Metagenomic assessment of nitrate-contaminated mine wastewaters and optimization of complete denitrification by indigenous enriched bacteria

Nitrate contamination in water remains to be on the rise globally due to continuous anthropogenic activities, such as mining and farming, which utilize high amounts of ammonium nitrate explosives and chemical-NPK-fertilizers, respectively. This study presents insights into the development of a bioremediation strategy to remove nitrates (NO3−) using consortia enriched from wastewater collected from a diamond mine in Lesotho and a platinum mine in South Africa. A biogeochemical analysis was conducted on the water samples which aided in comparing and elucidating their unique physicochemical parameters. The chemical analysis uncovered that both wastewater samples contained over 120 mg/L of NO3− and over 250 mg/L of sulfates (SO42-), which were both beyond the acceptable limit of the environmental surface water standards of South Africa. The samples were atypical of mine wastewaters as they had low concentrations of dissolved heavy metals and a pH of over 5. A metagenomic analysis applied to study microbial diversities revealed that both samples were dominated by the phyla Proteobacteria and Bacteroidetes, which accounted for over 40% and 15%, respectively. Three consortia were enriched to target denitrifying bacteria using selective media and then subjected to complete denitrification experiments. Denitrification dynamics and denitrifying capacities of the consortia were determined by monitoring dissolved and gaseous nitrogen species over time. Denitrification optimization was carried out by changing environmental conditions, including supplementing the cultures with metal enzyme co-factors (iron and copper) that were observed to promote different stages of denitrification. Copper supplemented at 50 mg/L was observed to be promoting complete denitrification of over 500 mg/L of NO3−, evidenced by the emission of nitrogen gas (N2) that was more than nitrous oxide gas (N2O) emitted as the terminal by-product. Modification and manipulation of growth conditions based on the microbial diversity enriched proved that it is possible to optimize a bioremediation system that can reduce high concentrations of NO3−, while emitting an environmentally-friendly N2 instead of N2O, that is, a greenhouse gas. Data collected and discussed in this research study can be used to model an upscale NO3− bioremediation system aimed to remove nitrogenous and other contaminants without secondary contamination

Evaluating the effects of pH and temperature on sulphate-reducing bacteria and modelling of their effects in stirred bioreactors

DATA AVAILABILITY STATEMENT : Data used and presented in this study can be made available upon requests to the corresponding authors.Sulphate (SO4) abundance in the earth’s crust contributes largely to industrial wastewater contamination lowering the pH, which exuberates the dissolution of metals forming acidic drainages. Biological sulphate reduction as a remediation process can be affected by factors such as pH, temperature and high sulphide concentrations. In this study, sulphate-reducing bacterial community enriched from mine wastewaters was applied in semi-automated bioreactors to assess the effects of these factors on microbial sulphate reduction capacities. Low pH (3.5) and temperature (10°C) were observed to promote the toxicity of sulphur-reduced species on the consortium while mesophilic temperature (25°C) and near neutral pH (6.2) were observed to induce optimum SO4 reduction attaining a maximum of 95% SO4 reduction. Obtained SO4 reduction dynamics data was then applied in formulating a unique non-competitive inhibition equation that models biogeochemical events during SO4 reduction under varied pH and temperature conditions and predicts the efficacy of a bioremediation system.http://www.tandfonline.com/journals/tcsb21am2024Chemical EngineeringSDG-12:Responsible consumption and productio

Whole genome comparison of Thermus sp. NMX2.A1 reveals principal carbon metabolism differences with closest relation Thermus scotoductus SA-01

Genome sequencing of the yellow-pigmented, thermophilic bacterium Thermus sp. NMX2.A1 resulted in a 2.29 Mb draft genome that encodes for 2312 proteins. The genetic relationship between various strains from the genus Thermus was assessed based on phylogenomic analyses using a concatenated set of conserved proteins. The resulting phylogenetic tree illustrated that Thermus sp. NMX2 A.1 clusters together with Thermus scotoductus SA-01, despite being isolated from vastly different geographical locations. The close evolutionary relationship and metabolic parallels between the two strains has previously been recognized; however, neither strain’s genome data were available at that point in time. Genomic comparison of the Thermus sp. NMX2.A1 and T. scotoductus SA-01, as well as other closely related Thermus strains, revealed a high degree of synteny at both the genomic and proteomic level, with processes such as denitrification and natural cell competence appearing to be conserved. However, despite this high level of similarity, analysis revealed a complete, putative Calvin–Benson–Bassham (CBB) cycle in NMX2.A1 that is absent in SA-01. Analysis of horizontally transferred gene islands provide evidence that NMX2 selected these genes due to pressure from its HCO3 - rich environment, which is in stark contrast to that of the deep subsurface isolated SA-01.The National Research Foundation and the Technology Innovation Agency, South Africa.http://www.g3journal.orgam2017Biochemistr

