Assessing microbial plastic degradation requires robust methods.

Plastics are versatile materials that have the potential to propel humanity towards circularity and ultimate societal sustainability. However, the escalating concern surrounding plastic pollution has garnered significant attention, leading to widespread negative perceptions of these materials. Here, we question the role microbes may play in plastic pollution bioremediation by (i) defining polymer biodegradability (i.e., recalcitrant, hydrolysable and biodegradable polymers) and (ii) reviewing best practices for evaluating microbial biodegradation of plastics. We establish recommendations to facilitate the implementation of rigorous methodologies in future studies on plastic biodegradation, aiming to push this field towards the use of isotopic labelling to confirm plastic biodegradation and further determine the molecular mechanisms involved.

Microbial hitchhikers harbouring antimicrobial-resistance genes in the riverine plastisphere.

BACKGROUND: The widespread nature of plastic pollution has given rise to wide scientific and social concern regarding the capacity of these materials to serve as vectors for pathogenic bacteria and reservoirs for Antimicrobial Resistance Genes (ARG). In- and ex-situ incubations were used to characterise the riverine plastisphere taxonomically and functionally in order to determine whether antibiotics within the water influenced the ARG profiles in these microbiomes and how these compared to those on natural surfaces such as wood and their planktonic counterparts. RESULTS: We show that plastics support a taxonomically distinct microbiome containing potential pathogens and ARGs. While the plastisphere was similar to those biofilms that grew on wood, they were distinct from the surrounding water microbiome. Hence, whilst potential opportunistic pathogens (i.e. Pseudomonas aeruginosa, Acinetobacter and Aeromonas) and ARG subtypes (i.e. those that confer resistance to macrolides/lincosamides, rifamycin, sulfonamides, disinfecting agents and glycopeptides) were predominant in all surface-related microbiomes, especially on weathered plastics, a completely different set of potential pathogens (i.e. Escherichia, Salmonella, Klebsiella and Streptococcus) and ARGs (i.e. aminoglycosides, tetracycline, aminocoumarin, fluoroquinolones, nitroimidazole, oxazolidinone and fosfomycin) dominated in the planktonic compartment. Our genome-centric analysis allowed the assembly of 215 Metagenome Assembled Genomes (MAGs), linking ARGs and other virulence-related genes to their host. Interestingly, a MAG belonging to Escherichia -that clearly predominated in water- harboured more ARGs and virulence factors than any other MAG, emphasising the potential virulent nature of these pathogenic-related groups. Finally, ex-situ incubations using environmentally-relevant concentrations of antibiotics increased the prevalence of their corresponding ARGs, but different riverine compartments -including plastispheres- were affected differently by each antibiotic. CONCLUSIONS: Our results provide insights into the capacity of the riverine plastisphere to harbour a distinct set of potentially pathogenic bacteria and function as a reservoir of ARGs. The environmental impact that plastics pose if they act as a reservoir for either pathogenic bacteria or ARGs is aggravated by the persistence of plastics in the environment due to their recalcitrance and buoyancy. Nevertheless, the high similarities with microbiomes growing on natural co-occurring materials and even more worrisome microbiome observed in the surrounding water highlights the urgent need to integrate the analysis of all environmental compartments when assessing risks and exposure to pathogens and ARGs in anthropogenically-impacted ecosystems. Video Abstract.

Assessing the impact of chronic and acute plastic pollution from construction activities and other anthropogenic sources: A case study from the coast of Antofagasta, Chile.

