Relevance

Cestodoses are infections caused by tapeworms, predominantly of the species Taenia spp. and Mesocestoides spp., which have a complex life cycle, which contributes to significant morbidity and mortality among wild and farm animals (humans), and also causes significant economic damage to livestock farming. Cestodoses are widespread in Kazakhstan, with widespread cases of detection in humans, dogs, livestock and wild animals. The study of the genetic characteristics of some types of cestodes in the territory of central and northern Kazakhstan is carried out with the aim of monitoring invasion and further developing diagnostic and preventive measures for this disease. There are only isolated data on the study of cestodoses in the study area; there is no information on the study of the species genetic diversity of the pathogens Taenia spp. and Mesocestoides spp., which makes it difficult to carry out diagnostics and comparative analysis of the epidemiological situation in the country as a whole.

The main idea of this project is to establish the Taenia spp. and Mesocestoides spp. species circulating in the territory of central and northern Kazakhstan at different life stages of development (intermediate and definitive hosts) in order to establish the main transmission routes of cestodes unusual for many animal species in ecological communities.

We carried out preliminary work with available open sources on the study region in order to understand the transmission routes of transmission of some types of cestodes that we identified that are not typical for local wild canines. However, due to the lack of many parasitological data that would be supported by genetic information, it is not possible to show the full life cycle of a particular type of cestode. These studies need to be completed. The question remains open: the newly discovered species of cestodes were not previously identified due to the imperfection of classical parasitological methods, or were they introduced from other regions or neighboring countries? These studies have not been conducted on the territory of Kazakhstan.

Target

The goal of the project is to determine the genetic identity of various types of pathogens Taenia spp. and Mesocestoides spp. circulating in the territory of central and northern Kazakhstan using bioinformation technologies and molecular genetics methods.

Expected results

– collection of biological material was carried out to identify pathogens Taenia spp. and Mesocestoides spp.in wild canines and other species of animals and birds included in their food chain;

– the taxonomic affiliation of the studied species of cestodes was established based on morphometric and morphological characteristics;

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

– PCR analysis was carried out to generate amplicons of marker regions of the genome of the pathogens Taenia spp. and Mesocestoides spp.;

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

– bioinformatics analysis was carried out and the species affiliation of the pathogens Taenia spp. and Mesocestoides spp. was established in various animals and birds circulating in the study area.

– the main transmission routes of the identified species of pathogens Taenia spp. and Mesocestoides spp.in wild canids have been established, taking into account their food chain.

The project will train highly qualified specialists in the field of molecular parasitology. Two theses and two dissertations will be prepared for the academic degree of Master of Science.

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

Smagulova Ainura Muratovna,Master of Engineering Science . H -index 1, 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

Lyalchenko Alexander Sergeevich, Master of Agricultural Sciences. ORCID: 0000-0002-3232-6342 .

