Evolution of prokaryotic colonisation of greenhouse plastics discarded into the environment.

Current knowledge on the capacity of plastics as vectors of microorganisms and their ability to transfer microorganisms between different habitats (i.e. air, soil and river) is limited. The objective of this study was to characterise the evolution of the bacterial community adhered to environmental plastics [low-density polyethylene (LDPE)] across different environments from their point of use to their receiving environment destination in the sea. The study took place in a typical Mediterranean intermittent river basin in Larnaka, Cyprus, characterised by a large greenhouse area whose plastic debris may end up in the sea due to mismanagement. Five locations were selected to represent the environmental fate of greenhouse plastics from their use, through their abandonment in soil and subsequent transport to the river and the sea, taking samples of plastics and the surrounding environments (soil and water). The bacterial community associated with each sample was studied by 16S rRNA metabarcoding; also, the main physicochemical parameters in each environmental compartment were analysed to understand these changes. The identification and chemical changes in greenhouse plastics were tracked using Attenuated Total Reflection Fourier Transform Infra-red spectroscopy (ATR-FTIR). Scanning Electron Microscope (SEM) analysis demonstrated an evolution of the biofilm at each sampling location. ß-diversity studies showed that the bacterial community adhered to plastics was significantly different from that of the surrounding environment only in samples taken from aqueous environments (freshwater and sea) (p-value p-value > 0.05). The environmental parameters (pH, salinity, total nitrogen and total phosphorus) explained the differences observed at each location to a limited extent. Furthermore, bacterial community differences among samples were lower in plastics collected from the soil than in plastics taken from rivers and seawater. Six genera (Flavobacterium, Altererythrobacter, Acinetobacter, Pleurocapsa, Georgfuchsia and Rhodococcus) were detected in the plastic, irrespective of the sampling location, confirming that greenhouse plastics can act as possible vectors of microorganisms between different environments: from their point of use, through a river system to the final coastal receiving environment. In conclusion, this study confirms the ability of greenhouse plastics to transport bacteria, including pathogens, between different environments. Future studies should evaluate these risks by performing complete sequencing metagenomics to decipher the functions of the plastisphere.

The small Ca2+-binding protein CSE links Ca2+ signalling with nitrogen metabolism and filament integrity in Anabaena sp. PCC 7120.

BACKGROUND: Filamentous cyanobacteria represent model organisms for investigating multicellularity. For many species, nitrogen-fixing heterocysts are formed from photosynthetic vegetative cells under nitrogen limitation. Intracellular Ca2+ has been implicated in the highly regulated process of heterocyst differentiation but its role remains unclear. Ca2+ is known to operate more broadly in metabolic signalling in cyanobacteria, although the signalling mechanisms are virtually unknown. A Ca2+-binding protein called the Ca2+ Sensor EF-hand (CSE) is found almost exclusively in filamentous cyanobacteria. Expression of asr1131 encoding the CSE protein in Anabaena sp. PCC 7120 was strongly induced by low CO2 conditions, and rapidly downregulated during nitrogen step-down. A previous study suggests a role for CSE and Ca2+ in regulation of photosynthetic activity in response to changes in carbon and nitrogen availability. RESULTS: In the current study, a mutant Anabaena sp. PCC 7120 strain lacking asr1131 (Δcse) was highly prone to filament fragmentation, leading to a striking phenotype of very short filaments and poor growth under nitrogen-depleted conditions. Transcriptomics analysis under nitrogen-replete conditions revealed that genes involved in heterocyst differentiation and function were downregulated in Δcse, while heterocyst inhibitors were upregulated, compared to the wild-type. CONCLUSIONS: These results indicate that CSE is required for filament integrity and for proper differentiation and function of heterocysts upon changes in the cellular carbon/nitrogen balance. A role for CSE in transmitting Ca2+ signals during the first response to changes in metabolic homeostasis is discussed.

Use of Cyanobacterial Luminescent Bioreporters to Report on the Environmental Impact of Metallic Nanoparticles.

Due to their ecological relevance, low cost, and easy maintenance, cyanobacteria have been used for bioreporter development. In this study, a battery of cyanobacterial bioreporters has been used to assess the ecotoxicity of four highly used metallic nanoparticles (NPs). The toxicity of these NPs was tested using the bioreporter Nostoc CPB4337 (Anabaena CPB4337). As oxidative stress is a primary toxic mechanism of metallic NPs, cyanobacterial reactive oxygen species (ROS)-detecting bioreporters were used. Metallic NPs release metal ions, which contribute to their toxic effect and the formation of ROS, so a metal-detecting bioreporter was also used to detect the bioavailable metals. The results confirm that ROS production by NPs was due to the NPs per se and not by released free-ions, which in fact were almost undetectable. Although the metal-detecting bioreporter could not detect the dissolved metal ions, it was able to detect the metallic NPs themselves, indicating that this bioreporter may be useful to detect them in the environment. ROS production varied depending on the growth medium or environmental matrices conditions and on the NP type. This work demonstrated the different levels of ROS production by metallic NPs and the importance of nanotoxicology studies in real matrices.

