Microsatellites – a separate class of molecular genetic markers, are widely used for the study of genetic variability, in particular in populations of animals bred by artificial selection under the influence of microevolutionary processes. The object of study is the gene pool of a population of animals of the Lebedyn cattle breed, which is under threat of extinction. The sample comprised 30 individuals from the farm "Komyshans'ke" in Sumy region. The analysis of population genetic structure was performed using 10 microsatellite loci recommended by FAO-ISAG: ETH225, BM2113, ETH3, BM1818, BM1824, ILSTS006, INRA023, TAGLA053, TAGLA12, ETH10. Amplification products were separated in polyacrylamide gels of different concentrations (5–8%), both native and denaturing. All studied loci were polymorphic. The number of detected alleles per locus ranged from 4 to 8 (on average 5 alleles per locus), the size of which ranged from 115 bp (ETH3) to 307 bp (ILSTS006). The majority of the investigated loci (except ETH3) belonged to valuable informative markers (PIC > 0.5). The most polymorphic TGLA053 (8 alleles), BM2113 (6) and ETH3 (6) loci have been identified. In general, the minimum number of alleles (4) was fixed in 50% loci. The main population genetic parameters for the studied loci have been calculated. The highest values of heterozygosity (He), and effective number of alleles (ne) was characterized for loci BM2113, ILSTS006, TGLA053 and ETH225. With the exception of ETH3 and VM1818 loci, the experimental group of animals is in a state of genetic equilibrium. The average value of the Wright fixation index indicates a tendency to increase in the number of homozygous individuals (inbreeding). Comparative analysis of genetic structure of breeds that have a common origin (Lebedyn (PJSC "Mykhaylivka"), Ukrainian grey (DPDG "Polyvanivka"), Red steppe (DPPR "Stepne"), etc.) has been carried out. The obtained results give grounds to assert that in the experimental population of the Lebedyn breed there are processes that lead to a decrease in genetic diversity. In order to overcome the negative effects of artificial reproduction in the gene pool of small populations of cattle, which include Lebedyn cattle, it is appropriate to use microsatellite markers in the selection and breeding work.
The aim of research was to analyze the contents of somatic cells (SC) in goat milk in the East of Ukraine, level of SC translocation in the process of milking, speed of SC evacuation in their secret, to conduct monitoring and ranging on the level of SC goat milk of herds in the Eastern region during 2015 seasonally. Somatic cell count was studied on a large number of animals in different zones of the East of Ukraine (1800 milk samples). Somatic cell count in goat milk of the Eastern region of Ukraine appears up to 800×10³ cells/ml at 65-71% of studied milk samples by the method of laser-running cytometry with the account of only those cells having DNA in the nucleus. While analyze of SC subpopulations in goat milk it is shown that at increase to 2 million cells/ml and more the concentration of neutrophils and lymphocytes does not change and the number of macrophages and eosinophils (р≤0,05) rises from 1,5 to 4 times accordingly. It is marked that SC contents in goat milk in the morning and evening yield of milk is different. In evening yield of milk, the somatic cell count is up to 30 % higher than in the morning one. A clear pattern of distribution of somatic cells in milk portions during milking was not detected. Average sample of yield is required to determine the somatic cell count in the milk of goats. Six of the eight goats over six months of lactation were given milk with a fairly constant somatic cell count. Low and fairly constant somatic cells content (15 to 63 × 103 cells / ml) is observed in the milk of primiparous goats.
