The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue

Katrine Toft; Marie Louise Honoré; Nichol E. Ripley; Martin K. Nielsen; Bastian Fromm; Maibritt Mardahl; Lise N. Nielsen; Peter Nejsum; Stig Milan Thamsborg; Susanna Cirera; Tina Holberg Pihl

doi:10.1016/j.ijpara.2024.10.005

The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue

Katrine Toft
, Marie Louise Honoré
, Nichol E. Ripley
, Martin K. Nielsen
, Bastian Fromm
, Maibritt Mardahl
, Lise N. Nielsen
, Peter Nejsum
, Stig Milan Thamsborg
, Susanna Cirera
, Tina Holberg Pihl

Veterinary Science

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

The equine bloodworm, Strongylus vulgaris, is a highly pathogenic parasite causing potentially fatal vascular and intestinal damage. Parasites express and release microRNAs (miRNAs) for internal regulation and to modulate host immunity. The complete set of miRNAs expressed by S. vulgaris (the S. vulgaris miRNAome) remains unannotated and the aim of this study was to annotate the miRNAome of L4 and L5 stages of S. vulgaris, and to examine differences in miRNA abundance between larval stages and sexes. Furthermore, we aimed to determine if miRNAs were detectable in excretory/secretory products (ESPs) from larvae and in arterial tissue from their predilection site, the cranial mesenteric artery (CMA). Larvae were collected from naturally infected foals, and categorized by sex and stage. A subset of larvae was snap-frozen, while those remaining were incubated and the (ESPs) collected. Arterial tissue samples were collected from the CMA. Small RNA sequencing, followed by a custom bioinformatic pipeline, was used for annotation. We identified 142 S. vulgaris miRNAs in larvae and 136 in ESPs. Significant differences in miRNA abundance were observed between larvae and ESPs, and between L5 females (L5Fs) and L5 males (L5Ms), L4s and L5Fs, and L4s and L5Ms. No differences were found between L4s and L5s overall. In ESPs, several miRNAs were differentially abundant across all groups. Validation through quantitative real-time PCR (qPCR) detected selected miRNAs and their differential abundance in larvae and ESPs. One parasite-derived miRNA was detected in some of the horse arterial tissue samples but at very low levels. This study provided the first annotation of the S. vulgaris miRNAome. Most of the annotated larval miRNAs were also detectable in ESPs, and differences in miRNA abundance between sexes were found for larvae, and between sexes and stages for ESPs. Parasite-derived miRNAs were, however, not consistently detectable in the surrounding host arterial tissue.

Original language	English
Pages (from-to)	45-58
Number of pages	14
Journal	International Journal for Parasitology
Volume	55
Issue number	1
DOIs	https://doi.org/10.1016/j.ijpara.2024.10.005
State	Published - Jan 2025

Bibliographical note

Publisher Copyright:
© 2024 The Authors

Funding

The authors want to acknowledge the expert help and guidance provided by laboratory technicians Minna Jakobsen and Tina Bahrt Neergaard Mahler at the Department of Veterinary and Animal Sciences, University of Copenhagen, Denmark, Maria Rhod and Tina Roust at the University of Copenhagen Large Animal Teaching Hospital and Holli Sullivan Gravatte, Gluck Equine Research Center, University of Kentucky, USA. Similarly, we would like to thank Dr. Bradley Whitehead for help with sample preparation and sharing his expert knowledge on parasite miRNA exploration. Finally, we would like to thank the devoted farm crew at the Gluck Equine Research Center, University of Kentucky for their skilled help with handling the horses involved in this study. This work was supported by the Independent Research Fund Denmark (case number 0136-00101B), the Danish Horse Levy Foundation, University of Copenhagen’s Graduate School, the Denmark America Foundation, the Hartmann Foundation, Denmark, the William Demant Foundation, Denmark and the Sveland Foundation, Sweden. Materials for sample collection and processing were received from E-vet, Denmark, Scanvet, Denmark and Eickemeyer, Denmark. The authors want to acknowledge the expert help and guidance provided by laboratory technicians Minna Jakobsen and Tina Bahrt Neergaard Mahler at the Department of Veterinary and Animal Sciences, University of Copenhagen, Denmark, Maria Rhod and Tina Roust at the University of Copenhagen Large Animal Teaching Hospital and Holli Sullivan Gravatte, Gluck Equine Research Center, University of Kentucky, USA. Similarly, we would like to thank Dr. Bradley Whitehead for help with sample preparation and sharing his expert knowledge on parasite miRNA exploration. Finally, we would like to thank the devoted farm crew at the Gluck Equine Research Center, University of Kentucky for their skilled help with handling the horses involved in this study. This work was supported by the Independent Research Fund Denmark (case number 0136-00101B), the Danish Horse Levy Foundation, University of Copenhagen's Graduate School, the Denmark America Foundation, the Hartmann Foundation, Denmark, the William Demant Foundation, Denmark and the Sveland Foundation, Sweden. Materials for sample collection and processing were received from E-vet, Denmark, Scanvet, Denmark and Eickemeyer, Denmark.

