Single-nucleotide polymorphism (SNP) was analysed for selected fragments of three genes - insulin-like growth factor 1 (IGF1), myosin-XV (MYO15A) and paired box homeotic gene 3 (PAX3) - in farm and wild red foxes from two continents. The study was undertaken in order to verify whether the SNP characteristics of these genes enable farm-bred foxes to be distinguished from free-living foxes. The greatest number of changes were detected in the IGF1 gene. For each of the genes investigated specific SNP profiles characteristic only for farm foxes and only for wild foxes were noted. At the same time, specific SNP profiles were noted for wild foxes from North America and from Europe. The frequency of SNP (bases per SNP) in the gene fragments examined was 22 bp for IGF1, 34 bp for PAX3 and 56 bp for MYO15A. Single-nucleotide polymorphism is a very good molecular marker enabling characterization of nucleotide variation in the genes investigated between wild and farm individuals
The gene IGF1 has been shown to have a significant influence on the size of individuals, including animals of the Canidae family. In this study we determined SNP mutations of the IGF1 gene in dogs, raccoon dogs and farmed and free-living red foxes from Poland and Canada. No SNP mutations were noted in dogs or raccoon dogs, but a total of 14 single nucleotide polymorphisms were identified in foxes, including 12 substitutions, as well as one new mutation missense variant (exon 6) in wild Polish foxes and one synonymous mutation variant in wild foxes from Canada. We identified specific SNP profiles characteristic only for farmed foxes and only for wild foxes, as well as specific SNP profiles or wild foxes from North America (Canada) and from Europe (Poland).
The gene MYO15A is involved in the production of a protein included in the group of motor proteins known as myosins. Myosin XVA is located in the inner ear, the pituitary gland and other tissues, and has a substantial influence on the hearing process. Mutations in this gene cause amino acid substitutions in the conserved motor domain of the myosin chain, leading to shortening of the stereocilia in the hair cells, so that the function of myosin XVA is impaired. A research hypothesis was put forth that mutations in the gene responsible for the hearing process in animals of the Canidae family can cause hypoacusis, as well as substantial behavioural changes in dogs (ranging from timidity to aggressive behaviour). The study determined SNP polymorphism in a fragment of the gene MYO15A, which can cause hearing disorders or hypoacusis, in wild and farmed individuals of the Canidae family.
Aleutian disease is one of the most serious disease entities affecting mink farms. The disease causes significant economic losses in mink breeding countries. The aim of the study was to optimize a diagnostic test based on duplex PCR to enable detection of Aleutian disease virus in biological and environmental samples.
Blood (n = 40) and spleen (n = 40) samples from animals with suspected infection, and swabs from cages in which infected animals were kept (n = 20) were used for analysis. DNA was isolated from the samples, followed by optimization of the duplex PCR reaction targeting sequences coding NS1 and VP2 proteins. The qPCR method was used to determine the sensitivity of the reaction. The specificity of the analysis was confirmed by the sequencing results.
Optimized duplex PCR enabled detection of Aleutian Mink Disease Virus (AMDV) genetic material in biological and environmental samples. Testing of the sensitivity of the method indicated clear amplification for both primer pairs at 102 copies of viral DNA in a reaction. Sequencing confirmed the specificity of the reaction, which in the case of both primer pairs indicated an over 90% agreement between the isolates and the variants of the virus from the databases.
The use of duplex PCR to detect two regions of the AMDV genome may increase the sensitivity and specificity of the method and significantly expand the possibilities of further analysis based on sequencing.
Canine parvovirus type 2 is one of the most common causes of death among puppies. Despite preventive vaccination, the disease continues to be diagnosed. The aim of the study was to provide a molecular characterization of CPV-2 isolates found in southeastern Poland. Genetic CPV-2 material was isolated from the blood (n=10) and feces (n=50) of infected dogs. The presence of CPV-2 was confirmed by amplification of sequences coding both VP1 and VP2 protein. The products of the PCR reaction with primers amplifying VP2 protein were sequenced and used for genotyping. Bioinformatics analysis of the sequenced PCR product was performed to determine the phylogenetic relationships with variants recorded in the public databases. Based on the analysis of polymorphism in the nucleotide sequence 7 nucleotide variants were detected and assigned into four amino acid groups. Representatives of three groups contained asparagine at amino acid position 426 of the VP2 protein, which is characteristic of CPV-2a. The variant from the fourth group belonged to type CPV-2b. CPV-2a is the dominant antigenic type of CPV-2 in Poland. The pathogen’s high degree of polymorphism is manifested not only by the presence of numerous variants within the type, but also by the presence of representatives of type CPV-2b. Further studies of the molecular epidemiology of CPV-2 are necessary to optimize the effectiveness of preventive measures.
