Haloalkane-utilizing Rhodococcus strains isolated from geographically distinct locations possess a highly conserved gene cluster encoding haloalkane catabolism

The sequences of the 16S rRNA and haloalkane dehalogenase (dhaA) genes of five gram-positive haloalkane-utilizing bacteria isolated from contaminated sites in Europe, Japan, and the United States and of the archetypal haloalkane-degrading bacterium Rhodococcus sp. strain NCIMB13064 were compared. The 16S rRNA gene sequences showed less than 1% sequence divergence, and all haloalkane degraders clearly belonged to the genus Rhodococcus. All strains shared a completely conserved dhaA gene, suggesting that the dhaA genes were recently derived from a common ancestor. The genetic organization of the dhaA gene region in each of the haloalkane degraders was examined by hybridization analysis and DNA sequencing. Three different groups could be defined on the basis of the extent of the conserved dhaA segment. The minimal structure present in all strains consisted of a conserved region of 12.5 kb, which included the haloalkane-degradative gene cluster that was previously found in strain NCIMB13064. Plasmids of different sizes were found in all strains. Southern hybridization analysis with a dhaA gene probe suggested that all haloalkane degraders carry the dhaA gene region both on the chromosome and on a plasmid (70 to 100 kb). This suggests that an ancestral plasmid was transferred between these Rhodococcus strains and subsequently has undergone insertions or deletions. In addition, transposition events and/or plasmid integration may be responsible for positioning the dhaA gene region on the chromosome. The data suggest that the haloalkane dehalogenase gene regions of these gram-positive haloalkane-utilizing bacteria are composed of a single catabolic gene cluster that was recently distributed world-wide

Biostimulation of jarosite and iron oxide-bearing mine waste enhances subsequent metal recovery

Novel resource recovery technologies are required for metals-bearing hazardous wastes in order to achieve circular economy outcomes and industrial symbiosis. Iron oxide and co-occurring hydroxysulphate-bearing wastes are globally abundant and often contain other elements of value. This work addresses the biostimulation of indigenous microbial communities within an iron oxide/ hydroxysulphate-bearing waste and its effect on the subsequent recoverability of metals by hydrochloric, sulphuric, citric acids, and EDTA. Laboratory-scale flow-through column reactors were used to examine the effect of using glycerol (10% w/w) to stimulate the in situ microbial community in an iron oxide/ hydroxysulphate-bearing mine waste. The effects on the evolution of leachate chemistry, changes in microbiological community, and subsequent hydrometallurgical extractability of metals were studied. Results demonstrated increased leachability and selectivity of Pb, Cu, and Zn relative to iron after biostimulation with a total of 0.027kg of glycerol per kg of waste. Biostimulation, which can be readily applied in situ, potentially opens new routes to metal recovery from globally abundant waste streams that contain jarosite and iron oxides

Genome sequences of two choline-utilising methanogenic archaea, Methanococcoides spp., isolated from marine sediments

The genomes of two Methanococcoides spp. that were isolated from marine sediments and are capable of carrying out methanogenesis from choline and other methylotrophic substrates were sequenced. The average nucleotide identity and in silico DNA-DNA hybridization analyses demonstrate that they represent species different from those previously described

The influence of migratory Paraburkholderia on growth and competition of wood-decay fungi

Certain bacteria are capable of migrating along fungal hyphae, using them as a dispersal mechanism to cross otherwise-prohibitory distances. Three strains of fungal-migratory Paraburkholderia were isolated from the mycelium of wood-decay fungi, and inoculated onto ten strains of wood-decay fungi growing on solid agar medium. Two of the three bacteria were able to migrate along the hyphae of all fungi, although to differing extents. No bacteria-associated growth inhibition was observed with eight of the ten fungi, but mycelial extension rate of two strains of Phanerochaete was significantly reduced. Bacteria were also introduced into fungus-fungus competitive pairings, and significantly reduced the competitive performance of one of the Phanerochaete strains. Additionally, in several cases, introducing bacteria into competitive interactions made the outcome unpredictable, whereas in the absence of bacteria one fungus was consistently dominant. This is the first time that bacteria have been shown to influence fungal inter-specific competition

Fungal control of early-stage bacterial community development in decomposing wood

The earliest stages of bacterial colonisation of wood have received little attention, particularly with respect to how the colonisation process may be affected by the presence of wood-decay fungi. This study used 16s rRNA gene sequencing to examine the bacterial community in wood that had been incubated in the field for 14 or 84 d, either in wood uncolonised by fungi or pre-colonised by Vuilleminia comedens, Trametes versicolor or Hypholoma fasciculare. All three fungal species significantly delayed bacterial colonisation of the wood. V. comedens and H. fasciculare also reduced bacterial OTU richness and altered bacterial community composition, increasing the relative abundance of Burkholderiales and reducing the proportion of Enterobacteriaceae and Bacteroidetes. Wood that had not been pre-colonised showed seasonal differences between autumn and spring: bacterial richness increased between 14 d and 84 d in the spring, but not in the autumn. Community composition at 84 d in spring was also different to the other time points, with reduced dominance of Gamma-proteobacteria. Archaea were also detected in nearly a third of samples, but with no apparent pattern, and always at low abundances

Towards passive bioremediation of dye-bearing effluents using hydrous ferric oxide wastes: Mechanisms, products and microbiology

