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. 2001 Dec;67(12):5585-92.
doi: 10.1128/AEM.67.12.5585-5592.2001.

Isolation, characterization, and polyaromatic hydrocarbon degradation potential of aerobic bacteria from marine macrofaunal burrow sediments and description of Lutibacterium anuloederans gen. nov., sp. nov., and Cycloclasticus spirillensus sp. nov

W K Chung  1 , G M King
Affiliations

Isolation, characterization, and polyaromatic hydrocarbon degradation potential of aerobic bacteria from marine macrofaunal burrow sediments and description of Lutibacterium anuloederans gen. nov., sp. nov., and Cycloclasticus spirillensus sp. nov

W K Chung et al. Appl Environ Microbiol. 2001 Dec.

Abstract

Two new polyaromatic hydrocarbon-degrading marine bacteria have been isolated from burrow wall sediments of benthic macrofauna by using enrichments on phenanthrene. Strain LC8 (from a polychaete) and strain M4-6 (from a mollusc) are aerobic and gram negative and require sodium chloride (>1%) for growth. Both strains can use 2- and 3-ring polycyclic aromatic hydrocarbons as their sole carbon and energy sources, but they are nutritionally versatile. Physiological and phylogenetic analyses based on 16S ribosomal DNA sequences suggest that strain M4-6 belongs to the genus Cycloclasticus and represents a new species, Cycloclasticus spirillensus sp. nov. Strain LC8 appears to represent a new genus and species, Lutibacterium anuloederans gen. nov., sp. nov., within the Sphingomonadaceae. However, when inoculated into sediment slurries with or without exogenous phenanthrene, only L. anuloederans appeared to sustain a significant phenanthrene uptake potential throughout a 35-day incubation. In addition, only L. anuloederans appeared to enhance phenanthrene degradation in heavily contaminated sediment from Little Mystic Cove, Boston Harbor, Boston, Mass.

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Figures

FIG. 1
FIG. 1
Time course of phenanthrene loss in N. virens burrow sediment slurries (○) and bulk surface sediment slurries (□). Arrows indicate the addition of phenanthrene. All data are means of results for triplicate determinations; error bars represent ±1 standard error.
FIG. 2
FIG. 2
Transmission electron micrographs of strain LC8 (A) and strain M4–6 (B) at ×15,500 magnification. Scale bars indicate 1 μm.
FIG. 3
FIG. 3
Phylogenetic tree based on maximum likelihood analysis of 16S rDNA gene sequences of strain LC8 and selected representatives of the Sphingomonadaceae and α-Proteobacteria. Numbers indicate the present bootstrap support for 100 resamplings of the data set.
FIG. 4
FIG. 4
Phylogenetic tree based on maximum likelihood analysis of 16S rDNA gene sequences of strain M4–6 and selected representatives of Cycloclasticus and the γ-Proteobacteria. Numbers indicate the percentages of bootstrap support for 100 resamplings of the data set.
FIG. 5
FIG. 5
Time course of phenanthrene concentrations in slurries of autoclaved Lowes Cove sediment (□), slurries containing no added strain LC8 (▵), and slurries incubated routinely with phenanthrene and inoculated with strain LC8 (○). All data are means of results for triplicate determinations; error bars represent ±1 standard error.
FIG. 6
FIG. 6
(A) Potential phenanthrene uptake rates for slurries from Lowes Cove (circles) and LMC (squares) inoculated with strain LC8 and incubated with (open symbols) or without (closed symbols) routine phenanthrene additions. All data are means of results for triplicate determinations; error bars represent ±1 standard error. (B) Potential phenanthrene uptake rates for uninoculated slurries from Lowes Cove incubated with (○) or without (●) routine phenanthrene additions and for autoclaved slurries (□). All data are means of results for triplicate determinations; error bars represent ±1 standard error.
FIG. 7
FIG. 7
Potential phenanthrene uptake rates for slurries from Lowes Cove (circles) and LMC (squares) inoculated with strain M4–6 and incubated with (open symbols) or without (closed symbols) routine phenanthrene additions. All data are means of results for triplicate determinations; error bars represent ±1 standard error.
FIG. 8
FIG. 8
Residual phenanthrene remaining in LMC sediment slurries incubated without routine phenanthrene additions and inoculated with strain LC8 (●) or strain M4–6 (■). Results for autoclaved control slurries (○) are shown, as well. All data are means of results for triplicate determinations; error bars represent ±1 standard error.
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