Diversity of microbial communities correlated to physiochemical parameters in a digestion basin of a zero-discharge mariculture system (original) (raw)
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Marine Ecology Progress Series, 2011
A culture-independent analysis of the bacterial assemblages in the sediments of a lugworm Arenicola marina L. bioturbated mesocosm was carried out using a molecular RNA-gene based approach. Clone libraries were constructed for 4 spatial locations in the mesocosm: deep yeast layer (DYL), mid sand (MS), new casts (NC) and old casts (OC) after 29 d of bioturbation. In total, 495 clones were analysed and grouped into 159 unique phylotypes. All libraries were dominated by Epsilonproteobacteria, with Alpha-, Delta-and Gammaproteobacteria, Verrucomicrobia and members of the Cytophaga-Flavobacteria-Bacteroides group also prevalent. Phyologenetic comparison of the sites with respect to evolutionary distance showed that the libraries fell into 2 groups: (1) DYL and MS and (2) NC and OC. There were significant differences in species composition between but not within the 2 groupings. Bacteria involved with sulphur cycling are most likely to be affected by lugworm bioturbation. An Epsilonproteobacterium phylotype with > 95% 16S similarity to Sulfurimonas denitrificans was abundant in the DYL sand layer but was not present in the egested sand (NC, OC). Promotion of the growth of sulphide-oxidising bacteria and their subsequent digestion would be consistent with the 'gardening' hypothesis and suggests that chemoautotrophic processes in shallow-water anaerobic sediments may be enhanced by the activities of the lugworm.
Systematic and Applied Microbiology, 1998
The phylogenetic and physiological diversity of sulphate-reducing bacteria inhabiting a salt marsh rhizosphere were investigated. Sulphate-reducing bacteria were isolated from a salt marsh rhizosphere using enrichment cultures with electron donors thought to be prevalent in the rhizosphere of Spartina alterniflora. The relationship between phylogeny and nutritional characteristics of 10 strains was investigated. None of the isolates had 16S rRNA sequences identical to other delta subclass sulphate-reducers, sharing 85.3 to 98.1 % sequence similarity with 16S rRNA sequences of their respective closest relatives. Phylogenetic analysis placed two isolates, obtained with ethanol as an electron donor, within the Desulfovibrionaceae. Seven isolates, obtained with acetate, butyrate, propionate, or benzoate, were placed within the Desulfobacteriaceae. One isolate, obtained with butyrate, fell within the Desulfobulbus assemblage, which is currently considered part of the Desulfobacteriaceae family. However, due to the phylogenetic breadth and physiological traits of this group, we propose that it be considered a new family, the "Desulfobulbusaceae." The isolates utilised an array of electron donors similar to their closest relatives with a few exceptions. As a whole, the phylogenetic and physiological data indicate isolation of several sulphate-reducing bacteria which might be considered as new species and representative of new genera. Comparison of the Desulfobacteriaceae isolates' 16S rRNA sequences to environmental clones originating from the same study site revealed that none shared more than 86% sequence similarity. The results provide further insight into the diversity of sulphate-reducing bacteria inhabiting the salt marsh ecosystem, as well as supporting general trends in the phylogenetic coherence of physiological traits of delta Proteobacteria sulphate reducers.
Diversity of Sulfur-Oxidizing Bacteria in Greenwater System of Coastal Aquaculture
Applied Biochemistry and Biotechnology, 2010
Reduced sulfur compounds produced by the metabolism are the one of the major problems in aquaculture. In the present study, herbivorous fishes have been cultured as biomanipulators for secretions of slime, which enhanced the production of greenwater containing beneficial bacteria. The genes encoding soxB which is largely unique to sulfuroxidizing bacteria (SOB) due to its hydrolytic function has been targeted for examining the diversity of SOB in the green water system of coastal aquaculture. Novel sequences obtained based on the sequencing of metagenomic clone libraries for soxB genes revealed the abundance of SOB in green water system. Phylogenetic tree constructed from aligned amino acid sequences demonstrated that different clusters have only 82-93% match with Roseobacter sp., Phaeobacter sp., Roseovarius sp., Sulfitobacter sp., Ruegeria sp., and Oceanibulbus sp. The level of conservation of the soxB amino acid sequences ranged from 42% to 71%. 16S rRNA gene analyses of enrichment culture from green water system revealed the presence of Pseudoxanthomonas sp., which has 97% similarity with nutritionally fastidious Indian strain of Pseudoxanthomonas mexicana-a sulfur chemolithotrophic γ-proteobacterium. Our results illustrate the relevance of SOB in the functioning of the green water system of coastal shrimp aquaculture for oxidation of reduced sulfur compounds, which in turn maintain the sulfide concentration well within the prescribed safe levels.
