{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T14:51:38Z","timestamp":1787496698724,"version":"build-2736575974"},"reference-count":54,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,4,9]],"date-time":"2021-04-09T00:00:00Z","timestamp":1617926400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Fondo Institucional de Fomento Regional para el Desarrollo Cient\u00edfico, Tecnol\u00f3gico y de Innovaci\u00f3n (FORDECYT)","award":["292399"],"award-info":[{"award-number":["292399"]}]},{"DOI":"10.13039\/501100003141","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda","doi-asserted-by":"publisher","award":["634647"],"award-info":[{"award-number":["634647"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Fusarium kuroshium is the fungal symbiont associated with the ambrosia beetle Euwallacea kuroshio, a plague complex that attacks avocado, among other hosts, causing a disease named Fusarium dieback (FD). However, the contribution of F. kuroshium to the establishment of this disease remains unknown. To advance the understanding of F. kuroshium pathogenicity, we profiled its exo-metabolome through metabolomics tools based on accurate mass spectrometry. We found that F. kuroshium can produce several key metabolites with phytotoxicity properties and other compounds with unknown functions. Among the metabolites identified in the fungal exo-metabolome, fusaric acid (FA) was further studied due to its phytotoxicity and relevance as a virulence factor. We tested both FA and organic extracts from F. kuroshium at various dilutions in avocado foliar tissue and found that they caused necrosis and chlorosis, resembling symptoms similar to those observed in FD. This study reports for first-time insights regarding F. kuroshium associated with its virulence, which could lead to the potential development of diagnostic and management tools of FD disease and provides a basis for understanding the interaction of F. kuroshium with its host plants.<\/jats:p>","DOI":"10.3390\/toxins13040268","type":"journal-article","created":{"date-parts":[[2021,4,12]],"date-time":"2021-04-12T21:47:33Z","timestamp":1618264053000},"page":"268","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Characterization of the Exo-Metabolome of the Emergent Phytopathogen Fusarium kuroshium sp. nov., a Causal Agent of Fusarium Dieback"],"prefix":"10.3390","volume":"13","author":[{"given":"Ang\u00e9lica","family":"Guti\u00e9rrez-S\u00e1nchez","sequence":"first","affiliation":[{"name":"Laboratorios de Fitopatolog\u00eda y Biolog\u00eda Molecular, Red de Estudios Moleculares Avanzados, Cl\u00faster BioMimic<sup>\u00ae<\/sup>, Instituto de Ecolog\u00eda, A. C. Xalapa, Veracruz 91073, Mexico"},{"name":"Laboratorio de Qu\u00edmica de Productos Naturales, Red de Estudios Moleculares Avanzados, Cl\u00faster BioMimic<sup>\u00ae<\/sup>, Instiuto de Ecolog\u00eda, A. C. Xalapa, Veracruz 91073, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2473-4503","authenticated-orcid":false,"given":"Javier","family":"Plasencia","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica, Facultad de Qu\u00edmica, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad de M\u00e9xico 04510, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2394-0524","authenticated-orcid":false,"given":"Juan L.","family":"Monribot-Villanueva","sequence":"additional","affiliation":[{"name":"Laboratorio de Qu\u00edmica de Productos Naturales, Red de Estudios Moleculares Avanzados, Cl\u00faster BioMimic<sup>\u00ae<\/sup>, Instiuto de Ecolog\u00eda, A. C. Xalapa, Veracruz 91073, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6692-8667","authenticated-orcid":false,"given":"Jos\u00e9 B.","family":"Rodr\u00edguez-Haas","sequence":"additional","affiliation":[{"name":"Laboratorios de Fitopatolog\u00eda y Biolog\u00eda Molecular, Red de Estudios Moleculares Avanzados, Cl\u00faster BioMimic<sup>\u00ae<\/sup>, Instituto de Ecolog\u00eda, A. C. Xalapa, Veracruz 91073, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose Abel","family":"L\u00f3pez-Buenfil","sequence":"additional","affiliation":[{"name":"Colegio de Postgraduados, Carretera M\u00e9xico-Texcoco Km 36.5 Texcoco, Estado de M\u00e9xico 56101, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clemente J.","family":"Garc\u00eda-\u00c1vila","sequence":"additional","affiliation":[{"name":"Centro Nacional de Referencia Fitosanitaria del Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria, Tec\u00e1mac, Estado de M\u00e9xico 55740, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1416-4973","authenticated-orcid":false,"given":"Eliel","family":"Ruiz-May","sequence":"additional","affiliation":[{"name":"Laboratorio de Prote\u00f3mica, Red de Estudios Moleculares Avanzados, Cl\u00faster BioMimic<sup>\u00ae<\/sup>, Instituto de Ecolog\u00eda, A. C. Xalapa, Veracruz 91073, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3206-0900","authenticated-orcid":false,"given":"Diana","family":"S\u00e1nchez-Rangel","sequence":"additional","affiliation":[{"name":"Laboratorios de Fitopatolog\u00eda y Biolog\u00eda Molecular, Red de Estudios Moleculares Avanzados, Cl\u00faster BioMimic<sup>\u00ae<\/sup>, Instituto de Ecolog\u00eda, A. C. Xalapa, Veracruz 91073, Mexico"},{"name":"C\u00e1tedra CONACyT en la Red de Estudios Moleculares Avanzados del Instituto de Ecolog\u00eda, A. C. 