ВПЛИВ РОЗВИТКУ ЛІНІЙНИХ ОЗНАК ЕКСТЕР'ЄРУ, ЯКІ ХАРАКТЕРИЗУЮТЬ СТАН РОЗВИТКУ ТУЛУБА, НА ЖИТТЄЗДАТНІСТЬКОРІВ УКРАЇНСЬКОЇ БУРОЇ МОЛОЧНОЇ ПОРОДИ В. І. ЛАДИКА, Л. М. ХМЕЛЬНИЧИЙ, С. Л. ХМЕЛЬНИЧИЙ Сумський національний аграрний університет (Суми, Україна) khmelnychy@ukr.net В аспекті пошуку предикторів щодо довголіття тварин молочної худоби досліджено залежність тривалості життя корів української бурої молочної породи від показників описових ознак екстер'єрного типу. Вивчали лінійні ознаки екстер'єру корів-первісток, які характеризують стан розвитку тулуба. Встановлена співвідносна залежність тривалості життя корів від оцінки описових ознак з різною мінливістю у межах дев'ятибальної шкали кожної конкретної статі будови тіла. Найвищою тривалістю життя характеризувались тварини оцінені у 7-9 балів за шириною грудей, глибиною тулуба, кутастістю та шириною заду. Дослідження ознаки нахилу заду засвідчило криволінійний характер зв'язку між оцінками за цією ознакою і тривалістю життя корів. Корови з оптимальною оцінкою статі у п'ять балів відрізнялися найвищою тривалістю життя -2796 днів, тоді як із підвищенням та зниженням оцінок кількість днів життя корів зменшувалася. За оцінкою вгодованості встановлено, що худорляві тварини живуть і використовуються значно довше. Різниця між максимальною (дев'ять балів) та мінімальною (один бал) оцінками склала 457 днів (Р < 0,001). Ключові слова: українська бура молочна порода, лінійні ознаки типу, тривалість життя In the aspect of search of predictors concerning longevity of animals of dairy cattle the dependence of cow's longevity of the Ukrainian brown dairy breed on indicators of descriptive conformation type traits has been investigated. The linear conformation type traits of the first-calf cows that characterize the state of body development were studied. The correlation dependence of cow's longevity was determined on the assessment of descriptive type traits with varying variability within the ninepoint scale of each specific type trait of the body structure. The highest longevity was characterized by animals estimated at 7-9 scores for: chest width, body depth, angularity and rear width. Studies of the rump angle trait testified to the curvilinear nature of relationship between the estimates on this trait and cow's longevity. Cows with an optimal five score were characterized by a high longevity of 2.796 days, while with an increasing and decreasing in estimates, the number of cow's days of life decreased. According to the estimation of body condition score, it was found that thin animals live and are used significantly longer. The difference between the maximum (9 scores) and the minimum (1 score) estimates was 457 days (P < 0.001).
Cytogenetic analysis of breeding animals makes it possible to identify animals carrying cytogenetic abnormalities, the rate of spontaneous and induced somatic mutagenesis and the level of general genomic instability of animals. Scientists emphasize that in breeding work it is desirable to use animals whose offspring inherit high productivity and do not carry a genetic burden. The main factors that contribute to chromosomal instability are environmental factors, enzyme failure, changes in the system of repair or replication of chromosomes, suppression of immunity. It is important that the methods of breeding farm animals also have an impact on the stability of their karyotype. The appearance of deconsolidation of heredity and destruction of gene adaptive complexes during animal crossing has been established. This also applies to the use of related breeds, but obtained in different environmental conditions. As a result, aneuploidy, polyploidy may occur. It is proved that with the help of cytogenetic methods it is possible to establish new sources of genetic variability and prevent the spread of harmful chromosomal aberrations in animal populations. Such work was carried out on the Ukrainian brown dairy breed, created by crossing the local swan breed and related Swiss breed of American and European selections. That is, a related breed was used in its creation, but it had a difference in the selection system and ecological conditions of detention, which in turn may affect the stability of the karyotype of the new breed. The aim of our work was to study the variability of the karyotype of cows of the Ukrainian brown dairy breed. Materials and methods. Cytogenetic analysis, which was aimed at establishing the karyotype variability of Ukrainian brown dairy cows, was performed on biological material obtained from animals kept in the breeding plant of the State Enterprise "Research Farm of the Institute of Agriculture of the Northeast NAAS NAAS Sumy region. 