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Open access

Sylwia Nisztuk-Pacek, Brygida Ślaska, Ludmiła Grzybowska-Szatkowska and Marek Babicz

Abstract

The aim of the study was to describe the mechanism of mitochondrial DNA inheritance in a group of farmed raccoon dogs. The study involved 354 individuals. Whole peripheral blood was the research material. DNA was isolated and PCR was performed for two fragments of mitochondrial genes: COX1 (cytochrome oxidase subunit 1 gene) and COX2 (cytochrome oxidase subunit 2 gene). The PCR products were sequenced and subjected to bioinformatics analyses. Three mitochondrial haplotypes were identified in the COX1 gene fragment and two in the COX2 gene fragment. The analysis of mtDNA inheritance in the paternal line confirmed the three cases of paternal mtDNA inheritance, i.e. the so-called “paternal leakage” in the analysed population. In two families, all offspring inherited paternal mitochondrial DNA, whereas in one family one descendant inherited paternal mtDNA and another one inherited maternal mtDNA. The lineage data indicated that one female which inherited maternal mitochondrial DNA transferred it onto the next generation. To sum up, the results of the study for the first time demonstrated the phenomenon of “paternal leakage” in farmed raccoon dogs, which facilitated description of mitochondrial DNA inheritance in the paternal line.

Open access

Krzysztof Kowal, Brygida Ślaska, Adam Bownik, Beata Horecka, Jan Gawor, Anna Śmiech and Angelika Tkaczyk

Abstract

The aim of the study was to find associations between the process of neoplastic transformation and mtDNA mutations/polymorphisms, i.e. factors with potential prognostic significance, and to determine their impact on the biochemical properties, as well as structural, and functional properties of proteins. Blood and neoplastic tissue samples were collected from a 9-year-old Labrador dog with a diagnosed malignant mammary tumour. Next-generation genome sequencing (NGS) of the entire mitochondrial genome was performed using Illumina technology, and bioinformatics analyses were carried out. This is the first report demonstrating the application of NGS in the analysis of the canine mtDNA genome in neoplastic disease. The proposed strategy is innovative and promising. For the first time in the literature, the sequence of 29 genes was analysed to determine their association with the prevalence of tumour. In total, 32 polymorphic loci and 15 mutations were identified. For the first time, as many as 26 polymorphisms and all the mutations have been described to be associated with the neoplastic process in dogs. Most polymorphisms/mutations were found in the D-loop (31% of the polymorphisms and 93% of the mutations) and the COX1 gene sequence (16% of the polymorphisms). Blood or cancer heteroplasmy was noted in 93% of the mutations. Four of the 18 polymorphisms detected in the protein-coding genes were non-synonymous polymorphisms that have not been described in the literature so far (m.T7593C in COX2, m.G8807A in COX3, m.A9911G in ND4L, and m.T13299A in ND5) but resulted in changes in amino acids in proteins. These mutations and polymorphisms can affect mitochondrial functions and may be a result of cell adaptation to the changes in the environment occurring during carcinogenesis. The replacement of “wild type” mtDNA by a mutated molecule may be an important phenomenon accompanying carcinogenesis.

Open access

Brygida Ślaska, Magdalena Surdyka, Adam Brodzki, Sylwia Nisztuk, Artur Gurgul, Monika Bugno-Poniewierska, Anna Śmiech, Dorota Różańska and Maciej Orzelski

Abstract

The aim of this study was to identify mutations in the D-loop region of mtDNA in head, neck, and limb tumours in dogs, and determination of their relationship with the process of neoplastic transformation. Blood and tumour tissue samples from 19 dogs with diagnosed sporadic malignant tumours were analysed. DNA extraction, amplification, and sequencing of the mtDNA D-loop, and bioinformatic analyses were performed. Five mutations and 19 polymorphisms were observed in 68.42% of all tumours. Polymorphic variants were noted in 42.86% of the head and neck tumours and in 58.33% of the limb tumours. Mutations were observed in 21.05% of dogs. The mutations were found in 28.57% of the head and neck tumours and in 16.66% of the limb tumours. The mutations were identified in 50% of the studied epithelial cancers. In the mesenchymal tumours, no mutations in the D-loop region were observed. Mitochondrial haplotype A17 was found in over 40% cases of limb tumours. No association between the age, breed, sex, type of tumour, and detected polymorphic variants were observed. Different mutational changes in the D-loop sequences of mtDNA identified in the blood and tumour tissues may indicate a relationship between the type of tumour and individual changes in the D-loop nucleotide sequences of mtDNA.

Open access

Paweł Różański, Brygida Ślaska and Dorota Różańska

Abstract

This study presents an analysis of the yeast-like fungi population in the healthy horses’ living environment. The study material was sampled from various elements and equipment of a stable. We compared the yeast-like fungi population from the breeding environment of five groups of horses (English Thoroughbred horses, Arabian horses, Polish Half-Bred horses, cold-blooded horses, and Hucul horses and Highland ponies). The samples were collected in the summer and winter period. The laboratory examination involved 260 samples and resulted in identification of yeast-like fungi belonging to 13 species. Over 50% of the samples collected from the horse environment in wintertime and fewer than 18% of the summer samples displayed fungal growth. The largest amounts of yeast fungi were isolated from samples collected from water and feed troughs. The laboratory analysis demonstrated qualitative diversity of the yeast-like fungi population depending on the breeding environment of the individual horse groups. Quantitative differences of isolates were additionally related to the season of the year.

