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Many of the methods used to stain semen result in very pronounced coloring of the sperm, but unfortunately they do not distinguish their individual structures, which play a key role in the fertilization process. Hence the aim of this study was to identify sperm structures using two staining techniques in the semen of roosters from breeding flocks. The subject of the study was the sperm of roosters from a Ross 308 breeding flock. To capture the differences in the dimensions of sperm subjected to the effect of different chemical substances in dyes, microscope slides were stained by two techniques: with an AgNO3 solution and by a differential method (eosin-nigrosin test). Assessment was made of the degree of coloration and the number of details that could be identified in the morphological structure of the sperm. The use of AgNO3 allowed accurate identification of the acrosome, nucleus, and midpiece, which were visible in the slides stained with eosin-nigrosin, but only in dead spermatozoa. The AGNO3 staining technique used in this study reveals the cell nucleus within the head and can be an alternative method to analysis with a scanning electron microscope. This staining technique can be used to stain sperm structures that cannot be seen in other methods of slide preparation, which means that it can be considered for routine use in assessing the fertility of roosters in breeder flocks.

Sperm morphometry has some value as an indicator of reproductive capacity in males. In laboratory practice a variety of slide-staining methods are used during morphological evaluation of semen to predict male fertility. The aim of this study was to determine the effect of staining of semen using four different techniques on the morphometry of the bull sperm cell. The material for the study consisted of semen collected from test bulls of the Black-and-White variety of Holstein-Friesians. The results obtained in the study indicate differences in the dimensions of bull sperm heads when different slide staining techniques were used. The most similar results for sperm head dimensions were obtained in the case of SpermBlue(®) and eosin+gentian violet complex, although statistically significant differences were found between all the staining techniques. Extreme values were noted for the other staining techniques - lowest for the Papanicolaou and highest for silver nitrate, which may indicate more interference in the cell by the reagents used in the staining process. However, silver nitrate staining was best at identifying the structures of the sperm cell. Hence it is difficult to determine which of the staining methods most faithfully reveals the dimensions and shape of the bull sperm.