Evaluation of sperm morphometry is an important criterion in the diagnosis of a male animal’s suitability for breeding. The aim of the study was to evaluate the morphometry of sperm from the epididymides of dogs subjected to routine castration using various staining methods. The study was carried out on semen collected from ten healthy dogs. Gonads were obtained from each dog during routine castration at a veterinary surgery. Then, the epididymides (caput, corpus, cauda) were isolated from the gonads, semen was collected from them and microscope slides were prepared. The slides for evaluation of sperm morphometry were prepared by four methods: DiffQuik, SpermBlue, eosin-nigrosin and eosin-gentian. A total of 2400 sperm were analyzed (240 sperm from the dog). The sperm collected from the caput and corpus of the epididymis were found to have larger heads and tails than those collected from the cauda of the epididymis. The staining method was shown to affect the morphometry of sperm taken from the epididymides of dogs. The staining methods differentiate the dimensions of the head of sperm in different parts of the epididymis but do not affect the length of the sperm tail. The occurrence of differences in the head dimensions of sperm may be linked to the use of different fixatives and chemical reagents in the staining procedure. Sperm stained by the EN method had the smallest head and tail dimensions. The greatest head area was noted in the sperm stained by the EG method. In the slides stained by the SB method, the sperm heads were relatively long but narrow. The methods used are suitable for the evaluation of sperm structure, and the possibility of using all four methods enables a full characterization of sperm collected from the caput, corpus and cauda of the epididymides of dogs.

The karyotype of the mallard duck, Anas platyrhynchos, was characterised on the basis of R and C bands. Chromosomal preparations obtained from in vitro blood lymphocyte cultures were RBG- and CBG-stained. The structures of nine and 14 pairs of chromosomes were analysed by the RBG and CBG chromosome banding techniques, respectively. The location of R bands, as well as the size and arrangement of constitutive heterochromatin blocks were determined. Ideograms of R and C banded patterns of the analysed chromosomes were drawn. The morphological makeup of the analysed chromosomes was assessed.

Nucleoli are the product of the activity of nucleolar organizer regions (NOR) in certain chromosomes. Their main functions are the formation of ribosomal subunits from ribosomal protein molecules and the transcription of genes encoding rRNA. The aim of the study was to determine the shape of nucleoli and analyse methylation in the gene RN28S in the spermatocytes of male Japanese quail (Coturnix japonica) in two age groups. Nucleoli were analysed in cells of the first meiotic prophase. Their number and shape were determined and they were classified as regular, irregular or defragmented. In the cells of the young birds no defragmented nucleoli were observed, with regular and irregular nucleoli accounting for 97% and 3%, respectively. In the cells of older birds no regular nucleoli were observed, while irregular and defragmented nucleoli accounted for 37% and 67%, respectively. MSP (methylation-specific PCR) showed that the gene RN28S is methylated in both 15-week-old and 52-week-old quails. In recent years an association has been established between nucleolus morphology and cellular ageing processes.

Morphological analysis of semen is a very important step in fertility assessment, but many semen defects are not detectable at the morphological level. These include pathological changes in sperm chromatin structure. During mammalian spermiogenesis, histone proteins associated with DNA structure are replaced by specific protamines, with which chromatin does not form nucleosomal complexes. In the fully developed, mature sperm, the histones are replaced with protamines. Disruptions of nucleoprotein structure can be treated as possible indicators of the biological value of spermatozoa. The experimental material consisted of sperm from one-and-a-half-year-old bulls, isolated post mortem from the tail of the epididymis. The smears were stained with silver nitrate (AgNO3), acridine orange (AO), aniline blue (AB) and chromomycin A3 (CMA3). Sperm dimensions largely depend on individual variability among the bulls. In most cases, differences in sperm dimensions were identified between individuals, which was confirmed in the statistics. Sperm with elevated, abnormal histone levels were proportionally quite scarce (1.4 %). Studies of nuclear proteins in the context of infertility demonstrate the important influence of normal chromatin structure on sperm functions.

A basic assay that detects genotoxic DNA damage disrupting DNA replication and repair mechanisms is the sister chromatid exchange test. The frequency

Chromosomal instability is a type of genome instability involving changes in genetic information at the chromosomal level. The basic tests used to identify this form of instability are sister chromatid exchange (SCE) tests and identification of fragile sites (FS). SCE is the process by which sister chromatids become fragmented as a result of DNA strand breakage and reassembly, followed by exchange of these fragments. FS can be observed in the form of breaks, gaps or constrictions on chromosomes, which often result from multiple nucleotide repeats in DNA that are difficult to replicate. The research material was the peripheral blood of ten breeds of cattle raised in Poland, including four native breeds covered by a genetic resources conservation programme, i.e. Polish Red, Polish Red-and-White, White-Backed, and Polish Black-and-White, as well as Polish Holstein-Friesian, Simmental, Montbéliarde, Jersey, Limousine and Danish Red. Two tests were performed on chromosomes obtained from in vitro cultures: SCE and FS. The average frequency of SCE was 5.08 ± 1.31, while the incidence of FS was 3.45 ± 0.94. Differences in the incidence of SCE and FS were observed between breeds. The least damage was observed in the Polish Red and White-Backed breeds, and the most in Polish Holstein-Friesians. The most damage was observed in the interstitial part of the chromosomes. Age was shown to significantly affect the incidence of SCE and FS. Younger cows showed less damage than older ones (SCE: 4.84 ± 1.25; 5.34 ± 1.24; FS: 3.10 ± 0.88, 3.80 ± 0.92).