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Bosma, T. N. P.

Publications and source records attributed to Bosma, T. N. P..

3 recordsLinked to original sources

Organohalide respiration by a Desulforhopalus-dominated community

Despite the fact that several potential organohalide-respiring bacteria (OHRB) were discovered in metagenome-assembled genomes (MAGs) in our previous study of marine sediments from Aarhus Bay, delineation of their roles and interactions are yet to be disentangled. Henceforth, obtaining corresponding pure cultures or more defined consortia would be highly instrumental for more detailed eco-physiological studies. To this end, we isolated a colony from an anaerobic slant tube culture inoculated with a stable PCE dehalogenating enrichment. Intriguingly, the derived culture exhibited debromination only, instead of PCE dechlorination, under sulfate-reducing conditions. The culture was capable of conserving energy for growth via debromination of 2,6-dibromophenol (2,6-DBP). Analysis of 16S rRNA gene sequence data extracted from shot gun metagenome sequences revealed that a strain belonging to Desulforhopalus was the predominant member of the consortium at a relative abundance of 29 %. Moreover, five bins (completeness > 85% and contamination < 3%) were assembled and all were identified as potentially new species (average nucleotide identity, ANI < 95%). Two bins from potential OHRB, bin.3 belonging to Desulfoplanes, and bin.4 belonging to Marinifilaceae, were found to encode reductive dehalogenase (RDase) genes, whereas bin.5 was found to contain a gene coding for thiolytic tetrachloro-p-hydroquinone (TPh-) RDase bearing 23.4 % identity to TPh-RDase of Sphingobium chlorophenolicum. The expression of all three RDase genes was strongly-induced after adding 2,6-DBP. Acetylene, a known inhibitor of different redox-active metalloenzymes, was found to inhibit methanogenesis as well as reductive dehalogenation without affecting gene expression, suggesting post-transcriptional inhibition. Phylogenomic analyses revealed the ecological importance of complementary roles of community members, including complete de novo vitamin B12 biosynthesis, which agreed with physiological data. Altogether, the findings presented here provided insight into the mutualism of the consortium and provided leads for synthetic OHR community optimization strategies for in situ bioremediation.

microbiology↗

Genome-resolved transcriptomics reveals novel organohalide-respiring bacteria from Aarhus Bay sediments

Organohalide-respiring bacteria (OHRB) are keystone microbes in bioremediation of sites contaminated with organohalides and in natural halogen cycling. Known OHRB belong to distinct genera within the phyla Chloroflexota, Proteobacteria and Firmicutes, whereas information about novel OHRB mediating natural halogen cycling remains scarce. In this study, we applied a genome-resolved transcriptomic approach to characterize the identity and activity of OHRB from PCE-respiring cultures previously enriched from sediments of Aarhus Bay. Combining short- and long-read sequencing approaches, we assembled 37 high quality bins with over 75 % completeness and less than 5 % contamination. Sixteen bins harbored RDase genes, and were affiliated taxonomically to the class of Bacilli, and phyla of Bacteroidota, Synergistota, and Spirochaetota, that have not been reported to catalyze reductive dehalogenation. Among the 16 bins, bin.26, phylogenetically closely related to the genus Vulcanibacillus, contained an unprecedented 97 RDase genes. Of these, 84 RDase genes of bin.26 were transcribed during PCE dechlorination in addition to RDase genes from members of Synergistales (bin.15 and bin.32) and Bacteroidales (bin.18 and bin.24). Moreover, metatranscriptome analysis suggested the RDase genes were likely under the regulation of transcriptional regulators not previously associated with OHR, such as HrcA and SigW, which are known to respond to abiotic environmental stresses, such as temperature changes. Combined application of genomic methods enabled us to pinpoint novel OHRB from pristine environments not previously known to mediate reductive dechlorination and to provide evidence towards the diversity, activity and regulation of reductive dehalogenases.

microbiology↗

Metagenomic- and cultivation-based exploration of anaerobic chloroform biotransformation in hypersaline sediments as natural source of chloromethanes

Chloroform (CF) is an environmental contaminant that can be naturally formed in various environments ranging from forest soils to salt lakes. Here we investigated CF removal potential in sediments obtained from hypersaline lakes in Western Australia. Reductive dechlorination of CF to dichloromethane (DCM) was observed in enrichment cultures derived from sediments of Lake Strawbridge, which has been reported as a natural source of CF. The lack of CF removal in the abiotic control cultures without artificial electron donors indicated that the observed CF removal is a biotic process. Metabolite analysis with 13C labelled CF in the sediment-free enrichment cultures (pH 8.5, salinity 5%) revealed that increasing the vitamin B12 concentration from 0.04 to 4 M enhanced CF removal, reduced DCM formation, and increased 13CO2 production, which is likely a product of CF oxidation. Known organohalide-respiring bacteria and reductive dehalogenase genes were neither detected by quantitative PCR nor metagenomic analysis. Rather, members of the order Clostridiales, known to co-metabolically transform CF to DCM and CO2, were detected in the enrichment cultures. Genome-resolved metagenome analysis indicated that their genomes encode enzymatic repertoires for the Wood-Ljungdahl pathway and cobalamin biosynthesis that are known to be involved in co-metabolic CF transformation. ImportanceMore than 90% of the global CF emission to the atmosphere originates from natural sources, including saline environments such as salt lake sediments. However, knowledge about the microbial metabolism of CF in such extreme environments is lacking. Here we showed CF transformation potential in a hypersaline lake that was reported as a natural source of CF production. Application of interdisciplinary approaches of microbial cultivation, stable isotope labelling, and metagenomics aided in defining potential chloroform transformation pathways. This study indicates that microbiota may act as a filter to reduce CF emission from hypersaline lakes to the atmosphere, and expands our knowledge of halogen cycling in extreme hypersaline environments.

microbiology↗