Assessment of Selenium Concentration in Selected Organs of Farmed Raccoon Dogs (Nyctereutes Procyonoides)
The aim of the study was to determine selenium concentrations in the liver, kidneys, lungs, heart and muscles of farmed raccoon dogs (Nyctereutes procyonoides) and to evaluate their impact on hair coat quality. Selenium concentration was determined using the modified Watkinson's spectrofluorometric method. Subjects were 20 farmed raccoon dogs (Nyctereutes procyonoides) at the age of 8-9 months, which were kept on a farm in south-eastern Poland. The results show that liver selenium content averaged 0.23±0.10 μg/g w.w. (wet weight). The concentrations ranged from 0.04 to 0.49 μg/g w.w. Kidney selenium concentration (0.49±0.17 μg/g w.w. on average) was over twice that of liver concentration. Animals with higher scores for hair coat quality had lower selenium concentrations in the kidneys and liver, and higher selenium concentrations in muscles, but the differences were not significant. When relating Se concentrations determined in the liver of raccoon dogs to the biochemical criteria, it is concluded that 80% of the analysed raccoon dogs were deficient in this element and 20% had marginal levels. The results obtained in our study suggest that the food used on the farm did not fully meet the Se requirement of the raccoon dogs.
The aim of the study was to determine the mechanical and geometric properties as well as bone tissue density of long bones in primiparous and multiparous dams of minks supplemented with β-hydroxy β-methylbutyrate (HMB) and/or 2-oxoketoglutarate (2-Ox) during gestation. Powdered 2-Ox was given at the daily dosage of 0.4 g/kg b.w. separately or simultaneously with HMB, which was administered at the daily dosage of 0.02 g/kg b.w. The study demonstrates for the first time that administration of 2-Ox and/or HMB to dams markedly influences bone tissue density and the mechanical and geometrical properties of mother`s bones in minks. Moreover, it was demonstrated that the supplementation was more effective in the thoracic limb, which was comprehensively used in contrast to the pelvic limb. The mechanical parameters and bone tissue density significantly increased in the humerus in multiparous minks. Only such diet may provide satisfactory production results in the animals. Nutritional deficiencies occurring during pregnancies may trigger body`s own reserves to cover the bone mass increase in developing foetuses and support milk production. This can prevent regeneration of dams’ organisms, which negatively affects their reproductive performance. 2-Ox or HMB may be regarded as a protective metabolite when administered orally to minks, counteracting the negative influences of pregnancy and lactation periods on bones condition. Both simultaneous treatment with 2-Ox and HMB and their separate administration were equally effective.
Introduction: The objective of the study was immunophenotypic and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture.
Material and Methods: The mesenchymal stem cells were obtained from equine bone marrow of three horses and from foal umbilical cords of six foals. The cells were cultured in CO2 incubators by standard procedure. Quantitative abnormalities of chromosomes, i.e. aneuploidy and polyploidy, and structural aberrations, i.e. breaks in chromosomes and chromatid, were taken into account during the study.
Results: The results of cytogenetic analysis of equine bone marrow mesenchymal stem cells at the third and fourth passages indicated that the level of karyotype variability of these cells corresponded to the spontaneous level of karyotype variability typical of the peripheral blood lymphocytes of this species. Equine bone marrow contained several clones of stem cells that differed in the expression of specific nuclear markers characteristic of proliferating cells.
Conclusion: Mesenchymal stem cells from foal umbilical cords during in vitro cultivation are characterised by quantitative abnormalities of the chromosomal apparatus.
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.
Creation of interspecific hybrids is widely common among plants and animals in order to improve economically important traits for humans. The studied material consisted of chromosomal preparations in the metaphase stage obtained from interspecies hybrids of arctic foxes (Alopex
lagopus) and red foxes (Vulpes vulpes). The aim of the study was to analyze the karyotype of the interspecific hybrids taking into account the number of chromosomes of sets A and B. With the use of techniques of classical cytogenetics (C bands, AgNOR bands) and molecular cytogenetics (FISH, PRINS) we carried out a genome analysis of Alopex-Vulpes hybrids. The results of this study showed that chromosomal markers of the interspecies hybrids are inherited from the parent species and are a result of combination of their two genomes. However, intraindividual differences are also observed which may result from aberrations of chromosome segregation during embryonic development. This may lead to the formation of different cell lines with different karyotypes (mosaicism). Moreover, chromosomes of the interspecies hybrids showed telomeric signals at the ends, in the centromers, as well as short chromosome arm rich in heterochromatin. The use of PRINS method led to identification of nucleolus organizer regions on 12 chromosomes of the interspecies hybrids. The hybridization signals obtained were characterized by different size and intensity. In addition, single copies of rDNA in the centromeric regions of several metacentric chromosomes were identified.