Metagenomic and Metatranscriptomic Responses of Chemical Dispersant Application during a Marine Dilbit Spill.

The global increase in marine transportation of dilbit (diluted bitumen) can increase the risk of spills, and the application of chemical dispersants remains a common response practice in spill events. To reliably evaluate dispersant effects on dilbit biodegradation over time, we set large-scale (1,500 mL) microcosms without nutrient addition using a low dilbit concentration (30 ppm). Shotgun metagenomics and metatranscriptomics were deployed to investigate microbial community responses to naturally and chemically dispersed dilbit. We found that the large-scale microcosms could produce more reproducible community trajectories than small-scale (250 mL) ones based on the 16S rRNA gene amplicon sequencing. In the early-stage large-scale microcosms, multiple genera were involved in the biodegradation of dilbit, while dispersant addition enriched primarily Alteromonas and competed for the utilization of dilbit, causing depressed degradation of aromatics. The metatranscriptomic-based metagenome-assembled genomes (MAG) further elucidated early-stage microbial antioxidation mechanism, which showed that dispersant addition triggered the increased expression of the antioxidation process genes of Alteromonas species. Differently, in the late stage, the microbial communities showed high diversity and richness and similar compositions and metabolic functions regardless of dispersant addition, indicating that the biotransformation of remaining compounds can occur within the post-oil communities. These findings can guide future microcosm studies and the application of chemical dispersants for responding to a marine dilbit spill. IMPORTANCE In this study, we employed microcosms to study the effects of marine dilbit spill and dispersant application on microbial community dynamics over time. We evaluated the impacts of microcosm scale and found that increasing the scale is beneficial for reducing community stochasticity, especially in the late stage of biodegradation. We observed that dispersant application suppressed aromatics biodegradation in the early stage (6 days), whereas exerting insignificant effects in the late stage (50 days), from both substance removal and metagenomic/metatranscriptomic perspectives. We further found that Alteromonas species are vital for the early-stage chemically dispersed oil biodegradation and clarified their degradation and antioxidation mechanisms. These findings help us to better understand microcosm studies and microbial roles for biodegrading dilbit and chemically dispersed dilbit and suggest that dispersant evaluation in large-scale systems and even through field trails would be more realistic after marine oil spill response.

Occurrence, Impact, Analysis and Treatment of Metformin and Guanylurea in Coastal Aquatic Environments of Canada, USA and Europe.

This review discusses the occurrence, impact, analysis and treatment of metformin and guanylurea in coastal aquatic environments of Canada, USA and Europe. Metformin, a biguanide in chemical classification, is widely used as one of the most effective first-line oral drugs for type 2 diabetes. It is difficult to be metabolized by the human body and exists in both urine and faeces samples in these regions. Guanylurea is metformin's biotransformation product. Consequently, significant concentrations of metformin and guanylurea have been reported in wastewater treatment plants (WWTPs) and coastal aquatic environments. The maximum concentrations of metformin and guanylurea in surface water samples were as high as 59,000 and 4502ngL-1, respectively. Metformin can be absorbed in non-target organisms by plants and in Atlantic salmon (Salmo salar). Guanylurea has a confirmed mitotic activity in plant cells. Analysis methods of metformin are currently developed based on high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). The removal of metformin from aquatic environments in the target regions is summarized. The review helps to fill a knowledge gap and provides insights for regulatory considerations. The potential options for managing these emerging pollutants are outlined too.

Screening of biosurfactant-producing bacteria from offshore oil and gas platforms in North Atlantic Canada.

From offshore oil and gas platforms in North Atlantic Canada, crude oil, formation water, drilling mud, treated produced water and seawater samples were collected for screening potential biosurfactant producers. In total, 59 biosurfactant producers belong to 4 genera, namely, Bacillus, Rhodococcus, Halomonas, and Pseudomonas were identified and characterized. Phytogenetic trees based on 16S ribosomal deoxyribonucleic acid (16S rDNA) were constructed with isolated strains plus their closely related strains and isolated strains with biosurfactant producers in the literature, respectively. The distributions of the isolates were site and medium specific. The richness, diversity, and evenness of biosurfactant producer communities in oil and gas platform samples have been analyzed. Diverse isolates were found with featured properties such as effective reduction of surface tension, producing biosurfactants at high rate and stabilization of water-in-oil or oil-in-water emulsion. The producers and their corresponding biosurfactants had promising potential in applications such as offshore oil spill control, enhancing oil recovery and soil washing treatment of petroleum hydrocarbon-contaminated sites.

