Biancalana, Florencia, Kopprio, Germán A., Lara, Rubén J. and Alonso, Cecilia (2017) A protocol for the simultaneous identification of chitin-containing particles and their associated bacteria. Systematic and Applied Microbiology, 40 (5). pp. 314-320. DOI

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Chitin is the second most abundant polymer on Earth, playing a crucial role in the biogeochemical cycles. A core issue for studying its processing in aquatic systems is the identification and enumeration of chitin-containing particles and organisms, ideally in a manner that can be directly linked to bulk chitin quantification. The aim of this study was the development of such a technique. We successfully combined the methodology of bulk chitin determination using wheat germ agglutinin (FITC-WGA) for staining chitin-containing particles and organisms along with CARD-FISH staining of either chitin-containing eukaryotic cells or bacteria associated with them. Environmental chitin staining was successfully applied to natural water samples. Fungal hyphae, diatoms, and dinoflagellates, sestonic aggregates and chitin-containing structures derived from metazoa were observed. Also, hybridized bacteria attached to chitinaceous debris were clearly visualized. Finally, as proof of principle, cultured yeast cells were simultaneously-targeted by FITC-WGA and the EUK516 probe without exhibiting any interference between both stains. The presented approach appears as a powerful tool to evaluate the contribution of different size classes and organisms to chitin production and consumption, opening the possibility for application of single-cell approaches targeting the ecophysiology of chitin transformations in aquatic systems.

Document Type: Article
Programme Area: UNSPECIFIED
Research affiliation: Biogeochemistry and Geology > Tropical Marine Microbiology
Refereed: Yes
Open Access Journal?: No
ISSN: 07232020
Date Deposited: 29 Mar 2019 12:08
Last Modified: 01 Oct 2020 12:58

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