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Bot."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"339","DOI":"10.3390\/insects3010339","article-title":"Ecological and Evolutionary Determinants of Bark Beetle -Fungus Symbioses","volume":"3","author":"Six","year":"2012","journal-title":"Insects"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1146\/annurev-ento-011019-024910","article-title":"Ecology and Evolution of Insect\u2013Fungus Mutualisms","volume":"65","author":"Biedermann","year":"2020","journal-title":"Annu. Rev. Entomol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1146\/annurev-ento-031616-035105","article-title":"The Ambrosia Symbiosis: From Evolutionary Ecology to Practical Management","volume":"62","author":"Hulcr","year":"2017","journal-title":"Annu. Rev. Entomol."},{"key":"ref_5","unstructured":"Eskalen, A. (2019, March 01). Invasive shot-hole borer and Fusarium dieback. Available online: http:\/\/eskalenlab.ucr.edu."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Gomez, D.F., Skelton, J., Steininger, M.S., Stouthamer, R., Rugman-Jones, P., Sittichaya, W., Rabaglia, R.J., and Hulcr, J. (2018). Species Delineation Within the Euwallacea fornicatus (Coleoptera: Curculionidae) Complex Revealed by Morphometric and Phylogenetic Analyses. Insect Syst. Divers., 2.","DOI":"10.1093\/isd\/ixy018"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Smith, S., Gomez, D., Beaver, R., Hulcr, J., and Cognato, A. (2019). Reassessment of the Species in the Euwallacea fornicatus (Coleoptera: Curculionidae: Scolytinae) Complex after the Rediscovery of the \u201cLost\u201d Type Specimen. Insects, 10.","DOI":"10.3390\/insects10090261"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1070","DOI":"10.1094\/PDIS-03-12-0276-PDN","article-title":"First report of a Fusarium sp. and its vector tea shot hole borer (Euwallacea fornicatus) causing fusarium dieback on avocado in California","volume":"96","author":"Eskalen","year":"2012","journal-title":"Plant Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1653\/024.099.0335","article-title":"First Report of Euwallacea nr.  fornicatus (Coleoptera: Curculionidae) in Mexico","volume":"99","author":"Carrillo","year":"2016","journal-title":"Florida Entomol."},{"key":"ref_10","unstructured":"SENASICA (2019). Complejo escarabajo barrenador pol\u00edfago Euwallacea sp.\u2013Fusarium euwallaceae. Serv. Nac. Sanidad, Inocuidad y Calid. Agroaliment. Gen. Sanid. Veg. Vigil. Epidemiol\u00f3gica Fitosanit, 62, 1\u201320."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1094\/PDIS-07-17-1042-RE","article-title":"Two novel fungal symbionts Fusarium kuroshium sp. Nov. and Graphium kuroshium sp. nov. of kuroshio shot hole borer (Euwallacea sp. nr. fornicatus) cause fusarium dieback on woody host species in California","volume":"102","author":"Na","year":"2018","journal-title":"Plant Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1094\/PDIS-01-19-0201-RE","article-title":"Probe-Based Multiplex Real-Time PCR as a Diagnostic Tool to Distinguish Distinct Fungal Symbionts Associated with Euwallacea kuroshio and Euwallacea whitfordiodendrus in California","volume":"104","author":"Carrillo","year":"2019","journal-title":"Plant Dis."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Avery, P.B., Bojorque, V., G\u00e1mez, C., Duncan, R.E., Carrillo, D., and Cave, R.D. (2018). Spore Acquisition and Survival of Ambrosia Beetles Associated with the Laurel Wilt Pathogen in Avocados after Exposure to Entomopathogenic Fungi. Insects, 9.","DOI":"10.3390\/insects9020049"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1146\/annurev-micro-092412-155650","article-title":"Fusarium pathogenomics","volume":"67","author":"Ma","year":"2013","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Perincherry, L., Lalak-K\u00e1nczugowska, J., and Stepi\u00e9n, L. (2019). Fusarium-produced mycotoxins in plant-pathogen interactions. Toxins, 11.","DOI":"10.3390\/toxins11110664"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e00881-17","DOI":"10.1128\/genomeA.00881-17","article-title":"Draft Genome Sequence of the Phytopathogenic Fungus Fusarium euwallaceae, the Causal Agent of Fusarium Dieback","volume":"5","year":"2017","journal-title":"Genome Announc."