15 cows were studied. Cytogenetic drugs were obtained from peripheral blood lymphocytes taken from the jugular vein using standard techniques. RPMI-1640 medium, bovine serum (preferably embryonic), antibiotic gentamicin, mitogen, a substance that stimulates mitotic division of lymphocytes in culture (phytohemagglutinin type P), were used to cultivate blood cells. The mixture was cultured in a thermostat at +37°C for 48 hours. Two hours before fixation, a solution of colchicine heated to 37°C at a final concentration of 0.3–0.5 μg/ml of culture medium was introduced into the culture. Freshly prepared 0.55% potassium chloride solution was used for hypotension. After hypotension, the culture was centrifuged, the supernatant was drained, and cooled to +4°C fixing fluid was added to the precipitate by mixing one part glacial acetic acid with three parts methyl (or ethyl) alcohol. The obtained preparations, after staining with the finished Giemsa dye, were analyzed for chromosomal variability under immersion magnification of the microscope by 1000 times and photographed. 100 metaphase plates were analyzed in each animal. The number of dinuclear lymphocytes (DJ), mononuclear lymphocytes with micronuclei (MJ), and mitotic index (MI) were counted on the same drugs. The frequency of DIA, IU, MI was calculated in ppm (number per 1000 cells). Analysis of karyotype variability of the studied animals showed the presence of quantitative and structural disorders of chromosomes. Quantitative disorders were detected in the form of aneuploidy, the average of which was 5.7%. Multiple chromosome enlargement (polyploidy) in the studied animals was not detected. The average value of asynchronous divergence of the centromeric regions of chromosomes was 0.5%. The limit of variability of ARCR in the studied cows was 0–5.0%, and structural chromosome abnormalities manifested themselves in the form of chromosomal breaks and amounted to 2.1%. According to the results of the micronucleus test, the proportion of lymphocytes with a micronucleus was 1.9‰, binuclear lymphocytes – 3.9‰, and the value of the mitotic index – 3.5‰. A positive relationship has been established between aneuploidy and ARCR, which are considered a prerequisite for chromosome loss; aneuploidy and mitotic index (P < 0.05), binuclear lymphocyte and mitotic index. A negative correlation was found between chromosome breakage and micronucleus lymphocyte. In animals of different blood levels according to the Swiss breed, chromosome set disorders in the form of aneuploidy were detected, the average values of which were 6.0 and 5.0%, respectively, which corresponds to the limit of spontaneous chromosomal variability. in the studied cows of brown dairy breed higher positive values were found between lymphocytes with a micronucleus and the duration of the service period and the interbody period. It was found that 43% of cows had stillborn calves during the life of the uterus with quantitative chromosome abnormalities (aneuploidy). In animals with undetected aneuploidy, only 25% of cows had stillborn calves during their lifetime. According to the results of cytogenetic testing of cows of the Ukrainian brown dairy breed of different linear affiliation, increased variability of the karyotype of animals of the Stretch line has been established. The results of the micronucleus test of brown dairy cows of different linear affiliation indicate the absence of mutagenic factors. Conclusions. Quantitative and structural chromosome disorders are characteristic of Ukrainian brown dairy cows. In animals of conditional blood status of 95% and more of the Swiss breed, the highest percentage of cells with aneuploidy (6.0%), the presence of asynchronous divergence of the centromere regions of chromosomes (0.63%) and chromosomal breaks (2.87%). According to the results of the micronucleus test, the cytogenetic parameters of cells (lymphocytes with a micronucleus, dinuclear lymphocytes and mitotic index) are 95% higher in animals of conditional blood status and more in the Swiss breed compared to animals of lower conditional blood status. karyotype (aneuploidy) on the percentage of stillbirth of calves and the proportion of lymphocytes with a micronucleus for the duration of the service and interstitial period. Differentiation of quantitative and structural chromosome disorders in brown dairy cows of different linear affiliation has been established. The fact of detection of chromosomal abnormalities indicates the need to significantly expand the scope of the survey of breeding stock, and especially in the leading breeding farms for the cultivation of Ukrainian brown dairy breed.
