Beyond the host: Unveiling the independent microbiome of equine gastrointestinal nematodes.
Abstract: Gastrointestinal nematode infections significantly impact equine health and welfare, with rising anthelmintic resistance demanding alternative control strategies. Emerging evidence suggests that parasitic nematodes harbour distinct microbiomes, potentially influencing host-parasite dynamics and parasite survival. This study aimed to characterize and compare the microbiomes of equine gastrointestinal nematodes and their hosts, focusing on differences in composition, diversity, and core microbiota structure across different intestinal sites, nematode subfamilies, and sexes. Faecal and nematode samples were collected from equids (Equus caballus and Equus asinus) at slaughterhouses. DNA was extracted, and the V3-V4 regions of the 16S rRNA gene were amplified and sequenced using the Illumina iSeq 100 platform. Bioinformatic analyses were performed with QIIME2 and MicrobiomeAnalyst, and statistical comparisons employed PERMANOVA, LEfSe, and alpha and beta diversity metrics. Nematodes exhibited a distinct microbiome dominated by Firmicutes, Proteobacteria, Bacteroidota, Verrucomicrobiota, and Actinobacteriota, differing significantly from the faecal microbiota. Alpha diversity analyses revealed lower richness in nematodes, while beta diversity indicated distinct community structures (p = 0.007). Microbial composition varied by gastrointestinal site, nematode subfamily, and sex. Proteobacteria were consistently enriched in nematodes, particularly in the caecum. Core microbiome analysis identified exclusive nematode-associated taxa such as Fusobacterium, Mesorhizobium, and Mycoplasma. Equine gastrointestinal nematodes harbour independent and structured microbiomes, distinct from those of their hosts. These findings underscore the ecological specialization of nematodes and highlight the potential of targeting parasite-associated microbiota for novel control strategies.
Copyright: © 2026 Gentilini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Publication Date: 2026-02-10 PubMed ID: 41666179PubMed Central: PMC12890152DOI: 10.1371/journal.pone.0339596Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- 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 research investigated the unique bacterial communities (microbiomes) that live inside equine gastrointestinal nematodes and how these differ from the microbiomes found in their host horses and donkeys.
- The study revealed that these nematodes harbor distinct, specialized microbiomes independent of their host’s gut, which may influence parasite survival and could offer new avenues for parasite control.
Introduction and Research Context
- Gastrointestinal nematodes are parasitic worms that infect horses and donkeys, causing significant health and welfare issues.
- Anthelmintic resistance (resistance to worm-killing drugs) is increasing, making traditional control methods less effective.
- There is emerging evidence that parasitic nematodes host their own unique microbiomes, which may impact how they survive and interact with their hosts.
- This study aimed to explore the nature of these nematode microbiomes and how they differ from those of their host animals.
Methods
- Samples were collected from the intestines and feces of horses (Equus caballus) and donkeys (Equus asinus) at slaughterhouses.
- Nematodes were isolated from different areas of the gastrointestinal tract, accounting for differences by intestinal site, nematode subfamily, and sex of the parasites.
- DNA was extracted from samples, and the bacterial 16S rRNA gene’s V3-V4 regions were amplified for sequencing, which helps identify bacterial species present.
- Sequencing was performed using the Illumina iSeq 100 platform, a next-generation DNA sequencing technology.
- Bioinformatics analyses were conducted using QIIME2 and MicrobiomeAnalyst software to process sequencing data and analyze microbial diversity and composition.
- Statistical tools like PERMANOVA (to test community differences), LEfSe (to detect biomarkers), and alpha/beta diversity metrics (to measure microbial richness and differences) were used for comparisons.
Key Findings
- The microbiomes found inside nematodes were distinctly different from those in the feces of their equine hosts.
- Major bacterial groups dominating the nematode microbiome included Firmicutes, Proteobacteria, Bacteroidota, Verrucomicrobiota, and Actinobacteriota.
- Alpha diversity (species richness) was lower inside nematodes compared to host fecal samples, indicating fewer types of bacteria internally.
- Beta diversity, measuring differences between bacterial communities, showed that nematode microbiomes were significantly distinct (p = 0.007) from host microbiomes.
- The specific bacterial makeup varied depending on the site of the intestine sampled, the nematode’s subfamily classification, and the parasite’s sex.
- Proteobacteria were particularly abundant in nematodes from the caecum, an important part of the horse intestine.
- Core microbiome analysis highlighted taxa unique to nematodes such as Fusobacterium, Mesorhizobium, and Mycoplasma, which were not found in host feces.
Conclusions and Implications
- Equine gastrointestinal nematodes have an independent and structured microbiome, clearly separated from their host’s gut bacteria.
