Analyzing Dehalochip: A Functional DNA Microarray for Reductive Dichlorination in Chloroethene-Contaminated Sites.

Interpreting high-throughput transcriptomic and metagenomic data from non-model microorganisms presents a challenge due to the significant number of genes with unknown functions and sequences. In this study, we applied an innovative microarray, Dehalochip, for detecting the expression of genes in various microorganisms, particularly focusing on genes involved in chloroethene degradation. Our results demonstrated that this approach can effectively identify dechlorination genes, such as 16S rRNA, tceA, bvcA, and vcrA, in Dehalococcoides mccartyi from samples of groundwater contaminated with chloroethene. Noticeably, the sensitivity and specificity of our Dehalochip are comparable to that of quantitative PCR. However, it stands out as a more viable option for in-situ applications due to its greater capacity to infer potential dechlorination genes. Consequently, we believe our dechlorination microarray offers valuable insights into the role of known microorganisms and their associated functional genes in chloroethene-contaminated environments. This contributes to a deeper understanding of the in-situ reductive dechlorination process.

Comparative genomics reveal unique markers to monitor by routine PCR assay bioinoculant of Sphingobium indicum B90A in hexachlorocyclohexane (HCH) contaminated soils.

Bioinoculants of Sphingobium indicum B90A have been used to decontaminate hexachlorocyclohexane (HCH)-contaminated soils in the past. There is no selective or convenient method available to track the added B90A in HCH-contaminated soils in the presence of several native sphingomonads. Here, we describe a method, BioMarkTrack, for tracking B90A bioinoculant by simple amplification of the B90A specific biomarker genes. Whole-genome sequence data of 120 different genera of sphingomonads (Sphingobium, Novosphingobium, Sphingomonas, Sphingopyxis, and Sphingosinicella) were retrieved from the NCBI database and annotated. Intra- and inter-genus similarity searches, including the genome of B90A as a reference was conducted. 122 unique gene sequences were identified in strain B90A, out of which 45 genes were selected that showed no similarity with the NCBI non-redundant (NR) database or gene sequences in the publicly available database. Primers were designed for amplification of 4 biomarkers. To validate the biomarkers B90A tracking efficacy in bioaugmented soils, a microcosm study was conducted in which sterile garden and HCH-contaminated dumpsite soils were amended with strain B90A. Amplification of the biomarker was observed both in sterile garden soil and HCH-contaminated dumpsite soil but not in control (lacking B90A) samples. Further, the primer set was used to track B90A in a bioremediation field trial soil, demonstrating the convenience and efficiency of the simple PCR-based method, which can be employed for tracking B90A in bioaugmented soils. The approach as presented here can be employed on different bioinoculants to identify unique biomarkers and then tracking these organisms during bioremediation. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01321-7.

Bioaugmentation: a strategy for enhanced degradation of pesticides in biobed.

Biopurification system (BPS) or biobeds are low-cost system for decontamination of on-farm generated pesticide waste. A biobed contains a mixture of soil, lignocellulosic biomass and organic matter source (compost/peat) and works on the principal of retention of pesticide in high organic matter matrix and its subsequent degradation by microbes. Bioaugmentation, a green technology, is defined as the improvement of the degradative capacity of biobeds by augmenting specific microorganisms. During last 20 years, several studies have evaluated pesticide degradation in biobeds augmented with bacterial and fungal species and prominent microorganism include genus Pseudomonas, Sphingomonas, Arthrobacter, Phanerochaete, Stereum, Delftia, Trametes, Streptomyces etc. Degradation of pesticides belonging to major classes have been studied in the bioaugmented biobeds. Studies suggested that some pesticides were degraded faster in the bioaugmented biobeds subject to survival and proliferation of degrading microbe. However, no effect of bioaugmentation was observed on degradation of some pesticides and no clear reason for the same was evident. Bioaugmentation with pesticide degrading microorganisms/consortium in combination with rhizosphere-assisted biodegradation could be an optimal strategy for accelerating the degradation of pesticides in biobeds.

Recent advances in methane and hydrogen production from lignocellulosic degradation with anaerobic fungi.