Plastic pollution is a critical environmental issue with far-reaching and not yet fully explored consequences. This study uncovered a significant source of plastic contamination arising from improper application and management of expanded polystyrene (EPS) utilised as expansion joints at a construction site near the coast of Antofagasta, Chile. Through meticulous field observations and calculations, we estimate that a staggering 82.9 million EPS spheres have the potential to be released into the environment from the 7.62 m3 of this material used for the construction of this coastal promenade, constituting a chronic source of pollution. Despite the ongoing construction, we have already evidenced mechanical fragmentation and dispersion of EPS microplastic pollution in the surrounding natural environment. To our knowledge, this is the first study that documents misused construction materials contributing to plastic pollution. In addition to the EPS pollution, our findings reveal an alarming accumulation of litter - an acute pollution source - including plastic cups, bottles, carrier bags, and several other construction materials (e.g. plastic nets, films) that are exacerbating the pollution problems within the region and potentially endangering marine and terrestrial organisms. These observations highlight the urgent need for mitigating measures and intervention policies targeting construction-related plastic and microplastic pollution, along with a more robust regulatory framework for construction activities as well as adequate surveillance and enforcement.

A mechanistic understanding of polyethylene biodegradation by the marine bacterium Alcanivorax.

Polyethylene (PE) is one of the most recalcitrant carbon-based synthetic materials produced and, currently, the most ubiquitous plastic pollutant found in nature. Over time, combined abiotic and biotic processes are thought to eventually breakdown PE. Despite limited evidence of biological PE degradation and speculation that hydrocarbon-degrading bacteria found within the plastisphere is an indication of biodegradation, there is no clear mechanistic understanding of the process. Here, using high-throughput proteomics, we investigated the molecular processes that take place in the hydrocarbon-degrading marine bacterium Alcanivorax sp. 24 when grown in the presence of low density PE (LDPE). As well as efficiently utilising and assimilating the leachate of weathered LDPE, the bacterium was able to reduce the molecular weight distribution (Mw from 122 to 83 kg/mol) and overall mass of pristine LDPE films (0.9 % after 34 days of incubation). Most interestingly, Alcanivorax acquired the isotopic signature of the pristine plastic and induced an extensive array of metabolic pathways for aliphatic compound degradation. Presumably, the primary biodegradation of LDPE by Alcanivorax sp. 24 is possible via the production of extracellular reactive oxygen species as observed both by the material's surface oxidation and the measurement of superoxide in the culture with LDPE. Our findings confirm that hydrocarbon-biodegrading bacteria within the plastisphere may in fact have a role in degrading PE.

Pili allow dominant marine cyanobacteria to avoid sinking and evade predation.

How oligotrophic marine cyanobacteria position themselves in the water column is currently unknown. The current paradigm is that these organisms avoid sinking due to their reduced size and passive drift within currents. Here, we show that one in four picocyanobacteria encode a type IV pilus which allows these organisms to increase drag and remain suspended at optimal positions in the water column, as well as evade predation by grazers. The evolution of this sophisticated floatation mechanism in these purely planktonic streamlined microorganisms has important implications for our current understanding of microbial distribution in the oceans and predator-prey interactions which ultimately will need incorporating into future models of marine carbon flux dynamics.

Marine Plastic Debris: A New Surface for Microbial Colonization.

Plastics become rapidly colonized by microbes when released into marine environments. This microbial community-the Plastisphere-has recently sparked a multitude of scientific inquiries and generated a breadth of knowledge, which we bring together in this review. Besides providing a better understanding of community composition and biofilm development in marine ecosystems, we critically discuss current research on plastic biodegradation and the identification of potentially pathogenic "hitchhikers" in the Plastisphere. The Plastisphere is at the interface between the plastic and its surrounding milieu, and thus drives every interaction that this synthetic material has with its environment, from ecotoxicity and new links in marine food webs to the fate of the plastics in the water column. We conclude that research so far has not shown Plastisphere communities to starkly differ from microbial communities on other inert surfaces, which is particularly true for mature biofilm assemblages. Furthermore, despite progress that has been made in this field, we recognize that it is time to take research on plastic-Plastisphere-environment interactions a step further by identifying present gaps in our knowledge and offering our perspective on key aspects to be addressed by future studies: (I) better physical characterization of marine biofilms, (II) inclusion of relevant controls, (III) study of different successional stages, (IV) use of environmentally relevant concentrations of biofouled microplastics, and (V) prioritization of gaining a mechanistic and functional understanding of Plastisphere communities.

