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Using buffered formalin to preserve the bent-tail characteristic of goatspermatozoa for the hypo-osmotic swelling test

Sakdichod Kimsakulvech^1*, Sookruetai Boonmasawai¹, Arpron Leesombun¹, ChowalitNakthong², Yupaporn Lanamtiang³, Waraluk Oupala³

¹Department of Pre-clinic and Applied Animal Sciences Faculty of Veterinary Science, Mahidol University, Salaya Campus, 999 Phuttamonthon 4 Road, Salaya, Nakhon Pathom, Thailand. 73170.
²Department of Clinical Science and Public Health, Faculty of Veterinary Science, Mahidol University, Salaya Campus, Nakhon Pathom, Thailand. 73170.
³Veterinary Medical Center for Livestock and Wildlife Animal Hospital, Faculty of Veterinary Science, Mahidol University, Kanchanaburi Campus, Sai-yok, Kanchanaburi, Thailand. 71150.

Malaysian J. Anim. Sci. 2021 24(1): 56-63

Click to view the abstract

The buffered formalin concentration and adding time for the hypo-osmotic swelling (HOS) test in
goat semen have never been recommended. This study examined the different buffered
concentrations for fixing bent-tail spermatozoa in HOS-treated goat semen for 5-120 minutes. In
experiment 1, nine HOS-treated goat semen samples were incubated at 37?C for 60 minutes and
bent-tail spermatozoa were evaluated as a control. At the same time, the different final
concentrations (0.99, 4.76, 9.09 and 13.04% (v/v)) of buffered formalin were added and compared
with the control. In experiment 2, nine HOS-treated goat semen samples were incubated at 37?C
for 5, 30, 60, 90 and 120 minutes before being evaluated for bent-tail spermatozoa. At the same
time, the different final concentrations (13.04, 16.67 and 20% (v/v)) of buffered formalin were
added to the HOS-treated semen. All buffered formalin groups, samples were placed at room
temperature for 24 hours before evaluation. The occurrence of bent-tail spermatozoa in the 0.99
and 4.76% (v/v) formalin groups was lower than the control and the 9.09% (v/v) trended to be
lower than the control. The 13.04% (v/v) formalin group showed some periods of a decrease in
bent-tail spermatozoa. The buffered formalin concentration of at least 16.67% (v/v) added to HOStreated goat semen had the relative stability of bent-tail spermatozoa for 5-90 minutes and was no
different from the non-fixative group. These results recommended the use of an optimal
concentration of buffered formalin at 16.67% (v/v) to fix the bent-tail spermatozoa in HOSs-treated
goat semen during 5-90 minutes of incubation. This technique will be suitable for goat semen
evaluation when there is a short period of incubation.