Sulfur bacteria in wastewater stabilization ponds periodically affected by the 'red-water'phenomenon
Applied Microbiology …, 2012
Several wastewater stabilization ponds (WSP) in Tunisia suffer periodically from the 'red-water' phenomenon due to blooming of purple sulfur bacteria, indicating that sulfur cycle is one of the main element cycles in these ponds. In this study, we investigated the microbial diversity of the El Menzeh WSP and focused in particular on the different functional groups of sulfur bacteria. For this purpose, we used denaturing gradient gel electrophoresis of PCR-amplified fragments of the 16S rRNA gene and of different functional genes involved in microbial sulfur metabolism (dsrB, aprA, and pufM). Analyses of the 16S rRNA revealed a relatively high microbial diversity where Proteobacteria, Chlorobi, Bacteroidetes, and Cyanobacteria constitute the major bacterial groups. The dsrB and aprA gene analysis revealed the presence of deltaproteobacterial sulfate-reducing bacteria (i.e., Desulfobacter and Desulfobulbus), while the analysis of 16S rRNA, aprA, and pufM genes assigned the sulfuroxidizing bacteria community to the photosynthetic representatives belonging to the Chlorobi (green sulfur bacteria) and the Proteobacteria (purple sulfur and non sulfur bacteria) phyla. These results point on the diversity of the metabolic processes within this wastewater plant and/or the availability of sulfate and diverse electron donors.
Abundance of sulphur-oxidizing bacteria in coastal aquaculture using soxB gene analyses
Aquaculture Research, 2010
Molecular techniques based on sequencing of metagenomic clone libraries provide an insight into the diversity of microbial populations. Using nucleic acid-based methods, the diversity of soxB genes was examined to detect and characterize sulphur-oxidizing bacteria in Indian coastal aquaculture environments. Gene-spe-ci¢c degenerate primers were used to amplify various fragments (710,753, 483^503, 280 and 239 bp) of soxB genes. Metagenomic clone libraries were constructed for 753, 483^503 and 239 bp fragments of soxB genes. The abundance of soxB revealed the presence of sulphur-oxidizing organisms. Amino acids in parts of the soxB-encoded proteins were aligned to known conserved amino acid residues. The level of conservation ranged from 23% to 30%. A phylogenetic tree constructed from aligned amino acid sequences of SoxB revealed di¡erent clusters associated with the branches of phototrophic aand g-proteobacteria. In general, soxB is widespread among the various phylogenetic groups, although this does not necessarily mean that the organism can use sulphur compounds. Our results suggest that the chemolithoautotrophy based on sulphur oxidation in coastal aquaculture is primarily sustained by the presence of sulphur oxidizers, which involve the soxB gene. This study aids identi¢cation of the phylogenetic characteristics related to sulphur bioremediation in poorly characterized coastal aquaculture environments.
