Sequential hydrotalcite precipitation, microbial sulfate reduction and in situ hydrogen sulfide removal for neutral mine drainage treatment.

This study proposed and examined a new process flowsheet for treating neutral mine drainage (NMD) from an open-pit gold mine. The process consisted of three sequential stages: (1) in situ hydrotalcite (HT) precipitation; (2) low-cost carbon substrate driven microbial sulfate reduction; and (3) ferrosol reactive barrier for removing biogenic dissolved hydrogen sulfide (H2S). For concept validation, laboratory-scale columns were established and operated for a 140-days period with key process performance parameters regularly measured. At the end, solids recovered from various depths of the ferrosol column were analysed for elemental composition and mineral phases. Prokaryotic microbial communities in various process locations were characterised using 16S rRNA gene sequencing. Results showed that the Stage 1 HT-treatment substantially removed a range of elements (As, B, Ba, Ca, F, Zn, Si, and U) in the NMD, but not nitrate or sulfate. The Stage 2 sulfate reducing bioreactor (SRB) packed with 70 % (v/v) Eucalyptus woodchip, 1 % (w/v) ground (<1 mm) dried Typha biomass, and 10 % (w/v) NMD-pond sediment facilitated complete nitrate removal and stable sulfate removal of ca. 50 % (50 g-SO4 m-3 d-1), with an average H2S generation rate of 10 g-H2S m-3d-1. The H2S-removal performance of the Stage 3 ferrosol column was compared with a synthetic amorphous Fe-oxyhydroxide-amended sand control column. Although both columns facilitated excellent (95-100 %) H2S removal, the control column only enabled a further ca. 10 % sulfate reduction, giving an overall sulfate removal of 56 %. In contrast, the ferrosol enabled an extra 99.9 % sulfate reduction in the SRB effluent, leading to a near complete sulfate removal. Overall, the process successfully eliminated a range of metal/metalloid contaminants, nitrate, sulfate (2500 mg-SO4 L-1 in the NMD to <10 mg-SO4 L-1 in the final effluent) and H2S (>95 % removal). Further optimisation is required to minimise release of ferrous iron from the ferrosol barrier into the final effluent.

Prokaryotic microbial ecology as an ecosurveillance tool for eukaryotic pathogen colonisation: Meiothermus and Naegleria fowleri.

Naegleria fowleri has been detected in drinking water distribution systems (DWDS) in Australia, Pakistan and the United States and is the causative agent of the highly fatal disease primary amoebic meningoencephalitis. Previous small scale field studies have shown that Meiothermus may be a potential biomarker for N. fowleri. However, correlations between predictive biomarkers in small sample sizes often breakdown when applied to larger more representative datasets. This study represents one of the largest and most rigorous temporal investigations of Naegleria fowleri colonisation in an operational DWDS in the world and measured the association of Meiothermus and N. fowleri over a significantly larger space and time in the DWDS. A total of 232 samples were collected from five sites over three-years (2016-2018), which contained 29 positive N. fowleri samples. Two specific operational taxonomic units assigned to M. chliarophilus and M. hypogaeus, were significantly associated with N. fowleri presence. Furthermore, inoculation experiments demonstrated that Meiothermus was required to support N. fowleri growth in field-collected biofilms. This validates Meiothermus as prospective biological tool to aid in the identification and surveillance of N. fowleri colonisable sites.

Simulating compatible solute biosynthesis using a metabolic flux model of the biomining acidophile, Acidithiobacillus ferrooxidans ATCC 23270.