Bag2 protects against doxorubicin-induced cardiotoxicity by maintaining Pink1-mediated mitophagy.

The clinical application of Doxorubicin (DOX) is limited due to its cardiotoxicity. Mitophagy dysfunction is the primary cause of DOX-induced cardiotoxicity (DIC). However, the precise mechanism by which DOX regulates mitophagy remains elusive. Bag2 (BCL2-associated athanogene 2) is a cochaperone implicated in multiple pathological states. The aim of this study was to investigate the potential cardio-protective effects of Bag2 in DIC. C57BL/6 mice and AC16 cells were used to establish DIC model. The expression of Bag2 were measured by western blotting and immunohistochemical. The effects of Bag2 on DIC were assessed through functional gain and loss experiments. Through in vitro and in vivo experiments, we found that Bag2 expression was significantly reduced after DOX treatment. Both Bag2 knockdown and DOX administration resulted in apoptosis, mitochondrial dysfunction, and impaired mitophagy. Conversely, Bag2 overexpression exerted protective effects against these phenotypes induced by DOX stimulation. Mechanistically, Bag2 maintained mitophagy activation by binding to Pink1 and protecting it from proteasome-dependent degradation, thereby preserving mitochondrial function and protecting against myocardial lesions. Our findings suggest that Bag2 may serve as a promising therapeutic target for the treatment of DIC.

Gradient of acid mine drainage regulates microbial community assembly and the diversity of species associated with native plants.

Acid mine drainage (AMD) is considered as one of the most important global environmental challenges. Therefore, understanding the impact of AMD on the diversity of microbial communities associated with native plants is important for phytoremediation. In this study, the community assembly and microbial diversity associated with native plants growing along an AMD impact gradient was investigated using metabarcoding and high throughput iChip technique. The study revealed that across both domains of bacteria and fungi, richness and species diversity decreased according to AMD impact. Bacterial species diversity was more stratified according to the pH gradient than fungi, and the AMD impact on the plant-associated microbial diversity decreased towards the plant roots. The microbial community composition of the undisturbed site was significantly different from the AMD impacted sites, and the communities in the AMD impacted sites were further stratified according to the degree of impact. The overall microbial diversity was mediated by the AMD impact, niche differences and plant species differences. Dispersal limitation was the most important community assembly process in the undisturbed site, while the homogenous selection of Burkholderia, Actinospica, Puia and Bradyrhizobium increased along the AMD impact gradient. Differential abundance analysis further revealed that Umbelopsis, Burkholderia and Sphingomonas were among the biomarkers of the AMD impacted sites. Several strains of some of these responsive genera were subsequently isolated using the iChip. Overall, this study presents novel insight into the ecology of plant-associated microbial communities that are relevant for environmental monitoring and for enhancing the revegetation of AMD impacted sites.

Acute Social Stress Influences Moral Decision-Making Under Different Social Distances in Young Healthy Men.

Acute social stress has been shown to influence social decision-making. This study aimed to examine how social distance modulates the influence of acute social stress on young male moral decision-making. Sixty healthy male college students were randomly divided to be exposed to the Trier Social Stress Test (TSST) or a placebo version of the TSST (P-TSST) before they performed moral decision-making tasks. The results showed that participants under acute social stress showed obvious increases in subjective stress perception, negative affect, salivary cortisol, and alpha-amylase and made more altruistic choices for others compared to the control group. However, social distance regulates the promotion of this acute social stress, with the promotion effect being stronger in socially distant others. Furthermore, the interpersonal order difference of different social distances in altruistic decisions is smaller in low-conflict dilemmas than in high-conflict dilemmas. In addition, an increase in salivary cortisol was positively correlated with altruistic choices toward both acquaintances and strangers, whereas an increase in salivary alpha-amylase was only positively correlated with altruistic choices toward friends. The results suggest that social distance modulates the promotion of acute social stress on moral decision-making, which might stem from the divergent effects of cortisol and alpha-amylase.

