of the Indian rhinoceros (Rhinoceros unicornis) oestrous cycle.

Longitudinal ultrasound, behaviour and endocrine evaluations were conducted, over 14 to 18 months, in two young female Indian rhinoceroses (Rhinoceros unicornis) to characterize the oestrous cycle. Both females showed the same pattern of follicular development producing a large follicle (10-12 cm diameter) on one of the ovaries that persisted for 8.5 +/- 4.68 days before spontaneously ovulating. Ovulation occurred in all eight cycles monitored in a 6- to 7-year-old female versus 10 out of 14 cycles monitored in a 5- to 6-year-old female. Ultrasound examinations confirmed follicular collapse 48 h following the onset of behavioural oestrus in ovulatory cycles, while anovulatory cycles were associated with the formation of a haemorrhagic follicle. The day of behavioural oestrus corresponded to peak urinary oestrogen conjugate concentrations for each cycle, but anovulatory cycles had lower concentrations on the day of behavioural oestrus compared with ovulatory cycles. A transient increase in urinary progesterone metabolite concentrations was detected 1 day prior to ovulation. Irregular urinary progesterone metabolite profiles followed anovulatory cycles, reflecting varying degrees of follicular luteinization. In an attempt to ensure that a cycle would result in ovulation in the 5- to 6-year-old female, a GnRH treatment was tested during two separate cycles. Administration of GnRH on the day of behavioural oestrus resulted in an increase in urinary luteinizing hormone concentrations 2 h following injection. Regardless, ovulation did not occur in response to treatment. This study provides the first ultrasound data on ovarian activity in the Indian rhinoceros and establishes normal physiologic and behavioural relationships during the oestrous cycle that may facilitate the breeding of this species in captivity.