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Experimental & applied acarology2024; 93(2); 439-458; doi: 10.1007/s10493-024-00933-4

Molecular and immunological studies on Theileria equi and its vector in Egypt.

Abstract: Equine piroplasmosis is not fully understood regarding pathogenicity, prophylaxis, host immune response expression, and specific vectors. Accurately identifying the parasite vector is crucial for developing an effective control plan for a particular infection. This study focused on morphologically identifying two Hyalomma species (H. anatolicum and H. marginatum) and one Rhipicephalus annulatus (R. annulatus) at the species level. The identification process was followed by phylogenetic analysis using the neighbor-joining method based on the cytochrome oxidase subunit 1 (COXI) gene as a specific vector for Theileria equi (T. equi) in horses. T. equi was diagnosed morphologically and molecularly from infected blood samples and crushed tick species using conventional PCR. Subsequently, phylogenetic analysis based on the amplification of the 18 S rRNA gene was conducted. The obtained sequence data were evaluated and registered in GenBank under accession numbers OR064161, OR067911, OR187727, and OR068139, representing the three tick species and the isolated T. equi, respectively. The study demonstrated that T. equi infection leads to immune system suppression by significantly increasing the levels of oxidative stress markers (CAT, GPx, MDA, and SOD) (P ≤ 0.0001), with this elevation being directly proportional to parasitemia levels in infected blood cells. Furthermore, a correlation was observed between parasitemia levels and the expression of immune response infection genes (IFN-gamma, TGF-β1, and IL-1β cytokines) in infected horses compared to non-infected equine. Common macroscopic symptoms indicating T. equi infection in horses include intermittent fever, enlarged lymph nodes (LN), and tick infestation.
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Publication Date: 2024-07-05 PubMed ID: 38967736PubMed Central: PMC11269342DOI: 10.1007/s10493-024-00933-4Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • This study investigates the molecular characteristics of the parasite Theileria equi and the tick vectors responsible for its transmission in Egypt, alongside the immune responses and oxidative stress in infected horses.

Background and Importance

  • Equine piroplasmosis is a disease caused by parasites such as Theileria equi, which affects horses.
  • There is limited knowledge about the disease’s mechanisms, including how pathogenic it is, how the horse’s immune system responds, potential preventive measures, and which tick species serve as vectors.
  • Identifying the specific tick species that transmit T. equi is crucial for creating an effective disease control strategy.

Objectives

  • To morphologically identify tick species that may serve as vectors of T. equi in Egypt.
  • To molecularly confirm the presence of T. equi in horses and ticks using PCR techniques.
  • To examine the phylogenetic relationships of the tick species and T. equi based on genetic sequences.
  • To analyze how T. equi infection impacts the horse’s immune response and oxidative stress levels.

Methodology

  • Tick Identification: Two Hyalomma tick species (H. anatolicum, H. marginatum) and one Rhipicephalus species (R. annulatus) were identified morphologically.
  • Genetic Analysis: Molecular identification was conducted by amplifying and sequencing the cytochrome oxidase subunit 1 (COXI) gene in ticks and the 18S rRNA gene in T. equi from blood and tick samples.
  • Phylogenetic Analysis: Used neighbor-joining method to analyze genetic relationships and registered the sequences in GenBank with specific accession numbers.
  • Diagnostics: Conventional PCR was used to detect T. equi infection morphologically and molecularly.
  • Immune and Oxidative Stress Markers Assessment: Measured oxidative stress markers (catalase (CAT), glutathione peroxidase (GPx), malondialdehyde (MDA), superoxide dismutase (SOD)) and expression levels of immune-related cytokines (IFN-gamma, TGF-β1, IL-1β) in infected versus non-infected horses.

Key Findings

  • The ticks identified (H. anatolicum, H. marginatum, R. annulatus) were confirmed as vectors capable of harboring T. equi.
  • Phylogenetic analysis provided detailed genetic relationships among the ticks and the parasite, enhancing understanding of their taxonomy and evolutionary position.
  • T. equi infection in horses significantly increased oxidative stress, as indicated by elevated markers CAT, GPx, MDA, and SOD levels (highly statistically significant, P ≤ 0.0001).
  • Oxidative stress marker levels correlated proportionally to the parasite load (parasitemia) in blood cells.
  • Expression of immune system genes (cytokines IFN-gamma, TGF-β1, IL-1β) was strongly correlated with parasitemia, indicating an active immune response to infection.
  • Typical clinical signs of T. equi infection included intermittent fever, enlargement of lymph nodes, and visible tick infestation.

Conclusions and Implications

  • The study clarifies specific tick species involved in transmitting T. equi in Egypt, essential for targeted control measures.
  • Molecular tools (PCR and sequencing) provide precise identification of both vectors and the parasite, allowing improved epidemiological tracking.
  • T. equi infection provokes immune system activation but also results in elevated oxidative stress, which may contribute to the pathology of equine piroplasmosis.
  • Understanding the relationship between parasitemia and host immune and oxidative responses could inform future therapies or preventive strategies.
  • Recognizing common clinical signs supports earlier diagnosis and treatment, improving horse health management.

