Concurrent Proteomic Fingerprinting and Molecular Analysis of Cyathostomins.
Abstract: Rapid, cost-effective, efficient, and reliable helminth species identification is of considerable importance to understand host-parasite interactions, clinical disease, and drug resistance. Cyathostomins (Nematoda: Strongylidae) are considered to be the most important equine parasites, yet research on this group is hampered by the large number of 50 morphologically differentiated species, their occurrence in mixed infections with often more than 10 species and the difficulties associated with conventional identification methods. Here, MALDI-TOF MS, previously successfully applied to identify numerous organisms, is evaluated and compared with conventional and molecular genetic approaches. A simple and robust protocol for protein extraction and subsequent DNA isolation allowing molecular confirmation of proteomic findings is developed, showing that MALDI-TOF MS can discriminate adult stages of the two closely related cyathostomin species Cylicostephanus longibursatus and Cylicostephanus minutus. Intraspecific variability of proteomic profiles within morphospecies demonstrated an identification of morphospecies with an accuracy of close to 100%. In contrast, three genospecies within C. minutus and sex-specific profiles within both morphospecies could not be reliably discriminated using MALDI-TOF MS. In conclusion, MALDI-TOF MS complemented by the molecular protocol is a reliable and efficient approach for cyathostomin species identification.
© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Publication Date: 2019-02-21 PubMed ID: 30786147DOI: 10.1002/pmic.201800290Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study investigates the use of mass spectrometry (specifically, MALDI-TOF MS) to identify different species of Cyathostomins, important equine parasites. This method proves reliable and efficient compared to conventional techniques, though it could not distinguish certain within-species variations.
Research on Cyathostomins
- Cyathostomins are a group of nematode parasites that heavily infest horses.
- They represent the largest challenge in equine parasitology, but identifying species has been difficult with traditional methods due to their similar morphologies and mixed infections.
- Accurate species identification is crucial for understanding host-parasite interactions, clinical disease, and potential drug resistance patterns.
MALDI-TOF MS Method
- The researchers evaluated a method called Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS).
- This technique is already successfully used to identify various types of organisms by analyzing their proteomic profiles (their complete set of proteins).
- The team developed a robust protocol that extracts the protein for MALDI-TOF MS analysis first, and then isolates the DNA for confirmatory molecular genetic analysis.
Findings and Implications
- MALDI-TOF MS was able to reliably distinguish between two closely related Cyathostomins species, Cylicostephanus longibursatus and Cylicostephanus minutus.
- Moreover, the method was able to identify morphospecies (species defined by their physical characteristics) with an accuracy close to 100%.
- However, it was not able to reliably differentiate the three genospecies within C. minutus or distinguish between sex-specific profiles within both species.
- The complement of the MALDI-TOF MS method with molecular genetic confirmation is suggested to be a more reliable and efficient approach for Cyathostomin species identification, providing a faster, more cost-effective tool for equine parasitology research and potential therapeutic intervention planning.
Cite This Article
APA
Bredtmann CM, Krücken J, Murugaiyan J, Balard A, Hofer H, Kuzmina TA, von Samson-Himmelstjerna G.
(2019).
Concurrent Proteomic Fingerprinting and Molecular Analysis of Cyathostomins.
Proteomics, 19(7), e1800290.
https://doi.org/10.1002/pmic.201800290 Publication
Researcher Affiliations
- Institute for Parasitology and Tropical Veterinary Medicine, Department of Veterinary Medicine, Freie Universität Berlin, 14163, Berlin, Germany.
- Institute for Parasitology and Tropical Veterinary Medicine, Department of Veterinary Medicine, Freie Universität Berlin, 14163, Berlin, Germany.
- Institute for Animal Hygiene and Environmental Health, Department of Veterinary Medicine, Freie Universität Berlin, 14163, Berlin, Germany.
- Department of Biotechnology, SRM University AP, 522502, Amaravati, India.
- Ecology and Evolution of Molecular Parasite Host Interactions, Molecular Parasitology, Institute for Biology, Humboldt University Berlin, 10115, Berlin, Germany.
- Ecology and Evolution of Molecular Parasite Host Interactions, Leibniz Institute for Zoo and Wildlife Research, 10315, Berlin, Germany.
- Institute for Parasitology and Tropical Veterinary Medicine, Department of Veterinary Medicine, Freie Universität Berlin, 14163, Berlin, Germany.
- Department of Ecological Dynamics, Leibniz Institute for Zoo and Wildlife Research, 10315, Berlin, Germany.
- Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195, Berlin, Germany.
- Department of Parasitology, I. I. Schmalhausen Institute of Zoology, 01030, Kiev, Ukraine.
- Institute for Parasitology and Tropical Veterinary Medicine, Department of Veterinary Medicine, Freie Universität Berlin, 14163, Berlin, Germany.
MeSH Terms
- Animals
- Cluster Analysis
- Horses
- Nematoda / pathogenicity
- Proteomics / methods
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
Citations
This article has been cited 7 times.- Ebersbach JC, Sato MO, de Araújo MP, Sato M, Becker SL, Sy I. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for differential identification of adult Schistosoma worms.. Parasit Vectors 2023 Jan 19;16(1):20.
- Sy I, Conrad L, Becker SL. Recent Advances and Potential Future Applications of MALDI-TOF Mass Spectrometry for Identification of Helminths.. Diagnostics (Basel) 2022 Dec 3;12(12).
- Gao Y, Zhang Z, Wang C, Zhao K. The Mitochondrial Genome of Cylicocyclus elongatus (Strongylida: Strongylidae) and Its Comparative Analysis with Other Cylicocyclus Species.. Animals (Basel) 2022 Jun 17;12(12).
- Feucherolles M, Nennig M, Becker SL, Martiny D, Losch S, Penny C, Cauchie HM, Ragimbeau C. Investigation of MALDI-TOF Mass Spectrometry for Assessing the Molecular Diversity of Campylobacter jejuni and Comparison with MLST and cgMLST: A Luxembourg One-Health Study.. Diagnostics (Basel) 2021 Oct 20;11(11).
- Wendel TP, Feucherolles M, Rehner J, Poppert S, Utzinger J, Becker SL, Sy I. Evaluating Different Storage Media for Identification of Taenia saginata Proglottids Using MALDI-TOF Mass Spectrometry.. Microorganisms 2021 Sep 22;9(10).
- Louro M, Kuzmina TA, Bredtmann CM, Diekmann I, de Carvalho LMM, von Samson-Himmelstjerna G, Krücken J. Genetic variability, cryptic species and phylogenetic relationship of six cyathostomin species based on mitochondrial and nuclear sequences.. Sci Rep 2021 Apr 15;11(1):8245.
- Feucherolles M, Poppert S, Utzinger J, Becker SL. MALDI-TOF mass spectrometry as a diagnostic tool in human and veterinary helminthology: a systematic review.. Parasit Vectors 2019 May 17;12(1):245.
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