Microplastics and non-natural cellulosic particles in Spanish bottled drinking water.

This investigation explored the presence of microplastics (MPs) and artificial cellulosic particles (ACPs) in commercial water marketed in single use 1.5 L poly(ethylene terephthalate) bottles. In this work we determined a mass concentration of 1.61 (1.10-2.88) µg/L and 1.04 (0.43-1.82) µg/L for MPs and ACPs respectively in five top-selling brands from the Spanish bottled water market. Most MPs consisted of white and transparent polyester and polyethylene particles, while most ACPs were cellulosic fibers likely originating from textiles. The median size of MPs and ACPs was 93 µm (interquartile range 76-130 µm) and 77 µm (interquartile range 60-96 µm), respectively. Particle mass size distributions were fitted to a logistic function, enabling comparisons with other studies. The estimated daily intake of MPs due to the consumption of bottled water falls within the 4-18 ng kg-1 day-1 range, meaning that exposure to plastics through bottled water probably represents a negligible risk to human health. However, it's worth noting that the concentration of plastic found was much higher than that recorded for tap water, which supports the argument in favour of municipal drinking water.

Leaching of herbicides mixtures from pre-exposed agricultural plastics severely impact microalgae.

Farmlands represent a source of aged plastics and pesticides to the surrounding environments. It has been shown that chemicals can be sorbed and desorbed from plastics, but the interaction between plastic and mixtures of pesticides and their effects on freshwater biota has not been assessed yet. The aim of the work was to assess the potential role of agricultural plastics as vectors for a mixture of two herbicides and the impact of the herbicide mixture lixiviated from them towards the freshwater microalga Chlamydomonas reinhardtii. Pristine and aged polyethylene plastics collected from agricultural areas were exposed to the herbicides, bifenox, oxyfluorfen and their mixtures. The microalgae were exposed for 72 h to the leachates desorbed from plastics and the effect was quantified in terms of total chlorophyll content and several physiological parameters assessed by flow cytometry. Our results showed that changes in physicochemical properties (hydroxyl and carbonyl index, hydrophobicity, texture) in aged plastics increased their capacity to retain and to desorb the herbicides. Microalgae exposed to leachates containing bifenox, oxyfluorfen, or their mixture showed reactive oxygen species overproduction, lipid peroxidation, membrane potential hyperpolarization, intracellular pH acidification, and a loss of metabolic activity. The toxicological interactions of the leachate mixture were assessed using the Combination Index (CI)-isobologram method showing antagonism at low effect levels turning to synergism when the effect increased. In this work, we proved the hypothesis that ageing increases the capacity of agricultural plastics to behave as vector for toxic chemicals to the biota.

Time-course biofilm formation and presence of antibiotic resistance genes on everyday plastic items deployed in river waters.

The plastisphere has been widely studied in the oceans; however, there is little information on how living organisms interact with the plastisphere in freshwater ecosystems, and particularly on how this interaction changes over time. We have characterized, over one year, the evolution of the eukaryotic and bacterial communities colonizing four everyday plastic items deployed in two sites of the same river with different anthropogenic impact. α-diversity analyses showed that site had a significant role in bacterial and eukaryotic diversity, with the most impacted site having higher values of the Shannon diversity index. ß-diversity analyses showed that site explained most of the sample variation followed by substrate type (i.e., plastic item) and time since first colonization. In this regard, core microbiomes/biomes in each plastic at 1, 3, 6 and 12 months could be identified at genus level, giving a global overview of the evolution of the plastisphere over time. The measured concentration of antibiotics in the river water positively correlated with the abundance of antibiotic resistance genes (ARGs) on the plastics. These results provide relevant information on the temporal dynamics of the plastisphere in freshwater ecosystems and emphasize the potential contribution of plastic items to the global spread of antibiotic resistance.

Biofilm formation strongly influences the vector transport of triclosan-loaded polyethylene microplastics.