Natural attenuation of oil in marine environments: A review.

Natural attenuation is an important process for oil spill management in marine environments. Natural attenuation affects the fate of oil by physical, chemical, and biological processes, which include evaporation, dispersion, dissolution, photo-oxidation, emulsification, oil particle aggregation, and biodegradation. This review examines the cumulative knowledge regarding these natural attenuation processes as well as their simulation and prediction using modelling approaches. An in-depth discussion is provided on how oil type, microbial community and environmental factors contribute to the biodegradation process. It describes how our understanding of the structure and function of indigenous oil degrading microbial communities in the marine environment has been advanced by the application of next generation sequencing tools. The synergetic and/or antagonist effects of oil spill countermeasures such as the application of chemical dispersants, in-situ burning and nutrient enrichment on natural attenuation were explored. Several knowledge gaps were identified regarding the synergetic and/or antagonistic effects of active response countermeasures on the natural attenuation/biodegradation process. This review highlighted the need for field data on both the effectiveness and potential detrimental effects of oil spill response options to support modelling and decision-making on their selection and application.

Machine learning-aided causal inference for unraveling chemical dispersant and salinity effects on crude oil biodegradation.

Chemical dispersants have been widely applied to tackle oil spills, but their effects on oil biodegradation in global aquatic systems with different salinities are not well understood. Here, both experiments and advanced machine learning-aided causal inference analysis were applied to evaluate related processes. A halotolerant oil-degrading and biosurfactant-producing species was selected and characterized within the salinity of 0-70 g/L NaCl. Notably, dispersant addition can relieve the biodegradation barriers caused by high salinities. To navigate the causal relationships behind the experimental data, a structural causal model to quantitatively estimate the strength of causal links among salinity, dispersant addition, cell abundance, biosurfactant productivity and oil biodegradation was built. The estimated causal effects were integrated into a weighted directed acyclic graph, which showed that overall positive effects of dispersant addition on oil biodegradation was mainly through the enrichment of cell abundance. These findings can benefit decision-making prior dispersant application under different saline environments.

Hinokitiol Protects Cardiomyocyte from Oxidative Damage by Inhibiting GSK3ß-Mediated Autophagy.

More and more attention has been paid to the use of traditional phytochemicals. Here, we first verified the therapeutic potential of a natural bioactive compound called Hinokitiol in myocardial ischemia reperfusion injury. Hinokitiol exerts cardioprotective effect through inhibition of GSK-3ß and subsequent elimination of excessive autophagy, tuning autophagic activity in moderate extent for remedial profit in acute myocardial infarction and myocardial ischemia reperfusion injury. Overall, our study establishes Hinokitiol as a novel available interventional treatment for myocardial ischemia reperfusion injury.

Responses of Alcanivorax species to marine alkanes and polyhydroxybutyrate plastic pollution: Importance of the ocean hydrocarbon cycles.

Understanding microbial responses to hydrocarbon and plastic pollution are crucial for limiting the detrimental impacts of environmental contaminants on marine ecosystems. Herein, we reported a new Alcanivorax species isolated from the North Atlantic Ocean capable of degrading alkanes and polyhydroxybutyrate (PHB) plastic (one of the emerging bioplastics that may capture the future plastic market). The whole-genome sequencing showed that the species harbors three types of alkane 1-monooxygenases (AlkB) and one PHB depolymerase (PhaZ) to initiate the degradation of alkanes and plastics. Growth profiling demonstrated that n-pentadecane (C15, the main alkane in the marine environment due to cyanobacterial production other than oil spills) and PHB could serve as preferential carbon sources. However, the cell membrane composition, PhaZ activity, and expression of three alkB genes were utterly different when grown on C15 and PHB. Further, Alcanivorax was a well-recognized alkane-degrader that participated in the ocean hydrocarbon cycles linking with hydrocarbon production and removal. Our discovery supported that the existing biogeochemical processes may add to the marine ecosystem's resilience to the impacts of plastics.

