Detection and enumeration of microbial cells within highly porous calcareous reef sands
Christian Wild A D F G , Christian Laforsch B E and Markus Huettel CA Max Planck Institute for Marine Microbiology, Celsiusstr. 1, 28359 Bremen, Germany.
B Biological Sciences Department, IENS, Lancaster University, Lancaster, LA1 4YQ, UK.
C Department of Oceanography, Florida State University, Tallahassee, FL 32306-4320, USA.
D Ocean Sciences Section, Intergovernmental Oceanographic Commission, UNESCO, 1 rue Miollis, 75015 Paris, France.
E Department Biology II, Ludwig-Maximilians University, Großhadernerstr. 2, 82152 Martinsried, Germany.
F Coral Reef Ecology Working Group, GeoBio-Center, Ludwig-Maximilians University, Richard-Wagner-Str. 10, 80333 Muenchen, Germany.
G Corresponding author. Email: c.wild@lrz.uni-muenchen.de
Marine and Freshwater Research 57(4) 415-420 https://doi.org/10.1071/MF05205
Submitted: 13 October 2005 Accepted: 21 March 2006 Published: 14 June 2006
Abstract
In order to assess and to compare the abundances of prokaryotes in coral sands from three different areas in the Indo-Pacific, a technique was developed and evaluated for enumeration of prokaryotes living on and within calcareous grains. Propidium iodide labelling of prokaryotes and consecutive confocal laser scanning microscopy showed microbial colonisation within pores and small fissures of the coral sands. This embedded microbial colonisation required at least four extractions with weak acetic acid to dissolve the grain surface layer in order to detach 97% of the prokaryotic cells. Microbial enumeration based on this technique revealed that the abundance of prokaryotes in the carbonate sands were not significantly different among the three sites, but were about one order of magnitude higher than reported for silicate sands of a similar grain size spectrum. A possible reason for this high abundance of prokaryotes is the complex surface structure of the biogenic calcareous grains, their correspondingly highly porous matrix and the associated ability of prokaryotes to penetrate into carbonate grains. Our results highlight the role of calcareous reef sands as a substratum with a large specific surface area for prokaryotic colonisation and emphasise the contribution of calcium carbonate reef sands for element cycles in subtropical and tropical ecosystems.
Extra keywords: acetic acid treatment, calcareous sands, coral reef sediments, embedded microbial cells, extraction method, microspatial distribution, prokaryotes abundance.
Acknowledgments
We thank B.B. Jørgensen for his support of this work and H. Schuette, L. Hönemann and V. Epple for help with the sample analyses. Thanks are also due to B. Cragg from University of Bristol for advice, J. Dent from University of Lancaster for assistance with the CLSM facilities and A. Rusch as well as two anonymous reviewers for helping to improve the manuscript.
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