Sediment-driven plastisphere community assembly on plastic debris in tropical coastal and marine environments.

Coastal habitats have been suggested to serve as a sink for unaccounted plastic debris, i.e., "missing plastic" in the sea, and hence, a hotspot of plastic pollution in the marine and coastal environments. Although the accumulation of plastic debris may pose significant threats to coastal ecosystems, we know little about the fate of these plastic debris and their ecological impacts due to the lack of studies on plastic-microbe interactions in coastal habitats, especially for the tropical marine and coastal environments. In this study, we collected plastic debris from 14 sites consisting of various coastal ecosystems (seagrass meadows, mangrove forests, and beaches), and marine ecosystem (coral reef) around Singapore and characterized the prokaryotic and eukaryotic microbial communities colonized on them. Our results showed that the composition of plastisphere communities in these intertidal ecosystems was predominantly influenced by the sediment than by the plastic materials. Compared with surrounding sediment and seawater, the plastic debris enriched potential plastic degraders, such as Muricauda, Halomonas, and Brevundimonas. The plastic debris was also found to host taxa that play significant roles in biogeochemical cycles (e.g., cyanobacteria, Erythrobacter), hygienically relevant bacteria (e.g., Chryseobacterium, Brevundimonas), and potential pathogens that may negatively impact the health of coastal ecosystems (e.g., Thraustochytriaceae, Labyrinthulaceae, Flavobacterium). Taken together, our study provides valuable insights into the plastic-microbe interactions in tropical coastal and marine ecosystems, highlighting the urgent need for plastisphere studies to understand the fate and ecological impacts of plastic debris accumulated in coastal habitats.

Early biofilm and streamer formation is mediated by wall shear stress and surface wettability: A multifactorial microfluidic study.

Biofilms are intricate communities of microorganisms encapsulated within a self-produced matrix of extra-polymeric substances (EPS), creating complex three-dimensional structures allowing for liquid and nutrient transport through them. These aggregations offer constituent microorganisms enhanced protection from environmental stimuli-like fluid flow-and are also associated with higher resistance to antimicrobial compounds, providing a persistent cause of concern in numerous sectors like the marine (biofouling and aquaculture), medical (infections and antimicrobial resistance), dentistry (plaque on teeth), food safety, as well as causing energy loss and corrosion. Recent studies have demonstrated that biofilms interact with microplastics, often influencing their pathway to higher trophic levels. Previous research has shown that initial bacterial attachment is affected by surface properties. Using a microfluidic flow cell, we have investigated the relationship between both wall shear stress (τw ) and surface properties (surface wettability) upon biofilm formation of two species (Cobetia marina and Pseudomonas aeruginosa). We investigated biofilm development on low-density polyethylene (LDPE) membranes, Permanox® slides, and glass slides, using nucleic acid staining and end-point confocal laser scanning microscopy. The results show that flow conditions affect biomass, maximum thickness, and surface area of biofilms, with higher τw (5.6 Pa) resulting in thinner biofilms than lower τw (0.2 Pa). In addition, we observed differences in biofilm development across the surfaces tested, with LDPE typically demonstrating more overall biofilm in comparison to Permanox® and glass. Moreover, we demonstrate the formation of biofilm streamers under laminar flow conditions within straight micro-channels.

High-throughput method development for in-situ quantification of aquatic phototrophic biofilms.

In the maritime field where biofouling has both economic and environmental impacts, in situ quantification methods of biofilm development are of outstanding importance. Indeed, it is challenging to temporally monitor biofilm formation due to the complexity of the marine ecosystem, common inaccessibility of sampling location and lack of standardized techniques. Here, an artificial polymeric surface was tested in situ and in vitro against natural phototrophic biofilms and monoculture biofilms using plate reader fluorescence. The suitability of the developed method was verified using fluorescence microscopy coupled with image analysis - a common quantification technique - demonstrating a strong correlation between the tested methods. The results indicated the efficiency of inherent chlorophyll fluorescence in quantifying undisturbed phototrophic biofilms in field and laboratory conditions using microplate reader. This work demonstrated that the suggested approach is promising for biofilm high-throughput testing, and therefore has the potential to be used in several research and industrial sectors for monitoring phototrophic biofilm development.

Priorities to inform research on marine plastic pollution in Southeast Asia.