},{"key":"ref_17","first-page":"1","article-title":"Environmental pH modulates transcriptomic responses in the fungus Fusarium sp. associated with KSHB Euwallacea sp. near fornicatus","volume":"19","year":"2018","journal-title":"BMC Genom."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s11306-007-0082-2","article-title":"Proposed minimum reporting standards for chemical analysis Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI)","volume":"3","author":"Sumner","year":"2007","journal-title":"Metabolomics"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Meir, Z., and Osherov, N. (2018). Vitamin Biosynthesis as an Antifungal Target. J. Fungi., 4.","DOI":"10.3390\/jof4020072"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Qian, C.-D., Fu, Y.-H., Jiang, F.-S., Xu, Z.-H., Cheng, D.-Q., Ding, B., Gao, C.-X., and Ding, Z.-S. (2014). Lasiodiplodia sp. ME4-2, an endophytic fungus from the floral parts of Viscum coloratum, produces indole-3-carboxylic acid and other aromatic metabolites. BMC Microbiol., 14.","DOI":"10.1186\/s12866-014-0297-0"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4039","DOI":"10.1128\/aem.62.11.4039-4043.1996","article-title":"Production of fusaric acid by Fusarium species","volume":"62","author":"Bacon","year":"1996","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1023\/A:1014946217539","article-title":"Characterization of gibberellin producing strains of Fusarium moniliforme based on DNA polymorphism","volume":"153","author":"Mitter","year":"2002","journal-title":"Mycopathologia"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1128\/aem.55.1.254-255.1989","article-title":"Mycotoxin production by Fusarium oxysporum and Fusarium sporotrichioides isolated from Baccharis spp. from Brazil","volume":"55","author":"Mirocha","year":"1989","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Carbou\u00e9, Q., Maresca, M., Herbette, G., Roussos, S., Hamrouni, R., and Bombarda, I. (2019). Naphtho-Gamma-Pyrones Produced by Aspergillus tubingensis G131: New Source of Natural Nontoxic Antioxidants. Biomolecules, 10.","DOI":"10.3390\/biom10010029"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.4155\/fmc-2016-0053","article-title":"Sphingolipids as targets for treatment of fungal infections","volume":"8","author":"Singh","year":"2016","journal-title":"Future Med. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1016\/S0031-9422(98)80053-8","article-title":"Naphthoquinone metabolites of the fungi","volume":"47","author":"Medentsev","year":"1998","journal-title":"Phytochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1016\/S0040-4039(00)85496-9","article-title":"Novel trichothecenes from Fusarium sporotrichioides","volume":"27","author":"Corley","year":"1986","journal-title":"Tetrahedron Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1021\/jo01012a027","article-title":"Fonsecin, a Pigment from an Aspergillus fonsecaeus Mutant","volume":"30","author":"Galmarini","year":"1965","journal-title":"J. Org. Chem."},{"key":"ref_29","unstructured":"CFM-ID (2019, July 22). Competitive Fragmentation Modeling for Metabolite Identification 2020. Available online: https:\/\/cfmid.wishartlab.com."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1080\/02652030500058403","article-title":"Effects of trichothecene mycotoxins on eukaryotic cells: A review","volume":"22","author":"Rocha","year":"2005","journal-title":"Food Addit. Contam."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e02414-18","DOI":"10.1128\/AEM.02414-18","article-title":"Genetic differentiation associated with fumonisin and gibberellin production in Japanese Fusarium Fujikuroi","volume":"85","author":"Suga","year":"2019","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e0164310","DOI":"10.1371\/journal.pone.0164310","article-title":"Diversity in Secondary Metabolites Including Mycotoxins from Strains of Aspergillus Section nigri Isolated from Raw Cashew Nuts from Benin, West Africa","volume":"11","author":"Lamboni","year":"2016","journal-title":"PLoS ONE"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/2191-2858-2-6","article-title":"Seven naphtho-\u03b3-pyrones from the marine-derived fungus Alternaria alternata: Structure elucidation and biological properties","volume":"2","author":"Shaaban","year":"2012","journal-title":"Org. Med. Chem. Lett."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.femsle.2004.10.005","article-title":"Endophytic naphthopyrone metabolites are co-inhibitors of xanthine oxidase, SW1116 cell and some microbial growths","volume":"241","author":"Song","year":"2004","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1081","DOI":"10.1007\/s00253-014-6295-1","article-title":"Fungal naphtho-\u03b3-pyrones\u2014secondary metabolites of industrial interest","volume":"99","author":"Choque","year":"2015","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_36","first-page":"1","article-title":"How fungi defend themselves against microbial competitors and animal predators","volume":"14","year":"2018","journal-title":"PLoS Pathog."