Anaerobic fungi (AF) efficiently degrade lignocellulosic biomass with unique pseudoroot system and enzymatic properties that can remove polysaccharides and some lignified components from plant cell walls, further releasing acetate, lactate, ethanol, hydrogen (H2), etc. As research on AF for bioengineering has become a hot topic, a review of lignocellulosic conversion with AF for methane (CH4) and H2 production is needed. Efficient degradation of lignocellulose with AF mainly relies on multiple free carbohydrate-active enzymes and cellulosomes in the free and bound state. Meanwhile, co-cultivation of AF and methanogens significantly improves the lignocellulose degradation and CH4 production, and the maximum CH4 yield reached 315 mL/g. Bioaugmentation of AF in anaerobic digestion increases the maximum CH4 yield by 330 %. Also, AF show H2 production potential, however, H2 yield from anaerobic fungal fermentation of lignocellulose remains low. Therefore, anaerobic fungi have great potential in the conversion of lignocellulosic biomass to CH4 and H2.

Improving microbial activity in high-salt wastewater: A review of innovative approaches.

The Zero discharge technology has become an important pathroute for sustainable development of high salt wastewater treatment. However, the cohabitation of organic and inorganic debris can cause serious problems such membrane clogging and the formation of hazardous impurity salts that further restrict the recovery of all salt varieties by evaporating and crystallizing. In highly salinized wastewater, biological treatments offer advantages in terms of cost and sustainability when used as a pre-treatment step to eliminate organic debris. On the other hand, high salinity is always a major obstacle to microbial diversity, abundance, and activity, which can result in low organic matter removal effectiveness or the failure of the microbial treatment system. Biofortification techniques can attenuate the negative effects of salt stress and other unfavourable conditions on microorganisms, while the regulation mechanisms of microbial and community collaboration by fortification methods have been an open question. Therefore, a comprehensive summary of the types, mechanisms, and effects of the major biofortification techniques is proposed. This review dialyzes the characteristics and sources of hypersaline wastewater and the main treatment methods. Then, the mechanisms of microbial salt tolerance are summarized and discussed based on microbial characteristics and the protective effects provided by the processes. Finally, the research and application of the main bioaugmentation methods are developed in detail, describing the characteristics, advantages and disadvantages of the different enhancement methods in their implementation. This review provides a more comprehensive perspective on the future engineering applications of bioaugmentation technology, and explores in depth the possibilities of applying biological methods to high-salinity wastewater treatment.

Bioaugmentation demonstrates similar outcomes and failure rates for arthroscopic revision rotator cuff repair compared to revision without bioaugmentation.

Background: Arthroscopic revision rotator cuff repairs (RCRs) exhibit lower healing rates and inferior outcomes compared to primary repairs. There is limited evidence regarding the use of bioaugmentation in the setting of revision RCRs. Autologous conditioned plasma (ACP) is a promising adjunct that has been shown to improve healing rates and patient-reported outcomes (PROs) in the primary setting. In addition, bioinductive patches such as collagen bovine patches have become a popular adjunct for stimulating healing in the primary setting. The aim of this study is to assess the outcomes after use of ACP and collagen bovine patch augmentation for revision arthroscopic RCR. We hypothesized improved PROs and higher healing rates would be observed with bioaugmentation for revision repair compared to without. Methods: This was an institutional review board-approved, retrospective case-control study from 2 fellowship-trained surgeons that included all consecutive patients undergoing arthroscopic revision RCR from 2010 to 2021. Reconstruction such as superior capsular reconstruction, partial revision repair, and less than 1-year follow-up were excluded. The bioaugmentation cohort received ACP and/or collagen bovine patch at the time of revision repair. PROs were collected from all patients including American Shoulder and Elbow Surgeons Standardized Assessment Form (ASES), visual analog scale for pain (VAS), Brophy score, and Patient-Reported Outcomes Measurement Information System (PROMIS) mental and physical scores. Failure of revision RCR was defined as an ASES postoperative total score less than 60 or a symptomatic retear confirmed on magnetic resonance imaging. Student's t-test was used for all comparisons of continuous variables. Chi-squared test used for comparison of all categorical variables. Statistical significance was set at <0.05. Results: Thirty-eight patients met inclusion criteria with average follow-up of 3.5 ± 1.7 years. There was no significant difference in follow-up between patients with and without bioaugmentation. Of the 38 patients, 14 patients met failure criteria. There was no significant difference in the rate of failure between the bioaugmentation cohort (6/19, 31.6%) vs. patients who did not receive bioaugmentation (8/19, 42.1%) (P = .74). In addition, no significant differences were identified for ASES (64.6 ± 20.1 vs. 57.5 ± 17.2, P = .32), Brophy (6.4 ± 5.2 vs. 6.0 ± 4.1, P = .84), PROMIS Mental (13.4 ± 3.9 vs. 11.7 ± 3.2), or PROMIS Physical (12.8 ± 3.1 vs. 11.9 ± 3.2) scores between the bioaugmentation vs. no bioaugmentation groups. Conclusion: Bioaugmentation with a bioinductive collagen patch or ACP demonstrated similar failure and PROs compared to without bioaugmentation in the setting of revision RCR.

