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Lomheim, L.

Publications and source records attributed to Lomheim, L..

9 recordsLinked to original sources

Thermodynamically controlled biokinetics modelling of a lindane-degrading anaerobic enrichment culture

The anaerobic biotransformation of lindane ({gamma}-hexachlorocyclohexane), a persistent legacy pesticide and challenging subsurface contaminant was modelled using an integrated Monod biokinetics framework. Calibrated against batch experimental data, this model accounted for the dissolution of solid phase lindane and the thermodynamics of reactions involved. The study focused on a methanogenic, lindane-degrading enrichment culture (GT2) that used ethanol as the electron donor, where ethanol fermentation supplied hydrogen to support reductive dechlorination of lindane. A Dehalobacter strain was identified as the lindane-degrading organism, while a member of bacterial order Geobacterales carried out ethanol fermentation. First-order decay constants for lindane-degrading Dehalobacter were estimated from experimental data and showed variation across different growth conditions. The biokinetics model captured interactions among key microbial processes, namely lindane dechlorination, ethanol fermentation, hydrogenotrophic methanogenesis, and acetoclastic methanogenesis. Thirteen biokinetics parameters including maximum substrate utilization rates (k), half-velocity constants (Ks), and decay constants for these microbial processes were estimated through a Bayesian optimization approach. Global sensitivity analysis showed that eight parameters, primarily k and Ks, had significant linear impact on the model output, whereas responses to decay constants were more variable and potentially non-linear. This study proposed an effective modelling approach for characterizing syntrophic microbial growth within enrichment cultures and provides valuable biokinetics parameters estimates for real-world applications of anaerobic lindane bioremediation.

microbiology↗

A novel virus lineage is abundant in metaviromes from Dehalococcoides-containing mixed cultures

Dehalococcoides mccartyi are obligately anaerobic organohalide-respiring bacteria that play important roles in the detoxification of chlorinated pollutants in groundwater and sediments. Despite having small genomes, they host a diverse set of mobile elements. Here we characterize a family of mobile elements, termed integrative and mobilizable element 1 or IME1, comprising 20 from Dehalococcoides and one from Dehalogenimonas alkenigignens. IME1s are 20,930 - 28,058 bp and are found both integrated in the genomes and as circular episomes. Bioinformatic characterization of IME1 encoded proteins revealed a highly conserved structure with 14 hierarchical orthologous groups (HOGs) found in all 21 IME1s. IME1s lack recognizable hallmark proteins of tailed bacterial viruses (or tailed phages) but encode proteins with similarities to those of filamentous bacterial viruses. In particular, one conserved HOG shows sequence similarity to the pI-like ATPase, the only conserved marker protein identified across filamentous bacterial viruses. Additionally, IME1s encode several small proteins with predicted transmembrane domains and signal peptides, another feature used to identify filamentous bacterial viruses. Both features are also found in budding archaeal viruses of various morphotypes. IME1s dominated metaviromes obtained from the Dehalococcoides-containing KB-1 mixed culture and electron micrographs of the corresponding viral fractions revealed abundant filamentous virus-like particles. We therefore propose that the IME1s represent a novel lineage of double stranded budding, likely filamentous, viruses. Database searches suggest IME1s are found in Dehalococcodia and other Chlorofexota but are so far restricted to this phylum.

microbiology↗

Dehalobacter converts 2,3-dichloroaniline to aniline via 2-chloroaniline in a methanogenic enrichment culture