Funders	Funder number
University of Copenhagen’s Graduate School
Department of Veterinary and Animal Sciences, University of Copenhagen
William Demant Fonden
Danish Horse Levy Foundation
University of Kentucky
University of Copenhagen's Graduate School
Denmark-America Foundation
Hartmann Fonden
Sveland Foundation
Independent Research Fund Denmark	0136-00101B

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Horse
MicroRNA Complement
Nematode
Parasite
Small RNA sequencing

ASJC Scopus subject areas

Parasitology
Infectious Diseases

Access to Document

10.1016/j.ijpara.2024.10.005

Cite this

Toft, K., Honoré, M. L., Ripley, N. E., Nielsen, M. K., Fromm, B., Mardahl, M., Nielsen, L. N., Nejsum, P., Thamsborg, S. M., Cirera, S., & Pihl, T. H. (2025). The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue. International Journal for Parasitology, 55(1), 45-58. https://doi.org/10.1016/j.ijpara.2024.10.005

@article{2d4c24cca0804c5bb755b99b81731b13,

title = "The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue",

abstract = "The equine bloodworm, Strongylus vulgaris, is a highly pathogenic parasite causing potentially fatal vascular and intestinal damage. Parasites express and release microRNAs (miRNAs) for internal regulation and to modulate host immunity. The complete set of miRNAs expressed by S. vulgaris (the S. vulgaris miRNAome) remains unannotated and the aim of this study was to annotate the miRNAome of L4 and L5 stages of S. vulgaris, and to examine differences in miRNA abundance between larval stages and sexes. Furthermore, we aimed to determine if miRNAs were detectable in excretory/secretory products (ESPs) from larvae and in arterial tissue from their predilection site, the cranial mesenteric artery (CMA). Larvae were collected from naturally infected foals, and categorized by sex and stage. A subset of larvae was snap-frozen, while those remaining were incubated and the (ESPs) collected. Arterial tissue samples were collected from the CMA. Small RNA sequencing, followed by a custom bioinformatic pipeline, was used for annotation. We identified 142 S. vulgaris miRNAs in larvae and 136 in ESPs. Significant differences in miRNA abundance were observed between larvae and ESPs, and between L5 females (L5Fs) and L5 males (L5Ms), L4s and L5Fs, and L4s and L5Ms. No differences were found between L4s and L5s overall. In ESPs, several miRNAs were differentially abundant across all groups. Validation through quantitative real-time PCR (qPCR) detected selected miRNAs and their differential abundance in larvae and ESPs. One parasite-derived miRNA was detected in some of the horse arterial tissue samples but at very low levels. This study provided the first annotation of the S. vulgaris miRNAome. Most of the annotated larval miRNAs were also detectable in ESPs, and differences in miRNA abundance between sexes were found for larvae, and between sexes and stages for ESPs. Parasite-derived miRNAs were, however, not consistently detectable in the surrounding host arterial tissue.",

keywords = "Horse, MicroRNA Complement, Nematode, Parasite, Small RNA sequencing",

author = "Katrine Toft and Honor{\'e}, \{Marie Louise\} and Ripley, \{Nichol E.\} and Nielsen, \{Martin K.\} and Bastian Fromm and Maibritt Mardahl and Nielsen, \{Lise N.\} and Peter Nejsum and Thamsborg, \{Stig Milan\} and Susanna Cirera and Pihl, \{Tina Holberg\}",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2025",

month = jan,

doi = "10.1016/j.ijpara.2024.10.005",

language = "English",

volume = "55",

pages = "45--58",

number = "1",

}

Toft, K, Honoré, ML, Ripley, NE, Nielsen, MK, Fromm, B, Mardahl, M, Nielsen, LN, Nejsum, P, Thamsborg, SM, Cirera, S & Pihl, TH 2025, 'The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue', International Journal for Parasitology, vol. 55, no. 1, pp. 45-58. https://doi.org/10.1016/j.ijpara.2024.10.005

The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue. / Toft, Katrine; Honoré, Marie Louise; Ripley, Nichol E. et al.
In: International Journal for Parasitology, Vol. 55, No. 1, 01.2025, p. 45-58.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue

AU - Toft, Katrine

AU - Honoré, Marie Louise

AU - Ripley, Nichol E.

AU - Nielsen, Martin K.

AU - Fromm, Bastian

AU - Mardahl, Maibritt

AU - Nielsen, Lise N.