Tracking atrazine degradation in soil combining 14C-mineralisation assays and compound-specific isotope analysis.

The quantification of pesticide dissipation in agricultural soil is challenging. In this study, we investigated atrazine biodegradation in both liquid and soil experiments bioaugmented with distinct atrazine-degrading bacterial isolates. This was achieved by combining 14C-mineralisation assays and compound-specific isotope analysis of atrazine. In liquid experiments, the three bacterial isolates mineralised over 40% of atrazine, demonstrating their potential for extensive degradation. However, the kinetics of mineralisation and degradation varied among the isolates. Carbon stable isotope fractionation was similar for Pseudomonas isolates ADPT34 and ADP2T0, but slightly higher for Chelatobacter SR27. In soil experiments, atrazine primarily degraded into atrazine-desethyl, while atrazine-hydroxy was mainly observed in experiments with SR27. Atrazine mineralisation in soil by ADPT34 and SR27 exceeded 40%, whereas ADP2T0 exhibited a mineralisation rate of 10%. In experiments with ADPT34 and SR27, atrazine 14C-residues were predominantly found in the non-extractable fraction, whereas they accumulated in the extractable fraction in the experiment with ADP2T0. Compound-specific isotope analysis (CSIA) relies on changes of stable isotope ratios and holds potential to evaluate herbicide transformation in soil. CSIA of atrazine indicated atrazine biodegradation in water and solvent extractable soil fractions and varied between 29% and 52%, depending on the bacterial isolate. Despite atrazine degradation in both soil fractions, a significant portion of atrazine residues persisted, depending on the bacterial degrader, initial cell concentration, and mineralisation and degradation rates. Overall, our approach can aid in quantifying atrazine persistence and degradation in soil, and in optimizing bioaugmentation strategies for remediating soils contaminated with persistent herbicides.

Efficient caproate production from lignocellulose via single-step electro-fermentation platform without organic electron donor.

Caproate production by microbial fermentation gained the advantages of sustainability and eco-friendliness, but challenged by sterile fermentation environment, necessity of organic electron donors. Here, a single-step electro-fermentation (EF) process of mixed culture was proposed for caprate production from rice straw. At the optimal potential of -0.8 V, caproate concentration, yield and selectivity in the neutral red (NR)-mediated EF system were 2.4 g/L, 0.2 g/g and 26.6%. Long-term operation accumulated 5.3 g/L caproate with the yield and selectivity of 0.2 g/g and 34.2% in the EF+NR system. Bioaugmentation by dosing chain-elongation microbial consortium further improved the caproate production, yield and selectivity to 9.1 g/L, 0.3 g/g and 41.5%, respectively. The improved caproate production in the bioaugmented EF+NR system was likely due to the enhanced interspecies electron transfer, reconstructed microbial community, multiple electron donors and suitable pH environment. Present study offers a feasible strategy for cost-effective caprate production directly from waste biomass.

Metagenomes, metagenome-assembled genomes, and metatranscriptomes from a chlorinated ethene-dechlorinating culture amended with biochar pyrolyzed at different temperatures.

We investigated the effects of biochar and pyrolysis temperature on a chlorinated ethene-dechlorinating anaerobic consortium. Sequencing of nucleic acids from suspended and biochar-attached cells yielded 9 metagenomes, 122 metagenome-assembled genomes, and 18 metatranscriptomes that provide insights into the structure, function, activity, and interactions of the dehalogenating consortium with biochar.