A novel, circular economy-inspired approach for the “passive” (non-powered and reagent-free) treatment of dye-bearing effluent is presented. The treatment utilises the biogeochemical interaction of dye-bearing wastewater with hydrous ferric oxide (HFO) bearing sludges. The work presented demonstrates for the first time the reuse of HFO-rich waste sludges from potable water and mine water treatment. The waste was used directly without modification or reagent addition, as media/substrate in simple flow-through reactors for the decolourisation and biodegradation of methyl orange (MO) and mixed dyes textile effluent. Three phases of exploratory proof of concept work were undertaken. Columns containing HFO sludges were challenged with solution of MO, and MO amended with glycerol (Phase I), MO in a synthetic textile effluent recipe (Phase II), and real mixed textile effluent containing a mixture of dyes (Phase III). After an initial lag period extensive decolourisation of dye was observed in all cases at rates comparable with pure strains and engineered bioreactor processes, with evidence of biodegradation beyond simple cleavage of the mono azo chromophore and mineralisation. The microbiology of the initial sludge samples in both cases exhibited a diverse range of iron oxidising and reducing bacteria. However, post experiment the microbiology of sludge evolved from being dominated by Proteobacteria to being dominated by Firmicutes. Distinct changes in the microbial community structure were observed in post-treatment MWTS and WTWS where genera capable of iron and sulphate reduction and/or aromatic amine degradation were identified. Average nitrogen removal rates for the columns ranged from 27.8 to 194 g/m3/day which is higher than engineered sequential anaerobic-aerobic bioreactor. Postulated mechanisms for the fast anaerobic decolourisation, biodegradation, and mineralisation of the dyes (as well nitrogen transformations) include various direct and indirect enzymatic and metabolic reactions, as well as reductive attack by continuously regenerated reductants such as Fe(II), HFO bound Fe(II), FeS, and HS−. The ability of iron reducers to degrade aromatic rings is also considered important in the further biodegradation and complete mineralisation of organic carbon. The study reveals that abundant and ubiquitous HFO-rich waste sludges, can be used without amendment, as a substrate in simple flow-through bioremediation system for the decolourisation and partial biodegradation of dyes in textile effluent

Methanogen activity and microbial diversity in Gulf of Cádiz mud volcano sediments

The Gulf of Cádiz is a tectonically active continental margin with over sixty mud volcanoes (MV) documented, some associated with active methane (CH4) seepage. However, the role of prokaryotes in influencing this CH4 release is largely unknown. In two expeditions (MSM1-3 and JC10) seven Gulf of Cádiz MVs (Porto, Bonjardim, Carlos Ribeiro, Captain Arutyunov, Darwin, Meknes, and Mercator) were analyzed for microbial diversity, geochemistry, and methanogenic activity, plus substrate amended slurries also measured potential methanogenesis and anaerobic oxidation of methane (AOM). Prokaryotic populations and activities were variable in these MV sediments reflecting the geochemical heterogeneity within and between them. There were also marked differences between many MV and their reference sites. Overall direct cell numbers below the SMTZ (0.2–0.5 mbsf) were much lower than the general global depth distribution and equivalent to cell numbers from below 100 mbsf. Methanogenesis from methyl compounds, especially methylamine, were much higher than the usually dominant substrates H2/CO2 or acetate. Also, CH4 production occurred in 50% of methylated substrate slurries and only methylotrophic CH4 production occurred at all seven MV sites. These slurries were dominated by Methanococcoides methanogens (resulting in pure cultures), and prokaryotes found in other MV sediments. AOM occurred in some slurries, particularly, those from Captain Arutyunov, Mercator and Carlos Ribeiro MVs. Archaeal diversity at MV sites showed the presence of both methanogens and ANME (Methanosarcinales, Methanococcoides, and ANME-1) related sequences, and bacterial diversity was higher than archaeal diversity, dominated by members of the Atribacterota, Chloroflexota, Pseudomonadota, Planctomycetota, Bacillota, and Ca. “Aminicenantes.” Further work is essential to determine the full contribution of Gulf of Cádiz mud volcanoes to the global methane and carbon cycles

Biostimulation of jarosite and iron oxide-bearing mine waste enhances subsequent metal recovery

Novel resource recovery technologies are required for metals-bearing hazardous wastes in order to achieve circular economy outcomes and industrial symbiosis. Iron oxide and co-occurring hydroxysulphate-bearing wastes are globally abundant and often contain other elements of value. This work addresses the biostimulation of indigenous microbial communities within an iron oxide/ hydroxysulphate-bearing waste and its effect on the subsequent recoverability of metals by hydrochloric, sulphuric, citric acids and EDTA. Laboratory-scale flow-through column reactors were used to examine the effect of using glycerol (10% w/w) to stimulate the in situ microbial community in an iron oxide/ hydroxysulphate-bearing mine waste. The effects on the evolution of leachate chemistry, changes in microbiological community and subsequent hydrometallurgical extractability of metals were studied. Results demonstrated increased leachability and selectivity of Pb, Cu, and Zn relative to iron after biostimulation with a total of 0.027 kg of glycerol per kg of waste. Biostimulation, which can be readily applied in situ , potentially opens new routes to metal recovery from globally abundant waste streams that contain jarosite and iron oxides