- This finding supports the idea that the nematodes have co-evolved with their own specific bacterial communities, possibly aiding their survival and adaptation within the host environment.
- The unique microbiome associated with nematodes could represent a target for new parasite control strategies that go beyond drug treatments, potentially exploiting the parasite’s bacterial partners to curb infections.
- This study contributes to a deeper ecological understanding of host-parasite-microbiome interactions in equine health.
Cite This Article
APA
Gentilini F, Ogundipe TG, Turba ME, Romagnoli N, Lambertini C, Pollera C, Cremonesi P, Stancampiano L.
(2026).
Beyond the host: Unveiling the independent microbiome of equine gastrointestinal nematodes.
PLoS One, 21(2), e0339596.
https://doi.org/10.1371/journal.pone.0339596 Publication
Researcher Affiliations
- Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Ozzano dell´Emilia (Bo) Italy.
- Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Ozzano dell´Emilia (Bo) Italy.
- Genefast srl, Forlì, Forlì-Cesena, Italy.
- Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Ozzano dell´Emilia (Bo) Italy.
- Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Ozzano dell´Emilia (Bo) Italy.
- Department of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, Lodi, Italy.
- Institute of Agricultural Biology and Biotechnology (IBBA), National Research Council (CNR), Lodi, Italy.
- Department of Veterinary Medical Sciences (DIMEVET), University of Bologna, Ozzano dell´Emilia (Bo) Italy.
MeSH Terms
- Animals
- Horses / parasitology
- Horses / microbiology
- Nematoda / microbiology
- Nematoda / genetics
- Gastrointestinal Microbiome / genetics
- Female
- Male
- RNA, Ribosomal, 16S / genetics
- Nematode Infections / veterinary
- Nematode Infections / parasitology
- Feces / microbiology
- Feces / parasitology
- Horse Diseases / parasitology
- Horse Diseases / microbiology
- Host-Parasite Interactions
- Bacteria / genetics
- Bacteria / classification
Conflict of Interest Statement
The authors have declared that no competing interests exist.
References
This article includes 33 references
- Mathewos M, Teshome D, Fesseha H. Study on Gastrointestinal Nematodes of Equines in and around Bekoji, South Eastern Ethiopia. J Parasitol Res 2022;2022:8210160.
- Wang T, Chen X, Yan X, Su Y, Gao W, Liu C. Progress in serology and molecular biology of equine parasite diagnosis: sustainable control strategies. Front Vet Sci 2025;12:1663577.
- Stear M, Preston S, Piedrafita D, Donskow-Łysoniewska K. The Immune Response to Nematode Infection. Int J Mol Sci 2023;24(3):2283.
- Prangthip P, Tummatorn J, Adisakwattana P, Uthailak N, Boonyuen U, Tipthara P. Anthelmintic efficacy evaluation and mechanism of N-methylbenzo[d]oxazol-2-amine. Sci Rep 2023;13(1):22840.
- Nielsen MK, Pyatt A, Perrett J, Tydén E, van Doorn D, Pihl TH. Global equine parasite control guidelines: Consensus or confusion?. Int J Parasitol Drugs Drug Resist 2025;28:100600.
- Boucher L, Leduc L, Leclère M, Costa MC. Current Understanding of Equine Gut Dysbiosis and Microbiota Manipulation Techniques: Comparison with Current Knowledge in Other Species. Animals 2024;14(5):758.
- Silva PA, Cernea M, Madeira de Carvalho L. Anthelmintic Resistance in Equine Nematodes – A Review on the Current Situation, with Emphasis in Europe. BUASVMCN-VM 2019;76(2):132–42.
- Shao T, Hsu R, Rafizadeh DL, Wang L, Bowlus CL, Kumar N. The gut ecosystem and immune tolerance. J Autoimmun 2023;141:103114.
- Kauter A, Epping L, Semmler T, Antao E-M, Kannapin D, Stoeckle SD. The gut microbiome of horses: current research on equine enteral microbiota and future perspectives. Anim Microbiome 2019;1(1):14.
- Al-Rashidi HS, El-Wakil ES. Parasites and Microbiota: Dual Interactions and Therapeutic Perspectives. Microorganisms 2024;12(10):2076.
- Rooney J, Northcote HM, Williams TL, Cortés A, Cantacessi C, Morphew RM. Parasitic helminths and the host microbiome - a missing “extracellular vesicle-sized” link?. Trends Parasitol 2022;38(9):737–47.
- Cain JL, Norris JK, Ripley NE, Suri P, Finnerty CA, Gravatte HS. The microbial community associated with Parascaris spp. infecting juvenile horses. Parasit Vectors 2022;15(1):408.
