title: Coelomocyte-derived fluorescence and DNA markers of composting earthworm species.
abstract: Supravital species identification of morphologically similar syntopic earthworms inhabiting dung and compost heaps or those from commercial cultures is difficult. The aim of the studies was to find out non-invasive species-specific markers for proper segregation of earthworm species from a dense mixed colony of waste decomposers. Worms were segregated according to external characteristics into Eisenia andrei, Eisenia fetida, and Dendrobaena veneta, and left for reproduction and analysis of non-invasively retrieved coelomocyte-containing coelomic fluid and/or species-specific partial sequences of cytochrome c oxidase subunit I (COI) gene in DNA extracted from amputated tail tips of adults and their offspring. Flow cytometric analysis of coelomocyte samples revealed that amount of nuclear DNA increases in order D. veneta ≪ E. andrei < E. fetida, and intensity of eleocyte-derived fluorescence is lower in D. veneta than in Eisenia spp. Spectrofluorimetry of coelomocyte lysates revealed that the amount of eleocyte-stored riboflavin is significantly lower in coelomocyte lysates from D. veneta than from Eisenia spp., and the emission peak of X-fluorophore is much more distinct in D. veneta than in Eisenia spp. Coelomic fluid of E. andrei exhibits a very distinct spectra of MUG fluorophore which are absent in D. veneta and in the majority of E. fetida, while some E. fetida possess MUG-like fluorophore. Sequences of the COI gene in the DNA of the worms from the mixed colony and their offspring confirmed species identity. In conclusion, species-specific coelomocyte-derived markers may be a useful complement to morphological and DNA-based taxonomy during studies on syntopic earthworms.
authors.affiliation: Institute of Environmental Engineering, Czestochowa University of Technology, Czestochowa, Poland; Laboratoire de Génie Civil et Géo-Environnement, University Lille Nord de France, Lille, France.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Rorat
authors.forename: Neli
authors.initials: N
authors.lastname: Kachamakova-Trojanowska
authors.forename: Alicja
authors.initials: A
authors.lastname: Jozkowicz
authors.forename: Jerzy
authors.initials: J
authors.lastname: Kruk
authors.forename: Claude
authors.initials: C
authors.lastname: Cocquerelle
authors.forename: Franck
authors.initials: F
authors.lastname: Vandenbulcke
authors.forename: Michal
authors.initials: M
authors.lastname: Santocki
authors.forename: Barbara
authors.initials: B
authors.lastname: Plytycz
mesh_terms: D000818:Animals; D001483:Base Sequence; D002965:Classification; D003576:Electron Transport Complex IV / Q000235:genetics*; D005453:Fluorescence; D005819:Genetic Markers / Q000235:genetics*; D009835:Oligochaeta / Q000235:genetics*; D013045:Species Specificity
chemical_list: D005819:Genetic Markers; D003576:Electron Transport Complex IV