Halotolerant, acidophilic, bioleaching microorganisms are crucial to biomining operations that utilize saline water. Compatible solutes play an important role in the adaptation of these microorganisms to saline environments. Acidithiobacillus ferrooxidans ATCC 23270, an iron- and sulfur-oxidizing acidophilic bacterium, synthesizes trehalose as its native compatible solute but is still sensitive to salinity. Recently, halotolerant bioleaching bacteria were found to use ectoine as their key compatible solute. Previously, bioleaching bacteria were recalcitrant to genetic manipulation; however, recent advancements in genetic tools and techniques allow successful genetic modification of A. ferrooxidans ATCC 23270. Therefore, this study aimed to test, in silico, the effect of native and synthetic compatible solute biosynthesis by A. ferrooxidans ATCC 23270 on its growth and metabolism. Metabolic network flux modelling was used to provide a computational framework for the prediction of metabolic fluxes during production of native and synthetic compatible solutes by A. ferrooxidans ATCC 23270, in silico. Complete pathways for trehalose biosynthesis by the bacterium are proposed and captured in the updated metabolic model including a newly discovered UDP-dependent trehalose synthesis pathway. Finally, the effect of nitrogen sources on compatible solute production was simulated and showed that using nitrogen gas as the sole nitrogen source enables the ectoine-producing 'engineered' microbe to oxidize up to 20% more ferrous iron in comparison to the native microbe that only produces trehalose. Therefore, the predictive outcomes of the model have the potential to guide the design and optimization of a halotolerant strain of A. ferrooxidans ATCC 23270 for saline bioleaching operations.

Biofilm formation on the surface of monazite and xenotime during bioleaching.

Microbial attachment and biofilm formation is a ubiquitous behaviour of microorganisms and is the most crucial prerequisite of contact bioleaching. Monazite and xenotime are two commercially exploitable minerals containing rare earth elements (REEs). Bioleaching using phosphate solubilizing microorganisms is a green biotechnological approach for the extraction of REEs. In this study, microbial attachment and biofilm formation of Klebsiella aerogenes ATCC 13048 on the surface of these minerals were investigated using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). In a batch culture system, K. aerogenes was able to attach and form biofilms on the surface of three phosphate minerals. The microscopy records showed three distinctive stages of biofilm development for K. aerogenes commencing with initial attachment to the surface occurring in the first minutes of microbial inoculation. This was followed by colonization of the surface and formation of a mature biofilm as the second distinguishable stage, with progression to dispersion as the final stage. The biofilm had a thin-layer structure. The colonization and biofilm formation were localized toward physical surface imperfections such as cracks, pits, grooves and dents. In comparison to monazite and xenotime crystals, a higher proportion of the surface of the high-grade monazite ore was covered by biofilm which could be due to its higher surface roughness. No selective attachment or colonization toward specific mineralogy or chemical composition of the minerals was detected. Finally, in contrast to abiotic leaching of control samples, microbial activity resulted in extensive microbial erosion on the high-grade monazite ore.

Klebsiella aerogenes Adhesion Behaviour during Biofilm Formation on Monazite.

The adsorption behaviour of micro-organisms during the initial attachment stage of biofilm formation affects subsequent stages. The available area for attachment and the chemophysical properties of a surface affect microbial attachment performance. This study focused on the initial attachment behaviour of Klebsiella aerogenes on monazite by measuring the ratio of planktonic against sessile subpopulations (P:S ratio), and the potential role of extracellular DNA (eDNA). eDNA production, effects of physicochemical properties of the surface, particle size, total available area for attachment, and the initial inoculation size on the attachment behaviour were tested. K. aerogenes attached to monazite immediately after exposure to the ore; however, the P:S ratio significantly (p = 0.05) changed in response to the particle size, available area, and inoculation size. Attachment occurred preferentially on larger-sized (~50 µm) particles, and either decreasing the inoculation size or increasing the available area further promoted attachment. Nevertheless, a portion of the inoculated cells always remained in a planktonic state. K. aerogenes produced lower eDNA in response to the changed surface chemical properties when monazite was replaced by xenotime. Using pure eDNA to cover the monazite surface significantly (p ≤ 0.05) hindered bacterial attachment due to the repulsive interaction between the eDNA layer and bacteria.

Integrating bioelectrochemical system with aerobic bioreactor for organics removal and caustic recovery from alkaline saline wastewater.

