Contribution of major bacterial groups to bacterial biomass production (thymidine and leucine incorporation) in the Delaware estuary

Assimilation of 3HH-thymidine and 3HH-leucine was examined at the single-cell level using a combination of microautoradiography and fluorescent in situ hybridization (Micro-FISH) to determine the contribution of various bacterial groups to bacterial production in aquatic systems. All of the major phylogenetic groups of bacteria examined along the salinity gradient of the Delaware estuary, including alpha-, beta-, and gamma-proteobacteria and Cytophaga-like bacteria, assimilated 3H-thymidine and 3H-leucine. However, groups differed substantially in their contribution to the assimilation of these compounds. Alpha-proteobacteria were the dominant substrate-active bacteria at salinities of .9 PSU, whereas beta-proteobacteria were more important in freshwater. At all salinities, Cytophaga-like bacteria comprised the second most important group, and gamma-proteobacteria were overall the least important. Bacterial abundance explained about half of the variation in 3H-thymidine and 3H-leucine assimilation by the major bacterial groups. The sizes of silver grains of active bacteria indicate no difference in singlecell activity for the bacterial groups, suggesting that the average growth rates of the groups we examined were similar. However, activity per cell was distributed differently in the phylogenetic groups. Our study suggests that estimates of bacterial production measured using 3H-thymidine and 3H-leucine include bacteria in all of the major phylogenetic groups found in aquatic systems and that growth rates within bacterial groups vary substantially.

[1]  J. Fuhrman,et al.  Thymidine incorporation as a measure of heterotrophic bacterioplankton production in marine surface waters: Evaluation and field results , 1982 .

[2]  J. Fuhrman,et al.  Determination of Active Marine Bacterioplankton: a Comparison of Universal 16S rRNA Probes, Autoradiography, and Nucleoid Staining , 1997, Applied and environmental microbiology.

[3]  R. Neihof,et al.  Improved Microautoradiographic Method to Determine Individual Microorganisms Active in Substrate Uptake in Natural Waters , 1982, Applied and environmental microbiology.

[4]  P. D. Del Giorgio,et al.  Linking the physiologic and phylogenetic successions in free‐living bacterial communities along an estuarine salinity gradient , 2002 .

[5]  R. Amann,et al.  Culturability and In Situ Abundance of Pelagic Bacteria from the North Sea , 2000, Applied and Environmental Microbiology.

[6]  John C. Russ,et al.  The Image Processing Handbook , 2016, Microscopy and Microanalysis.

[7]  R. Amann,et al.  Comparison of Cellular and Biomass Specific Activities of Dominant Bacterioplankton Groups in Stratified Waters of the Celtic Sea , 2001, Applied and Environmental Microbiology.

[8]  J. Fuhrman,et al.  Microbial Desulfurization of a Crude Oil Middle-Distillate Fraction: Analysis of the Extent of Sulfur Removal and the Effect of Removal on Remaining Sulfur , 1999, Applied and Environmental Microbiology.

[9]  D. Kirchman The ecology of Cytophaga-Flavobacteria in aquatic environments. , 2002, FEMS microbiology ecology.

[10]  M. Cottrell,et al.  Natural Assemblages of Marine Proteobacteria and Members of the Cytophaga-Flavobacter Cluster Consuming Low- and High-Molecular-Weight Dissolved Organic Matter , 2000, Applied and Environmental Microbiology.

[11]  A. J. Ramsay The Use of Autoradiography to Determine the Proportion of Bacteria Metabolizing in an Aquatic Habitat , 1974 .

[12]  K. Schleifer,et al.  Phylogenetic Oligodeoxynucleotide Probes for the Major Subclasses of Proteobacteria: Problems and Solutions , 1992 .

[13]  R. Hodson,et al.  Leucine incorporation and its potential as a measure of protein synthesis by bacteria in natural aquatic systems , 1985, Applied and environmental microbiology.

[14]  J. Fuhrman Bacterioplankton Roles in Cycling of Organic Matter: The Microbial Food Web , 1992 .

[15]  R. Amann,et al.  Bacterioplankton Compositions of Lakes and Oceans: a First Comparison Based on Fluorescence In Situ Hybridization , 1999, Applied and Environmental Microbiology.

[16]  R. Amann,et al.  Growth Patterns of Two Marine Isolates: Adaptations to Substrate Patchiness? , 2001, Applied and Environmental Microbiology.

[17]  R. Amann,et al.  Changes in community composition during dilution cultures of marine bacterioplankton as assessed by flow cytometric and molecular biological techniques. , 2000, Environmental microbiology.

[18]  R. Amann,et al.  Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations , 1990, Applied and environmental microbiology.

[19]  M. Sieracki,et al.  Measurement of marine picoplankton cell size by using a cooled, charge-coupled device camera with image-analyzed fluorescence microscopy , 1992, Applied and environmental microbiology.

[20]  F. Azam,et al.  Protein content and protein synthesis rates of planktonic marine bacteria , 1989 .

[21]  R Amann,et al.  Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment. , 1996, Microbiology.

[22]  David C. Smith,et al.  A simple, economical method for measuring bacterial protein synthesis rates in seawater using 3H-leucine , 1992 .

[23]  R. Amann,et al.  Succession of Pelagic Marine Bacteria during Enrichment: a Close Look at Cultivation-Induced Shifts , 2000, Applied and Environmental Microbiology.

[24]  S. Giovannoni,et al.  Evolution, diversity, and molecular ecology of marine prokaryotes , 2000 .

[25]  T. Bouvier,et al.  Compositional changes in free‐living bacterial communities along a salinity gradient in two temperate estuaries , 2002 .

[26]  M. Cottrell,et al.  Carbon versus iron limitation of bacterial growth in the California upwelling regime , 2000 .

[27]  A. Entwistle,et al.  Confocal microscopy of surface‐ and cytoplasmically‐labelled bacteria immobilized by APS‐centrifugation , 1991 .

[28]  D. Hahn,et al.  Analysis of bacterial community structure in bulk soil by in situ hybridization , 1997, Archives of Microbiology.

[29]  B. Methé,et al.  Diversity of bacterial communities in Adirondack lakes : do species assemblages reflect lake water chemistry? , 1999 .

[30]  J. Novitsky,et al.  Microautoradiography-based enumeration of bacteria with estimates of thy-midine-specific growth and production rates , 1987 .

[31]  G. Budd Techniques of Autoradiography.Andrew W. Rogers , 1968 .

[32]  R. Amann,et al.  Seasonal Community and Population Dynamics of Pelagic Bacteria and Archaea in a High Mountain Lake , 1998, Applied and Environmental Microbiology.

[33]  Farooq Azam,et al.  Microbial Control of Oceanic Carbon Flux: The Plot Thickens , 1998, Science.

[34]  R. Amann,et al.  Linking the composition of bacterioplankton to rapid turnover of dissolved dimethylsulphoniopropionate in an algal bloom in the North Sea. , 2001, Environmental microbiology.