Ammonia-stressed anaerobic digestion: Sensitivity dynamics of key syntrophic interactions and methanogenic pathways-A review.

The problematic anaerobic digestion (AD) of protein-rich substrates owing to their high ammonia content continues to hinder optimum methanation despite their high potential for offsetting greenhouse gas (GHG) emissions. This review focuses on the analyses of the sensitivity dynamics of key AD processes as well as the microbial interactions and exchanges that occur with them. Aside from the apparent increased risk associated with thermophilic ammonia-rich substrate AD, the marginally higher energy generation compared to mesophilic systems is not commensurate to the energy requirement. Moreover, while comparable FAN thresholds have been confirmed, TAN thresholds are susceptible to physical chemistry and so vary greatly. Profiling of the metabolic capability of front-end AD microbiome revealed Bacteroidetes, Firmicutes, and Synergistetes as some of the ammonia-resilient bacteria groups while Proteobacteria and Actinobacteria were the most fragile taxa. Besides the predominance of incomplete propionate oxidizing bacteria under ammonia stress conditions, syntrophic propionate oxidation (SPO) is usually shifted from the methylmalonyl CoA to the dismutation pathway. Furthermore, besides their different recoverability potentials, distinct methanogenic groups are differentially impacted by different ammonia species. Prevailing literature evidence suggests that conductive material assisted bioaugmentation with SAO-HM consortia, and in-situ H2 supplementation are the most effective for expediting electron transfer and relieving ammonia stress. These valuable insights should inform the design of targeted ammonia inhibition mitigation strategies.

Efficient production of hydrogen through bioaugmentation of the organic fraction of municipal solid waste by the newly isolated Clostridium sartagoforme SA1.

Bio-hydrogen from organic waste holds promise as renewable energy. However, its large-scale production is limited by technical challenges, with low H2 yields and the absence of robust microbial strains being the major ones. To address these limitations, H2-producing microbes have been isolated from a full-scale anaerobic digestor treating complex organic waste. Clostridium sartagoforme SA1 was selected because of high H2 yields from glucose, soluble starch, and carboxymethylcellulose. The strain was then tested for H2 production from the Organic Fraction of Municipal Solid Waste (OFMSW), rich in starch and cellulose, with productions up to 55 mLH2 g/VS. Additionally, C. sartagoforme SA1 confirmed high H2 performances even in the presence of OFMSW's indigenous microflora, increasing the H2 yield by 38 % and highlighting its robustness in a highly competitive environment. This is the first report describing the efficient adoption of a C. sartagoforme strain for bioaugmentation of non-sterile OFMSW towards high H2 yields.

Construction of a synthetic anaerobic dechlorination microbiome to degrade chlorinated ethenes by application of metabolic interactions principle.

Bioaugmentation is a bioremediation approach to treat groundwater contaminated with chlorinated ethenes, but currently it faces challenges such as poor microbiome stability and effectiveness, due to blind species integration and metabolic inhibition. The objective of this study was to create a controllable and functionally stable microbial community for dichlorination application. For this, we utilized targeted screening to identify dechlorinating bacteria from contaminated groundwater, that in combination would form an anaerobic dechlorination microbial community with stabilizing metabolic interactions between the constituents. The results showed that two organohalide-respiring bacterial (OHRB) species were isolated, and these were identified as Enterobacter bugandensis X4 and Enterobacter sichuanensis C4. Upon co-cultivation with lactic acid as the carbon source, the strains demonstrated metabolic interactions and synergistic dehalogenation ability towards trichloroethene (TCE). It was further demonstrated that the functional microbiome constructed with the strains was stable over time and exhibited a robust TCE degradation rate of 80.85% at 13.13 mg/L TCE within 10 days. Additionally, the complete conversion of TCE was achieved through microbiome bioaugmentation, this augmented microbiome increased the degradation rate towards 52.55 mg/L TCE by approximately 30% within 6 days. Additionally, bioaugmentation stimulated the growth of indigenous OHRB, such as Dehalobacter and Desulfovibrio. It also promoted a positive succession of the microbial community. These findings offer valuable insights into the microbial remediation of chlorinated ethenes-contaminated groundwater and offers novel ideas for the construction of an artificial functional microbiome.

Rapid screening of indigenous degrading microorganisms for enhancing in-situ bioremediation of organic pollutants-contaminated soil.