Co-bioaugmentation with microalgae and probiotic bacteria: Sustainable solutions for upcycling of aquaculture wastewater and agricultural residues into microbial-rice bran complexes.

Aquaculture farming generates a significant amount of wastewater, which has prompted the development of creative bioprocesses to improve wastewater treatment and bioresource recovery. One promising method of achieving these aims is to directly recycle pollutants into microbe-rice bran complexes, which is an economical and efficient technique for wastewater treatment that uses synergetic interactions between algae and bacteria. This study explores novel bioaugmentation as a promising strategy for efficiently forming microbial-rice bran complexes in unsterilized aquaculture wastewater enriched with agricultural residues (molasses and rice bran). Results found that rice bran serves a dual role, acting as both an alternative nutrient source and a biomass support for microalgae and bacteria. Co-bioaugmentation, involving the addition of probiotic bacteria (Bacillus syntrophic consortia) and microalgae consortiums (Tetradesmus dimorphus and Chlorella sp.) to an existing microbial community, led to a remarkable 5-fold increase in microbial-rice bran complex yields compared to the non-bioaugmentation approach. This method provided the most compact biofloc structure (0.50 g/L) and a large particle diameter (404 µm). Co-bioaugmentation significantly boosts the synthesis of extracellular polymeric substances, comprising proteins at 6.5 g/L and polysaccharides at 0.28 g/L. Chlorophyta, comprising 80% of the total algal phylum, and Proteobacteria, comprising 51% of the total bacterial phylum, are emerging as dominant species. These microorganisms play a crucial role in waste and wastewater treatment, as well as in the formation of microbial-rice bran complexes that could serve as an alternative aquaculture feed. This approach prompted changes in both microbial community structure and nutrient cycling processes, as well as water quality. These findings provide valuable insights into the transformative effects of bioaugmentation on the development of microbial-rice bran complexes, offering potential applications in bioprocesses for waste and wastewater management.

SIP-metagenomics reveals key drivers of rhizospheric Benzo[a]pyrene bioremediation via bioaugmentation with indigenous soil microbes.

Rhizoremediation and bioaugmentation have proven effective in promoting benzo[a]pyrene (BaP) degradation in contaminated soils. However, the mechanism underlying bioaugmented rhizospheric BaP degradation with native microbes is poorly understood. In this study, an indigenous BaP degrader (Stenotrophomonas BaP-1) isolated from petroleum-contaminated soil was introduced into ryegrass rhizosphere to investigate the relationship between indigenous degraders and rhizospheric BaP degradation. Stable isotope probing and 16S rRNA gene amplicon sequencing subsequently revealed 15 BaP degraders, 8 of which were directly associated with BaP degradation including Bradyrhizobium and Streptomyces. Bioaugmentation with strain BaP-1 significantly enhanced rhizospheric BaP degradation and shaped the microbial community structure. A correlation of BaP degraders, BaP degradation efficiency, and functional genes identified active degraders and genes encoding polycyclic aromatic hydrocarbon-ring hydroxylating dioxygenase (PAH-RHD) genes as the primary drivers of rhizospheric BaP degradation. Furthermore, strain BaP-1 was shown to not only engage in BaP metabolism but also to increase the abundance of other BaP degraders and PAH-RHD genes, resulting in enhanced rhizospheric BaP degradation. Metagenomic and correlation analyses indicated a significant positive relationship between glyoxylate and dicarboxylate metabolism and BaP degradation, suggesting a role for these pathways in rhizospheric BaP biodegradation. By identifying BaP degraders and characterizing their metabolic characteristics within intricate microbial communities, our study offers valuable insights into the mechanisms of bioaugmented rhizoremediation with indigenous bacteria for high-molecular-weight PAHs in petroleum-contaminated soils.

Field test of a bioaugmentation agent for the bioremediation of chlorinated ethene contaminated sites.