Bioelectrochemical systems (BES) are increasingly being explored as an auxiliary unit process to enhance conventional waste treatment processes. This study proposed and validated the application of a dual-chamber bioelectrochemical cell as an add-on unit for an aerobic bioreactor to facilitate reagent-free pH-correction, organics removal and caustic recovery from an alkaline and saline wastewater. The process was continuously fed (hydraulic retention time (HRT) of 6 h) with a saline (25 g NaCl/L) and alkaline (pH 13) influent containing oxalate (25 mM) and acetate (25 mM) as the target organic impurities present in alumina refinery wastewater. Results suggested that the BES concurrently removed the majority of the influent organics and reduced the pH to a suitable range (9-9.5) for the aerobic bioreactor to further remove the residual organics. Compared to the aerobic bioreactor, the BES enabled a faster removal of oxalate (242 ± 27 vs. 100 ± 9.5 mg/L.h), whereas similar removal rates (93 ± 16 vs. 114 ± 23 mg/L.h, respectively) were recorded for acetate. Increasing catholyte HRT from 6 to 24 h increased the caustic strength from 0.22% to 0.86%. The BES enabled caustic production at an electrical energy demand of 0.47 kWh/kg-caustic, which is a fraction (22%) of the electrical energy requirement for caustic production using conventional chlor-alkali processes. The proposed application of BES holds promise to improve environmental sustainability of industries in managing organic impurities in alkaline and saline waste streams.

Bioelectrochemical extraction of ammonium from low-strength wastewater with concomitant generation of high-purity hydrogen.

Bioelectrochemical systems (BES) are emerging environmental biotechnology for recovering ammonia from waste streams. It has been tested extensively for treating ammonium-rich wastewater. This study examined the suitability of BES to facilitate carbon removal and ammonium extraction from a low ammonium liquor (3.7 mM) that mimics municipal wastewater, and concomitant production of high-purity hydrogen gas, which could potentially be harnessed as a fuel or internally recycled for ammonia stripping. Results showed that a two-chamber cation exchange membrane-equipped BES enabled a high hydrogen yield (22.8 m3 H2 m-3 d-1; > 98% cathodic efficiency) and chemical oxygen demand (COD) removal (80%; 2.43 kg COD m-3 d-1 at a hydraulic retention time of 4.4 h). However, for the treatment of wastewater, the system demanded high energy (2.3 kWh kg COD-1; 152 kWh kg-1 N removed) and base for pH adjustment. The technology may be more suitable for recovering ammonium from wastewaters with molar ammonium to BOD ratio closer to the desired stoichiometric ratio of four, and for waste streams containing sufficient alkalinity or pH-buffering capacity, eliminating the need for dosing cation-bearing alkali.

Three synthetic biology applications and their paths to impact in Australia: Cane toads, bacteriophages, and biomining microbes.

Synthetic biology [synbio] applications have the potential to assist in addressing significant global health and environmental challenges. Australian research institutes are investing in formative research to develop synbio technologies capable of meeting these challenges. Alongside the laboratory research, investigating the broader social, institutional, and ethical considerations that synbio presents has been a priority. We conducted targeted qualitative research to uncover the barriers and opportunities for a range of multisectoral stakeholders identified as potential end-users of the science under development. The research provides insights into the research implementation environment for three synthetic biology applications: (1) gene editing cane toads (Rhinella marina) to reduce their environmental impact; (2) engineering bacteriophages to combat antimicrobial resistance in humans; and (3) engineering microbes to improve biomining efficiency in the mining industry. In-depth interviews (N = 23) with government, research and civil society representatives revealed key challenges in the impact pathway for each application. The strongest themes uncovered during interviews related to perceived negative public attitudes towards genetic technologies, a lack of investment in critical research infrastructure, unclear regulatory pathways and the presence of a strong social and environmental imperative underpinning technology development. These findings reveal specific entry points for further engagement with the most immediate end-users of synbio. Separate from research on public attitudes to synbio, the cases highlight the various hurdles to achieving research impact, according to experts who will likely use, approve or invest in these applications in the future. The themes uncovered inform avenues for strengthening engagement and research coordination in Australia and elsewhere.

Quorum sensing inhibitors applications: A new prospect for mitigation of microbiologically influenced corrosion.

Quorum sensing (QS) is a process of bacterial communication that involves the use of biochemical signals and adjusts the expression of specific genes as a response to the bacterial cell density within an environment. This process is employed by both Gram-positive and Gram-negative bacteria to regulate different physiological functions. In both cases, QS involves production, detection and responses to signalling chemicals, termed auto-inducers. Expression of virulence factors and formation of biofilms are the typical processes controlled by QS, which, therefore, inspires the exploration of QS as a plausible solution to mitigating the increasing microbial resistance to antibiotics. QS inhibitors (QSIs) from different origins have been recognised as a promising solution to biofilm related challenges in a large variety of applications. Though QSIs have demonstrated some strength in tackling biofouling, a key focus in the literature on QSIs based strategies has been to control microbially influenced corrosion. This article reviews the principles of QS, its mechanistic roles in biofilm formation and the feasibility of QSIs to mitigate biofilm related challenges in a number of commercial applications. The potential of QSIs in microbially influenced corrosion for future applications is also discussed.