12,3-dichloroaniline (2,3-DCA) has widespread use in chemical manufacturing and remains a persistent groundwater contaminant. To better understand the pathway and kinetics of its reductive dechlorination, we conducted a laboratory kinetic experiment using an anaerobic enrichment culture dominated by Dehalobacter. At an initial field-relevant concentration of 40 mg/L, complete and stoichiometric dechlorination of 2,3-DCA to aniline via 2-CA was achieved. The intermediates, 2-chloroaniline (2-CA) and 3-chloroaniline, were transiently formed in a ratio of 8:1. The growth yields of Dehalobacter on 2,3-DCA and 2-CA were 1.2 {+/-} 0.1 x 108 and 1.3 {+/-} 0.1 x 108 16S rRNA gene copies/mol chloride, respectively. The maximum specific growth rate for 2,3-DCA, max, was 0.18 {+/-} 0.03 day-{superscript 1} with a half-saturation constant, Ks, at 45 {+/-} 16 mg /L. The first-order decay constant for Dehalobacter when starved of chlorinated electron acceptors was estimated at 0.017 {+/-} 0.001 day-{superscript 1}. Lactate fermentation, acetogenesis from ethanol, syntrophic propionate oxidation, and hydrogenotrophic methanogenesis were observed during dechlorination. This work provides insights into the organohalide respiration of 2,3-DCA to aniline and advances the understanding of microbial interactions during anaerobic dechlorination. These results offer guidance for developing stable dechlorinating microbial ecosystems and key kinetic parameters for predictive modeling of groundwater 2,3-DCA fate and transport. 3 SynopsisThis study demonstrated stoichiometric dechlorination of 2,3-dichloroaniline to aniline, characterized syntrophic interactions in a mixed anaerobic dechlorinating culture, and determined growth yield, first-order decay and Monod constants for Dehalobacter. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=141 SRC="FIGDIR/small/639126v3_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@5d59c4org.highwire.dtl.DTLVardef@b05a3aorg.highwire.dtl.DTLVardef@1f39f6borg.highwire.dtl.DTLVardef@1676753_HPS_FORMAT_FIGEXP M_FIG C_FIG Created in BioRender. Wang, S. (2025) https://BioRender.com/f47t993

microbiology↗

Dehalobacter dechlorinates dichloroanilines and contributes to the natural attenuation of dichloronitrobenzenes at a complex industrial site

The potential for bioremediation of dichloroanilines (2,3- and 3,4-DCA) and dichloronitrobenzenes (3,4-, 2,5- and 2,3-DCNB) was investigated using inocula from an industrial site in northeast of Brazil. Anaerobic biotransformation of these chlorinated compounds was observed in microcosms simulating site conditions, particularly when electron donor was added. To disentangle specific transformation reactions, sub-cultures from active anaerobic microcosms were enriched with individual DCA or DCNB isomers. In these single-compound enrichment cultures, DCNB isomers were reduced to the corresponding DCA. Each DCA isomer was stoichiometrically dechlorinated to the corresponding monochloroaniline (CA) isomers. Further dechlorination of CA to aniline was noted. The reduction of DCNB to DCA was sustained by known fermenters and possibly Desulfitobacterium, while Dehalobacter was clearly responsible for dechlorination of DCAs. These findings helped to attribute function to microbial community data retrieved from field samples to inform the conceptual site model and enable identification of bioactive zones and rate-limiting conditions at a complex site. More generally, these data broaden the understanding of the metabolic repertoire of Dehalobacter and provide a model for incorporating microbial community data into site remediation efforts. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/638192v2_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@1667948org.highwire.dtl.DTLVardef@a4a128org.highwire.dtl.DTLVardef@507067org.highwire.dtl.DTLVardef@103a155_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical AbstractC_FLOATNO C_FIG Short synopsis statementA diverse microbial community contributes to natural attenuation at a complex contaminated site. Notably, dichloroanilines produced by biologically-mediated nitroreduction of dichloronitrobenzenes were dechlorinated by Dehalobacter.

microbiology↗

Identification of a fully dechlorinated product of chlordecone in soil microcosms and enrichment cultures

Anaerobic microcosms constructed with soil from Guadeloupe, amended with electron donor (ethanol and acetone) and incubated for more than a decade, transformed chlordecone (CLD) into a suite of progressively more dechlorinated products, including a fully dechlorinated carboxylated indene product. This fully dechlorinated transformation product has never before been observed and indicates that complete dechlorination of CLD is possible. The carboxylated indene was detected by LC-MS and structure was confirmed by LC-MS/MS using a Q-Exactive Orbitrap mass spectrometer.

microbiology↗

Biotransformation of lindane (γ-hexachlorocyclohexane) to non-toxic end products by sequential treatment with three mixed anaerobic microbial cultures