Chlorinated ethenes are toxic compounds that were widely used in the past, and their improper handling and storage caused notable pollutions worldwide. In situ bioremediation by reductive dechlorination of bacteria is a cost-effective and ecologically friendly way to eliminate these pollutions. During the present study, the efficiency of a previously developed bioaugmentation agent combined with biostimulation was tested under field conditions in contaminated soil. Furthermore, the preservation of dechlorinating ability was also investigated in a long-term experiment. Initially, aerobic conditions were present in the groundwater with possible presence of anaerobic micro-niches providing habitat for Brocadia related anammox bacteria. "Candidatus Omnitrophus" was also identified as a dominant member of community then. Significant changes were detected after the biostimulation, anaerobic conditions established and most of the dominant OTUs were related to fermentative taxa (e.g. Clostridium, Trichococcus and Macillibacteroides). Dominant presence of vinyl-chloride coupled with the lack of vinyl-chloride reductase gene was observed. The most notable change after the bioaugmentation was the significant decrease in the pollutant quantities and the parallel increase in the vcrA gene copy numbers. Similar to post-biostimulation state, fermentative bacteria dominated the community. Bacterial community composition transformed considerably with time after the treatment, dominance of fermentative-mainly Firmicutes related-taxa decreased and chemolithotrophic bacteria became abundant, but the dechlorinating potential of the community remained and could be induced by the reappearance of the pollutants even after 4 years.

Fruit and vegetable waste used as bacterial growth media for the biocementation of two geomaterials.

This paper investigates the feasibility of using randomly collected fruit and vegetable (FV) waste as a cheap growing medium of bacteria for biocementation applications. Biocementation has been proposed in the literature as an environmentally-friendly ground improvement method to increase the stability of geomaterials, prevent erosion and encapsulate waste, but currently suffers from the high costs involved, such as bacteria cultivation costs. After analysis of FV waste of varied composition in terms of sugar and protein content, diluted FV waste was used to grow ureolytic (S. pasteurii, and B.licheniformis) and also an autochthonous heterotrophic carbonic anhydase (CA)-producing B.licheniformis strain, whose growth in FV media had not been attempted before. Bacterial growth and enzymatic activity in FV were of appropriate levels, although reduced compared to commercial media. Namely, the CA-producing B.licheniformis had a maximum OD600 of 1.799 and a CA activity of 0.817 U/mL in FV media. For the ureolytic pathway, B. licheniformis reached a maximum OD600 of 0.986 and a maximum urease activity of 0.675 mM urea/min, and S. pasteurii a maximum OD600 = 0.999 and a maximum urease activity of 0.756 mM urea/min. Biocementation of a clay and locomotive ash, a geomaterial specific to UK railway embankments, using precultured bacteria in FV was then proven, based on recorded unconfined compressive strengths of 1-3 MPa and calcite content increases of up to 4.02 and 8.62 % for the clay and ash respectively. Scanning Electron Microscope (SEM) and energy dispersive X-ray spectroscopy (EDS), attested the formation of bioprecipitates with characteristic morphologies and elementary composition of calcite crystals. These findings suggest the potential of employing FV to biocement these problematic geomaterials and are of wider relevance for environmental and geoenvironmental applications involving bioaugmentation. Such applications that require substrates in very large quantities can help tackle the management of the very voluminous fruit and vegetable waste produced worldwide.

Bioaugmentation protocols involving Methanobrevibacter thaueri and Pecoramyces ruminantium for investigating lignocellulose degradation and methane production from alfalfa stalks.

Two protocols involving batch cultures were used to investigate the bioaugmentation of methane production by Pecoramyces ruminantium, and Methanobrevibacter thaueri. Protocol I examined the effect of altering the proportion of the microbial constituents in inoculum on alfalfa stalk fermentations and showed a 25 % improvement in dry matter loss in cultures where the inoculum contained just 30 % of co-culture and 70 % of fungal monoculture. Protocol II involved consecutive cultures and alternating inoculations. This protocol resulted in 17-22 mL/g DM methane production with co-cultures a 30 % increase in methane relative to the fungal monoculture. Both protocols indicate that the co-culture rapidly dominated and was more resilient than the monoculture. Synergistic interaction between fungus and methanogen, promoted more efficient lignocellulose degradation and higher methane yield. This study highlighted the potential of microbial co-cultures for enhancing methane production from lignocellulosic biomass, offering a promising bioaugmentation strategy for improving biogas yields and waste valorization.

A novel simultaneous short-course nitrification, denitrification and fermentation process: bio-enhanced phenol degradation and denitrification in a single reactor.