Sequential removal of selenate, nitrate and sulfate and recovery of elemental selenium in a multi-stage bioreactor process with redox potential feedback control.

Bioreduction can facilitate oxyanions removal from wastewater. However, simultaneously removing selenate, nitrate and sulfate and recovering high-purity elemental selenium (Se0) from wastewater by a single system is difficult and may lead to carcinogenic selenium monosulfide (SeS) formation. To solve this issue, a two-stage biological fluidized bed (FBR) process with ethanol dosing based on oxidation-reduction potential (ORP) feedback control was developed in this study. FBR1 performance was first evaluated at various ORP setpoints (between -520 and -360 mV vs. Ag/AgCl) and elevated sulfate concentration. Subsequently, ethanol-fed FBR2 was used to reduce sulfate from FBR1 effluent, followed by an aerated sulfide oxidation reactor (SOR). At - 520 mV≤ ORPs≤ -480 mV, FBR1 removed 100 ±â€¯0.1% nitrate and 99.7 ±â€¯0.3% selenate without sulfate reduction. At ORPs ≥ -440 mV, selenate reduction was incomplete, whereas nitrate removal remained stable. Se0 recovery efficiency from FBR1 effluent was 37.5% with 71% Se purity. FBR2 converted 86% of the remaining sulfate in FBR1 effluent to hydrogen sulfide, but the over-oxidation of dissolved sulfide in SOR decreased the overall sulfate removal efficiency to ~46.3%. Overall, the two-stage FBR process with ORP feedback dosing of ethanol was effective for sequentially removing selenate, nitrate and sulfate and recovering Se0 from wastewater.

Potential of Acidithiobacillus ferrooxidans to Grow on and Bioleach Metals from Mars and Lunar Regolith Simulants under Simulated Microgravity Conditions.

The biomining microbes which extract metals from ores that have been applied in mining processes worldwide hold potential for harnessing space resources. Their cell growth and ability to extract metals from extraterrestrial minerals under microgravity environments, however, remains largely unknown. The present study used the model biomining bacterium Acidithiobacillus ferrooxidans to extract metals from lunar and Martian regolith simulants cultivated in a rotating clinostat with matched controls grown under the influence of terrestrial gravity. Analyses included assessments of final cell count, size, morphology, and soluble metal concentrations. Under Earth gravity, with the addition of Fe3+ and H2/CO2, A. ferrooxidans grew in the presence of regolith simulants to a final cell density comparable to controls without regoliths. The simulated microgravity appeared to enable cells to grow to a higher cell density in the presence of lunar regolith simulants. Clinostat cultures of A. ferrooxidans solubilised higher amounts of Si, Mn and Mg from lunar and Martian regolith simulants than abiotic controls. Electron microscopy observations revealed that microgravity stimulated the biosynthesis of intracellular nanoparticles (most likely magnetite) in anaerobically grown A. ferrooxidans cells. These results suggested that A. ferrooxidans has the potential for metal bioleaching and the production of useful nanoparticles in space.

Treatment of neutral gold mine drainage by sequential in situ hydrotalcite precipitation, and microbial sulfate and cyanide removal.