The {gamma} isomer of hexachlorocyclohexane (HCH), also known as lindane, is a carcinogenic persistent organic pollutant. Lindane was used worldwide as an agricultural insecticide. Legacy soil and groundwater contamination with lindane and other HCH isomers is still a big concern. The biotic reductive dechlorination of HCH to non-desirable and toxic lower chlorinated compounds such as monochlorobenzene (MCB) and benzene, among others, has been broadly documented. Here, we demonstrate for the first time that complete biotransformation of lindane to non-toxic end products is attainable using a sequential treatment approach with three mixed anaerobic microbial cultures referred to as culture I, II, and III. Biaugmentation with culture I achieved dechlorination of lindane to MCB and benzene. Culture II was able to dechlorinate MCB to benzene, and finally, culture III carried out methanogenic benzene degradation. Distinct Dehalobacter populations, corresponding to different 16S rRNA amplicon sequence variants in culture I and culture II, were responsible for lindane and MCB dechlorination, respectively. This study continues to highlight key roles of Dehalobacter spp. as chlorobenzene- and HCH-organohalide-respiring bacteria and demonstrates that sequential treatment with specialized anaerobic cultures may be explored at field sites in order to address legacy soil and groundwater contamination with HCH. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=82 SRC="FIGDIR/small/354597v2_ufig1.gif" ALT="Figure 1"> View larger version (11K): org.highwire.dtl.DTLVardef@16ebd4corg.highwire.dtl.DTLVardef@14dfa8dorg.highwire.dtl.DTLVardef@7c5637org.highwire.dtl.DTLVardef@100634a_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

Microbial Communities Associated with Sustained Anaerobic Reductive Dechlorination of α-, β-, γ-, and δ-Hexachlorocyclohexane Isomers to Monochlorobenzene and Benzene

Intensive historical and worldwide use of the persistent pesticide technical-grade hexachlorocyclohexane (HCH), composed of the active ingredient {gamma}-HCH (called lindane) along with several other HCH isomers, has led to widespread contamination. We derived four anaerobic enrichment cultures from HCH-contaminated soil capable of sustainably dechlorinating each of -, {beta}-, {gamma}-, and {delta}-HCH isomers stoichiometrically and completely to benzene and monochlorobenzene (MCB). For each isomer, the dechlorination rates increased progressively from <3 {micro}M/day to [~]12 {micro}M/day over two years. The molar ratio of benzene to MCB produced was a function of the substrate isomer, and ranged from {beta} (0.77{+/-}0.15), (0.55{+/-}0.09), {gamma} (0.13{+/-}0.02) to {delta} (0.06{+/-}0.02) in accordance with pathway predictions based on prevalence of antiperiplanar geometry. Cultivation with a different HCH isomer resulted in distinct bacterial communities, but similar archaeal communities. Data from 16S rRNA gene amplicon sequencing and quantitative PCR revealed significant increases in the absolute abundance of Pelobacter and Dehalobacter, especially in the -HCH and {delta}-HCH cultures. This study provides the first direct comparison of shifts in anaerobic microbial communities induced by the dechlorination of distinct HCH isomers. It also uncovers candidate microorganisms responsible for the dechlorination of -, {beta}-, {gamma}-, and {delta}-HCH, a key step towards better understanding and monitoring of natural attenuation processes and improving bioremediation technologies for HCH-contaminated sites.

microbiology↗

Evidence for extensive anaerobic dechlorination and transformation of the pesticide chlordecone (C10Cl10O) in soil from Guadeloupe