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. 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. Uakhit R, Smagulova A , Syzdykova A, Abdrakhmanov S, Kiyan V (2022) Genetic diversity of Echinococcus spp. in wild carnivorous animals in Kazakhstan, Veterinary World, 15(6): 1489–1496. Percentile – 79, doi : www.doi.org/10.14202/vetworld.2022.1489-1496 .
11. Kiyan V.S. , Aitmagambetova M., Smagulova A.M. , Aitmagambetova S. Development of a method for extracting opisthorchid DNA from biological material. Bulletin of the Shakarim State University of Semey, Semey. – 2018. – pp. 173-177.
12. Nikitina M.I., Leader L.A., Kiyan V.S. , Smagulova A.M. Ecological and biological features and distribution of Opisthorchis felineus in the Korgalzhyn district of Akmola region. Bulletin of SemGU . Biological series, Semey, 2018, No. 1 (81). – pp. 259-263.
13. Kiyan V.S. , Smagulova A.M. , Katokhin A.V. Methorchiasis in Northern Kazakhstan: state of knowledge and distribution // Experimental Bulletin Biology , Al-Farabi Kazakh National University . – Almaty. – 2019 No. 3(80). – pp. 170-179. https://doi.org/10.26577/eb-2019-3-b15
14. Smagulova A.M. , Kiyan V.S. , Bekenova A.B. Efficiency of different methods for DNA extraction from Opisthorchis felineus . KATU Bulletin. Veterinary Series, Nur-Sultan, 2019, No. 3 (102). – pp. 269-278.
15. Smagulova A.M. , Bekenova A.B., Katokhin A.V., Kiyan V.S. Molecular markers in the diagnosis of opisthorchiasis. KATU Bulletin. Veterinary Series, Nur-Sultan, 2020, No. 2 (105). – pp. 190-199. https://doi.org/10.47100/herald.v1i2.55
16. Bekenova AB, Smagulova AM , Kiyan VS Veterinary-sanitary expertise of the fish from the Shagalaly river // Bulletin KazNU . – 2021. – Vol. No. 3 (88). – R . 87 -95. https://doi.org/10.26577/eb.2021.v88.i3.09
17. Uakhit RS , Smagulova AM , Lider LA, Leont’ev SV, Berber AP, Kiyan VS Species composition of wolf ( Canis lupus ) helminth fauna in Kazakhstan // Eurasian Journal of Applied Biotechnology. – 2022. – Vol. No.1 , – P. 49-58 . https://doi.org/10.11134/btp.1.2022.6
18. Smagulova A . M. _ , Uakhit RS , Lider L. A. , Lyalchenko A.S. _ , Kiyan VS Distribution and genetic features of the causative agent of Alaria alata in wild carnivores of the steppe and forest-steppe zones of Kazakhstan. // Eurasian Journal of Applied Biotechnology. – 2022. – Vol . No.2 ,– P. 38-43 . https://doi.org/10.11134/btp.2.2022.6
19. Uakhit RS, Kan M. Advantages Of The Staining Method For Studying The Morphological Characteristics Of Cestodes // International Scientific – Practical Conferences ” Seifullinsky” Readings – 18(2): “ Science of the XXI Century – Epoch Transformations .” Astana, 2022. – pp. 205-206.
20. Zhumalin A.Kh., Uakhit R.S. , Syzdykova A.S., Smagulova A.M. , Leontyev S.V. Helminth fauna of wild carnivores in the steppe zone of Kazakhstan // International Scientific and Practical Conference “ Seifullin Readings – 17 Modern Agrarian Science: Digital Transformation”. Astana, 2021.- T.1, Part 1 – P.316-318.

Results achieved

2023

During the first year of the project, samples of biological material were collected from 6 regions of Kazakhstan from wild canids (wolf, fox, corsac fox), wild ungulates, game birds, rodents and farm animals. Cestodoses during development go through several stages of development, sequentially passing into one another and are localized in different animals. We collected samples of pathogens of the genus Taeniasis belonging to various stages of development.

As a result of the activities carried out, biological material was collected from intermediate and definitive hosts of cestodes in the study area. A total of 2,741 samples were collected from various animal species in compliance with high biosafety standards . The obtained samples were examined by the parasitological method, and the primary identification of species of helminths of the family Taeniidae . Wolves were found to have – Taenia saginata, Taenia hydatigena, Taenia krabbei; the fox has Taenia crassiceps, Taenia multiceps, Taenia krabbei; in corsacs – Taenia krabbei; in wild ungulates and farm animals – Eimeria spp., free-living nematodes, strongylate eggs, oocysts, cystode eggs were not found; in game birds – Diorchis spp., Hymenolepis spp.; rodents – cysticercosis cysts.

All samples are stored in a refrigerator at 4 °C in separate containers and fixed in 70% ethyl alcohol for further use.

As a result of the research, biological material was collected to identify causative agents of cestodiasis in intermediate and definitive hosts in central and northern Kazakhstan. The extent and intensity of invasion have been established. A parasitological collection has been collected.