Southeast Asia is considered to have some of the highest levels of marine plastic pollution in the world. It is therefore vitally important to increase our understanding of the impacts and risks of plastic pollution to marine ecosystems and the essential services they provide to support the development of mitigation measures in the region. An interdisciplinary, international network of experts (Australia, Indonesia, Ireland, Malaysia, the Philippines, Singapore, Thailand, the United Kingdom, and Vietnam) set a research agenda for marine plastic pollution in the region, synthesizing current knowledge and highlighting areas for further research in Southeast Asia. Using an inductive method, 21 research questions emerged under five non-predefined key themes, grouping them according to which: (1) characterise marine plastic pollution in Southeast Asia; (2) explore its movement and fate across the region; (3) describe the biological and chemical modifications marine plastic pollution undergoes; (4) detail its environmental, social, and economic impacts; and, finally, (5) target regional policies and possible solutions. Questions relating to these research priority areas highlight the importance of better understanding the fate of marine plastic pollution, its degradation, and the impacts and risks it can generate across communities and different ecosystem services. Knowledge of these aspects will help support actions which currently suffer from transboundary problems, lack of responsibility, and inaction to tackle the issue from its point source in the region. Being profoundly affected by marine plastic pollution, Southeast Asian countries provide an opportunity to test the effectiveness of innovative and socially inclusive changes in marine plastic governance, as well as both high and low-tech solutions, which can offer insights and actionable models to the rest of the world.

Marine biofilms on different fouling control coating types reveal differences in microbial community composition and abundance.

Marine biofouling imposes serious environmental and economic impacts on marine applications, especially in the shipping industry. To combat biofouling, protective coatings are applied on vessel hulls which are divided into two major groups: biocidal and non-toxic fouling release. The current study aimed to explore the effect of coating type on microbial biofilm community profiles to better understand the differences between the communities developed on fouling control biocidal antifouling and biocidal-free coatings. Biocidal (Intersmooth® 7460HS SPC), fouling release (Intersleek® 900), and inert surfaces were deployed in the marine environment for 4 months, and the biofilms that developed on these surfaces were investigated using Illumina NGS sequencing, targeting the prokaryotic 16S rRNA gene. The results confirmed differences in the community profiles between coating types. The biocidal coating supported communities dominated by Alphaproteobacteria (Loktanella, Sphingorhabdus, Erythrobacter) and Bacteroidetes (Gilvibacter), while other taxa, such as Portibacter and Sva0996 marine group, proliferated on the fouling-release surface. Knowledge of these marine biofilm components on fouling control coatings will serve as a guide for future investigations of marine microfouling as well as informing the coatings industry of potential microbial targets for robust coating formulations.

The oceans are changing: impact of ocean warming and acidification on biofouling communities.

Climate change (CC) is driving modification of the chemical and physical properties of estuaries and oceans with profound consequences for species and ecosystems. Numerous studies investigate CC effects from species to ecosystem levels, but little is known of the impacts on biofilm communities and on bioactive molecules such as cues, adhesives and enzymes. CC is induced by anthropogenic activity increasing greenhouse emissions leading to rises in air and water temperatures, ocean acidification, sea level rise and changes in ocean gyres and rainfall patterns. These environmental changes are resulting in alterations within marine communities and changes in species ranges and composition. This review provides insights and synthesis of knowledge about the effect of elevated temperature and ocean acidification on microfouling communities and bioactive molecules. The existing studies suggest that CC will impact production of bioactive compounds as well as the growth and composition of biofouling communities. Undoubtedly, with CC fouling management will became an even greater challenge.

Biofilm Inhibition by Novel Natural Product- and Biocide-Containing Coatings Using High-Throughput Screening.

The use of natural products (NPs) as possible alternative biocidal compounds for use in antifouling coatings has been the focus of research over the past decades. Despite the importance of this field, the efficacy of a given NP against biofilm (mainly bacteria and diatoms) formation is tested with the NP being in solution, while almost no studies test the effect of an NP once incorporated into a coating system. The development of a novel bioassay to assess the activity of NP-containing and biocide-containing coatings against marine biofilm formation has been achieved using a high-throughput microplate reader and highly sensitive confocal laser scanning microscopy (CLSM), as well as nucleic acid staining. Juglone, an isolated NP that has previously shown efficacy against bacterial attachment, was incorporated into a simple coating matrix. Biofilm formation over 48 h was assessed and compared against coatings containing the NP and the commonly used booster biocide, cuprous oxide. Leaching of the NP from the coating was quantified at two time points, 24 h and 48 h, showing evidence of both juglone and cuprous oxide being released. Results from the microplate reader showed that the NP coatings exhibited antifouling efficacy, significantly inhibiting biofilm formation when compared to the control coatings, while NP coatings and the cuprous oxide coatings performed equally well. CLSM results and COMSTAT analysis on biofilm 3D morphology showed comparable results when the NP coatings were tested against the controls, with higher biofilm biovolume and maximum thickness being found on the controls. This new method proved to be repeatable and insightful and we believe it is applicable in antifouling and other numerous applications where interactions between biofilm formation and surfaces is of interest.

Anti-biofilm performance of three natural products against initial bacterial attachment.