AU - Nejsum, Peter

AU - Thamsborg, Stig Milan

AU - Cirera, Susanna

AU - Pihl, Tina Holberg

PY - 2025/1

Y1 - 2025/1

N2 - The equine bloodworm, Strongylus vulgaris, is a highly pathogenic parasite causing potentially fatal vascular and intestinal damage. Parasites express and release microRNAs (miRNAs) for internal regulation and to modulate host immunity. The complete set of miRNAs expressed by S. vulgaris (the S. vulgaris miRNAome) remains unannotated and the aim of this study was to annotate the miRNAome of L4 and L5 stages of S. vulgaris, and to examine differences in miRNA abundance between larval stages and sexes. Furthermore, we aimed to determine if miRNAs were detectable in excretory/secretory products (ESPs) from larvae and in arterial tissue from their predilection site, the cranial mesenteric artery (CMA). Larvae were collected from naturally infected foals, and categorized by sex and stage. A subset of larvae was snap-frozen, while those remaining were incubated and the (ESPs) collected. Arterial tissue samples were collected from the CMA. Small RNA sequencing, followed by a custom bioinformatic pipeline, was used for annotation. We identified 142 S. vulgaris miRNAs in larvae and 136 in ESPs. Significant differences in miRNA abundance were observed between larvae and ESPs, and between L5 females (L5Fs) and L5 males (L5Ms), L4s and L5Fs, and L4s and L5Ms. No differences were found between L4s and L5s overall. In ESPs, several miRNAs were differentially abundant across all groups. Validation through quantitative real-time PCR (qPCR) detected selected miRNAs and their differential abundance in larvae and ESPs. One parasite-derived miRNA was detected in some of the horse arterial tissue samples but at very low levels. This study provided the first annotation of the S. vulgaris miRNAome. Most of the annotated larval miRNAs were also detectable in ESPs, and differences in miRNA abundance between sexes were found for larvae, and between sexes and stages for ESPs. Parasite-derived miRNAs were, however, not consistently detectable in the surrounding host arterial tissue.

AB - The equine bloodworm, Strongylus vulgaris, is a highly pathogenic parasite causing potentially fatal vascular and intestinal damage. Parasites express and release microRNAs (miRNAs) for internal regulation and to modulate host immunity. The complete set of miRNAs expressed by S. vulgaris (the S. vulgaris miRNAome) remains unannotated and the aim of this study was to annotate the miRNAome of L4 and L5 stages of S. vulgaris, and to examine differences in miRNA abundance between larval stages and sexes. Furthermore, we aimed to determine if miRNAs were detectable in excretory/secretory products (ESPs) from larvae and in arterial tissue from their predilection site, the cranial mesenteric artery (CMA). Larvae were collected from naturally infected foals, and categorized by sex and stage. A subset of larvae was snap-frozen, while those remaining were incubated and the (ESPs) collected. Arterial tissue samples were collected from the CMA. Small RNA sequencing, followed by a custom bioinformatic pipeline, was used for annotation. We identified 142 S. vulgaris miRNAs in larvae and 136 in ESPs. Significant differences in miRNA abundance were observed between larvae and ESPs, and between L5 females (L5Fs) and L5 males (L5Ms), L4s and L5Fs, and L4s and L5Ms. No differences were found between L4s and L5s overall. In ESPs, several miRNAs were differentially abundant across all groups. Validation through quantitative real-time PCR (qPCR) detected selected miRNAs and their differential abundance in larvae and ESPs. One parasite-derived miRNA was detected in some of the horse arterial tissue samples but at very low levels. This study provided the first annotation of the S. vulgaris miRNAome. Most of the annotated larval miRNAs were also detectable in ESPs, and differences in miRNA abundance between sexes were found for larvae, and between sexes and stages for ESPs. Parasite-derived miRNAs were, however, not consistently detectable in the surrounding host arterial tissue.

KW - Horse

KW - MicroRNA Complement

KW - Nematode

KW - Parasite

KW - Small RNA sequencing

UR - https://www.scopus.com/pages/publications/85209585382

UR - https://www.scopus.com/pages/publications/85209585382#tab=citedBy

U2 - 10.1016/j.ijpara.2024.10.005

DO - 10.1016/j.ijpara.2024.10.005

M3 - Article

C2 - 39510492

AN - SCOPUS:85209585382

SN - 0020-7519

VL - 55

SP - 45

EP - 58

JO - International Journal for Parasitology

JF - International Journal for Parasitology

IS - 1

ER -

The microRNAome of Strongylus vulgaris larvae and their excretory/secretory products with identification of parasite-derived microRNAs in horse arterial tissue

Abstract

Bibliographical note

Funding

UN SDGs

Keywords

ASJC Scopus subject areas

Access to Document

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