Marker-dependent breeding is one of the most relevant areas for improving the dairy productivity of cattle. It allows the selection of parent forms at the gene level. The growing demands of the market for milk quality make it necessary to select dairy herds using genetic markers associated with qualitative characteristics of milk productivity. A topical issue in dairy cattle breeding today is the study of the relationship between hereditary factors that determine the types of proteins in milk. Casein is the main component of milk proteins and is represented by three fractions – alpha (CSN1S1), beta (CSN2) and kappa (CSN3). The content of individual casein fractions depends on the breed of cows. Genetic variants of beta-casein significantly affect human health, while kappa-casein is associated with the quality of milk and the greater suitability of milk for processing and cheese production. The aim of this work is to evaluate the genotype of sires approved for use in Ukrainian farms in 2020 based on beta- and kappa-casein genes. Materials and methods. The research was conducted by analyzing the catalog of bulls of dairy and dairy-meat breeds for breeding stock reproduction in 2020. Bulls were analyzed according to the genotype of beta – and kappa-casein of Holstein (n = 872), Simmental (n = 17), Montbellard (n = 3), (wro.synel.net; motbeliarde.org) Jersey (n = 59), Red Danish (n = 8), Red Norwegian (n = 7), Schwyz (n = 17), Aishir (n = 8) breeds, evaluated by offspring quality and genomic assessment. The website DairyBulls.com is used to analyze the results of the evaluation of bulls-producers on a set of characteristics. The research results were processed by mathematical statistics methods using the "Statistica-6.1" package in a Windows PC environment. It was found that the majority of bulls-producers approved for use were evaluated based on these genotypes. Depending on the breed, the number of bulls estimated by the studied characteristics ranged from 43 to 85%. Depending on the breed, from 2 to 16 different combinations of beta- and kappa-casein genotypes were identified. The frequency of occurrence of the desired complex genotype A2A2/BB, depending on the breed, was in the range of 0.0 to 75.0%. Holstein bulls of the desired A2A2/BB genotype were estimated to be superior to bulls of some other complex genotypes in terms of daughter milk yield, the amount of milk fat, milk protein, lifetime profit indices, cheese profit, and milk profit. Therefore, the use of sperm production of bulls with the A2A2/BB genotype will improve the economically useful characteristics of offspring, and will further increase the number of livestock with the desired complex genotype. Summary. 1. Taking into account the data of many domestic and foreign scientists who claim that the genotype of animals by the beta – and kappa-casein polymorphism affects the qualitative composition of milk, it is necessary to have in herds the necessary number of animals carrying the A2 allele of beta-casein and the B allele of the kappa-casein gene. The desired breeding parameters of the herd can be achieved by using bulls with BB genotypes for kappa-casein and A2A2 for beta-casein. Correction of the bulls-producers selection system can only be carried out if the breeding stock of the herd is genotyped according to the polymorphism of the beta-and kappa – casein genotypes. Holding this event will increase the protein content and will improve cheese suitability of milk of cattle of future generations. The use of bulls-producers without taking into account their genotypes for the casein fractions described above and the lack of polymorphism control of these genes among cows can lead to a decrease in the frequency of desired genotypes in the herd and a decrease in the technological qualities of milk as raw materials. 2. The created database of bulls-producers approved for use in Ukraine, evaluated by the beta-and kappa – casein genotypes, allows us to establish a positive perspective on the possibility of forming staffed herds of animals with the desired complex A2A2/BB genotype in Ukraine. 3. It was found that bulls of dairy breeds have different frequencies of beta – and kappa casein alleles. The part of bulls-producers of the desired complex genotype A2A2/BB of Holstein breed is 8.49%, Holstein Red-pockmarked – 2.94%, Schwyz – 75%, Jersey – 13.79%, Red Danish – 12.5%, Red Norwegian – 20.0%, Montbellard – 25%. Among animals of Simmental and Aishir breeds – bulls with the desired complex genotype A2A2/BB were not detected. 4. Bulls of the desired complex genotype A2A2/BB according to the indicators of milk yield of daughters, the amount of milk fat, milk protein, indices of lifetime profit, profit for cheese, profit for milk were dominated bulls with separate complex genotypes CSN2-CSN3