A critical review on the environmental application of lipopeptide micelles.

The importance of lipopeptide micelles in environmental applications has been highlighted. These vessels exhibit various sizes, shapes, and surface properties under different environmental conditions. An in-depth understanding of the tunable assembling behavior of biosurfactant micelles is of great importance for their applications. However, a systematic review of such behaviors with assorted micro/nano micellar structures under given environmental conditions, particularly under low temperature and high salinity, remains untapped. Such impacts on their environmental applications have yet to be summarized. This review tried to fill the knowledge gaps by providing a comprehensive summary of the recent knowledge advancement in genetically regulated lipopeptides production, micelles associated decontamination mechanisms in low temperature and high salinity environments, and up-to-date environmental applications. This work is expected to deliver valuable insights to guide lipopeptide design and discovery. The mechanisms concluded in this study could inspire the forthcoming research efforts in the advanced environmental application of lipopeptide micelles.

A cross-comparison of biosurfactants as marine oil spill dispersants: Governing factors, synergetic effects and fates.

Biosurfactant-based dispersants (BBDs) may be more effective, cost-efficient and environmentally friendly than dispersants currently used for oil spill response. An improved understanding of BBD performance is needed to advance their development and commercial use. In this study, the ability of four BBDs, i.e. sufactins, trehalose lipids, rhamnolipids and exmulsins, alone and as various combinations to disperse Arabian light crude oil and weathered Alaska North Slope crude oil was compared to a widely used commercial oil dispersant (Corexit 9500A). Surfactin and trehalose lipids, which have balanced surface activity/emulsification ability, showed dispersion efficacy comparable to Corexit 9500A. Rhamnolipids (primarily a surface-active agent) and exmulsins (primarily an emulsifier) when used alone had significantly lower efficacy. However, blends of these surfactants had excellent dispersion performance because of synergistic effects. Balanced surface activity and emulsification ability may be key to formulate effective BBDs. Of the BBDs evaluated, surfactins with an effective dispersant-to-oil ratio as low as 1:62.3 and trehalose lipids with high oil affinity, biodegradation rate, and low toxicity characteristics show the most promise for commercial development.

The effect of pressure variation on droplet size distribution of dispersed oil under simulated deep-water conditions.

Droplet size distribution of dispersed oil in deep-water is critical to the transport and biodegradation of spilled oil in deep-sea. Few studies have focused on the effects of pressure on chemically dispersed oil through experiments. This study thus simulated how the crude oil homogenously pre-dispersed by Corexit 9500A using baffled flasks would behave after being exposed to deep-water conditions. Key factors included dispersant-to-oil ratio (DOR), mixing energy (energy dissipation rate and Kolmogorov microscale), and pressure (up to 150 bar). The variations of pressure were demonstrated to have insignificant effects on the size distribution of pre-dispersed oil. Both the average and medium droplet sizes were correlated negatively with DOR and mixing energy in an established model with a p-value ≤ 0.0011. The log-normal and log-logistic distributions provided a reasonable fit to simulate the droplet size distribution. The two parameters of log-logistic distribution were dependent on DOR and mixing energy with a p-value < 0.005. The results would be valuable to advance the understanding of the behaviours and trajectories of chemically dispersed oil under deep-water conditions. The research helped provide more scientific evidence to improve the understanding of dispersed oil behaviours under high pressure and support deep-sea oil spill research and potential extension of the existing results from shallow water to deep water conditions.

Towards sulfide removal and sulfate reducing bacteria inhibition: Function of biosurfactants produced by indigenous isolated nitrate reducing bacteria.