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Srivastava, L.M. (2002). Plant Growth and Development. Chapter 6\u2014Auxins, Academic Press.","DOI":"10.1016\/B978-012660570-9\/50147-7"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1016\/j.funbio.2016.06.002","article-title":"Indole-3-acetic acid in Fusarium graminearum: Identification of biosynthetic pathways and characterization of physiological effects","volume":"120","author":"Luo","year":"2016","journal-title":"Fungal Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1292","DOI":"10.1007\/s12010-012-0037-6","article-title":"Indole-3-Acetic Acid Biosynthesis in Fusarium delphinoides strain GPK, a causal agent of wilt in Chickpea","volume":"169","author":"Kulkarni","year":"2013","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.fgb.2012.05.010","article-title":"Identification of gene clusters associated with fusaric acid, fusarin, and perithecial pigment production in Fusarium verticillioides","volume":"49","author":"Brown","year":"2012","journal-title":"Fungal Genet. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1111\/mpp.12536","article-title":"Fusaric acid contributes to virulence of Fusarium oxysporum on plant and mammalian hosts","volume":"19","author":"Rahjoo","year":"2018","journal-title":"Mol. Plant Pathol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s40529-014-0066-2","article-title":"Fusaric acid induced cell death and changes in oxidative metabolism of Solanum lycopersicum L.","volume":"55","author":"Singh","year":"2014","journal-title":"Bot. Stud."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Wang, M., Ling, N., Dong, X., Liu, X., Shen, Q., and Guo, S. (2014). Effect of fusaric acid on the leaf physiology of cucumber seedlings. Eur. J. Plant Pathol., 138.","DOI":"10.1007\/s10658-013-0306-4"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1007\/s00425-013-1928-7","article-title":"Fusaric acid induction of programmed cell death modulated through nitric oxide signalling in tobacco suspension cells","volume":"238","author":"Jiao","year":"2013","journal-title":"Planta"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.plaphy.2017.06.028","article-title":"Role of fusaric acid in the development of \u2018Fusarium wilt\u2019 symptoms in tomato: Physiological, biochemical and proteomic perspectives","volume":"118","author":"Singh","year":"2017","journal-title":"Plant Physiol. Biochem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"913","DOI":"10.1111\/nph.16193","article-title":"Fusaric acid instigates the invasion of banana by Fusarium oxysporum f. sp. cubense TR4","volume":"225","author":"Liu","year":"2020","journal-title":"New Phytol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.foodchem.2019.01.136","article-title":"Endorsing and extending the repertory of nutraceutical and antioxidant sources in mangoes during postharvest shelf life","volume":"285","author":"Aluja","year":"2019","journal-title":"Food Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e86","DOI":"10.1002\/cpbi.86","article-title":"Using MetaboAnalyst 4.0 for Comprehensive and Integrative Metabolomics Data Analysis","volume":"68","author":"Chong","year":"2019","journal-title":"Curr. Protoc. Bioinforma."},{"key":"ref_49","unstructured":"METLIN (2019, December 05). Available online: https:\/\/metlin.scripps.edu."},{"key":"ref_50","unstructured":"MassBank Europe Mass Spectral DataBase (2019, December 08). Available online: https:\/\/massbank.eu."},{"key":"ref_51","unstructured":"FooDB (2019, November 10). Available online: https:\/\/foodb.ca."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"6487","DOI":"10.1021\/jf0709193","article-title":"Structure\u2212activity relationships of trichothecene toxins in an Arabidopsis thaliana leaf assay","volume":"55","author":"Desjardins","year":"2007","journal-title":"J. Agric. Food Chem."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Rueden, C.T., Schindelin, J., Hiner, M.C., DeZonia, B.E., Walter, A.E., Arena, E.T., and Eliceiri, K.W. (2017). ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinform., 18.","DOI":"10.1186\/s12859-017-1934-z"},{"key":"ref_54","unstructured":"R Core Team R (2019, May 09). The R Project for Statistical Computing 2013. Available online: https:\/\/www.r-project.org\/."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/4\/268\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:59:07Z","timestamp":1760363947000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/4\/268"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,9]]},"references-count":54,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["toxins13040268"],"URL":"https:\/\/doi.org\/10.3390\/toxins13040268","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,9]]}}}