Open access

Anna Śmiech, Wojciech Łopuszyński, Brygida Ślaska, Kamila Bulak and Agnieszka Jasik

Abstract

Introduction: Breed predisposition to cutaneous mast cell tumours (MCT) in a population of dogs in Poland affected by various skin tumours was assessed, and the distribution of MCT characteristics such as histological grading, sex, age, and location, in predisposed breeds was evaluated.

Material and Methods: The retrospective epidemiological study included 550 dogs affected by cutaneous MCTs with a reference group of 2,557 dogs diagnosed with other skin tumours.

Results: A univariable logistic regression analysis was performed to determine the odds ratios (ORs) with 95% confidence intervals. The risk of high-grade MCTs was the highest for Shar-Peis (OR: 26.394) and American Staffordshire Terriers (OR: 2.897). Boxers (OR: 6.619), Labrador Retrievers (OR: 2.630), French Bulldogs (OR: 2.050), Golden Retrievers (OR: 1.949), and American Staffordshire Terriers (OR: 2.592) were mainly affected by low-grade MCTs. The high risk of MCT was calculated to be at the age of 4–6 years for Labrador Retrievers (OR: 2.686) and 7–10 years for Boxers (OR: 2.956) and French Bulldogs (OR: 9.429). MCTs were significantly more often located on the trunk in French Bulldogs (OR: 4.680), American Staffordshire Terriers (OR: 2.520), and Labrador Retrievers (OR: 1.948). There was no statistically significant correlation between gender and the occurrence of MCTs in the breeds.

Conclusions: The breed-predicated differences in the clinical course of MCTs suggest a genetic background for the tumours.

Open access

B. Ślaska, L. Grzybowska-Szatkowska, M. Bugno-Poniewierska, A. Gurgul, A. Śmiech, D. Różańska and J. Dudka

Abstract

The aim of the study was to identify polymorphisms and mutations in the mitochondrial ND4 gene and to analyse the associations between the occurrence of molecular changes in mtDNA and phenotypic traits in tumours in German Shepherd dogs. Fifty samples obtained from blood and tumour tissues of German Shepherd dogs with diagnosed tumours were analysed. DNA extraction, amplification, and sequencing of the mtDNA ND4 gene, and bioinformatics, statistical, and in silico protein coding SNP analyses were performed. ND4 mutations and/or polymorphisms were noted in eleven nucleotide positions in nearly half of the examined dogs. All the changes were substitution mutations. A majority of the changes identified were homoplasmic. In one dog with osteosarcoma, blood heteroplasmy was detected. In two positions of the ND4 gene, presence of non-synonymous mutations leading to amino acid changes in the ND4 protein was reported. Analyses carried out to determine the deleterious effect of mutations indicated an almost 97 and 62% probability that a single amino acid substitution (p.G239V and p.I401T, respectively) in the protein has a negative impact on its function. The results of statistical analyses indicate a significant association between the occurrence of mutations in three loci of the ND4 gene and the location of tumours. The mutations identified may be a result of cell adaptation to the changes in the environment occurring during carcinogenesis. The high frequency of mutations in the tumours may indicate genetic instability of mtDNA, which may also play a role in carcinogenesis.

Open access

Kornel Kasperek, Beata Horecka, Andrzej Jakubczak, Brygida Ślaska, Magdalena Gryzińska, Monika Bugno-Poniewierska, Małgorzata Piórkowska and Grażyna Jeżewska-Witkowska

Abstract

The aim of this study was to detect possible differences between farmed and wild-living raccoon dogs. Analysis of polymorphism in 15 microsatellite sequences led to the conclusion that raccoon dogs raised on Polish farms and wild raccoon dogs living in Poland are two genetically distinct groups of animals. Wild Polish raccoon dogs are genetically more similar to the population of wild animals from the Kaliningrad Region than to farmed animals. The analysis of microsatellite loci showed clear genetic differences between farmed and wild-living populations of raccoon dog, despite only 50 years of isolation of the two groups of animals. The farmed population was characterized by higher genetic variation than the wild-living population. On the basis of the analyses three microsatellite loci (INU014, Ren13J22 and Ren41D20) were proposed for determination of the origin of animals that have escaped from farms.

Open access

Grażyna Jeżewska-Witkowska, Beata Horecka, Andrzej Jakubczak, Kornel Kasperek, Brygida Ślaska, Monika Bugno-Poniewierska and Małgorzata Piórkowska

Abstract

This study was designed to determine the degree of genetic distinctiveness between farmed and wild foxes (Vulpes vulpes). Analysis of polymorphism in 16 microsatellite sequences led to the conclusion that red foxes raised on Polish farms and wild foxes living in Poland are two groups of genetically distinct animals. Farmed Polish foxes are genetically more similar to the population of wild animals from North America than they are to the free-living population in Poland, as confirmed by the fact that the farmed animals are descended from animals raised in Canada.

The small genetic distance between wild Canadian foxes (indicated as the progenitor of farmed Polish foxes) and farmed Polish foxes possibly suggests that the differences between the farmed and wild Polish populations may result from the fact that Canadian and Polish foxes took separate evolutionary paths.