AR14869645 “Establishment of the genetic affiliation of various types of echinococcosis pathogen to haplotypes circulating in the territory of Kazakhstan”

Relevance

The main idea of this project is to establish all haplotypes and species of the pathogen Echinococcus spp. circulating in Kazakhstan at different life stages of development (intermediate and definitive hosts) in order to establish the main transmission routes between natural reservoirs, farm animals and humans.

We have made attempts to study a complete map of the distribution and transmission of invasion in the ecosystem, a detailed analysis and identification of all available haplotypes common in Kazakhstan. However, due to the lack of genetic information in the GenBank database on pathogen haplotypes isolated from different hosts, completion of this work is not possible. The question remains open about the haploid genotypes of the pathogen circulating among domestic and farm animals, as well as humans. These studies have not been conducted on the territory of Kazakhstan.

Goal

Determination of the genetic affiliation of various types of echinococcosis pathogen to haplotypes using methods of molecular genetics and bioinformatics

Expected results

– collection of biological material will be carried out to identify the causative agent of echinococcosis in intermediate and definitive hosts on the territory of Kazakhstan;

– the taxonomic affiliation of echinococcosis was established based on morphometric and morphological characteristics;

– protocols and methods for isolating genomic DNA of echinococcosis were developed depending on the stage of the life cycle and a collection was created;

– PCR analysis was carried out and amplicons of mitochondrial regions of the genome of the Echinococcus spp. were generated;

– nucleotide sequences of marker genes were deciphered using the Sanger sequencing method and deposited in the GenBank database;

– bioinformatics analysis was carried out and haploid genotypes of various types of echinococcosis pathogen circulating in the study area were established.

– the main transmission routes of haplotypes and types of the causative agent of echinococcosis between natural reservoirs, farm animals and humans have been established.

The project will train highly qualified specialists in the field of molecular parasitology. Two dissertations will be prepared for the academic degree of Master of Science and the preparation of at least one PhD will begin.

Project leader

Kiyan Vladimir Sergeevich, PhD , associate professor. H -index 4, Researcher ID: O-7403-2017, ORCID: 0000-0001-9787-9151, Scopus Author ID: 6701646393.

Research team members:

Abdybekova Aida Makenovna, Doctor of Veterinary Sciences, Professor. H -index 7, Researcher ID: AAQ-4561-2020, ORCID: 0000-0002-3307-7237, Scopus Author ID: 6507171000

Leader Lyudmila Aleksandrovna, candidate of veterinary sciences, associate professor. H -index 2, Researcher ID: O-8442-2017, ORCID: 0000-0001-5842-0751, Scopus Author ID: 56058488900

Smagulova Ainura Muratovna,Master of Engineering Science. H -index 2, Researcher ID: P-7618-2017, ORCID: 0000-0002-3067-3666, Scopus Author ID: 57213811809

Uakhit Rabiga Seitbattalkyzy, Master of Engineering Science. H -index 1, ORCID: 0000-0001-7737-7162, Scopus Author ID: 57226673682

Leontyev Sergey Valerievich, Master of Agricultural Sciences. ORCID: 0000-0001-7737-7162, Scopus Author ID: 5722667797.

Publications and documents of protection of the scientific director of the project and members of the research group concerning the topic of the project

1. Chien -Hung Chen, Vladimir Kiyan , AA Zhylkibayev , Dubek Kazyken , Olga Bulgakova , Kent E. Page, Rakhmet I. Bersimbaev , Eric Spooner and Dos D. Sarbassov . Autoregulation of the mTOR Complex 2 integrity is controlled by the ATP-dependent mechanism. J Biol. Chem. – 2013. – V. 288 (38). – P. 27019-27030. IF-4 .65 . Q1. Percentile – 90. DOI: 10.1074/jbc.M113.498055 .

2. Kazyken D., Kiyan V. , Kaz Y., Zhylkibayev AA, Chen CH, Agarwal NK, Sarbassov DD The nuclear import of ribosomal proteins is regulated by mTOR . Oncotarget . – 2014. – Vol. 5 (23). – P. 9577-9593. IF-6.359. Q1. Percentile – 69. DOI: 10.18632/oncotarget.2473 .

3. Bulashev AK, Borovikov SN, Serikova SS, SuranshievZh.A ., Kiyan VS , Eskendirova SZ Development of ELISA using anti- idiotic antibodies for diagnosis of opisthorchiasis . Folia Parasitologica . – 2016. – R. 63: 025. IF-1.271. Q3. Percentile – 47. DOI: 10.14411/fp.2016.025 .

4. Vladimir S. Kiyan , Aitbay K. Bulashev , Aleksey V. Katokhin . Opisthorchis felineus and Metorchis bilis Metacercariae in Cyprinid Fish Leuciscus idus in Nura-Sarysu River, Kazakhstan. Korean J Parasitol . – 2018. – Vol. 56(3). – P. 267-274. IF-1.207. Q3. Percentile – 39. DOI: 10.3347/kjp.2018.56.3.267.