Cite This Article

APA
Ramadan RM, Taha NM, Auda HM, Elsamman EM, El-Bahy MM, Salem MA. (2024). Molecular and immunological studies on Theileria equi and its vector in Egypt. Exp Appl Acarol, 93(2), 439-458. https://doi.org/10.1007/s10493-024-00933-4

Publication

Experimental & applied acarology
ISSN: 1572-9702
NlmUniqueID: 8507436
Country: Netherlands
Language: English
Volume: 93
Issue: 2
Pages: 439-458

Researcher Affiliations

Ramadan, Reem M
  • Department of Parasitology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt. reem.montaser@cu.edu.eg.
Taha, Noha Madbouly
  • Department of Parasitology, Faculty of Medicine, Cairo University, Giza, Egypt.
Auda, Hend M
  • Department of Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt.
Elsamman, Eslam M
  • Faculty of Veterinary Medicine, Cairo University (Equine Veterinarian), Giza, 12211, Egypt.
El-Bahy, Mohamed M
  • Department of Parasitology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.
Salem, Mai A
  • Department of Parasitology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.

MeSH Terms

  • Animals
  • Theileria / genetics
  • Egypt
  • Theileriasis / parasitology
  • Horse Diseases / parasitology
  • Horses
  • Ixodidae / physiology
  • Phylogeny
  • Arachnid Vectors / parasitology
  • Rhipicephalus / physiology
  • Female
  • RNA, Ribosomal, 18S / analysis

Conflict of Interest Statement

The authors declare no competing interests.

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  3. Salem MA, Mahdy OA, El-Saied MA, Kamel MS, Mohammed FF, Ramadan RM. Molecular and pathological insights into gene expression and oxidative stress in Clinostomum complanatum and Euclinostomum heterostomum. Sci Rep 2025 Oct 28;15(1):37586.
    doi: 10.1038/s41598-025-16469-5pubmed: 41152306google scholar: lookup
  4. Berdikulov M, Maikhin K, Karibayev T, Kalkabayev K, Kazybay B, Nissanova R, Makhmutov A, Rametov N, Abdikalyk A, Abdrakhmanov S, Yang DK, Mussayeva G. Genetic evidence of regional circulation of Crimean-Congo hemorrhagic fever virus in ixodid ticks from southern Kazakhstan. Front Vet Sci 2025;12:1623822.
    doi: 10.3389/fvets.2025.1623822pubmed: 41112156google scholar: lookup
  5. Hacilarlioglu S, Bilgic HB, Karagenc T, Aydin HB, Toker H, Kanlioglu H, Pekagirbas M, Bakirci S. Molecular Detection and Prevalence of Equine Piroplasmosis and Other Blood Parasites in Equids of Western Aegean Türkiye. Vet Sci 2025 Aug 27;12(9).
    doi: 10.3390/vetsci12090826pubmed: 41012752google scholar: lookup
  6. Chen X, Li Z, Zhang X, Duo H, Shen X, Ma Y, Fu Y, Guo Z. The genetic diversity of tick species in selected areas of Qinghai Province. Parasitol Res 2025 Sep 19;124(9):107.
    doi: 10.1007/s00436-025-08556-3pubmed: 40970977google scholar: lookup
  7. Salem MA, El-Gameel SM, Kamel MS, Elsamman EM, Ramadan RM. Innovative diagnostic strategies for equine habronemiasis: exploring molecular identification, gene expression, and oxidative stress markers. Parasit Vectors 2025 Aug 2;18(1):325.
    doi: 10.1186/s13071-025-06970-1pubmed: 40753398google scholar: lookup
  8. Ramadan RM, Khalifa MM, Youssef FS, Fouad EA, Kamel M, El-Bahy MM, Taha NM. A paradigm shift in trichinellosis management: curcumin-olive oil nanocomposite's multi-faceted therapeutic approach. BMC Vet Res 2025 May 22;21(1):370.
    doi: 10.1186/s12917-025-04821-wpubmed: 40405173google scholar: lookup
  9. Ramadan RM, Wahby AM, Bakry NM, Auda HM, Mohammed FF, El-Bahy MM, Hekal SHA. Targeted pre-partum strategies to suppress Toxocara vitulorum hypobiotic larvae: Reducing transmission to calves and genotypic insights into buffalo infections. Vet World 2025 Feb;18(2):329-340.
    doi: 10.14202/vetworld.2025.329-340pubmed: 40182831google scholar: lookup
  10. Taha NM, Salem MA, El-Saied MA, Mohammed FF, Kamel M, El-Bahy MM, Ramadan RM. Multifaceted analysis of equine cystic echinococcosis: genotyping, immunopathology, and screening of repurposed drugs against E. equinus protoscolices. BMC Vet Res 2025 Mar 17;21(1):178.
    doi: 10.1186/s12917-025-04616-zpubmed: 40098107google scholar: lookup
  11. Ramadan RM, Bakr AF, Fouad E, Mohammed FF, Abdel-Wahab AM, Abdel-Maogood SZ, El-Bahy MM, Salem MA. Novel insights into antioxidant status, gene expression, and immunohistochemistry in an animal model infected with camel-derived Trypanosoma evansi and Theileria annulata. Parasit Vectors 2024 Nov 18;17(1):474.
    doi: 10.1186/s13071-024-06564-3pubmed: 39558410google scholar: lookup
  12. Salem MA, Taha NM, El-Bahy MM, Ramadan RM. Phylogenetic position of the pigeon mite, Ornithonyssus sylviarum, with amplification of its immunogenetic biomarkers in Egypt. Sci Rep 2024 Sep 25;14(1):22026.
    doi: 10.1038/s41598-024-72433-9pubmed: 39322649google scholar: lookup
  13. Mahdy OA, Ramadan RM, Salem MA. Innovative molecular and immunological approaches of heterophyiasis infecting some Egyptian marketed fishes. BMC Vet Res 2024 Aug 30;20(1):385.
    doi: 10.1186/s12917-024-04226-1pubmed: 39215340google scholar: lookup
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