Marine bacteria contribute significantly towards the fouling consortium, both directly (modern foul release coatings fail to prevent "slime" attachment) and indirectly (biofilms often excrete chemical cues that attract macrofouling settlement). This study assessed the natural product anti-biofilm performance of an extract of the seaweed, Chondrus crispus, and two isolated compounds from terrestrial sources, (+)-usnic acid and juglone, against two marine biofilm forming bacteria, Cobetia marina and Marinobacter hydrocarbonoclasticus. Bioassays were developed using quantitative imaging and fluorescent labelling to test the natural products over a range of concentrations against initial bacterial attachment. All natural products affected bacterial attachment; however, juglone demonstrated the best anti-biofilm performance against both bacterial species at a concentration range between 5-20 ppm. In addition, for the first time, a dose-dependent inhibition (hormetic) response was observed for natural products against marine biofilm forming bacteria.

Marine biofilms on artificial surfaces: structure and dynamics.

The search for new antifouling (AF) coatings that are environmentally benign has led to renewed interest in the ways that micro-organisms colonize substrates in the marine environment. This review covers recently published research on the global species composition and dynamics of marine biofilms, consisting mainly of bacteria and diatoms found on man-made surfaces including AF coatings. Marine biofilms directly interact with larger organisms (macrofoulers) during colonization processes; hence, recent literature on understanding the basis of the biofilm/macrofouling interactions is essential and will also be reviewed here. Overall, differences have been identified in species composition between biofilm and planktonic forms for both diatoms and bacteria at various exposure sites. In most studies, the underlying biofilm was found to induce larval and spore settlement of macrofoulers; however, issues such as reproducibility, differences in exposure sites and biofilm composition (natural multispecies vs. monospecific species) may influence the outcomes.

Life under flow: A novel microfluidic device for the assessment of anti-biofilm technologies.

In the current study, we have developed and fabricated a novel lab-on-a-chip device for the investigation of biofilm responses, such as attachment kinetics and initial biofilm formation, to different hydrodynamic conditions. The microfluidic flow channels are designed using computational fluid dynamic simulations so as to have a pre-defined, homogeneous wall shear stress in the channels, ranging from 0.03 to 4.30 Pa, which are relevant to in-service conditions on a ship hull, as well as other man-made marine platforms. Temporal variations of biofilm formation in the microfluidic device were assessed using time-lapse microscopy, nucleic acid staining, and confocal laser scanning microscopy (CLSM). Differences in attachment kinetics were observed with increasing shear stress, i.e., with increasing shear stress there appeared to be a delay in bacterial attachment, i.e., at 55, 120, 150, and 155 min for 0.03, 0.60, 2.15, and 4.30 Pa, respectively. CLSM confirmed marked variations in colony architecture, i.e.,: (i) lower shear stresses resulted in biofilms with distinctive morphologies mainly characterised by mushroom-like structures, interstitial channels, and internal voids, and (ii) for the higher shear stresses compact clusters with large interspaces between them were formed. The key advantage of the developed microfluidic device is the combination of three architectural features in one device, i.e., an open-system design, channel replication, and multiple fully developed shear stresses.

Designing biomimetic antifouling surfaces.

Marine biofouling is the accumulation of biological material on underwater surfaces, which has plagued both commercial and naval fleets. Biomimetic approaches may well provide new insights into designing and developing alternative, non-toxic, surface-active antifouling (AF) technologies. In the marine environment, all submerged surfaces are affected by the attachment of fouling organisms, such as bacteria, diatoms, algae and invertebrates, causing increased hydrodynamic drag, resulting in increased fuel consumption, and decreased speed and operational range. There are also additional expenses of dry-docking, together with increased fuel costs and corrosion, which are all important economic factors that demand the prevention of biofouling. Past solutions to AF have generally used toxic paints or coatings that have had a detrimental effect on marine life worldwide. The prohibited use of these antifoulants has led to the search for biologically inspired AF strategies. This review will explore the natural and biomimetic AF surface strategies for marine systems.

A galvanic sensor for monitoring the corrosion condition of the concrete reinforcing steel: relationship between the galvanic and the corrosion currents.

This work reports a study carried out on the design and performance of galvanic and polarization resistance sensors to be embedded in concrete systems for permanent monitoring of the corrosion condition of reinforcing steel, aiming to establish a correlation between the galvanic currents, I(gal), and the corrosion currents, I(corr), estimated from the polarization resistance, R(p). Sensors have been tested in saturated Ca(OH)(2) aqueous solutions, under a variety of conditions, simulating the most important parameters that can accelerate the corrosion of concrete reinforcing steel, such as carbonation, ingress of chloride ions, presence or absence of O(2). For all the conditions, the influence of temperature (20 to 55 °C) has also been considered. From this study, it could be concluded that the galvanic currents are sensitive to the various parameters following a trend similar to that of the R(p) values. A relationship between the galvanic and the corrosion current densities was obtained and the limiting values of the I(gal), indicative of the state condition of the reinforcing steel for the designed sensor, were established.