5. Abetov DA , Kiyan VS , Zhylkibayev AA , Sarbassova DA , Alybayev SD , Spooner E , Song MS , Bersimbaev RI , Sarbassov DD . Formation of mammalian pre- ribosomes proceeds from intermediate to composed state during ribosome maturation. J Biol. Chem. – 2019. – V. 292 (28). – P. 10746-10757. IF-5.157. Q2. Percentile – 84. DOI: 10.1074/jbc.AC119.008378.

6. Vladimir Kiyan , Aitbay Bulashev , Aibek Zhumalin , Ainura Smagulova , Ludmila Leader . Immunogenicity and Antigenicity of Opisthorchis felineus Proteins. Adv. Anim. Vet. Sci. 2020, 8(9): 933-939. Percentile – 31 . http://dx.doi.org/10.17582/journal.aavs/2020/8.9.933.939 .

7. Aiganym Bekenova , Ainura Smagulova , Alexey Katokhin , Sergei Borovikov , Vladimir Kiyan . Molecular Differential Diagnosis between Opisthorchis felineus and Metorchis bilis. Adv. Anim. Vet. Sci. 2020, 8(s3): 27-32. Percentile – 31. http://dx.doi.org/10.17582/journal.aavs/2020/8.s3.27.32.

8. Smagulova A. , Katokhin A., Mambetpayeva B., Kulmaganbetova N., Kiyan V. A multiplex PCR assay for the differential detection of Opisthorchis felineus and Metorchis bilis . Georgian Medical News No. 1 (310) 2021. 176-182. Percentile – 20. PMID: 33658428.

9. Uakhit RS , Lider LA , Smagulova AM , Leontyev SV , Abdrakhmanov SK, Kiyan VS. First Molecular Identification of Dirofilaria repens Found in a Wolf Heart in Kazakhstan. Adv. Anim. Vet. Sci. 2021, 9(9): 1342-1346. Percentile – 31 . http://dx.doi.org/10.17582/journal.aavs/2021/9.9.1342.1346.

10. Torgerson PR, Shaikenov BS, Baitursinov KK, Abdybekova AM . The emerging epidemic of echinococcosis in Kazakhstan. Trans R Soc Trop Med Hyg . 2002 Mar-Apr ;96 (2):124-8. IF-5.157. Q2. Percentile – 84. doi: 10.1016/s0035-9203(02)90276-2.PMID: 12055797.

11. Shaikenov BS, Torgerson PR, Usenbayev AE, Baitursynov KK, Rysmukhambetova AT, Abdybekova AM , Karamendin KO. The changing epidemiology of echinococcosis in Kazakhstan due to transformation of farming practices. Acta Trop. 2003 Feb ;85 (2):287-93. IF-3.112. Q1. Percentile – 79. doi : 10.1016/s0001-706x(02)00236-x. PMID: 12606108.

12. Torgerson PR, Shaikenov BS, Rysmukhambetova AT, Ussenbayev AE, Abdybekova AM, Burtisurnov KK. Modeling the transmission dynamics of Echinococcus granulosus in dogs in rural Kazakhstan. Parasitology. 2003 May ;126 (Pt 5):417-24. IF-3.234. Q1. Percentile – 74. doi : 10.1017/s0031182003002932. PMID: 12793645.

13. Torgerson PR, Burtisurnov KK, Shaikenov BS, Rysmukhambetova AT, Abdybekova AM, Ussenbayev AE. Modeling the transmission dynamics of Echinococcus granulosus in sheep and cattle in Kazakhstan. Vet Parasitol. 2003 May 30 ;114 (2):143-53. IF-2.738. Q2. Percentile – 71. doi : 10.1016/s0304-4017(03)00136-5. PMID: 12781476.

14. Abdybekova AM , Torgerson PR. Frequency distributions of helminths of wolves in Kazakhstan. Vet Parasitol . 2012 Mar 23 ;184 (2-4):348-51. IF-2.738. Q2. Percentile – 71. doi : 10.1016/j.vetpar.2011.09.004. Epub 2011 Sep 12. PMID: 21962968.

15. Abdybekova A., Sultanov A., Karatayev B., Zhumabayeva A., Shapiyeva Z., Yeshmuratov T., Toksanbayev D., Shalkeevand R., Torgerson PR Epidemiology of echinococcosis in Kazakhstan: an update //Journal of Helminthology (2015) 89. – P .647-650. IF-1.6. Q2. Percentile – 68. DOI: 10.1017/S0022149X15000425.

16. Abdybekova A., Sultanov A., Dzhusupbekova N., Abdibayeva A., Zhaksylykova A., Kerimbaeva R., Akhmetova G., Torgerson PR . Parasites of farmed marals in Kazakhstan // Small Ruminant Research, 153, 2017. – P.142-145. IF-0.947. Q2. Percentile – 71. DOI: 10.1016/j.smallrumres.2017.06.011.

17. Abdybekova A.M., Zhuangzhi Zhang, Sultanov A.A., Abdibayeva A.A., Zhaksylykova A.A. , Junisbayeva S., Aubakirov M., Akhmetova GD, Torgerson PR Genotypes of Echinococcus isolated from domestic livestock in Kazakhstan //Journal of Helminthology , 2019. – P. 1-3. IF-1.436. Q2. Percentile – 68. DOI: 10.1017/S0022149X19000634.

