Establishment of semen collection technique using electroejaculator and semen cryopreservation of Javan leopard (Panthera pardus melas Cuvier, 1809)

Bongot Huaso Mulia; Ardyta Widianti; Jansen Manansang; Dedi Rahmat Setiadi; Vincentia Trisna Yoelinda; Taufiq Purna Nugraha; Ni Wayan Kurniani Karja; Raden Iis Arifiantini

doi:10.14202/vetworld.2021.3156-3163

Establishment of semen collection technique using electroejaculator and semen cryopreservation of Javan leopard (Panthera pardus melas Cuvier, 1809)

Vet World. 2021 Dec;14(12):3156-3163. doi: 10.14202/vetworld.2021.3156-3163. Epub 2021 Dec 25.

Authors

Bongot Huaso Mulia^{1

2}, Ardyta Widianti², Jansen Manansang², Dedi Rahmat Setiadi³, Vincentia Trisna Yoelinda¹, Taufiq Purna Nugraha⁴, Ni Wayan Kurniani Karja³, Raden Iis Arifiantini³

Affiliations

¹ Biology Reproductive Program, Post Graduate School, IPB University, Jl. Raya Dramaga, Dramaga Campus, Bogor 16680, Indonesia.
² Department of Life Sciences, Taman Safari Indonesia, Jalan Kapten Harun Kabir No.724 Cisarua 16750, Bogor, West Java, Indonesia.
³ Department of Clinic, Reproduction, and Pathology, Faculty of Veterinary Medicine, IPB University, Jl. Agatis, Dramaga Campus, Bogor 16680, Indonesia.
⁴ Zoological Division, Research Center for Biology, Indonesian Institute of Science, Cibinong 16911 West Java, Indonesia.

Abstract

Background and aim: The Javan leopard (Panthera pardus melas Cuvier, 1809) is a subspecies of Panthera pardus spp., spread across the African and Asian regions. Information on reproductive aspects is crucial for wild animals, including the Javan leopard. In this study, we aimed to developelectroejaculator (EE) techniques and evaluate cryopreservation success in Javan leopard semen.

Materials and methods: The semen of four adult Javan leopards was collected once a week using EE. Placement of the EE probe in the rectum was performed after ultrasound imaging (ultrasonography) to determine the prostate body location. The semen obtained was then evaluated macroscopically and microscopically. Three Javan leopards were used for cryopreservation. The ejaculate was divided into two parts [i.e., one part diluted with AndroMed^® (Minitüb, Tiefenbach, Germany) and the other part with Steridyl^®(Minitüb, Tiefenbach, Germany)] at a 1:1 ratio immediately after collection and evaluation. The semen was then packed in a 0.25 mL MiniStraw^® (Minitüb, Tiefenbach, Germany) then equilibrated at 4°C for 2 h. After equilibration, the straw was then frozen in liquid nitrogen vapor. Frozen semen was then stored in containers until further evaluation.

Results: The results showed that ejaculation response occurred at all levels of stimulation, while erections did not always occur. The fastest ejaculation and erection occurred at the fourth voltage. The macroscopic evaluation showed that the semen volume was 0.80±0.26 mL, cloudy white, pH 7.44±0.14, and with watery semen consistency. The microscopic evaluation showed that the sperm motility was 66.98±0.39%, with sperm viability of 75.6±1.79%. Sperm concentration was 62.17±46.95×10⁶ mL^-1 with a total concentration of 42.14±23.51×10⁶ cells. Normal sperm morphology is only 40.72±6.26%.

Conclusion: This study concluded that the development of a semen collection technique using an EE preceded by imaging of the EE probe location using ultrasound was effective for the ejaculation of Javan leopards. The characteristics of the semen of the Javan leopard showed moderate semen volume, sperm motility, and viability. Javan leopard showed low sperm concentration and normal sperm morphology.

Keywords: Javan leopard; electroejaculator; semen characteristics; semen cryopreservation.