Novel lichen-dominated hypolithic communities in the Namib Desert

The ventral surfaces of translucent rocks from hot desert pavements often harbor hypolithic microbial communities, which are mostly dominated by cyanobacteria. The Namib Desert fog belt supports extensive hypolithic colonization of quartz rocks, which are also colonized by lichens on their dorsal surfaces. Here, we aim to evaluate whether lichens colonize the ventral surface of the rocks (i.e., show hypolithic lifestyle) and compare the bacterial composition of these coastal hypolithic communities with those found inland. Fungal DNA barcoding and fungal and bacterial Illumina metabarcoding were combined with electron microscopy to characterize the composition and spatial structure of hypolithic communities from two (coastal and inland) areas in the Namib Desert. We report, for the first time, the structure and composition of lichen-dominated hypolithic communities found in the coastal zone of the Namib Desert with extensive epilithic lichen cover. Lichen modified areoles with inverted morphology of the genus Stellarangia (three lineages) and Buellia (two lineages) were the main components of these hypolithic communities. Some of these lineages were also found in epilithic habitats. These lichen-dominated hypolithic communities differed in structural organization and bacterial community composition from those found in inland areas. The hypolithic lichen colonization characterized here seems not to be an extension of epilithic or biological soil crust lichen growths but the result of specific sublithic microenvironmental conditions. Moisture derived from fog and dew could be the main driver of this unique colonization.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. DC acknowledges the financial support of the University of Pretoria and the National Research Foundation of South Africa. AV was supported by the project “CLU-2019–05 – IRNASA/CSIC Unit of Excellence,” funded by the Junta de Castilla y León and co-financed by the European Union (ERDF “Europe drives our growth”). This work was supported by grants CTM2015-64728-C2- 2-R (MINECO/FEDER, EU) and PID2019-105469RB-C22 (MICINN).http://link.springer.com/journal/248am2022BiochemistryGeneticsMicrobiology and Plant Patholog

New ecosystems in the deep subsurface follow the flow of water driven by geological activity

Eukarya have been discovered in the deep subsurface at several locations in South Africa, but how organisms reach the subsurface remains unknown. We studied river-subsurface fissure water systems and identified Eukarya from a river that are genetically identical for 18S rDNA. To further confirm that these are identical species one metazoan species recovered from the overlying river interbred successfully with specimen recovered from an underlying mine at −1.4 km. In situ seismic simulation experiments were carried out and show seismic activity to be a major force increasing the hydraulic conductivity in faults allowing organisms to create ecosystems in the deep subsurface. As seismic activity is a non-selective force we recovered specimen of algae and Insecta that defy any obvious other explanation at a depth of −3.4 km. Our results show there is a steady flow of surface organisms to the deep subsurface where some survive and adapt and others perish. As seismic activity is also present on other planets and moons in our solar system the mechanism elucidated here may be relevant for future search and selection of landing sites in planetary exploration.publishedVersio

Biomineralization and Bioaccumulation of Europium by a Thermophilic Metal Resistant Bacterium

Rare earth metals are widely used in the production of many modern technologies. However, there is concern that supply cannot meet the growing demand in the near future. The extraction from low-grade sources such as geothermal fluids could contribute to address the increasing demand for these compounds. Here we investigated the interaction and eventual bioaccumulation of europium (Eu) by a thermophilic bacterium, Thermus scotoductus SA-01. We demonstrated that this bacterial strain can survive in high levels (up to 1 mM) of Eu, which is hundred times higher than typical concentrations found in the environment. Furthermore, Eu seems to stimulate the growth of T. scotoductus SA-01 at low (0.01–0.1 mM) concentrations. We also found, using TEM-EDX analysis, that the bacterium can accumulate Eu both intracellularly and extracellularly. FT-IR results confirmed that carbonyl and carboxyl groups were involved in the biosorption of Eu. Infrared and HR-XPS analysis demonstrated that Eu can be biomineralized by T. scotoductus SA-01 as Eu2(CO3)3. This suggests that T. scotoductus SA-01 can potentially be used for the biorecovery of rare earth metals from geothermal fluids