18. Brusentsov II, Katokhin AV, Brusentsova IV, Shekhovtsov SV, Borovikov SN, Goncharenko GG, Lider LA , Romashov BV et al. Low genetic diversity in wide-spread Eurasian liver fluke Opisthorchis felineus suggests special demographic history of this trematode species. PLoS One. 2013; 8(4):e62453. IF-3.24. Q2. Percentile – 87. DOI: 10.1371/journal.pone.0062453.

19. Ibrayev B., Lider L. , Bauer C. Gasterophilus spp. infections in horses from northern and central Kazakhstan. Vet Parasitol . 2015; 207(1-2):94-98. IF-2.738. Q1. Percentile – 96. DOI: 10.1016/j.vetpar.2014.11.015.

20. Akibekov OS, Leader LA, Odoevskiĭ IM, Tokpan SS, Ospanova AZ. [Immunochemical properties of the excretory- secretory antigen of Trichinella spiralis ]. Med Parazitol ( Mosk ). 2015 Jan- Mar; (1):43-6. Russian. Percentile – 30. PMID: 25850317.

21. Lider L. , Leontyev S. , Ussenbayev A., Kiyan V. , Akibekov O., Seitkamzina D., ZhanabayevA . Intestinal helminths of wild Canidae from the Kazakhstan steppe ecosystems Veterinaria. 2020; 69(2): pp. 111-119. URI: http://repository.kazatu.kz/jspui/handle/123456789/1442

Results achieved

2022

Biological material was collected from intermediate and definitive hosts of echinococcosis in Kazakhstan; 392 samples were collected in compliance with high biosafety standards. The obtained samples were examined by the parasitological method, and the primary identification of Echinococcus spp.

Publications in domestic publications recommended by the Committee for Quality Assurance in Science and Higher Education:

  1. Abdybekova, A., Yalysheva, S., Zhaksylykova, A., Uakhit, R., & Kiyan, V. (2022). Gastrointestinal helminthiases of dogs with molecular genetic identification of echinococcosis. Eurasian Journal of Applied Biotechnology, (3), 51–57. https://doi.org/10.11134/btp.3.2022.6

2023

As a result of collecting biological material, 53 positive human samples of cysts were selected from farm animals: cattle – 1044, small cattle – 693 positive samples of hydatid cysts from the affected organs of the liver and lungs. From 725 dog fecal samples collected, 30 of them were positive for taeniid eggs. Also, as a result of autopsy, 120 wild carnivores (foxes, wolves and corsacs) were infected with echinococcosis larvae.

Echinococcus spp. samples was established, protocols for isolating genomic DNA and performing PCR analysis have been developed, and amplicons of specific marker genes have been developed, which will be sequenced for a more in-depth analysis.

Participation in international congresses:

Project employees took part in the XXIX World Congress on Echinococcosis, held at the I.K. Akhunbaev Kyrgyz State Medical Academy (Bishkek, Kyrgyzstan) from August 27 to 30, 2023. Project manager Kiyan V.S. gave an oral presentation on the topic “Genetic features of echinococcosis common in the Republic of Kazakhstan”, leading researcher Abdybekova A.M. made an oral report on the topic “Epizootological characteristics of the territory of the Republic of Kazakhstan regarding echinococcosis”, researcher. Uakhit R.S. gave a poster presentation on the topic “Molecular genetic identification of Echinococcus spp. found in wolves of Kazakhstan”.

Publications in international conferences:

  1. Uakhit R.S., Mannapova N.E., Jazina K.T. PRELIMINARY STUDY OF ECHINOCOCCUS SPECIES DISTRIBUTION IN KAZAKHSTAN // International scientific and theoretical conference “SEIFULLIN READINGS – 19”, Astana, 2023. Vol. 1, part 2, p. 88-90.
  2. Kiyan V.S., Uakhit R.S., Smagulova A.M. Genetic identification of helminthiases in wild carnivores. Materials of the International Scientific Conference “Zoological Research in Kazakhstan in the 21st Century: Results, Problems and Prospects”, dedicated to the 90th anniversary of the RSE “Institute of Zoology” of the National Science Ministry of Education and Science of the Republic of Kazakhstan. Almaty , 2023. – pp. 101-107.
  3. Kiyan V.S., Leader L.A., Uakhit R.S., Smagulova A.M. Genetic diversity of echinococcosis in the Republic of Kazakhstan // Abstracts of the VII All-Russian Congress on Medical Microbiology, Clinical Mycology and Immunology ( XXVI Kashkin Readings). – T. 25, No. 2. – St. Petersburg, 2023. – P. 123.

Guidelines:

1 Uakhit R.S., Kiyan V.S. Methodological recommendations for genetic identification of the causative agent of echinococcosis // National Center of Biotechnology LLP, Ministry of Health of the Republic of Kazakhstan. – Astana, 2023. – 24 p.