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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Three in one: fixing marine nematodes for ecological, molecular, and morphological studies

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Author(s):
Fonseca, Gustavo [1] ; Fehlauer-Ale, Karin H. [1]
Total Authors: 2
Affiliation:
[1] Univ Sao Paulo, Ctr Biol Marinha, BR-11600000 Sao Paulo - Brazil
Total Affiliations: 1
Document type: Journal article
Source: LIMNOLOGY AND OCEANOGRAPHY-METHODS; v. 10, p. 516-523, JUL 2012.
Web of Science Citations: 13
Abstract

Multidisciplinary benthic studies are still hindered by the lack of a unique fixative that satisfactorily preserves morphology and DNA, and that is simultaneously adequate for ecological surveys. The objective of this study is to test the performance of five fixatives: formalin, ethanol, dimethylsulfoxide with EDTA and NaCl salts (DESS), methanol with acetic acid (METHAC), and ethanol with acetic acid (ETHAC), for the preservation of estuarine and exclusively marine nematode assemblages for morphological, molecular, and ecological studies. The presence of the stain rose bengal in each fixative was also evaluated in the yield of PCR reactions. For molecular analyses, one species of each habitat was considered. Results revealed that fixative performance for morphological studies is habitat-and species-dependent. For studies of estuarine sediment nematodes, we recommend the use of pure ethanol, because it caused little morphological distortion (<10% of the assemblage), preserved all the species for ecological studies, and yielded high quality DNA sequences. For studies of exclusively marine environments, METHAC or DESS are the most adequate. The first performed better for morphological and ecological surveys, whereas the second was more appropriate for molecular research. For ecological studies, DESS should be used in comparison with formalin, in order to cross check the results. Finally, staining of samples with rose bengal is not recommended, because it hindered DNA amplification regardless of the fixative used. (AU)

FAPESP's process: 09/14019-0 - Free living marine nematodes: taxonomy, systematics and phylogeny
Grantee:Gustavo Fernandes Camargo Fonseca
Support Opportunities: Research Grants - Young Investigators Grants