This study proposed and validated a method integrating in situ hydrotalcite precipitation (Virtual Curtain™ (VC) technology) with bioprocess for treating a cyanide (CN)-augmented (ca. 5 mg-CN L-1) sulfate-laden neutral mine drainage, from a waste rock dump (WD2) of an Australian gold mine. Efficacies of various carbon (C) sources (ethanol, lactate, and two natural substrates; Eucalyptus wood sawdust (EW) and Typha biomass (TB)) for promoting microbial reduction in both: CN-augmented WD2 water and VC-treated CN-augmented WD2 water were assessed in a 60-days microcosms study at 30 °C. The microcosms were monitored over time for pH, redox potential, dissolved hydrogen sulfide, chloride, nitrite, nitrate, sulfate, phosphate, biogas production, dissolved organic carbon, total dissolved nitrogen, and dissolved CN. The VC treatment removed a range of metals (Mg, Ni and Zn) and metalloid Se from the CN-augmented WD2 water to below detection. Other elements substantially reduced in concentration included Ba, F, Si and U. However, the VC treatment did not remove substantial nitrate, sulfate or CN. Microcosm trials revealed that the indigenous microbial community in WD2 could effectively denitrify and reduce sulfate, with TB was the most efficient C source for promoting sulfate and CN removal; whereas, EW facilitated only marginally higher sulfate reduction compared with controls. The highest sulfate reduction rate (76 g-SO42- m-3 d-1) was achieved with VC-treated water amended with TB, indicating that VC pre-treatment was beneficial. Further, all treatments amended with external C, facilitated 100% removal of dissolved CN after 60 days, whereas only partial (65%) CN removal was recorded in the control. Overall, the proposed integrated method appears a viable option for treating neutral gold mine drainage.

Effect of ferrous iron loading on dewaterability, heavy metal removal and bacterial community of digested sludge by Acidithiobacillus ferrooxidans.

Acidithiobacillus ferrooxidans ILS-2 was adapted in digested sludge and used to treat sludge for dewaterability improvement. Results showed that increasing ferrous iron loading increased sludge dewaterability, but the inoculation of the bioleaching strain had little effect on sludge dewaterability compared to controls without the strain. The total extracellular polymeric substances (EPS) contents of sludges with and without bioleaching treatment were similar except for bioleaching treatment at 10% ferrous iron loading (on sludge total solids) where total EPS was higher with bioleaching treatment. However, bioleaching treatment for 48 h had a notable effect on removal of heavy metals, such as Mn, Ni and Zn, especially at the high loadings of ferrous iron. In the presence of A. ferrooxidans, the removal of Ni, Mn and Zn reached 93%, 88% and 80%, respectively, at a ferrous iron loading of 21%. The sequencing of 16S rRNA genes indicated that increasing ferrous iron loadings to 15% and 21% increased the relative abundance of Acidithiobacillus, Acidocella (with A. ferrooxidans) and Carboxylicivirga (without A. ferrooxidans) but decreased the abundance of Pseudomonas and Acinetobacter after 48 h treatment. This study enhanced the understanding of the correlations between bioleaching treatment of digested sludge, sludge dewaterability, heavy metal removal and bacterial communities.

Optimization of nitrate and selenate reduction in an ethanol-fed fluidized bed reactor via redox potential feedback control.

Electron donors are a major cost-factor in biological removal of oxyanions, such as nitrate and selenate from wastewater. In this study, an online ethanol dosing strategy based on feedback control of oxidation-reduction potential (ORP) was designed to optimize the performance of a lab-scale fluidized bed reactor (FBR) in treating selenate and nitrate (5 mM each) containing wastewater. The FBR performance was evaluated at various ORP setpoints ranging between -520 mV and -240 mV (vs. Ag/AgCl). Results suggested that both nitrate and selenate were completely removed at ORPs between -520 mV and -360 mV, with methylseleninic acid, selenocyanate, selenosulfate and ammonia being produced at low ORPs between -520 mV and -480 mV, likely due to overdosing of ethanol. At ORPs between -300 mV and -240 mV, limited ethanol dosing resulted in an apparent decline in selenate removal whereas nitrate removal remained stable. Resuming the ORP to -520 mV successfully restored complete selenate reduction. An optimal ORP of -400 mV was identified for the FBR, whereby selenate and nitrate were nearly completely removed with a minimal ethanol consumption. Overall, controlling ORP via feedback-dosing of the electron donor was an effective strategy to optimize FBR performance for reducing selenate and nitrate in wastewater.

Contrasting effects and mode of dredging and in situ adsorbent amendment for the control of sediment internal phosphorus loading in eutrophic lakes.