Endophytic seed mycobiome of six sorghum (Sorghum bicolor) cultivars from commercial seedlots using an Illumina sequencing approach

18 páginas, 6 figuras, 2 tablasPathogen-free seeds are important for the establishment of young seedlings, prevention of health problems upon consumption by animals and livestock, and inadvertent movement of pathogens into and out of countries. However, testing for the presence of fungi, including pathogens from seeds is a time consuming and difficult process. In this study, we characterized the seed fungal microbiome (mycobiome) of six commercial sorghum cultivars from South Africa using a deep amplicon next generation sequencing approach based on the Internal Transcribed Spacer (ITS) region of the ribosomal operon. Sorghum is the fifth most important crop in the world, and widely used by African farmers. We found that the fungi present in each of the seedlots were similar to those reported by cultural studies. By comparing phylotypes of certain key families and genera to phylotypes used in established phylogenies and reputed sequences from public databases, the diagnostic value of the NGS method was also investigated. We showed that a number of molecular operational taxonomic units (MOTUs) could be identified at the species level and established that certain known pathogens are not present in the tested seeds, for instance in the Aspergillus group. Other groups could not be identified, not even to genus level. While acknowledging the shortcomings of using partial ITS data, we demonstrated that deep amplicon sequencing is a valuable diagnostic tool for seed disease control and prevention in some cases.Research funding was provided by the Agricultural Research Council Collaborative Consortium to Broaden the Food Base.Peer reviewe

Biodegradation of chloroethene compounds under microoxic conditions

14 páginas, 3 tablas, 9 figurasThe biodegradation of chloroethene compounds under oxic and anoxic conditions is well established. However, the biological reactions that take place under microoxic conditions are unknown. Here, we report the biostimulated (BIOST: addition of lactate) and natural attenuated (NAT) degradation of chloroethene compounds under microoxic conditions by bacterial communities from chloroethene compounds-contaminated groundwater. The degradation of tetrachloroethene was significantly higher in NAT (15.14% on average) than in BIOST (10.13% on average) conditions at the end of the experiment (90 days). Sporomusa, Paracoccus, Sedimentibacter, Pseudomonas, and Desulfosporosinus were overrepresented in NAT and BIOST compared to the source groundwater. The NAT metagenome contains phenol hydrolase P1 oxygenase (dmpL), catechol-1,2-dioxygenase (catA), catechol-2,3-dioxygenases (dmpB, todE, and xylE) genes, which could be involved in the cometabolic degradation of chloroethene compounds; and chlorate reductase (clrA), that could be associated with partial reductive dechlorination of chloroethene compounds. Our data provide a better understanding of the bacterial communities, genes, and pathways potentially implicated in the reductive and cometabolic degradation of chloroethene compounds under microoxic conditions.This research was conducted with financial support from the TIA/BIOSAENSE group, University of the Free State, Republic of South Africa.Peer reviewe

Characterization of Microbial Diversity of Two Tomato Cultivars through Targeted Next-Generation Sequencing 16S rRNA and ITS Techniques

Even though the nutritional and economic values of Solanum lycopersicum (tomato) are substantially impacted by microbial spoilage, the available data on its microbial community, particularly during spoilage, are limited and have primarily been characterized using conventional culture-dependent methods. This study employed a targeted high-throughput next-generation sequencing method to longitudinally characterize the microbial diversity of two South African tomato cultivars (jam and round) at varied storage intervals (1, 6, and 12 days). Throughout the storage period, the bacterial communities of the two cultivars were more diverse than the fungal communities. The microbial diversity of both bacteria and fungi was greater and comparable between the cultivars on day 1, but becomes distinct as the storage period increases, with round tomatoes being more diverse than jam tomato, though, on day 12, jam tomato develops greater diversity than round tomato. Overall, the most abundant phyla (though Proteobacteria was most dominant) were Proteobacteria, Firmicutes, and Bacteriodota in the bacterial communities, while Ascomycota and Basidiomycota formed most fungal communities with Ascomycota being dominant. At the genus level, Pantoea and Klebsiella (bacteria), Hanseniaspora, Stemphylium, and Alternaria (fungi) were prevalent. Taken together, this study casts light on a broad microbial diversity profile thus, confirms the cultivars’ diversity and abundance differences