SCIENTIFIC RESULTS, December, 21, 2020
Missions successfully accomplished within the Project:
1. The breeding stock of 300 Kabardian horses was picked out of 13 best studs of the North Caucasus region (in the Kabardino-Balkarian Republic, the Karachay-Cherkess Republic, the Republic of Adygea and the Krasnodarsky Krai). During the field expeditions the zootechnical specification of these horses and collection of genetic material (blood) was carried out.
2. The DNA extraction from the 300 blood samples was executed at the Medico-Biological Center of the Kh.M.Berbekov Kabardino-Balkarian State University, Nalchik, Russia (KBSU). The DNA was divided into three equal portions, two of them were delivered to the All-Russian Research Institute of Horses, Russia (ARRIH) and the Humboldt University of Berlin, Germany (HUB), the third was used in ARRIH. Preliminarily, the protocol of DNA extraction from horse blood was worked in thoroughly.
3. In Germany, hair samples from 60 Kabardian horses were taken for DNA preparation. These samples were used for comparative genetic analysis with the breeding stock in the Caucasus.
4.The DNA samples of 300 horses are analyzed availing the standard panel from the 17 microsatellites at the Molecular-Genetic Laboratory of ARRIH. The same work was done for the 60 horses from Germany.
5. The original electronic database «Kabardian horse Base» is composed allowing one to perform population statistical analysis and archiving of the genetic information. This electronic database and its governing system are legalized with the Russian Federation State Certificate of Registration.
6. The genetic population analysis performed with help of the electronic database «Kabardian horse Base» reveals the high degree of microsatellite polymorphism within the studied population of Kabardian horses. The genetic diversity is greater than that in all other investigated horses breed in Russia. The microsatellite profile of the Kabardian breed was composed. The comparison of the Kabardian, Arab and Thoroughbred microsatellite profiles leads to a deduction of some specific characteristics. A paper about the microsatellite structure of Kabardian horse was published in the journal «Konevodstvo i konnyy sport».
7. The training of the graduate students A. Khaudov and A. Duduev at the Molecular-Genetic Laboratory of ARRIH was finished.
8. The aim to study the interrelations between DNA and endurance of Kabardian horses was one of the project purposes. That is why several distance tests of horses were attempted at Malkinsky Stud (Russia). 12 specially trained horses participated in the last of the trials – «Mountain Test» on stamina. Representatives from HUB and ARRIH took part in realization of it. The test was divided into two stages each over 35 kilometers with general height drop of about 500 meters. The programmed complex «Smart Trainer» was specially designed and approbated for this event on the basis of a smartphone and an electronic pulsometer «Zephyr». The system allows to detect the kinematic (trajectory and speed of motion, height over the sea level), electrophysiological (heart beat frequency) and biochemical (level of biolactate in blood) parameters of horses. The complex also allows to study the biomechanics of horse movement. A publication about the adaptive suppression of motion artifacts was made (elibrary.ru) and two about the use of the project for further selection and the development of the breed.
9. For the first time in Russia, the original system of the electronic timing of horse competitions «Ride-2» was devised in collaboration with Institute of radio engineering system and management of the Southern Federal University (IREM SFU) as well as the mobile electrocardiograph adopted to study horses.
10. In the frameworks of delegation from HUB to KBSU, there were organized (i) a scientific seminar, (ii) an open lecture of Dr. Monika Reissmann for students and lectors of KBSU, (iii) visits of a number of pedigree studs aimed on inspection of Kabardian horses under natural conditions, and (iiii) visiting of the Sevkavrentgen plant.
11. The 3-month training and several scientific enterprises of graduate students A. Khaudov and A. Duduev from KBSU the Moleculargenetic laboratory of HUB under the guidance of Dr. M. Reissmann was carried out.
12. The study of mitochondrial D-loop variation for a detection of the maternal lines in the Kabardian breed was carried out at HUB with participation of the PhD students A. Khaudov and A. Duduev. The high level of the mitochondrial diversity of the Kabardian horses agrees totally with the results of the microsatellite analysis conducted in cooperation with ARRIH.
13. Additional to the maternal lines, mutations in the Y chromosome were detected for an identification of the paternal lines. The three common haplotypes of riding horses were found in the Kabardian stallions. The article about the maternal (mutation in mitochondrial DNA) and paternal (mutation in Y chromosome) lines in the Kabardian horse is published.
14. A Genome-wide association study (GWAS) with 72 horses was done at HUB using the Equine 70K SNP chip. The output of this analysis was used for (i) a generation of a dendogram showing the genetic distance within the Kabardian breed and in relation to some other breeds like Thoroughbred and (ii) an association study between SNPs and performance data. One SNP with significant association to endurance was found. Two genes near to this SNP are analyzed by sequencing of horses with different performance looking for causal mutations (HUB).
15. The frequency of known coat color associated genes was estimated. Beside the most common alleles for the basic colors, some two rare dilution alleles were found in the population. DNA from Kabardian horse is used for bay-brown mutation search a coat color often found in Kabardian horses (HUB).
16. In addition to GWAS, genes working in the pathway of performance traits were selected as candidate genes for mutation analyses in good and bad endurance horses.
17. RNA samples were taken from good and bad endurances horses during the mountain test ride. The analysis of these RNA samples should give an inside into the different expression of performance associated genes. This investigation is still in progress.
18. TV-shows, newspaper articles and articles in horse breeding journals.
19. The dissertations of the graduate students A. Khaudov and A. Duduev dealt with aspects of the project are in time. The first proposals are submitted to the ARRIH.
20. The recommendations for pedigree work with the Kabardian horses are elaborated.
21. The results of research on the project are published in 9 articles in Russian and foreign scientific journals and received 2 certificates of state registration of the Database.
22. A summarizing seminar about the results of the scientific activities and results throughout the project was executed.