Dredging and in situ adsorbent inactivation are two methods which are frequently used in eutrophic water bodies such as ponds, lakes and estuaries to control internal phosphorus (P) loading from sediments. However, their effects and modes on the control of sediment P loading has been seldom compared. In this study, a long-term sediment core incubation experiment in the field was undertaken to investigate changes in sediment P loading (P fluxes, supply ability and forms of P and transformation) comparing two remediation techniques, that of lanthanum-modified bentonite (LMB) addition or dredging to a control. A 360-day field investigation indicated that LMB addition more effectively reduced pore water P concentrations and sediment P fluxes than dredging in comparison with the control. On average, dredging and in situ LMB inactivation reduced the P flux by 82% and 90%, respectively relative to the control sediment. Whilst both the LMB inactivation and dredging can reduce the mobile P concentration, the impact of LMB in reducing mobile P was demonstrated to be more prolonged than that of dredging after 360 days. The P fraction composition in the LMB inactivated sediment differed significantly from the dredged and control sediment. Contrary to physical removal of dredging, chemical transformation of sediment mobile P and Al-P into Ca-P is the main function mode of LMB for sediment internal P control. Both LMB addition and dredging caused changes in the composition of sediment bacterial communities. Whilst LMB addition increased bacterial diversity, dredging temporarily reduced it. This study indicates that in situ inactivation by LMB is superior to dredging in the long-term control of sediment P loading.

High-rate microbial selenate reduction in an up-flow anaerobic fluidized bed reactor (FBR).

Wastewater contaminated with high concentrations of selenium oxyanions requires treatment prior to discharge. Biological fluidized bed reactors (FBRs) can be an option for removing selenium oxyanions from wastewater by converting them into elemental selenium, which can be separated from the treated effluent. In this study, a lab-scale FBR was constructed with granular activated carbon as biofilm carrier and inoculated with a consortium of selenate reducing bacteria enriched from environmental samples. The FBR was loaded with an influent containing ethanol (10 mM) and selenate (10 mM) as the microbial electron donor and acceptor, respectively. The performance of the FBR in reducing selenate was evaluated under various hydraulic retention times (HRTs) (120 h, 72 h, 48 h, 24 h, 12 h, 6 h, 3 h, 1 h and 20 min). After process acclimatization, selenate was completely removed with no notable selenite produced when the HRT was stepwise decreased from 120 h to 6 h. However, decreasing the HRT to 3 h resulted in selenite accumulation (0.17 ± 0.023 mM) in the effluent although selenate removal efficiency remained at 99.8 ± 0.20%. At 1 h HRT, the FBR removed 90.8 ± 1.4% of the selenate at a rate of 9.6 ± 0.15 mM h-1, which is the highest selenate reduction rate reported in the literature so far. However, 1 h HRT resulted in notable selenite accumulation (up to 2.4 ± 0.27 mM). Further decreasing the HRT to 20 min resulted in a notable decline in selenate reduction. Selenate reduction recovered from the "shock loading" after the HRT was increased back to 3 h. However, selenite still accumulated until the FBR was operated in batch mode for 6 days. This study affirmed that FBR is a promising treatment option for selenate-rich wastewater, and the process can be efficiently operated at low HRTs.

Unlocking Survival Mechanisms for Metal and Oxidative Stress in the Extremely Acidophilic, Halotolerant Acidihalobacter Genus.

Microorganisms used for the biohydrometallurgical extraction of metals from minerals must be able to survive high levels of metal and oxidative stress found in bioleaching environments. The Acidihalobacter genus consists of four species of halotolerant, iron-sulfur-oxidizing acidophiles that are unique in their ability to tolerate chloride and acid stress while simultaneously bioleaching minerals. This paper uses bioinformatic tools to predict the genes and mechanisms used by Acidihalobacter members in their defense against a wide range of metals and oxidative stress. Analysis revealed the presence of multiple conserved mechanisms of metal tolerance. Ac. yilgarnensis F5T, the only member of this genus that oxidizes the mineral chalcopyrite, contained a 39.9 Kb gene cluster consisting of 40 genes encoding mobile elements and an array of proteins with direct functions in copper resistance. The analysis also revealed multiple strategies that the Acidihalobacter members can use to tolerate high levels of oxidative stress. Three of the Acidihalobacter genomes were found to contain genes encoding catalases, which are not common to acidophilic microorganisms. Of particular interest was a rubrerythrin genomic cluster containing genes that have a polyphyletic origin of stress-related functions.

Bioleaching of Gold from Sulfidic Gold Ore Concentrate and Electronic Waste by Roseovarius tolerans and Roseovarius mucosus.

