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The micronucleus (MN) test is a common tool used to evaluate cellular genetic instability at the chromosomal level. It determines the effect of physical, chemical and environmental factors on DNA, and thus the body's individual resistance to harmful substances. The karyotypes of blue and silver foxes and their interspecific hybrids are characterized by morphological and structural variation. This variation is partly attributable to the presence of chromosomal polymorphism, which may significantly influence the stability of genetic material in the cells of these species. The objective of the study was to evaluate genetic material stability in selected Canidae species. To this end, analyses using the MN test were performed. Binucleated cells (BNCs) were analysed in microscopic preparations, and the number of micronuclei was determined within these cells. For the proportions of both MN and BNCs, highly significant differences were observed between the fox species. The interspecific hybrids differed from the other fox species in MN percentage. The lowest average was noted in blue foxes (3.33) and the highest in interspecific hybrids (15.21). [ABSTRACT FROM AUTHOR]

Simple Summary: The paper describes the karyotypes of blue and silver foxes and their hybrids, in terms of the numbers of A and B chromosomes and the frequency of fragile sites on chromosomes. Genome stability in these species is affected by Robertson translocations in the karyotype of the blue fox and by B chromosomes in the silver fox. The fragile sites assay was used as a biomarker to assess genome stability in foxes. This test enables the identification of breaks, chromatid gaps, and deletions. In healthy individuals, the number of these instabilities remains low. The test can be used to select individuals with the most stable genome for breeding of blue and silver foxes. The fewer an individual's susceptible sites, the more likely it is to have good reproductive performance. This factor is extremely important in the case of blue foxes, which are an endangered species. A cytogenetic assay based on fragile sites (FS) enables the identification of breaks, chromatid gaps, and deletions. In healthy individuals, the number of these instabilities remains low. Genome stability in these species is affected by Robertsonian translocations in the karyotype of the blue fox and by B chromosomes in the silver fox. The aims of the study were to characterise the karyotype of blue foxes, silver foxes, and their hybrids and to identify chromosomal fragile sites used to evaluate genome stability. The diploid number of A chromosomes in blue foxes ranged from 48 to 50, while the number of B chromosomes in silver foxes varied from one to four, with a constant number of A chromosomes (2n = 34). In interspecific hybrids, both types of karyotypic variation were identified, with the diploid number of A chromosomes ranging from 40 to 44 and the number of B chromosomes varying from 0 to 3. The mean frequency of FS in foxes was 4.06 ± 0.19: 4.61 ± 0.37 in blue foxes, 3.46 ± 0.28 in silver foxes, and 4.12 ± 0.22 in hybrids. A relationship was identified between an increased number of A chromosomes in the karyotype of the hybrids and the frequency of chromosomal breaks. The FS assay was used as a biomarker for the evaluation of genomic stability in the animals in the study. [ABSTRACT FROM AUTHOR]

Simple Summary: The present study describes a detailed morphometric analysis of the sperm of the blue fox (Alopex lagopus) and silver fox (Vulpes vulpes), together with determination of the shape indices of the sperm head. Staining with silver nitrate enables precise identification of the acrosome and reveals structural details of the sperm tail, so that they can be accurately measured. Statistically significant differences were found for most of the morphometric parameters of the two fox species. The blue fox sperm were generally larger, but the acrosome area and coverage were greater in the silver fox. There are no clear recommendations regarding sperm staining techniques for foxes, and no reference values for morphometric parameters of the sperm of foxes or for canines in general. Staining with silver nitrate for evaluation of the morphometry of fox sperm can be used as an independent technique or an auxiliary technique in routine analysis of canine semen. The results presented in this study are the first such extensive characterization of the sperm morphometry of the blue fox (Alopex lagopus) and silver fox (Vulpes vulpes), as representatives of the family Canidae. Canine spermatozoa, especially the sperm of farmed foxes, are not often described in studies on reproduction. The aim of the study was a detailed comparison of the morphometric dimensions and shape of the sperm of two fox species: silver fox and blue fox. Semen collected from 10 silver foxes and 10 blue foxes was used for the study. The specimens were stained with silver nitrate. Measurements were performed of the length, width, perimeter, and area of the head; the area of the acrosome and its coverage; the length of the midpiece and its coverage; the length of the tail; and the length of the end piece of the tail. In addition, four head shape indices were calculated: ellipticity, elongation, roughness and regularity. The following values for the morphometric parameters and shape indices were obtained for blue fox and silver fox, respectively: head length—6.72 µm and 6.33 µm; head width—4.54.µm and 4.21 µm; head perimeter—18.11 µm and 17.37 µm; head area—21.94 µm2 and 21.11 µm2; acrosome area—11.50 µm2 and 10.92 µm2; midpiece length—12.85 µm and 12.79 µm; tail end piece length—3.44 µm and 3.28 µm; tail length—65.23 µm and 65.09 µm; acrosome coverage—52.43% and 52.83%; midpiece coverage—19.71% and 19.65%; sperm length—71.95 µm and 71.42 µm; ellipticity—1.49 and 1.52; elongation—0.19 and 0.20; roughness—0.84 and 1.88; regularity—1.09 and 0.99. The significance of differences between species was verified by Tukey's test at p ≤ 0.05. Statistically significant differences between species were found for the following parameters: head length, width, perimeter and area; acrosome area; tail, end piece, and total sperm length; roughness and regularity. The differences in the size and shape of sperm can be used to establish reference patterns for fox sperm enabling more accurate species identification. [ABSTRACT FROM AUTHOR]

• Semen analysis is a way to predict male fertility. • Genetic material of the sperm of silver foxes was more stable, than arctic foxes. • Analysis of chromatin stability should complement morphological and molecular evaluation of semen. The latest reports indicate that sperm function is significantly influenced by the chromatin structure of the sperm and the integrity of its DNA. The aim of the study was to determine the level of condensation in the chromatin structure of fresh semen and the degree of damage to the DNA chain in fresh and chilled semen of farmed blue and silver foxes. The sperm of 15 blue foxes and 18 silver foxes at the age of one year was used in the study. The chromatin structure was evaluated by three staining techniques: aniline blue, chromomycin A3 and acridine orange. A comet assay was used to assess sperm DNA integrity in fresh and chilled semen. The average percentage of head DNA changed over time: in blue foxes from 99.12% to 97.83%, and in silver foxes from 99.37% to 98.05%. Highly significant differences (P < 0.01) in head DNA were found between species. Within each species, highly significant differences (P < P < 0.01) were found in the chilled semen (0 h/24 h/48 h/72 h). After staining with aniline blue (AB), chromomycin A3 (CMA3) and acridine orange (AO), most sperm had normal histone retention and a stable, native DNA structure. In blue foxes, the average percentage of sperm with elevated, abnormal histone content, sperm with impaired protamination, and sperm with DNA fragmentation was 1–13%; 1–16% and 0%, respectively. The percentages for silver foxes were similar (3–17%, 5–17% and 2%). [ABSTRACT FROM AUTHOR]