The feasibility of a simultaneous nitrification, denitrification and fermentation process (SNDF) under electric stirrer agitation conditions was verified in a single reactor. Enhanced activated sludge for phenol degradation and denitrification in pharmaceutical phenol-containing wastewater under low dissolved oxygen conditions, additional inoculation with Comamonas sp. BGH and optimisation of co-metabolites were investigated. At a hydraulic residence time (HRT) of 28 h, 15 mg/L of substrate as strain BGH co-metabolised substrate degraded 650 ± 50 mg/L phenol almost completely and was accompanied by an incremental increase in the quantity of strain BGH. Strain BGH showed enhanced phenol degradation. Under trisodium citrate co-metabolism, strain BGH combined with activated sludge treated phenol wastewater and degraded NO2--N from 50 ± 5 to 0 mg/L in only 7 h. The removal efficiency of this group for phenol, chemical oxygen demand (COD) and TN was 99.67%, 90.25% and 98.71%, respectively, at an HRT of 32 h. The bioaugmentation effect not only promotes the degradation of pollutants, but also increases the abundance of dominant bacteria in activated sludge. Illumina MiSeq sequencing research showed that strain BGH promoted the growth of dominant genera (Acidaminobacter, Raineyella, Pseudarcobacter) and increased their relative abundance in the activated sludge system. These genera are resistant to toxicity and organic matter degradation. This paper provides some reference for the activated sludge to degrade high phenol pharmaceutical wastewater under the action of biological enhancement.

Biodegradation and bioaugmentation of tetracycline by Providencia stuartii TX2: Performance, degradation pathway, genetic background, key enzymes, and application risk assessment.

The antibiotic tetracycline (TC) is an emerging pollutant frequently detected in various environments. Biodegradation is a crucial approach for eliminating TC contamination. However, only a few efficient TC-degrading bacteria have been isolated, and the molecular mechanisms of TC degradation, as well as their application potential, remain poorly understood. This study isolated a novel TC-degrading bacterium, Providencia stuartii TX2, from the intestine of black soldier fly larvae. TX2 exhibited remarkable performance, degrading 72.17 % of 400 mg/L TC within 48 h. Genomic analysis of TX2 unveiled the presence of antibiotic resistance genes and TC degradation enzymes. Transcriptomic analysis highlighted the roles of proteins related to efflux pumps, enzymatic transformation, adversity resistance, and unknown functions. Three TC degradation pathways were proposed, with TC being transformed into 27 metabolites through epimerization, hydroxylation, oxygenation, ring opening, and de-grouping, reducing TC toxicity. Additionally, TX2 significantly enhanced TC biodegradation in four TC-contaminated environmental samples and reduced antibiotic resistance genes and mobile genetic elements in chicken manure. This research provides insights into the survival and biodegradation mechanisms of Providencia stuartii TX2 and evaluates its potential for environmental bioremediation.

Enhancing soil petrochemical contaminant remediation through nutrient addition and exogenous bacterial introduction.

Biostimulation (providing favorable environmental conditions for microbial growth) and bioaugmentation (introducing exogenous microorganisms) are effective approaches in the bioremediation of petroleum-contaminated soil. However, uncertainty remains in the effectiveness of these two approaches in practical application. In this study, we constructed mesocosms using petroleum hydrocarbon-contaminated soil. We compared the effects of adding nutrients, introducing exogenous bacterial degraders, and their combination on remediating petroleum contamination in the soil. Adding nutrients more effectively accelerated total petroleum hydrocarbon (TPH) degradation than other treatments in the initial 60 days' incubation. Despite both approaches stimulating bacterial richness, the community turnover caused by nutrient addition was gentler than bacterial degrader introduction. As TPH concentrations decreased, we observed a succession in microbial communities characterized by a decline in copiotrophic, fast-growing bacterial r-strategists with high rRNA operon (rrn) copy numbers. Ecological network analysis indicated that both nutrient addition and bacterial degrader introduction enhanced the complexity and stability of bacterial networks. Compared to the other treatment, the bacterial network with nutrient addition had more keystone species and a higher proportion of negative associations, factors that may enhance microbial community stability. Our study demonstrated that nutrient addition effectively regulates community succession and ecological interaction to accelerate the soil TPH degradation.

Genetically modified indigenous Pseudomonas aeruginosa drove bacterial community to change positively toward microbial enhanced oil recovery applications.

