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Home / Equine Research Bank / Parasit Vectors / Innovative diagnostic strategies for equine habronemiasis: e...
Parasites & vectors2025; 18(1); 325; doi: 10.1186/s13071-025-06970-1

Innovative diagnostic strategies for equine habronemiasis: exploring molecular identification, gene expression, and oxidative stress markers.

Abstract: Equine habronemiasis, caused by Habronema (H.) muscae, H. microstoma, and Draschia megastoma, is a parasitic disease that presents in both gastric and cutaneous forms. Conventional diagnostic methods often lack sensitivity due to intermittent egg shedding and nonspecific clinical signs. This study aimed to enhance diagnostic accuracy by integrating molecular identification, oxidative stress profiling, and cytokine gene expression analysis. Methods: A total of 100 horses from a private farm in Giza, Egypt, were clinically examined for signs of habronemiasis. Fecal and skin samples were examined using parasitological techniques alongside polymerase chain reaction (PCR) targeting the mitochondrial cytochrome c oxidase subunit I (COXI) gene. The serum levels of oxidative stress biomarkers, including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and total antioxidant capacity (TAC), were quantified. Additionally, the expression of cytokines (interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 beta (IL-1β) and interleukin-6 (IL-6))was assessed via real-time PCR. Results: Habronema spp. eggs were detected in 62% of fecal samples, with molecular analysis confirming H. muscae as the predominant species. Infected horses exhibited significantly elevated oxidative stress markers compared with those in healthy controls. Cytokine gene expression analysis demonstrated a marked upregulation of proinflammatory markers, indicating a Th1-dominated immune response. Conclusions: This study underscores the value of molecular diagnostics combined with immunological profiling for the detection and characterization of equine habronemiasis. The integration of oxidative stress and cytokine biomarkers provides important insights into host-pathogen interactions and may contribute to the development of improved diagnostic and therapeutic strategies.
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Publication Date: 2025-08-02 PubMed ID: 40753398PubMed Central: PMC12318383DOI: 10.1186/s13071-025-06970-1Google Scholar: Lookup
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Summary

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Innovative diagnostic strategies were developed to improve detection of equine habronemiasis, a parasitic disease in horses, by using molecular identification, oxidative stress markers, and cytokine gene expression analysis alongside traditional methods.

Background

  • Equine habronemiasis is caused by three types of parasites: Habronema muscae, H. microstoma, and Draschia megastoma.
  • The disease manifests in two primary forms: gastric (affecting the stomach) and cutaneous (affecting the skin).
  • Common diagnostic approaches rely on detecting parasite eggs in feces or observing clinical signs, but these methods often lack sensitivity due to intermittent egg shedding and nonspecific symptoms.
  • This low sensitivity can lead to underdiagnosis or misdiagnosis, hindering appropriate treatment.

Objectives and Aims

  • The study aimed to improve diagnostic accuracy for equine habronemiasis.
  • This was pursued by combining molecular diagnostic techniques with assessments of oxidative stress and immune response biomarkers.
  • The authors intended to better understand the host-pathogen interaction through immunological profiling, aiding both diagnosis and potential treatment development.

Methodology

  • A total of 100 horses on a private farm in Giza, Egypt, were clinically examined for signs of habronemiasis.
  • Parasitological techniques were employed to detect parasite eggs in fecal and skin samples.
  • Molecular identification was performed using polymerase chain reaction (PCR) targeting the mitochondrial cytochrome c oxidase subunit I (COXI) gene, allowing species-specific identification of the parasites.
  • Serum oxidative stress biomarkers were measured, including:
    • Malondialdehyde (MDA) – a marker for lipid peroxidation and oxidative damage.
    • Superoxide dismutase (SOD) – an antioxidant enzyme indicating oxidative stress defense.
    • Glutathione (GSH) – a key antioxidant molecule involved in cellular protection.
    • Total antioxidant capacity (TAC) – overall ability of serum to counteract oxidative damage.
  • Gene expression of cytokines was assessed using real-time PCR to analyze immune responses. The cytokines measured were:
    • Interferon-gamma (IFN-γ)
    • Tumor necrosis factor-alpha (TNF-α)
    • Interleukin 1 beta (IL-1β)
    • Interleukin 6 (IL-6)

Key Findings

  • Eggs of Habronema species were detected in 62% of fecal samples, highlighting the relatively high prevalence among these horses.
  • Molecular analysis confirmed Habronema muscae as the predominant parasite species infecting the horses on this farm.
  • Infected horses showed significantly elevated levels of oxidative stress biomarkers (increased MDA and altered antioxidant defenses such as SOD, GSH, and TAC) compared to healthy controls, indicating oxidative stress as a key feature of infection.
  • Cytokine gene expression analysis revealed upregulation of proinflammatory cytokines (IFN-γ, TNF-α, IL-1β, and IL-6), reflecting a strong localized immune response dominated by Th1-type immunity.

