The Acrosome Integrity Examination of Post-thawed Spermatozoa of Several Ongole Grade Bull in Indonesia Using Giemsa Staining Method

Abstract

Cement cryopreservation is a preserving technique of spermatozoa cells that the spermatozoa cells can be used in a long period of time. Frozen semen is produced by the National Artificial Insemination Center (AIC) and the Regional Artificial Insemination Center (RAIC) in Indonesia. The indicators in predicting spermatozoa fertilizing ability include individual motility and movement scores, but the indicators have not been able to accurately predict the spermatozoa fertilizing ability, Acrosome integrity of spermatozoa cells is an important indicator of the success of the fertilization. The study aimed at evaluating the quality of the acrosome integrity of Ongole Grade Bull post-thawed spermatozoa from several Artificial Insemination Centers (AIC) in Indonesia using Giemsa staining method. It used 60 straw samples of 6 PO cattle aged 6 to 8 years with healthy and normal reproductive organs obtained from 3 different Artificial Insemination Centers (AICs) in Indonesia. The spermatozoa acrosome integrity was evaluated using Trypan Blue-Giemsa (TBG) and Giemsa stainings combined with fixation using methanol. The results showed that the motility (43.60±1.65%) and the viability of the spermatozoa (55.75±1.83%) of Ongole grade bull in the AIC A was higher than those in the other AIC (P<0.05). The results of the evaluation of the acrosome integrity of the Ongole grade bull from the AIC A using the TBG staining showed that the acrosome integrity of the bull from the AIC A was higher than that of the bull from the other AIC (P<0.05) (77.00±1.52%). They also showed that the acrosome integrity of the Ongole grade bull from the AIC A evaluated using Giemsa staining was also higher than that of the bull from the other AIC (P<0.05) (76.84±1.57%). Spermatozoa acrosomes can be evaluated using TBG or Giemsa stainings. However, the TBG staining is more complex and it takes a long time for the staining to give expected results. Meanwhile, the Giemsa staining combined with fixation using methanol is a simple method and it does not take a long time for the staining to give satisfying results.