Systematic and Applied Microbiology, 2008
In leather tanning industrial areas sulphide management represents a major problem. However, biological sulphide oxidation to sulphur represents a convenient solution to this problem. Elemental sulphur is easy to separate and the process is highly efficient in terms of energy consumption and effluent quality. As the oxidation process is performed by specialized bacteria, selection of an appropriate microbial community is fundamental for obtaining a good yield. Sulphur oxidizing bacteria (SOB) represent a wide-ranging and highly diversified group of microorganisms with the capability of oxidizing reduced sulphur compounds. Therefore, it is useful to select new microbes that are able to perform this process efficiently. For this purpose, an experimental membrane bioreactor for sulphide oxidation was set up, and the selected microbial community was characterized by constructing 16S rRNA gene libraries and subsequent screening of clones. Fluorescence in situ hybridization (FISH) was then used to assess the relative abundance of different bacterial groups. Sulphide oxidation to elemental sulphur proceeded in an efficient (up to 79% conversion) and stable way in the bioreactor. Both analysis of clone libraries and FISH experiments revealed that the dominant operational taxonomic unit (OTU) in the bioreactor was constituted by Gammaproteobacteria belonging to the Halothiobacillaceae family. FISH performed with the specifically designed probe tios_434 demonstrated that this OTU constituted 90.671.3% of the bacterial community. Smaller fractions were represented by bacteria belonging to the classes Betaproteobacteria, Alphaproteobacteria, Deltaproteobacteria, Clostridia, Mollicutes, Sphingobacteria, Bacteroidetes and Chlorobia. Phylogenetic analysis revealed that clone sequences from the dominant OTU formed a stable clade (here called the TIOS44 cluster), within the Halothiobacillaceae family, with sequences from many organisms that ARTICLE IN PRESS www.elsevier.de/syapm 0723-2020/$ -see front matter
Environmental Microbiology, 1999
In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization is a valuable tool for a more realistic assessment of SRB abundance in the natural environment. The distribution of SRB was investigated in a coastal marine sediment by hybridization of membrane-immobilized rRNA with oligonucleotide probes. As represented by general probe-target groups, SRB rRNA contributed between 18% and 25% to the prokaryotic rRNA pool. The dominant SRB were related to complete oxidizing genera (Desulphococcus, Desulphosarcina and Desulphobacterium), while Desulphobacter could not be detected. The vertical profile and quantity of rRNA from SRB was compared with sulphate reduction rates (SRR) measured with 35 SO 4 2¹ tracer in whole-core incubations. While SRB abundance was highest near the surface, peaking at around 1.5 cm, measured sulphate reduction rates were lowest in this region. A second peak of SRB rRNA was observed at the transition zone from oxidized to reduced sediment, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 × 10 8 cells cm ¹3 wet sediment. Cellular sulphate reduction rates calculated on the basis of these estimated cell numbers were between 0.01 and 0.09 fmol SO 4 2¹ cell ¹1 day ¹1 , which is below the rates that have been determined for pure cultures (0.2-50 fmol SO 4 2¹ cell ¹1 day ¹1 ) growing exponentially at nearoptimal temperature with a surplus of substrates.
Applied and Environmental Microbiology, 2010
New primers were designed for the amplification of dsrAB genes by nested PCR to investigate the diversity of sulfate-reducing prokaryotes (SRP) in environments with low bacterial cell density. The success of the nested PCR for the determination of SRP diversity was estimated by terminal-restriction fragment length polymorphism analysis in the Reigous, a small creek at an inactive mine (Carnoulès, France), which constitutes an extreme acidic arsenic-rich environment. Nested PCR limits were evaluated in dsrAB-rich sediments, and this technique was compared to direct PCR using either known primers (DSR1F/DSR4R) or new primers (dsr619AF/dsr1905BR). The comparison of clone libraries revealed that, even if the levels of diversity observed were not identical, nested PCR did not reduce the diversity compared to that of direct DSR1F/DSR4R PCR. Clone sequences were affiliated mainly with the Desulfobacteraceae and Desulfohalobiaceae families. Many sequences (ϳ30%) were related to a deeply branching lineage unaffiliated with any cultured SRP. Although this dsrAB cluster was found in all libraries, the new primers better amplified this lineage, providing more information on this unknown bacterial group. Thanks to these new primers in nested PCR, the SRP community from Carnoulès could be characterized. Specific SRP populations were obtained according to environmental characteristics. Desulfomicrobiaceae-related sequences were recovered in samples with low pH, low levels of dissolved oxygen, and high As content, while sequences belonging to the deeply branching group were found in a less extreme sample. Furthermore, for the first time, dsrAB sequences related to the latter group were recovered from freshwater.
Aquatic biosystems, 2013
Background: The Cariaco Basin is characterized by pronounced and predictable vertical layering of microbial communities dominated by reduced sulfur species at and below the redox transition zone. Marine water samples were collected in May, 2005 and, at the sampling stations A (10°30′ N, 64°40′ W), B (10°40′ N, 64°45′ W) and D (10°43'N, 64°32'W) from different depths, including surface, redox interface, and anoxic zones. In order to enrich for sulfate reducing bacteria (SRB), water samples were inoculated into anaerobic media amended with lactate or acetate as carbon source. To analyze the composition of enrichment cultures, we performed DNA extraction, PCR-DGGE, and sequencing of selected bands.