Conclusions and Implications

  • The integration of molecular diagnostics such as PCR targeting COXI gene with traditional parasitological methods improves detection sensitivity and specificity for equine habronemiasis.
  • Assessment of oxidative stress biomarkers provides complementary information on the physiological impact of infection and helps indicate disease status.
  • Elevated proinflammatory cytokine expression profiles reveal key aspects of the host immune response, suggesting potential targets for immune modulation therapies or prognostic markers.
  • Together, these diagnostic strategies contribute to a more comprehensive understanding of equine habronemiasis and support the development of improved diagnostic and therapeutic interventions.
  • This study exemplifies the benefit of a multidisciplinary approach combining parasitology, molecular biology, immunology, and biochemistry in veterinary infectious disease research.

Cite This Article

APA
Salem MA, El-Gameel SM, Kamel MS, Elsamman EM, Ramadan RM. (2025). Innovative diagnostic strategies for equine habronemiasis: exploring molecular identification, gene expression, and oxidative stress markers. Parasit Vectors, 18(1), 325. https://doi.org/10.1186/s13071-025-06970-1

Publication

Parasites & vectors
ISSN: 1756-3305
NlmUniqueID: 101462774
Country: England
Language: English
Volume: 18
Issue: 1
Pages: 325
PII: 325

Researcher Affiliations

Salem, Mai A
  • Department of Parasitology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.
El-Gameel, Sohila M
  • Department of Parasitology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.
Kamel, Mohamed S
  • Department of Medicine and Infectious Diseases, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.
Elsamman, Eslam M
  • Faculty of Veterinary Medicine, Cairo University (Equine Veterinarian), Giza, 12211, Egypt.
Ramadan, Reem M
  • Department of Parasitology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt. reem.montaser@cu.edu.eg.

MeSH Terms

  • Animals
  • Oxidative Stress
  • Horse Diseases / diagnosis
  • Horse Diseases / parasitology
  • Horses
  • Biomarkers / blood
  • Cytokines / genetics
  • Feces / parasitology
  • Spirurida Infections / veterinary
  • Spirurida Infections / diagnosis
  • Spirurida Infections / parasitology
  • Egypt
  • Female
  • Male
  • Gene Expression

Conflict of Interest Statement

Declarations. Ethics approval and consent to participate: This study was conducted and approved in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC-VET-CU-110520251130) of the Faculty of Veterinary Medicine, Cairo University. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests.

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Citations

This article has been cited 3 times.
  1. Abdel-Radi S, Salem MA, Youssef FS, Kamel MS, El-Bahy MM, Ramadan RM. Acaricidal activity of Astragalus polysaccharides nanoemulsion against camel tick, Hyalomma dromedarii.. Exp Appl Acarol 2025 Dec 2;95(4):63.
    doi: 10.1007/s10493-025-01085-9pubmed: 41329442google scholar: lookup
  2. Ullah A, Geng M, Chen W, Zhu Q, Shi L, Zhang X, Akhtar MF, Wang C, Khan MZ. Effect of Parasitic Infections on Hematological Profile, Reproductive and Productive Performance in Equines.. Animals (Basel) 2025 Nov 14;15(22).
    doi: 10.3390/ani15223294pubmed: 41302002google scholar: lookup
  3. Vasconcelos AB, França DA, Prado ACD, Yamauchi DH, Silva ACAD, Barros IO, Valença SRFA, Lucheis SB, Bosco SMG. Molecular Detection of Pythium insidiosum in Cutaneous Lesions of Horses from Northeastern Brazil.. Animals (Basel) 2025 Sep 30;15(19).
    doi: 10.3390/ani15192863pubmed: 41096457google scholar: lookup
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