AIMS: Microbial enhanced oil recovery (MEOR) is cost-effective and eco-friendly for oil exploitation. Genetically modified biosurfactants-producing high-yield strains are promising for ex-situ MEOR. However, can they survive and produce biosurfactants in petroleum reservoirs for in-situ MEOR? What is their effect on the native bacterial community? METHODS AND RESULTS: A genetically modified indigenous biosurfactants-producing strain Pseudomonas aeruginosa PrhlAB was bioaugmented in simulated reservoir environments. Pseudomonas aeruginosa PrhlAB could stably colonize in simulated reservoirs. Biosurfactants (200 mg l-1) were produced in simulated reservoirs after bio-augmenting strain PrhlAB. The surface tension of fluid was reduced to 32.1 mN m-1. Crude oil was emulsified with an emulsification index of 60.1%. Bio-augmenting strain PrhlAB stimulated the MEOR-related microbial activities. Hydrocarbon-degrading bacteria and biosurfactants-producing bacteria were activated, while the hydrogen sulfide-producing bacteria were inhibited. Bio-augmenting P. aeruginosa PrhlAB reduced the diversity of bacterial community, and gradually simplified the species composition. Bacteria with oil displacement potential became dominant genera, such as Shewanella, Pseudomonas, and Arcobacter. CONCLUSIONS: Culture-based and sequence-based analyses reveal that genetically modified biosurfactants-producing strain P. aeruginosa PrhlAB are promising for in-situ MEOR as well.

Optimal conditions and nitrogen removal performance of aerobic denitrifier Comamonas sp. pw-6 and its bioaugmented application in synthetic domestic wastewater treatment.

To assess the possibility of using aerobic denitrification (AD) bacteria with high NO2--N accumulation for nitrogen removal in wastewater treatment, conditional optimization, as well as sole and mixed nitrogen source tests involving AD bacterium, Comamonas sp. pw-6 was performed. The results showed that the optimal carbon source, pH, C/N ratio, rotational speed, and salinity for this strain were determined to be succinate, 7, 20, 160 rpm, and 0%, respectively. Further, this strain preferentially utilized NH4+-N, NO3--N, and NO2--N, and when NO3--N was its sole nitrogen source, 92.28% of the NO3--N (150 mg·L-1) was converted to NO2--N. However, when NH4+-N and NO3--N constituted the mixed nitrogen source, NO3--N utilization by this strain was significantly lower (p < 0.05). Therefore, a strategy was proposed to combine pw-6 bacteria with traditional autotrophic nitrification to achieve the application of pw-6 bacteria in NH4+-N-containing wastewater treatment. Bioaugmented application experiments showed significantly higher NH4+-N removal (5.96 ± 0.94 mg·L-1·h-1) and lower NO3--N accumulation (2.52 ± 0.18 mg·L-1·h-1) rates (p < 0.05) than those observed for the control test. Thus, AD bacteria with high NO2--N accumulation can also be used for practical applications, providing a basis for expanding the selection range of AD strains for wastewater treatment.

Mechanism of self-supporting montmorillonite composite material for bio-enhanced degradation of chlorotetracycline: Electron transfer and microbial response.

The efficient degradation of antibiotics holds significant implications for mitigating environmental pollution. This study synthesized a montmorillonite chitosan composite material (MMT-CS) using the gel template method. Subsequently, a bio-enhanced reactor was constructed to facilitate the degradation of chlorotetracycline (CTC). The addition of MMT-CS composite material enables the degradation of different concentrations of CTC. MMT-CS, a conductive carrier, effectively promotes microbial adhesion and boosts the metabolic activity of functional microorganisms. Additionally, it facilitates the maintenance of microbial activity under CTC pressure by promoting the secretion of extracellular polymeric substances, increasing critical enzyme activity, and enhancing the electron transfer capacity within the system. In this MMT-CS bio-enhanced process, Paracoccus (11.4%) and Bacillus (3.9%) are utilized as essential bacteria genes. The results of metabolic pathways prediction indicated significant enhancements in membrane-transport, nucleotide-metabolism, replication-repair, and lipid-metabolism. Thus, the developed self-supporting MMT-CS bio-enhanced process ensured the stability of the system during the removal of antibiotics.