Gold bioleaching mediated by iodide oxidizing bacteria (IOB) has been proposed as a sustainable alternative to conventional technologies such as cyanidation. This study evaluated the ability of two IOB sourced from a commercial culture collection, Roseovarius (R.) tolerans DSM 11457T and R. mucosus DSM 17069T, to bioleach gold from electronic waste (e-waste) (1030 ppm gold) and sulfidic gold ore concentrate (45 ppm gold) using one-step, two-step and spent medium leaching at 1% pulp density over 10 days. Two-step bioleaching of ore concentrate resulted in the highest gold leaching yields (approximately ~100% and 34% for R. tolerans and R. mucosus, respectively), followed by spent medium leaching and one-step leaching. The yields remained low for e-waste with both strains (maximum 0.93% and 1.6% for R. tolerans and R. mucosus, respectively) and decreased over time, likely due to the instability of the solubilized gold at relatively low redox potentials (<300 mV vs. Ag/AgCl). Another limiting factor may be the partial inhibition of bacterial growth in the presence of the ore concentrate and e-waste. Therefore, future studies should evaluate the pre-treatment of the ore concentrate and e-waste to remove inhibitory and oxidant consuming compounds before bioleaching with IOB to optimize leaching yields.

Genome-based classification of Acidihalobacter prosperus F5 (=DSM 105917=JCM 32255) as Acidihalobacter yilgarnensis sp. nov.

The genus Acidihalobacter has three validated species, Acidihalobacter ferrooxydans, Acidihalobacter prosperus and Acidihalobacter aeolinanus, all of which were isolated from Vulcano island, Italy. They are obligately chemolithotrophic, aerobic, acidophilic and halophilic in nature and use either ferrous iron or reduced sulphur as electron donors. Recently, a novel strain was isolated from an acidic, saline drain in the Yilgarn region of Western Australia. Strain F5T has an absolute requirement for sodium chloride (>5 mM) and is osmophilic, growing in elevated concentrations (>1 M) of magnesium sulphate. A defining feature of its physiology is its ability to catalyse the oxidative dissolution of the most abundant copper mineral, chalcopyrite, suggesting a potential role in biomining. Originally categorized as a strain of A. prosperus, 16S rRNA gene phylogeny and multiprotein phylogenies derived from clusters of orthologous proteins (COGS) of ribosomal protein families and universal protein families unambiguously demonstrate that strain F5T forms a well-supported separate branch as a sister clade to A. prosperus and is clearly distinguishable from A. ferrooxydans DSM 14175T and A. aeolinanus DSM14174T. Results of comparisons between strain F5T and the other Acidihalobacter species, using genome-based average nucleotide identity, average amino acid identity, correlation indices of tetra-nucleotide signatures (Tetra) and genome-to-genome distance (digital DNA-DNA hybridization), support the contention that strain F5T represents a novel species of the genus Acidihalobacter. It is proposed that strain F5T should be formally reclassified as Acidihalobacter yilgarnenesis F5T (=DSM 105917T=JCM 32255T).

Carbon steel corrosion by bacteria from failed seal rings at an offshore facility.

Corrosion of carbon steel by microorganisms recovered from corroded seal rings at an offshore floating production facility was investigated. Microbial diversity profiling revealed that communities in all sampled seal rings were dominated by Pseudomonas genus. Nine bacterial species, Pseudomonas aeruginosa CCC-IOB1, Pseudomonas balearica CCC-IOB3, Pseudomonas stutzeri CCC-IOB10, Citrobacter youngae CCC-IOB9, Petrotoga mobilis CCC-SPP15, Enterobacter roggenkampii CCC-SPP14, Enterobacter cloacae CCC-APB1, Cronobacter sakazakii CCC-APB3, and Shewanella chilikensis CCC-APB5 were isolated from corrosion products and identified based on 16S rRNA gene sequence. Corrosion rates induced by the individual isolates were evaluated in artificial seawater using short term immersion experiments at 40 °C under anaerobic conditions. P. balearica, E. roggenkampii, and S. chilikensis, which have not been associated with microbiologically influenced corrosion before, were further investigated at longer exposure times to better understand their effects on corrosion of carbon steel, using a combination of microbiological and surface analysis techniques. The results demonstrated that all bacterial isolates triggered general and localised corrosion of carbon steel. Differences observed in the surface deterioration pattern by the different bacterial isolates indicated variations in the corrosion reactions and mechanisms promoted by each isolate.