The effectiveness of nitrate-mediated souring control highly depends on the interactions of sulfate reducing bacteria (SRB) and nitrate reducing bacteria (NRB). Biosurfactants produced by natural NRB are promising bio-agents for enhancing NRB competence towards SRB. However, the function of NRB-produced biosurfactants in NRB-SRB interactions remains unexplored due to the rarely successful isolation of natural biosurfactant-producing NRB. Hereby, biosurfactant-aided inhibitory control of SRB strain Desulfomicrobium escambiense ATCC 51164 by biosurfactant-producing NRB strain Pseudomonas stutzeri CX3, reported in our previous work, was investigated. Under non-sour conditions, insufficient nitrate injection resulted in limited SRB inhibition. Phospholipid fatty acid (PLFA) biomarkers traced the overall bacterial responses. Compositional PLFA patterns revealed biosurfactant addition benefitted both SRB and NRB towards stressful conditions. Under sour conditions, nitrite oxidation of sulfide proved to be the primary mechanism for sulfide removal. The subsequent elevation of redox potential and pH inhibited SRB activities. NRB-produced biosurfactants significantly enhanced SRB inhibition by NRB through more efficient sulfide removal and effective duration of nitrate in the microcosms. Biosurfactants specially produced by the NRB strain are for the first time reported to significantly strengthen SRB inhibition by NRB via reduced nitrate usage and prolonged effective duration of nitrate, which has encouraging potential in nitrate-dependent souring control.

Fish Waste Based Lipopeptide Production and the Potential Application as a Bio-Dispersant for Oil Spill Control.

There is a growing acceptance worldwide for the application of dispersants as a marine oil spill response strategy. The development of more effective dispersants with less toxicity and higher biodegradability would be a step forward in improving public acceptance and regulatory approvals for their use. By applying advances in environmental biotechnology, a bio-dispersant agent with a lipopeptide biosurfactant produced by Bacillus subtilis N3-1P as the key component was formulated in this study. The economic feasibility of producing biosurfactant (a high-added-value bioproduct) from fish waste-based peptone as a nutrient substrate was evaluated. Protein hydrolyzate was prepared from cod liver and head wastes obtained from fish processing facilities. Hydrolysis conditions (i.e., time, temperature, pH and enzyme to substrate level) for preparing protein hydrolyzates were optimized by response surface methodology using a factorial design. The critical micelle dilution (CMD) value for biosurfactant produced from the fish liver and head waste generated peptones was 54.72 and 47.59 CMD, respectively. Biosurfactant product generated by fish liver peptone had a low critical micelle concentration of 0.18 g L-1 and could reduce the surface tension of distilled water to 27.9 mN/m. Structure characterization proved that the generated biosurfactant product belongs to the lipopeptide class. An alternative to the key surfactant dioctyl sulfosuccinate sodium (DOSS) used in Corexit 9500 has been proposed based on a binary mixture of lipopeptides and DOSS that exhibited synergistic effects. Using the standard baffled flask test, a high dispersion efficiency of 76.8% for Alaska North Slope oil was achieved at a biodispersant composition of 80/20 (v/v) of lipopeptides/DOSS. The results show that fish waste can be utilized to produce a more effective, environmentally acceptable and cost-efficient biodispersant that can be applied to oil spills in the marine environment.

Microbial eco-physiological strategies for salinity-mediated crude oil biodegradation.

Salinity variability strongly affects the behaviors of oil degrading bacteria for spilled oil biodegradation in the marine environment. However, limited studies explored the strategies of microbes on salinity-mediated crude oil biodegradation. In this study, a halotolerant bio-emulsifier producer, Exiguobacterium sp. N41P, was examined as a model strain for Alaska North Slope (ANS) crude oil (0.5%, v/v) biodegradation. Results indicated that Exiguobacterium sp. N41P could tolerant a wide range of salinity (0-120 g/L NaCl) and achieve the highest degradation efficiency under the salinity of 15 g/L NaCl due to the highest biofilm formation ability. Moreover, increased salinity induced decreased cell surface hydrophobicity and a migration of microbial growth from oil phase to aqueous phase, leading to limited bio-emulsifier productivity and depressed degradation of insoluble long-chain n-alkanes while enhancing the degradation of relative soluble naphthalene. Research findings illustrated the microbial eco-physiological mechanism for spilled oil biodegradation under diverse salinities and advanced the understanding of sophisticated marine crude oil biodegradation process.

