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Journal of the South African Veterinary Association

versión On-line ISSN 2224-9435
versión impresa ISSN 1019-9128

J. S. Afr. Vet. Assoc. vol.80 no.3 Pretoria  2009




Assessment of viability and acrosomal status of Asian elephant (Elephas maximus) sperm after treatment with calcium ionophore and heparin



M Sa-ardritI, II; N ThongtipII, III; K KornkaewratIII; T FaisaikarmIV; Y KitiyanantIV, V; S MahasawangkulVI; A PinyopumminIII; K SaikhunIV

INational Laboratory Animal Center, Mahidol University, Nakorn Pathom 73170, Thailand
IICenter for Agricultural Biotechnology, Kasetsart University, Nakorn Pathom 73140, Thailand
IIIFaculty of Veterinary Medicine, Kasetsart University, Nakorn Pathom 73140, Thailand
IVInstitute of Science and Technology for Research and Development, Mahidol University, Nakorn Pathom 73170, Thailand
VDepartment of Anatomy, Faculty of Science, Mahidol University, Nakorn Pathom 73170, Thailand
VIThe Thai Elephant Conservation Center, The Forest Industry Organization, Lampang 52000, Thailand




Knowledge about the acrosomal status of Asian elephant (Elephas maximus) sperm is extremely limited. The objective of this study was to evaluate the viability and acrosomal status of Asian elephant sperm following induction by calcium ionophore and heparin using propidium iodide (PI) and fluorescein isothiocyanate conjugated peanut agglutinin (FITC-PNA). Semen samples were collected from elephant bulls by manual stimulation. Semen was diluted with extender, cooled to 4 °C and transported to a laboratory for the experiment. Sperm cells were incubated in modified Tyrode's medium containing either 1 mM calcium ionophore or 10 mg/ml heparin for 5 h at 39°C. Sperm recovered at the onset (0 h), 1, 2, 3,4and5hof incubation were simultaneously assessed for the viability and acrosomal status using dual staining of FITC-PNA and PI. Results were confirmed by transmission electron microscopy. A progressive increase in the proportion of live-acrosome reacted sperm was observed within3hofincubation in both treatment groups which slightly decreased at 4 to5hof incubation. At 1 to3hof incubation, the percentage of live-acrosome reacted sperm induced by calcium ionophore was higher (P < 0.05) than those induced by heparin and the control. However, there were no statistical differences at 4 to 5 h of incubation. A progressive reduction of the percentage of motile sperm was observed in the control as well as both treatment groups. Sperm motility decreased sharply when they were incubated in calcium ionophore compared with incubation in heparin and control groups. These results indicate that the occurrence of live-acrosome reacted sperm in the Asian elephant was induced by calcium ionophore at a rate higher than that induced by heparin.

Keywords: acrosome reaction, calcium ionophore, elephant, heparin, sperm



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Received: January 2009
Accepted: June 2009



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