This study aimed at evaluating the influence of biofilm in the role of microplastics (MPs) as vectors of pollutants and their impact on Daphnia magna. To do this, virgin polyethylene MPs, (PE-MPs, 40-48 µm) were exposed for four weeks to wastewater (WW) from influent and effluent to promote biofouling. Then, the exposed PE-MPs were put in contact with triclosan. Finally, the toxicity of TCS-loaded and non-TCS loaded PE-MPs were tested on the survival of D. magna adults for 21 days. Results from metabarcoding analyses indicated that exposure to TCS induced shifts in the bacterial community, selecting potential TCS-degrading bacteria. Results also showed that PE-MPs were ingested by daphnids. The most toxic virgin PE-MPs were those biofouled in the WW effluent. The toxicity of TCS-loaded PE-MPs biofouled in the WW effluent was even higher, reporting mortality in all tested concentrations. These results indicate that biofouling of MPs may modulate the adsorption and subsequent desorption of co-occurring pollutants, hence affecting their potential toxicity towards aquatic organisms. Future studies on realistic environmental plastic impact should include the characterization of biofilms growing on plastic. Since inevitably plastic biofouling occurs over time in nature, it should be taken into account as it may modulate the sorption of co-occurring pollutants.

Tracking nanoplastics in freshwater microcosms and their impacts to aquatic organisms.

In this work, we used palladium-doped polystyrene NPLs (PS-NPLs with a primary size of 286 ± 4 nm) with an irregular surface morphology which allowed for particle tracking and evaluation of their toxicity on two primary producers (cyanobacterium, Anabaena sp. PCC7120 and green algae, Chlamydomonas reinhardtii) and one primary consumer (crustacean, Daphnia magna). the concentration range for Anabaena and C. reinhardtii was from 0.01 to 1000 mg/L and for D. magna, the range was from 7.5 to 120 mg/L.EC50 s ranged from 49 mg NPLs/L for D. magna (48hEC50 s) to 248 mg NPLs/L (72hEC50 s for C. reinhardtii). PS-NPLs induced dose-dependent reactive oxygen species overproduction, membrane damage and metabolic alterations. To shed light on the environmental fate of PS-NPLs, the short-term distribution of PS-NPLs under static (using lake water and sediments) and stirring (using river water and sediments) conditions was studied at laboratory scale. The results showed that most NPLs remained in the water column over the course of 48 h. The maximum percentage of settled particles (∼ 30 %) was found under stirring conditions in comparison with the ∼ 10 % observed under static ones. Natural organic matter increased the stability of the NPLs under colloidal state while organisms favored their settlement. This study expands the current knowledge of the biological effects and fate of NPLs in freshwater environments.

A nationwide monitoring of atmospheric microplastic deposition.

Plastic production continues to increase every year, yet it is widely acknowledged that a significant portion of this material ends up in ecosystems as microplastics (MPs). Among all the environmental compartments affected by MPs, the atmosphere remains the least well-known. Here, we conducted a one-year simultaneous monitoring of atmospheric MPs deposition in ten urban areas, each with different population sizes, economic activities, and climates. The objective was to assess the role of the atmosphere in the fate of MPs by conducting a nationwide quantification of atmospheric MP deposition. To achieve this, we deployed collectors in ten different urban areas across continental Spain and the Canary Islands. We implemented a systematic sampling methodology with rigorous quality control/quality assurance, along with particle-oriented identification and quantification of anthropogenic particle deposition, which included MPs and industrially processed natural fibres. Among the sampled MPs, polyester fibres were the most abundant, followed by acrylic polymers, polypropylene, and alkyd resins. Their equivalent sizes ranged from 22 µm to 398 µm, with a median value of 71 µm. The particle size distribution of MPs showed fewer large particles than expected from a three-dimensional fractal fragmentation pattern, which was attributed to the higher mobility of small particles, especially fibres. The atmospheric deposition rate of MPs ranged from 5.6 to 78.6 MPs m-2 day-1, with the higher values observed in densely populated areas such as Barcelona and Madrid. Additionally, we detected natural polymers, mostly cellulosic fibres with evidence of industrial processing, with a deposition rate ranging from 6.4 to 58.6 particles m-2 day-1. There was a positive correlation was found between the population of the study area and the median of atmospheric MP deposition, supporting the hypothesis that urban areas act as sources of atmospheric MPs. Our study presents a systematic methodology for monitoring atmospheric MP deposition.

Ageing and fragmentation of marine microplastics.

