{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T21:20:03Z","timestamp":1790716803460,"version":"4.1.0"},"reference-count":37,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,12,31]],"date-time":"2020-12-31T00:00:00Z","timestamp":1609372800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["Emmanuel O. Ngbede was supported by the Cooperation Visits Programme of The World Academy of Sciences and Deutsche Forschungsgemeinschaft (TWAS-DFG)."],"award-info":[{"award-number":["Emmanuel O. Ngbede was supported by the Cooperation Visits Programme of The World Academy of Sciences and Deutsche Forschungsgemeinschaft (TWAS-DFG)."]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Fascioliasis is a neglected trematode infection caused by Fasciola gigantica and Fasciola hepatica. Routine diagnosis of fascioliasis relies on macroscopic identification of adult worms in liver tissue of slaughtered animals, and microscopic detection of eggs in fecal samples of animals and humans. However, the diagnostic accuracy of morphological techniques and stool microscopy is low. Molecular diagnostics (e.g., polymerase chain reaction (PCR)) are more reliable, but these techniques are not routinely available in clinical microbiology laboratories. Matrix-assisted laser\/desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a widely-used technique for identification of bacteria and fungi; yet, standardized protocols and databases for parasite detection need to be developed. The purpose of this study was to develop and validate an in-house database for Fasciola species-specific identification. To achieve this goal, the posterior parts of seven adult F. gigantica and one adult F. hepatica were processed and subjected to MALDI-TOF MS to create main spectra profiles (MSPs). Repeatability and reproducibility tests were performed to develop the database. A principal component analysis revealed significant differences between the spectra of F. gigantica and F. hepatica. Subsequently, 78 Fasciola samples were analyzed by MALDI-TOF MS using the previously developed database, out of which 98.7% (n = 74) and 100% (n = 3) were correctly identified as F. gigantica and F. hepatica, respectively. Log score values ranged between 1.73 and 2.23, thus indicating a reliable identification. We conclude that MALDI-TOF MS can provide species-specific identification of medically relevant liver flukes.<\/jats:p>","DOI":"10.3390\/microorganisms9010082","type":"journal-article","created":{"date-parts":[[2020,12,31]],"date-time":"2020-12-31T10:10:37Z","timestamp":1609409437000},"page":"82","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Identification of Adult Fasciola spp. Using Matrix-Assisted Laser\/Desorption Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9103-8861","authenticated-orcid":false,"given":"Issa","family":"Sy","sequence":"first","affiliation":[{"name":"Institute of Medical Microbiology and Hygiene, Saarland University, 66421 Homburg\/Saar, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lena","family":"Margardt","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology and Hygiene, Saarland University, 66421 Homburg\/Saar, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8951-876X","authenticated-orcid":false,"given":"Emmanuel O.","family":"Ngbede","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology and Hygiene, Saarland University, 66421 Homburg\/Saar, Germany"},{"name":"College of Veterinary Medicine, Federal University of Agriculture, Makurdi 970213, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed I.","family":"Adah","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Federal University of Agriculture, Makurdi 970213, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saheed T.","family":"Yusuf","sequence":"additional","affiliation":[{"name":"Kubwa Abattoir, Public Health Section, Department of Veterinary Services, Federal Capital Territory Administration, Abuja 901101, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0290-3521","authenticated-orcid":false,"given":"Jennifer","family":"Keiser","sequence":"additional","affiliation":[{"name":"Swiss Tropical and Public Health Institute, CH-4002 Basel, Switzerland"},{"name":"University of Basel, CH-4003 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacqueline","family":"Rehner","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology and Hygiene, Saarland University, 66421 Homburg\/Saar, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00fcrg","family":"Utzinger","sequence":"additional","affiliation":[{"name":"Swiss Tropical and Public Health Institute, CH-4002 Basel, Switzerland"},{"name":"University of Basel, CH-4003 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sven","family":"Poppert","sequence":"additional","affiliation":[{"name":"Swiss Tropical and Public Health Institute, CH-4002 Basel, Switzerland"},{"name":"University of Basel, CH-4003 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3634-8802","authenticated-orcid":false,"given":"S\u00f6ren L.","family":"Becker","sequence":"additional","affiliation":[{"name":"Institute of Medical Microbiology and Hygiene, Saarland University, 66421 Homburg\/Saar, Germany"},{"name":"Swiss Tropical and Public Health Institute, CH-4002 Basel, Switzerland"},{"name":"University of Basel, CH-4003 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1128\/CMR.00012-09","article-title":"Food-borne trematodiases","volume":"22","author":"Keiser","year":"2009","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1186\/s13071-016-1898-2","article-title":"Identity of Fasciola spp. in sheep in Egypt","volume":"9","author":"Amer","year":"2016","journal-title":"Parasit. Vectors"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.ijpara.2019.12.002","article-title":"Which species is in the faeces at a time of global livestock movements: Single nucleotide polymorphism genotyping assays for the differentiation of Fasciola spp","volume":"50","author":"Calvani","year":"2020","journal-title":"Int. J. Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1186\/s13071-018-3248-z","article-title":"Patterns of Fasciola hepatica infection in Danish dairy cattle: Implications for on-farm control of the parasite based on different diagnostic methods","volume":"11","author":"Denwood","year":"2018","journal-title":"Parasit. Vectors"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"597","DOI":"10.21037\/hbsn.2019.04.02","article-title":"Fasciola hepatica in a country of low incidence: A tricky diagnosis","volume":"8","author":"Perrodin","year":"2019","journal-title":"Hepatobiliary Surg. Nutr."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.vetpar.2015.09.024","article-title":"DNA sequence analyses reveal co-occurrence of novel haplotypes of Fasciola gigantica with F. hepatica in South Africa and Zimbabwe","volume":"214","author":"Mucheka","year":"2015","journal-title":"Vet. Parasitol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Chougar, L., Mas-Coma, S., Artigas, P., Harhoura, K., Aissi, M., Agramunt, V.H., and Bargues, M.D. (2020). Genetically \u201cpure\u201d Fasciola gigantica discovered in Algeria: DNA multimarker characterization, trans-Saharan introduction from a Sahel origin and spreading risk into north-western Maghreb countries. Transbound. Emerg. Dis.","DOI":"10.1111\/tbed.13572"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/S0065-308X(09)69002-3","article-title":"Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control","volume":"69","author":"Valero","year":"2009","journal-title":"Adv. Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Jean-Richard, V., Crump, L., Abicho, A.A., Nar\u00e9, N.B., Greter, H., Hattendorf, J., Schelling, E., and Zinsstag, J. (2014). Prevalence of Fasciola gigantica infection in slaughtered animals in south-eastern Lake Chad area in relation to husbandry practices and seasonal water levels. BMC Vet. Res., 10.","DOI":"10.1186\/1746-6148-10-81"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1292\/jvms.16-0331","article-title":"High prevalence of cattle fascioliasis in coastal areas of Thua Thien Hue province, Vietnam","volume":"79","author":"Nguyen","year":"2017","journal-title":"J. Vet. Med. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.4102\/ojvr.v86i1.1706","article-title":"Phylogenetic analysis of Fasciola spp. isolated from slaughtered cattle in KwaZulu-Natal and Mpumalanga provinces of South Africa based on the cytochrome c oxidase subunit I mitochondrial marker","volume":"86","author":"Chikowore","year":"2019","journal-title":"Onderstepoort J. Vet. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S1567-1348(01)00019-3","article-title":"European Lymnaeidae (Mollusca: Gastropoda), intermediate hosts of trematodiases, based on nuclear ribosomal DNA ITS-2 sequences","volume":"1","author":"Bargues","year":"2001","journal-title":"Infect. Genet. Evol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1017\/S0031182007004076","article-title":"The distribution of Fasciola hepatica and Fasciola gigantica within southern Tanzania--constraints associated with the intermediate host","volume":"135","author":"Walker","year":"2008","journal-title":"Parasitology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1389","DOI":"10.1007\/s00436-007-0656-y","article-title":"Pseudosuccinea columella (Say 1817) (Gastropoda, Lymnaeidae), snail host of Fasciola hepatica: First record for France in the wild","volume":"101","author":"Pointier","year":"2007","journal-title":"Parasitol. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s13059-015-0632-2","article-title":"The Fasciola hepatica genome: Gene duplication and polymorphism reveals adaptation to the host environment and the capacity for rapid evolution","volume":"16","author":"Cwiklinski","year":"2015","journal-title":"Genome Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e8597","DOI":"10.7717\/peerj.8597","article-title":"Fasciola gigantica, F. hepatica and Fasciola intermediate forms: Geometric morphometrics and an artificial neural network to help morphological identification","volume":"8","author":"Sumruayphol","year":"2020","journal-title":"PeerJ"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1186\/1756-3305-4-101","article-title":"Genetic characterization, species differentiation and detection of Fasciola spp. by molecular approaches","volume":"4","author":"Ai","year":"2011","journal-title":"Parasit. Vectors"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"S83","DOI":"10.1016\/j.nmni.2018.08.020","article-title":"MALDI-TOF MS and point of care are disruptive diagnostic tools in Africa","volume":"26","author":"Chabriere","year":"2018","journal-title":"New Microbes New Infect"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1128\/CMR.00072-12","article-title":"Matrix-assisted laser desorption ionization-time of flight mass spectrometry: A fundamental shift in the routine practice of clinical microbiology","volume":"26","author":"Clark","year":"2013","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.mimet.2016.09.003","article-title":"Matrix assisted laser desorption time of flight mass spectrometry (MALDI-TOF MS) in clinical microbiology","volume":"138","author":"Angeletti","year":"2017","journal-title":"J. Microbiol. Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s12936-016-1657-z","article-title":"MALDI-TOF MS as an innovative tool for detection of Plasmodium parasites in Anopheles mosquitoes","volume":"16","author":"Laroche","year":"2017","journal-title":"Malar. J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3148","DOI":"10.1002\/pmic.201600287","article-title":"Standardization of sample homogenization for mosquito identification using an innovative proteomic tool based on protein profiling","volume":"16","author":"Nebbak","year":"2016","journal-title":"Proteomics"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Diarra, A.Z., Almeras, L., Laroche, M., Berenger, J.-M., Kon\u00e9, A.K., Bocoum, Z., Dabo, A., Doumbo, O., Raoult, D., and Parola, P. (2017). Molecular and MALDI-TOF identification of ticks and tick-associated bacteria in Mali. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0005762"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1186\/s13071-019-3493-9","article-title":"MALDI-TOF mass spectrometry as a diagnostic tool in human and veterinary helminthology: A systematic review","volume":"12","author":"Feucherolles","year":"2019","journal-title":"Parasit. Vectors"},{"key":"ref_25","first-page":"95","article-title":"BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95\/98\/NT","volume":"Volume 41","author":"Hall","year":"1999","journal-title":"Proceedings of the Nucleic Acids Symposium Series"},{"key":"ref_26","unstructured":"(2020, November 18). National Center for Biotechnology Information Nucleotide BLAST: Search Nucleotide Databases Using A Nucleotide Query, Available online: https:\/\/blast.ncbi.nlm.nih.gov\/Blast.cgi?PROGRAM=blastn&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3143","DOI":"10.1002\/pmic.201100049","article-title":"Proteomics for routine identification of microorganisms","volume":"11","author":"Welker","year":"2011","journal-title":"Proteomics"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.anaerobe.2018.07.001","article-title":"Routine use of MALDI-TOF MS for anaerobic bacterial identification in clinical microbiology","volume":"54","author":"Shannon","year":"2018","journal-title":"Anaerobe"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1186\/s13071-020-04029-x","article-title":"Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviae","volume":"13","author":"Nebbak","year":"2020","journal-title":"Parasit Vectors"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1111\/mve.12250","article-title":"Comparative analysis of storage conditions and homogenization methods for tick and flea species for identification by MALDI-TOF MS","volume":"31","author":"Nebbak","year":"2017","journal-title":"Med. Vet. Entomol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1051\/parasite\/2020026","article-title":"Development of MALDI-TOF mass spectrometry for the identification of lice isolated from farm animals","volume":"27","author":"Ouarti","year":"2020","journal-title":"Parasite"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"107890","DOI":"10.1016\/j.exppara.2020.107890","article-title":"Species identification of Trichinella originated from various host and different geographical location by MALDI-TOF","volume":"213","author":"Karadjian","year":"2020","journal-title":"Exp. Parasitol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Marzano, V., Pane, S., Foglietta, G., Levi Mortera, S., Vernocchi, P., Onetti Muda, A., and Putignani, L. (2020). Mass spectrometry-based proteomic analysis of Anisakis spp.: A preliminary study towards a new diagnostic tool. Genes, 11.","DOI":"10.3390\/genes11060693"},{"key":"ref_34","unstructured":"Sy, I., Wendel, T., Feucherolles, M., Nimmesgern, A., Stuermann, A., Endriss, Y., Utzinger, J., Poppert, S., and Becker, S.L. (2020, January 18\u201321). Application of MALDI-TOF MS for identification of helminths in clinical samples. Abstract no. 6730. Proceedings of the European Congress of Clinical Microbiology and Infectious Diseases, Paris, France. Cancelled Due to the COVID-19 Pandemic."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"453","DOI":"10.3347\/kjp.2018.56.5.453","article-title":"Distribution status of hybrid types in large liver flukes, Fasciola species (Digenea: Fasciolidae), from ruminants and humans in Vietnam","volume":"56","author":"Nguyen","year":"2018","journal-title":"Korean J. Parasitol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1186\/1756-3305-7-150","article-title":"Complete mitochondrial genomes of the \u201cintermediate form\u201d of Fasciola and Fasciola gigantica, and their comparison with F. hepatica","volume":"7","author":"Liu","year":"2014","journal-title":"Parasit. Vectors"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1645\/19-66","article-title":"Molecular confirmation of a Fasciola gigantica \u00d7 Fasciola hepatica hybrid in a Chadian bovine","volume":"106","author":"Evack","year":"2020","journal-title":"J. Parasitol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/1\/82\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:48:24Z","timestamp":1760179704000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/1\/82"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12,31]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["microorganisms9010082"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9010082","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,12,31]]}}}