Determination of the seminal leucocytary profile in cattle and buffalo semen using the Leishman, and Papanicolaou staining protocols

Abstract

Despite indications that seminal leucocytes can alter spermatozoa quality, investigations on the qualitative and quantitative parameters of such cells in bull semen, as well as methods for determining them are scarce. Therefore, the objective of this study was to assess the concentration of leucocytes in different breeds and age groups using the staining methods viz. Leishman and Papanicolaou stain. Semen ejaculates from two indigenous breeds (Sahiwal, Murrah) and one cross breed named Vrindavani were randomly collected. For the studies on effects of age, the ejaculates were divided into 2 groups, namely Group I, ejaculates from bulls less than 8 years of age (5 bulls x 6 ejaculates) and Group II, ejaculates from bulls more than 8 years of age (5 x 6 x 2 groups = 60 samples). All the samples are stained with Leishman and Papanicolaou stain for leucocytary counts. For the studies on effect of breed data was divided into 6 ejaculates x 3 bulls x 3 breeds (n=54). The concentration of leucocytes comprising mostly macrophages were found and was shown to be significantly higher in the Sahiwal breed (3.05±0.18) than other breeds and in Group II, more than 8 years old bulls (3.16±0.16) with Leishman stain. The study finds Leishman staining procedure better in terms of ease of doing and results obtained for leucocyte count than the Papanicolaou staining protocol.