The use of two staining methods for identification of spermatozoon structure in roosters.

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.

Identification of chromosome instability in Japanese quail (Coturnix japonica).

1. A study of the incidence of chromosome instability in the Japanese quail as assessed by sister chromatid exchange (SCE) and fragile site identification in chromosomes was conducted in two parent breeds and their F1 and F2 generations. 2. The mean incidence of SCEs was 6.02 ± 0.45 and the frequency of fragile sites was 1.17 ± 0.79. 3. There were moderately negative correlations of 0.51-0.64 between chromosome instability and fertility in the F1 and 0.10-0.23 in the F2. The hatch of fertilised eggs was negatively correlated with the number of SCE in male (0.31) and female (0.33) F1 and was lower in P (0.18 and 0.19, respectively), whereas the correlations were similar for the number of fragile sites in both generations (0.51-0.62).

Sister chromatid exchange as an index of chromosome instability in chondrodystrophic chickens (Gallus domesticus).

Commercially bred broiler chickens reared in inadequate breeder-created habitat conditions often suffer from a disease that afflicts their lower limbs, chondrodystrophy. The sister chromatid exchange (SCE) test makes it possible to detect chromosome instability that corresponds with elevated vulnerability of the organism to genotoxic factors of a mutagenic and carcinogenic nature. The frequency of SCE in chromosomes of chondrodystrophic and healthy broilers was analyzed. Chromosome preparations were obtained from our in vitro culture of peripheral blood lymphocytes stained using the fluorescence plus Giemsa technique. The SCE/cell mean of the population under analysis was 6.8 ± 1.6. The SCE/cell mean in the chromosomes of the chondrodystrophic chickens was 8.5 ± 1.0. The healthy chickens had an SCE/cell mean of 5.1 ± 1.3. Statistically significant differences were identified between both chicken groups. Moreover, a higher SCE/cell mean was observed in the males: 6.9 ± 1.7 compared with 6.7 ± 1.7 in the females. However, this difference was not statistically significant. Additionally, SCE incidence in the first 3 biggest autosome pairs (1, 2, 3) was analyzed in detail. The number of identified SCE was proportional to chromosome length. The most exchanges were observed in the proximal region of the chromosomes under analysis.

Sister chromatid exchange in Greenleg Partridge and Polbar hens covered by the gene-pool protection program for farm animals in Poland.

A basic assay that detects genotoxic DNA damage disrupting DNA replication and repair mechanisms is the sister chromatid exchange test. The frequency of sister chromatid exchanges was analyzed in chromosomes of the following hen breeds: Greenleg Partridge and Polbar. Chromosome preparations were obtained from our in vitro culture of peripheral blood lymphocytes stained using the fluorescence plus Giemsa (FPG) technique. The sister chromatid exchange (SCE)/cell mean of the hens under analysis was 7.83 ± 1.76 (7.22 ± 1.70 in the Greenleg Partridge and 8.43 ± 1.61 in the Polbar population). Statistically significant differences were identified between the hen breeds. A higher mean number of SCE/cell was observed in the group of hens producing fewer eggs (8.55 ± 1.51) compared with the group with a better egg yield (7.10 ± 1.65). The differences were statistically significant. Additionally, SCE frequency in the first, second, and third chromosome was analyzed in detail. The highest number of SCE was observed in the first and the lowest in the third chromosome. The SCE distribution in the particular regions of the analyzed chromosomes was also studied. The most numerous exchanges were observed in the proximal region, followed by the interstitial and distal areas.