Beyond oil degradation: enzymatic potential of Alcanivorax to degrade natural and synthetic polyesters.

Pristine marine environments are highly oligotrophic ecosystems populated by well-established specialized microbial communities. Nevertheless, during oil spills, low-abundant hydrocarbonoclastic bacteria bloom and rapidly prevail over the marine microbiota. The genus Alcanivorax is one of the most abundant and well-studied organisms for oil degradation. While highly successful under polluted conditions due to its specialized oil-degrading metabolism, it is unknown how they persist in these environments during pristine conditions. Here, we show that part of the Alcanivorax genus, as well as oils, has an enormous potential for biodegrading aliphatic polyesters thanks to a unique and abundantly secreted alpha/beta hydrolase. The heterologous overexpression of this esterase proved a remarkable ability to hydrolyse both natural and synthetic polyesters. Our findings contribute to (i) better understand the ecology of Alcanivorax in its natural environment, where natural polyesters such as polyhydroxyalkanoates (PHA) are produced by a large fraction of the community and, hence, an accessible source of carbon and energy used by the organism in order to persist, (ii) highlight the potential of Alcanivorax to clear marine environments from polyester materials of anthropogenic origin as well as oils, and (iii) the discovery of a new versatile esterase with a high biotechnological potential.

Distribution of plastic polymer types in the marine environment; A meta-analysis.

Despite growing plastic discharge into the environment, researchers have struggled to detect expected increases of marine plastic debris in sea surfaces, sparking discussions about "missing plastics" and final sinks, which are hypothesized to be coastal and deep-sea sediments. While it holds true that the highest concentrations of plastic particles are found in these locations (103-104 particles m-3 in sediments vs. 0.1-1 particles m-3 in the water column), our meta-analysis also highlights that in open oceans, microplastic polymer types segregated in the water column according to their density. Lower density polymers, such as polypropylene and polyethylene, dominated sea surface samples (25% and 42%, respectively) but decreased in abundance through the water column (3% and 2% in the deep-sea, respectively), whereas only denser polymers (i.e. polyesters and acrylics) were enriched with depth (5% in surface seawater vs. 77% in deep-sea locations). Our meta-analysis demonstrates that some of the most abundant and recalcitrant manufactured plastics are more persistent in the sea surface than previously anticipated and that further research is required to determine the ultimate fate of these polymers as current knowledge does not support the deep sea as the final sink for all polymer types.

Microbial community composition and trophic role along a marked salinity gradient in Laguna Puilar, Salar de Atacama, Chile.

The geological, hydrological and microbiological features of the Salar de Atacama, the most extensive evaporitic sedimentary basin in the Atacama Desert of northern Chile, have been extensively studied. In contrast, relatively little attention has been paid to the composition and roles of microbial communities in hypersaline lakes which are a unique feature in the Salar. In the present study biochemical, chemical and molecular biological tools were used to determine the composition and roles of microbial communities in water, microbial mats and sediments along a marked salinity gradient in Laguna Puilar which is located in the "Los Flamencos" National Reserve. The bacterial communities at the sampling sites were dominated by members of the phyla Bacteroidetes, Chloroflexi, Cyanobacteria and Proteobacteria. Stable isotope and fatty acid analyses revealed marked variability in the composition of microbial mats at different sampling sites both horizontally (at different sites) and vertically (in the different layers). The Laguna Puilar was shown to be a microbially dominated ecosystem in which more than 60% of the fatty acids at particular sites are of bacterial origin. Our pioneering studies also suggest that the energy budgets of avian consumers (three flamingo species) and dominant invertebrates (amphipods and gastropods) use minerals as a source of energy and nutrients. Overall, the results of this study support the view that the Salar de Atacama is a heterogeneous and fragile ecosystem where small changes in environmental conditions may alter the balance of microbial communities with possible consequences at different trophic levels.