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Journal of Biotechnology in Livestock Production

In vitro embryo production by lower than 40% motility frozen Red American Brahman semen

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Malee Apimeteetumrong Anone Thuangsanthia Sarawut Chayprasart Narong Liengchareon

Abstract
The aim of this study was to evaluate the efficacy of lower than 40% motility frozen Red American
Brahman (RAB) semen on embryo production following insemination in vitro with oocytes collected from live

American Brahman donors by ovum pick up. Frozen semen samples from two RAB bulls (RAB 280 and RAB 102)
with low (5-35%) motility, produced by Cattle Semen Production Center Inthanon Royal Project, were used in this

study. Sperm quality parameters such as sperm motility, concentration, viability and acrosomal status were
evaluated before and after sperm processing (swim-up) for in vitro fertilization (IVF). Embryos were cultured for 7-

8 days and recorded the development until reached blastocyst stage. Differences were analyzed by SPSS. Data
were expressed as % means ± S.E., before and after swim-up. For RAB 280 bull, the swim-up treatment

significantly improved (P < 0.05) the proportions of motility (26.76 ± 2.18 % vs 47.65 ± 1.97 %, before and after
swim-up) and live spermatozoa (31.45 ± 3.24 % vs 64.34 ± 3.26 %, before and after swim-up). The percentages

of membrane integrity (26.87 ± 1.65 % vs 32.41 ± 1.99 %) and normal intacted acrosome (27.66 ± 1.37 % vs
41.82 ± 2.47 %) were not significantly different after swim-up (P>0.05).After IVF, 24 blastocyst embryos were

produced with this bull semen (12 replicates). For RAB 102 bull, the swim-up treatment increased but there was
no significant difference in the proportions of all sperm quality parameters tested (20.00 ± 5.77 % vs 36.67 ± 3.33

% motility,21.72 ± 3.11% vs 44.05 ± 5.25 % live spermatozoa, 27.11 ± 2.01% vs 34.89 ± 5.79 % membrane
integrity and 25.83 ± 2.13 % vs 32.22 ± 2.22% normal intacted acrosome), for before and after swim-up treatment.

No cleaved and blastocyst embryo was produced from 3 experiments, due to contamination occurred during
insemination process. Results from this study indicate that frozen Red American Brahman (RAB) semen with lower

than 40% motility can produced blastocyst-stage embryos in vitro.

Keywords: in vitro fertilization, bull, frozen semen, embryos

Registered No. : 52 (1)-0208-087

1/ Embryo Transfer Technology and Animal Germplasm Research Center, Nakhon Ratchasima Province
2/ Chiang Mai Artificial Insemination and Biotechnology Research Center, Chiang Mai Province

3/ Bureau of Biotechnology in Livestock Production, Pathum Thani Province

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