- Jorge F, Dheilly NM, Poulin R. Persistence of a Core Microbiome Through the Ontogeny of a Multi-Host Parasite. Front Microbiol 2020;11:954.
- Lichtenfels JR, Kharchenko VA, Dvojnos GM. Illustrated identification keys to strongylid parasites (Strongylidae: Nematoda) of horses, zebras and asses (Equidae). Vet Parasitol 2008;156(1–2):4–161.
- Dirksen P, Marsh SA, Braker I, Heitland N, Wagner S, Nakad R. The native microbiome of the nematode Caenorhabditis elegans: gateway to a new host-microbiome model. BMC Biol 2016;14:38.
- White EC, Houlden A, Bancroft AJ, Hayes KS, Goldrick M, Grencis RK. Manipulation of host and parasite microbiotas: Survival strategies during chronic nematode infection. Sci Adv 2018;4(3):eaap7399.
- Hogan G, Walker S, Turnbull F, Curiao T, Morrison AA, Flores Y. Microbiome analysis as a platform R&D tool for parasitic nematode disease management. ISME J 2019;13(11):2664–80.
- Bhat AH, Malik IM, Tak H, Ganai BA, Bharti P. Host, parasite, and microbiome interaction: Trichuris ovis and its effect on sheep gut microbiota. Vet Parasitol 2025;333:110356.
- Dao MC, Everard A, Aron-Wisnewsky J, Sokolovska N, Prifti E, Verger EO. Akkermansia muciniphila and improved metabolic health during a dietary intervention in obesity: relationship with gut microbiome richness and ecology. Gut 2016;65(3):426–36.
- Menjivar C, Pagella E, Biswas I. Akkermansia muciniphila. Trends Microbiol 2024;32(11):1143–4.
- Jenkins TP, Brindley PJ, Gasser RB, Cantacessi C. Helminth Microbiomes - A Hidden Treasure Trove?. Trends Parasitol 2019;35(1):13–22.
- Corrêa PS, Fernandes MA, Jimenez CR, Mendes LW, Lima P de MT, Abdalla AL. Interaction between methanotrophy and gastrointestinal nematodes infection on the rumen microbiome of lambs. FEMS Microbiol Ecol 2024;100(6):fiae083.
- Drew GC, Stevens EJ, King KC. Microbial evolution and transitions along the parasite-mutualist continuum. Nat Rev Microbiol 2021;19(10):623–38.
- Costa MC, Silva G, Ramos RV, Staempfli HR, Arroyo LG, Kim P. Characterization and comparison of the bacterial microbiota in different gastrointestinal tract compartments in horses. Vet J 2015;205(1):74–80.
- Ericsson AC, Johnson PJ, Lopes MA, Perry SC, Lanter HR. A Microbiological Map of the Healthy Equine Gastrointestinal Tract. PLoS One 2016;11(11):e0166523.
- Jorge F, Froissard C, Dheilly NM, Poulin R. Bacterial community dynamics following antibiotic exposure in a trematode parasite. Int J Parasitol 2022;52(5):265–74.
- Tee MZ, Er YX, Easton AV, Yap NJ, Lee IL, Devlin J. Gut microbiome of helminth-infected indigenous Malaysians is context dependent. Microbiome 2022;10(1):214.
- Mate L, Alvarez LI, Lloberas M, Imperiale F, Lanusse CE, Liron JP. Interaction between bacterial microbiota and nematode parasite communities in sheep’s gastrointestinal tract. PLoS One 2024;19(6):e0306390.
- Shin N-R, Whon TW, Bae J-W. Proteobacteria: microbial signature of dysbiosis in gut microbiota. Trends Biotechnol 2015;33(9):496–503.
- Rizzatti G, Lopetuso LR, Gibiino G, Binda C, Gasbarrini A. Proteobacteria: A Common Factor in Human Diseases. Biomed Res Int 2017;2017:9351507.
- Jorge F, Dheilly NM, Froissard C, Wainwright E, Poulin R. Consistency of Bacterial Communities in a Parasitic Worm: Variation Throughout the Life Cycle and Across Geographic Space. Microb Ecol 2022;83(3):724–38.
- Sheth RU, Cabral V, Chen SP, Wang HH. Manipulating Bacterial Communities by in situ Microbiome Engineering. Trends Genet 2016;32(4):189–200.
- Bouchery T, Lefoulon E, Karadjian G, Nieguitsila A, Martin C. The symbiotic role of Wolbachia in Onchocercidae and its impact on filariasis. Clin Microbiol Infect 2013;19(2):131–40.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists