Extreme weather events as an important factor for the evolution of plastisphere but not for the degradation process.

Marine plastics, with their negative effects on marine life and the human health, have been recently recognized as a new niche for the colonization and development of marine biofilms. Members of the colonizing communities could possess the potential for plastic biodegradation. Thus, there is an urgent need to characterize these complex and geographically variable communities and elucidate the functionalities. In this work, we characterize the fungal and bacterial colonizers of 5 types of plastic films (High Density Polyethylene, Low Density Polyethylene, Polypropylene, Polystyrene and Polyethylene Terepthalate) over the course of a 242-day incubation in the south-eastern Mediterranean and relate them to the chemical changes observed on the surface of the samples via ATR-FTIR. The 16s rRNA and ITS2 ribosomal regions of the plastisphere communities were sequenced on four time points (35, 152, 202 and 242 days). The selection of the time points was dictated by the occurrence of a severe storm which removed biological fouling from the surface of the samples and initiated a second colonization period. The bacterial communities, dominated by Proteobacteria and Bacteroidetes, were the most variable and diverse. Fungal communities, characterized mainly by the presence of Ascomycota, were not significantly affected by the storm. Neither bacterial nor fungal community structure were related to the polymer type acting as substrate, while the surface of the plastic samples underwent weathering of oscillating degrees with time. This work examines the long-term development of Mediterranean epiplastic biofilms and is the first to examine how primary colonization influences the microbial community re-attachment and succession as a response to extreme weather events. Finally, it is one of the few studies to examine fungal communities, despite them containing putative plastic degraders.

Intertwined synergistic abiotic and biotic degradation of polypropylene pellets in marine mesocosms.

The accumulation of plastic waste in the oceans has caused growing concern for its effects on marine life. The interactions of plastics with environmental factors have been linked to fragmentation to micro- and nanoparticles with different properties and consequences, but the mechanism of fragmentation has not been fully understood yet. In this work, we investigate the combined effect of marine communities and ultraviolet (UV) radiation towards the degradation of virgin and artificially weathered polypropylene (PP) pellets after a long-term incubation period in marine mesocosms. The surface chemical alterations and deterioration of the polymer, in conjunction with the attachment and evolution of marine bacterial communities, the development of biofilm and exopolymeric substances (EPS), as well as the colloidal properties (zeta-potential and hydrodynamic diameter) of the mesocosms were studied. The surface area of both types of pellets decreased over time, despite no concrete weight change being observed. Cell growth, EPS production and colloid particle size were correlated to the loss of area. Therefore, we propose that surface area could be effectively monitored, instead of weight loss, as an alternative indicator of polymer degradation in biodegradation experiments. Changes in the chemical structure of the polymer, in addition to the evolution of the biological factors, implied that a complex degradation process alternated between two phases: an abiotic phase, when UV irradiation contributes to the deterioration of the polymer surface layers and a biotic phase, when marine communities degrade the weathered polymer surface to reveal the underlying layer of virgin polymer. Finally, microscopic particles, produced as a result of the decrease in pellet area, promoted the aggregation of colloidal particles. The role and impacts of these colloidal particles in marine ecosystems are yet as unidentified as that of micro- and nano-sized plastic particles and call for further investigation.

Relationship between the Carbonyl Index (CI) and Fragmentation of Polyolefin Plastics during Aging.

The durability of plastics in the marine environment has emerged as a crucial environmental issue. However, the contribution of several factors and the threshold point after which a plastic product generates secondary micro- and nanoplastics is still unclear. To investigate the interaction of environmental parameters with the physicochemical properties of polyethylene (PE) and polypropylene (PP) films in the marine environment, polyolefin films were subjected to weathering in emulated coastal and marine environments for 12 months, focusing on the relationship between radiation load, alteration on the surface, and subsequent generation of microplastics (MPs). The weight average molecular weight (Mw) was found to be strongly correlated with the generated particles and the Feret diameter, implying the generation of secondary microplastics at decreased Mw. A significant and strong relationship between the carbonyl index (CI) and the Feret diameter for PP films weathered on beach sand was identified. This CI-fragmentation relationship involves three sequential stages and suggests that spontaneous fragmentation occurs at CI values above 0.7.

Sinking characteristics of microplastics in the marine environment.

Plastic pollution is presently one of the most widespread and minimally understood problems. Vast quantities of plastics that have entered the marine environment should be detected floating on the sea surface are seemingly missing from the global budget. A vertical transfer process should be able to explain the imbalance in mass, as well as the findings of buoyant plastics at the bottom of the sea. These processes are of paramount importance to modelling efforts on the fate of plastics and microplastics in the marine environment. In order to fill this gap and develop correlations that could be used in modelling activities, we have designed and performed a 300-day long field experiment to monitor the interactions between microplastics (pellets and films) and the marine environment for five types of plastic polymers. Fouling, changes in diameter, gravimetric weight and sinking velocity were monitored and the correlations between them were studied using principal component analysis (PCA). Density, fouling and sample form (strip or pellet) were found to greatly affect the sinking characteristics of the polymers, leading to an increase or decrease in the sinking velocity. Finally, mathematical expressions for the estimation of fouling attachment and the sinking velocity with respect to time for each type of plastic were determined from the experimental data.

Biodegradation of mixture of plastic films by tailored marine consortia.

This work sheds light on the physicochemical changes of naturally weathered polymer surfaces along with changes of polymer buoyancy due to biofilm formation and degradation processes. To support the degradation hypothesis, a microcosm experiment was conducted where a mixture of naturally weathered plastic pieces was incubated with an indigenous pelagic community. A series of analyses were employed in order to describe the alteration of the physicochemical characteristics of the polymer (FTIR, SEC and GPC, sinking velocity) as well as the biofilm community (NGS). At the end of phase II, the fraction of double bonds in the surface of microbially treated PE films increased while changes were also observed in the profile of the PS films. The molecular weight of PE pieces increased with incubation time reaching the molecular weight of the virgin pieces (230,000 g mol-1) at month 5 but the buoyancy displayed no difference throughout the experimental period. The number-average molecular weight of PS pieces decreased (33% and 27% in INDG and BIOG treatment respectively), implying chain scission; accelerated (by more than 30%) sinking velocities compared to the initial weathered pieces were also measured for PS films with biofilm on their surface. The orders Rhodobacterales, Oceanospirillales and Burkholderiales dominated the distinct platisphere communities and the genera Bacillus and Pseudonocardia discriminate these assemblages from the planktonic counterpart. The functional analysis predicts overrepresentation of adhesive cells carrying xenobiotic and hydrocarbon degradation genes. Taking these into account, we can suggest that tailored marine consortia have the ability to thrive in the presence of mixtures of plastics and participate in their degradation.

Plastic pellets, meso- and microplastics on the coastline of Northern Crete: Distribution and organic pollution.

Plastic pollution in the marine environment is one of the foremost environmental problems of our time, as it affects wildlife and human health both directly and indirectly through the effects of contaminants carried by microplastics. This study investigates the temporal and spatial distribution of plastic pellets and fragments in sandy beaches along the coastline of Northern Crete, during 2013. Their densities varied throughout the year in each beach, with highest densities during the summer and towards the upper parts of the beaches. The concentrations of 16 polycyclic aromatic hydrocarbons (PAHs) sorbed on microplastics sampled from nine sandy beaches of Northern Crete was quantified using Gas chromatography - Ion Trap Mass Spectrometry (GC-ITMS). PAHs concentrations ranged from non-detectable levels to 1592 ng/g and fluctuated between sampling periods. Based on the observed patterns of meso- and microplastics distribution, practical guidelines are proposed to minimize the entrance of microplastics into the seawater wherefrom they are exceptionally difficult to collect, if mitigation actions are to be applied.

Biodegradation of weathered polystyrene films in seawater microcosms.

A microcosm experiment was conducted at two phases in order to investigate the ability of indigenous consortia alone or bioaugmented to degrade weathered polystyrene (PS) films under simulated marine conditions. Viable populations were developed on PS surfaces in a time dependent way towards convergent biofilm communities, enriched with hydrocarbon and xenobiotics degradation genes. Members of Alphaproteobacteria and Gammaproteobacteria were highly enriched in the acclimated plastic associated assemblages while the abundance of plastic associated genera was significantly increased in the acclimated indigenous communities. Both tailored consortia efficiently reduced the weight of PS films. Concerning the molecular weight distribution, a decrease in the number-average molecular weight of films subjected to microbial treatment was observed. Moreover, alteration in the intensity of functional groups was noticed with Fourier transform infrared spectrophotometry (FTIR) along with signs of bio-erosion on the PS surface. The results suggest that acclimated marine populations are capable of degrading weathered PS pieces.

Development of tailored indigenous marine consortia for the degradation of naturally weathered polyethylene films.

This study investigated the potential of bacterial-mediated polyethylene (PE) degradation in a two-phase microcosm experiment. During phase I, naturally weathered PE films were incubated for 6 months with the indigenous marine community alone as well as bioaugmented with strains able to grow in minimal medium with linear low-density polyethylene (LLDPE) as the sole carbon source. At the end of phase I the developed biofilm was harvested and re-inoculated with naturally weathered PE films. Bacteria from both treatments were able to establish an active population on the PE surfaces as the biofilm community developed in a time dependent way. Moreover, a convergence in the composition of these communities was observed towards an efficient PE degrading microbial network, comprising of indigenous species. In acclimated communities, genera affiliated with synthetic (PE) and natural (cellulose) polymer degraders as well as hydrocarbon degrading bacteria were enriched. The acclimated consortia (indigenous and bioaugmented) reduced more efficiently the weight of PE films in comparison to non-acclimated bacteria. The SEM images revealed a dense and compact biofilm layer and signs of bio-erosion on the surface of the films. Rheological results suggest that the polymers after microbial treatment had wider molecular mass distribution and a marginally smaller average molar mass suggesting biodegradation as opposed to abiotic degradation. Modifications on the surface chemistry were observed throughout phase II while the FTIR profiles of microbially treated films at month 6 were similar to the profiles of virgin PE. Taking into account the results, we can suggest that the tailored indigenous marine community represents an efficient consortium for degrading weathered PE plastics.