Organic pollutants (OPs) have caused severe environmental contaminations in the world and aroused wide public concern. Autochthonous bioaugmentation (ABA) is considered a reliable bioremediation approach for OPs contamination. However, the rapid screening of indigenous degrading strains from in-situ environments remains a primary challenge for the practical application of ABA. In this study, 3,5,6-Trichloro-2-pyridinol (TCP, an important intermediate in the synthesis of various pesticides) was selected as the target OPs, and DNA stable isotope probing (DNA-SIP) combined with high-throughput sequencing was employed to explore the rapid screening of indigenous degrading microorganisms. The results of DNA-SIP revealed a significant enrichment of OTU557 (Cupriavidus sp.) in the 13C-TCP-labeled heavy DNA fractions, indicating that it is the key strain involved in TCP metabolism. Subsequently, an indigenous TCP degrader, Cupriavidus sp. JL-1, was rapidly isolated from native soil based on the analysis of the metabolic substrate spectrum of Cupriavidus sp. Furthermore, ABA of strain JL-1 demonstrated higher remediation efficacy and stable survival compared to the exogenous TCP-degrading strain Cupriavidus sp. P2 in in-situ TCP-contaminated soil. This study presents a successful case for the rapid acquisition of indigenous TCP-degrading microorganisms to support ABA as a promising strategy for the in-situ bioremediation of TCP-contaminated soil.

Methanosarcina thermophila bioaugmentation with biochar growth support for valorisation of food waste via thermophilic anaerobic digestion.

Methanosarcina thermophila bioaugmentation on biochar as the growth support particle has previously been shown to enhance biomethane production of anaerobic digestion of food waste. In this paper, the duration of the beneficial effects is examined by a semi-continuous thermophilic regime starting from pooled digestate from a previous batch digestion. An additional experiment is performed to decouple the solids retention time, mitigating the washout effect and resulting in improved methane yield for 17 days. The second experiment is extended incorporating various permutations of biochar amendment, and the findings suggest that liquid soluble supplements are essential for prolonging the advantages. Experimental and microbiological analyses indicate that the biochar's enhancement is likely due to microbial factors like direct interspecies electron transfer (DIET) or syntrophic interactions, rather than physicochemical mechanisms.

The Influence of Activated Sludge Augmentation on Its Ability to Degrade Paracetamol.

Paracetamol is one of the most commonly used painkillers. Its significant production and consumption result in its presence in the environment. For that reason, paracetamol has a negative impact on the organisms living in ecosystems. Therefore, it is necessary to develop effective methods to remove paracetamol from sewage. One of the methods is the bioaugmentation of activated sludge with organisms with increased degradation potential in relation to paracetamol. This study determined the effectiveness of paracetamol degradation by activated sludge augmented with a free or immobilised Pseudomonas moorei KB4. To immobilise the strain, innovative capsules made of cellulose acetate were used, the structure of which provides an optimal environment for the development of bacteria. Augmentation with both a free and immobilised strain significantly improves the efficiency of paracetamol biodegradation by activated sludge. Over a period of 30 days, examined systems allowed ten doses of paracetamol decomposition, while the unaugmented system degraded only four. At the same time, using the immobilised strain does not significantly affect the functioning of the activated sludge, which was reflected in the stability of processes such as nitrification. Due to the high stability of the preparation, it can become a valuable tool in wastewater treatment processes.

Enhanced tetracycline removal in sequencing batch reactors by bioaugmentation using tetX-carrying strains: Efficiency and mechanisms.

Tetracyclines antibiotics (TCs) pose notable environmental challenges due to their persistence in the effluent of wastewater treatment systems. Bioaugmentation offers a promising strategy for their removal, yet information is still very limited. This study aimed to assess the efficacy of bioaugmentation using wild-type (Sphingobacterium sp. WM1) and engineered tetX-carrying (PUC-tetX) strains for enhancing tetracycline (TC) removal in sequencing batch reactors (SBRs). Bioaugmentation mitigated TC's inhibitory effects on denitrification and phosphorus removal processes within SBR systems. Specifically, strain WM1 outperformed strain PUC-tetX in removing TC from sludge and maintained a longer viability. TC addition (500 µg/L, at an environmentally relevant concentration) and bioaugmentation did not significantly impact overall microbial community diversity. Notably, the introduction of these exogenous bacteria markedly increased the abundance of the tetX gene, correlating with the increase in TC degradation. Interestingly, MAGs associated with the Chloroflexi phylum in bioaugmented reactors showed the transfer of the tetX gene to autochthonous bacterial species, promoting TC removal capability. These findings underscored the potential of bioaugmentation to enhance antibiotic removal and provided insights into the dynamics of ARGs and tetX gene within activated sludge systems.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.