Chlordecone (C10Cl10O) is a bishomocubane molecule, that has been used as pesticide, in many countries in Europe, America, and Africa, from the 1960s to 1990s. In the French West Indies, the historic use of chlordecone to control banana weevil infestations has resulted in pollution of large land areas. Although currently banned, chlordecone persists because it adsorbs strongly to soil and its complex structure is stable, particularly under aerobic conditions. A leaching model established that CLD pollution will last in French west indies soils several decades to half a millennium depending on soil type. However, abiotic chemical transformation catalyzed by reduced vitamin B12 has been shown to break down chlordecone by opening the cage structure to produce C9 polychloroindenes, and more recently these C9 polychloroindenes were also observed as products of anaerobic microbiological transformation by Citrobacter. To assess the potential for bioremediation, the anaerobic biotransformation of chlordecone by microbes native to soils from the French West Indies was investigated. Anaerobic microcosms were constructed from chlordecone impacted Guadeloupe soil and sludge to mimic natural attenuation and eletron donor-stimulated reductive dechlorination. Original microcosms and transfers were incubated over a period of 8 years, during which they were repeatedly amended with chlordecone and electron donor (ethanol and acetone). Using LC/MS, chlordecone and degradation products were detected in all the biologically active microcosms. Observed products in active incubations included monohydro-, dihydro- and trihydrochlordecone derivatives (C10Cl10-nO2Hn, n= 1,2,3), as well as \"open cage\" C9 polychloroindene compounds (C9Cl5-nH3+n, n=0,1,2) and C10 carboxylated polychloroindene derivatives (C10Cl4-nO2H4+n, n=0-3). Products with as many as 9 chlorine atoms removed were detected. These products were not observed in sterile incubations. Chlordecone concentrations decreased in active microcosms as concentrations of products increased, indicating that anaerobic dechlorination processes have occurred. An crude estimation of partitioning coefficients between soil and water showed that carboxylated intermediates sorb poorly, and as a consequence may be flushed away while polychlorinated indenes sorb strongly to soil. Microbial community analysis in microcosms showed enrichment of anaerobic fermenting and acetogenic microbes possibly involved in anaerobic chlordecone biotransformation. It thus should be possible to stimuilate anaerobic dechlorination through donor amendment to contaminated soils, particularly as some metabolites (in particular pentachloroindene) were already detected in field samples as a result of intrinsic processes. Extensive dechlorination in the microcosms, with evidence for up to 9 Cl atoms removed from the parent molecule is game-changing, giving hope to the possibility of using bioremediation to reduce the impact of CLD contamination.

microbiology↗

Eight new genomes of organohalide-respiring Dehalococcoides mccartyi reveal evolutionary trends in reductive dehalogenase enzymes

BackgroundBioaugmentation is now a well-established approach for attenuating toxic groundwater and soil contaminants, particularly for chlorinated ethenes and ethanes. The KB-1 and WBC-2 consortia are cultures used for this purpose. These consortia contain organisms belonging to the Dehalococcoidia, including strains of Dehalococcoides mccartyi in KB-1 and of both D. mccartyi and Dehalogenimonas in WBC-2. These tiny anaerobic bacteria couple respiratory reductive dechlorination to growth and harbour multiple reductive dehalogenase genes (rdhA) in their genomes, the majority of which have yet to be characterized.\n\nResultsUsing a combination of Illumina mate-pair and paired-end sequencing we closed the genomes of eight new strains of Dehalococcoides mccartyi found in three related KB-1 sub-cultures that were enriched on trichloroethene (TCE), 1,2-dichloroethane (1,2-DCA) and vinyl chloride (VC), bringing the total number of genomes available in NCBI to 24. A pangenome analysis was conducted on 24 Dehalococcoides genomes and five Dehalogenimonas genomes (2 in draft) currently available in NCBI. This Dehalococcoidia pangenome generated 2875 protein families comprising of 623 core, 2203 accessory, and 49 unique protein families. In Dehalococcoides mccartyi the complement of reductive dehalogenase genes varies by strain, but what was most surprising was how the majority of rdhA sequences actually exhibit a remarkable degree of synteny across all D. mccartyi genomes. Several homologous sequences are also shared with Dehalogenimonas genomes. Nucleotide and predicted protein sequences for all reductive dehalogenases were aligned to begin to decode the evolutionary history of reductive dehalogenases in the Dehalococcoidia.\n\nConclusionsThe conserved synteny of the rdhA genes observed across Dehalococcoides genomes indicates that the major differences between strain rdhA gene complement has resulted from gene loss rather than recombination. These rdhA have a long evolutionary history and trace their origin in the Dehalococcoidia prior to the speciation of Dehalococcoides and Dehalogenimonas. The only rdhA genes suspected to have been acquired by lateral gene transfer are protein-coding rdhA that have been identified to catalyze dehalogenation of industrial pollutants. Sequence analysis suggests that evolutionary pressures resulting in new rdhA genes involve adaptation of existing dehalogenases to new substrates, mobilization of rdhA between genomes or within a genome, and to a lesser degree manipulation of regulatory regions to alter expression.

bioinformatics↗