| As a result of molecular-genetic analysis of two cow populations -Ukrainian Brown Dairy breed (UBD) and Sumy interbreed type of Ukrainian Black-and-White breed (SITUBWD) in the technique of ISSR analysis with the use of the fragment of microsatellite loci with trinucleotide core motif (AGC) 6 C (S1) as an amplification primer, there were identified 23 DNA-patterns. It was determined that genetic locus with the identified size of 1350 bp was monomorphic in animals of SITUBWD occurring in all study samples, however, in UBD breed cows the amplicons of this size were not detected at all (p<0.001). The population-genetic analysis of the two studied animal groups (taking into account maternal origin) by molecular-genetic testing in the ISSR-PCR technique revealed essential statistically significant differences. The total average number of PCR products obtained from the cows of the newly developed Sumy interbreed type of Ukrainian Black-and-White breed was 15,050 being 12,450 in Ukrainian Brown Dairy breed (p <0,001). A rather high level of genetic similarity between the animals of Ukrainian Brown Dairy and Sumy interbreed type of Ukrainian Black-and-White Dairy (0,768) is quite possibly connected with the influence of the unique aboriginal Lebedynsky breed on which maternal base they were developed. citation | Ladyka V, Skliarenko Y, Pavlenko Y, Metlytska O, Ivankova I (2019). Molecular-genetic analysis of cows genetic structure and determination of genealogical relatedness level of bulls of modern dairy breeds. Adv. Anim. Vet. Sci. 7(5): 405-411. doI | http://dx.
To study the polymorphism of β-casein gene and its effecton the quality of cow's milk the research was conducted in the breeding herds of Ukrainian Black-and-White Dairy, Lebedynand Simmental breeds. Genotyping of 200 head of cattle wascarried out. Determination of β-casein gene polymorphismwas performed in the genetic laboratory of Bohomolets Institute of Physiology. The TagMan@Genotyping system and aset of primers and probes were used for allelic discrimination.It was found that the frequencies of alleles A1 (0.294-0.380) and A2 in the locus of β-casein gene differed in animals of different breeds. The highest frequency of the desiredallele A2 had the population of Lebedyn breed cows (0.706),while the lowest was in Simmental ones (0.620). The frequency of A1 allele was the lowest in the cows of Lebedyn breed(0.294), and the highest in Ukrainian Black-and-White Dairybreed (0.380). Accordingly, the frequencies of genotypesA1A1, A1A2 and A2A2 differed depending on the origin. Thedesired genotype was more common in animals of UkrainianBlack-and-White Dairy and Lebedyn breeds (53-52%, respectively). Heterozygous genotype occurred more frequently in animals of Lebedyn and Simmental breeds (37%). Thehighest frequency of A1A1 genotype was characteristic ofanimals of Ukrainian Black-and-White Dairy and Simmentalbreeds (20%). According to the genetic and statistical analysis, there was an excess of homozygous variants of A1A1 andA2A2 in β-casein locus, and a lack of heterozygous A1A2.At the same time in animals of Ukrainian Black-and-WhiteDairy breed, the difference between the actual and expecteddistribution of genotypes was statistically significant.There was a difference in the quality characteristics ofmilk between animals of different genotypes. In animals ofdifferent breeds, the content of certain components of milkdid not vary equally depending on theβ-casein genotype, butthis difference was statistically insignificant.Cows of Ukrainian Black-and-White Dairy breed withhomozygous A1A1 genotype exceeded others in terms offat content, those with heterozygous A1A2 genotype had anadvantage in terms of protein, lactose and dried skimmedmilk remainder (DSMR). Animals of Lebedyn breed with theA2A2 genotype predominated animals of other genotypes inall investigated indicators of milk quality. Animals of Simmental breed with the desired A2A2 genotype had a lower fatcontent in milk compared to animals of other genotypes andthe average values in the sample. These animal together withA1A1 homozygotes had higher protein content in milk thanheterozygous animals and average values in the sample. Inour opinion the small number of experimental animals wasone of the reasons for the lack of a definite difference betweenthe average values of the milk components of cows of different β-casein genotypes. Key words: breed, fatcontent, proteincontent, kappacasein, genotype, allele, sire.
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