Oil-in-water emulsion breaking marine bacteria for demulsifying oily wastewater.

Oily wastewater is a large waste stream produced by a number of industries. This wastewater often forms stable oil-in-water (O/W) emulsion. These emulsions require demulsification in order to effectively treat the water prior to release. Although biological demulsification of O/W emulsion has advantages over traditional approaches, its development is at a preliminary stage with few demulsifying bacteria reported and a need for effective screening methods for such bacteria. In this study, thirty-seven marine O/W emulsion demulsifying bacterial strains belonging to 5 genera and 15 species were reported. Cell hydrophobicity and interfacial activity played key roles in the emulsion breaking. One of the highly effective demulsifying bacteria, Halomonas venusta strain N3-2A was identified and characterized. Both its extracellular biosurfactant and cell surface contributed to demulsification resulting in breaking of 92.5% of the emulsion within 24 h. A high throughput and effective screening strategy targeting O/W emulsion breaking bacteria using oil spreading test coupled with cell hydrophobicity test was proposed. In addition, the 37 demulsifying bacteria showed a certain degree of species/genus specific patterns of surface activity and cell hydrophobicity. The reported bacteria and the screening strategy have promising potential for the biological demulsification of O/W emulsions and oily wastewater treatment.

Effective removal of coordinated copper from wastewater using a new dithiocarbamate-type supramolecular heavy metal precipitant.

A new dithiocarbamate-type heavy metal precipitant, sodium 1,3,5-hexahydrotriazinedithiocarbamate (HTDC), was prepared and used to remove coordinated copper from wastewater. In the reported dithiocarbamate-type precipitants, HTDC possesses the highest percentage of the effective functional groups. It could effectively precipitate copper to less than 0.5mgl(-1) from both synthetic and actual industrial wastewater containing CuEDTA in the range of pH 3-9. UV-vis spectral investigation and elemental analysis suggested that the precipitate was a kind of coordination supramolecular compound, [Cu(3)(HTDC)(2)](n). The toxicity characteristic leaching procedure (TCLP) and semi-dynamic leaching test (SDLT) indicated that the supramolecular precipitate was non-hazardous and stable in weak acid and alkaline conditions. Tests of an anion exchange resin D231 provided a clue to simultaneously remove excess HTDC and residual CuEDTA in practical process of wastewater treatment.

Complete Genome Sequence of Exiguobacterium sp. Strain N4-1P, a Psychrophilic Bioemulsifier Producer Isolated from a Cold Marine Environment in North Atlantic Canada.

Here, we present the complete genome sequence of Exiguobacterium sp. strain N4-1P, a psychrophilic bacterium that produces bioemulsifier, isolated for the first time from petroleum hydrocarbon-contaminated sediment samples from shoreline Newfoundland, Canada. Many strains of the genus Exiguobacterium are extremophiles and have properties of biotechnological interest.

Microbial degradation of four crude oil by biosurfactant producing strain Rhodococcus sp.

Rhodococcus erythropolis M-25, one of the representative biosurfactant producers, performed effectively during the biodegradation of four crude oil. The microbial degradation efficiency is positively relevant to the API of the crude oil. The chemical dispersant Corexit 9500A did not enhance the biodegradation of the petroleum hydrocarbons during the experimental period. 70.7% of the N-4 oil was degraded after 30days, while in the Corexit 9500A plus sample the biodegradation removal was 42.8%. The Corexit-derived compounds were metabolized by M-25 at the same time of the petroleum hydrocarbons biodegrading. Neither biodegradation nor chemical dispersion process has almost no effect on the biomarker (m/z=231). The saturated methyl-branched fatty acids increased from 37.3%, to 49.4%, when M-25 was exposed with the N-4 crude oil. Similarly, the saturated methyl-branched fatty acids in the membrane of N3-2P increased from 20.25% to 44.1%, when exposed it with the N-4 crude oil.