Abstract
Determining the biomass of microbial plankton is central to the study of fluxes of energy and materials in aquatic ecosystems. This is typically accomplished by applying proper volume-to-carbon conversion factors to group-specific abundances and biovolumes. A critical step in this approach is the accurate estimation of biovolume from two-dimensional (2D) data such as those available through conventional microscopy techniques or flow-through imaging systems. This paper describes a simple yet accurate method for the assessment of the biovolume of planktonic microorganisms, which works with any image analysis system allowing for the measurement of linear distances and the estimation of the cross sectional area of an object from a 2D digital image. The proposed method is based on Archimedes’ principle about the relationship between the volume of a sphere and that of a cylinder in which the sphere is inscribed, plus a coefficient of ‘unellipticity’ introduced here. Validation and careful evaluation of the method are provided using a variety of approaches. The new method proved to be highly precise with all convex shapes characterised by approximate rotational symmetry, and combining it with an existing method specific for highly concave or branched shapes allows covering the great majority of cases with good reliability. Thanks to its accuracy, consistency, and low resources demand, the new method can conveniently be used in substitution of any extant method designed for convex shapes, and can readily be coupled with automated cell imaging technologies, including state-of-the-art flow-through imaging devices.
Highlights
The problem of estimating the biomass of microbial plankton is paramount to any ecological study concerned with fluxes of energy and materials in an aquatic ecosystem, and has been addressed many times [1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17]
This paper describes a simple yet accurate method for the assessment of the biovolume of planktonic microorganisms, which works with any image analysis system allowing for the measurement of linear distances and the estimation of the cross sectional area of an object from a 2D digital image
Display an approximate radial symmetry, and the typical conditions under which images are acquired allow approximating these cells by solids of revolution
Summary
The problem of estimating the biomass of microbial plankton is paramount to any ecological study concerned with fluxes of energy and materials in an aquatic ecosystem, and has been addressed many times [1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17]. Cell counts per se are inadequate as a measure of microbial biomass [18, 19] since in a microbial community, microorganism size spectrum encompasses several orders of magnitude, and even within lower taxa—such as genera and species—size variability may be significant. A volume-to-carbon relationship is applied to determine the amount of organic matter, in carbon units, to be ascribed to PLOS ONE | DOI:10.1371/journal.pone.0151955 May 19, 2016
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