The generation of small fragments from the environmental ageing of microplastics (MPs) is still a poorly known process. This work addresses the fragmentation of MPs obtained from marine debris consisting of polyethylene and polypropylene (PE and PP in environmental mixture) and polystyrene (PS) after exposure to accelerated ageing by irradiation and mechanical stirring. Number particle size distribution in the 1-100 µm range was assessed by combining laser diffractometry with particle counts from flow cytometry. The results showed the generation of a high number of small MP particles, which reached 105-106 items/mg of plastic with most fragments <2 µm. The results showed that environmentally aged MPs give rise to a larger number of small MPs in a pattern consistent with progressive fragmentation in the three spatial dimensions. The proportion of small MPs was much higher than that found in current sampling campaigns, suggesting a severe underestimation of the environmental presence of small MPs. We also demonstrated the generation of nanoplastics (NPs) in the fraction <1 µm from irradiated runs. The results showed that the mechanism that produced nanoplastics (NPs) from MPs was irradiation, which yielded up to 1011-1013 NPs/g with particle size in the few hundreds of nm range. Our results are relevant for the assessment of fate and risk of plastic debris in the environment showing that the number of small plastic fragments produced during the ageing of MPs is much larger than expect from the extrapolation of larger size populations.

Polystyrene nanoplastics and wastewater displayed antagonistic toxic effects due to the sorption of wastewater micropollutants.

The knowledge about the interaction of nanoplastics with other aquatic pollutants and their combined effects on biota is very scarce. In this work, we studied the interaction between polystyrene nanoplastics (PS NPs) (30 nm) and the micropollutants in a biologically treated wastewater effluent (WW). The capacity of PS NPs to sorb micropollutants was studied as well as their single and combined toxicity towards three freshwater organisms: the recombinant bioluminescent cyanobacterium, Anabaena sp. PCC 7120 CPB4337; the duckweed, Spirodela polyrhiza and the cladoceran, Daphnia magna. The endpoints were the inhibition of bioluminescence, the growth inhibition of the aquatic plant and the immobilization of D. magna after 24, 72 and 48 h of exposure, respectively. Combination Index (CI)-isobologram method was used to quantify mixture toxicity and the nature of interactions. PS NPs sorbed a variety of chemicals present in WW as micropollutants in a range of tens of ng/L to µg/L. It was found that those pollutants with positive charge were the main ones retained onto PS NPs, which was attributed to the electrostatic interaction with the negatively charged PS NPs. Regarding the toxicological effects, single exposure to PS NPs affected the three tested organisms. However, single exposure to WW only had a negative impact on the cyanobacterium and S. polyrhiza with no observed toxicity to D. magna. Regarding PS NPs-WW combined exposure, a reduction of toxicity in comparison with single exposure was observed probably due to the sorption of micropollutants onto PS NPs, which resulted in lower bioavailability of the micropollutants. In addition, the formation of PS NPs-WW heteroaggregates was observed which could result in lower bioavailability of PS NPs and sorbed micropollutants, thus lowering toxicity. This study represents a near-realistic scenario approach to the potential sorption of wastewater pollutants onto nanoplastics that could alter the toxicological effect on the biota.

Identification and toxicity towards aquatic primary producers of the smallest fractions released from hydrolytic degradation of polycaprolactone microplastics.

Bioplastics are thought as a safe substitute of non-biodegradable polymers. However, once released in the environment, biodegradation may be very slow, and they also suffer abiotic fragmentation processes, which may give rise to different fractions of polymer sizes. We present novel data on abiotic hydrolytic degradation of polycaprolactone (PCL), tracking the presence of by-products during 132 days by combining different physicochemical techniques. During the study a considerable amount of two small size plastic fractions were found (up to âˆ¼ 6 mg of PCL by-product/g of PCL beads after 132 days of degradation); and classified as submicron-plastics (sMPs) from 1 µm to 100 nm and nanoplastics (NPs, <100 nm) as well as oligomers. The potential toxicity of the smallest fractions, PCL by-products < 100 nm (PCL-NPs + PCL oligomers) and the PCL oligomers single fraction, was tested on two ecologically relevant aquatic primary producers: the heterocystous filamentous nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120, and the unicellular cyanobacterium Synechococcus sp. PCC 7942. Upon exposure to both, single and combined fractions, Reactive Oxygen Species (ROS) overproduction, intracellular pH and metabolic activity alterations were observed in both organisms, whilst membrane potential and morphological damages were only observed upon PCL-NPs + PCL oligomers exposure. Notably both PCL by-products fractions inhibited nitrogen fixation in Anabaena, which may be clearly detrimental for the aquatic trophic chain. As conclusion, fragmentation of bioplastics may render a continuous production of secondary nanoplastics as well as oligomers that might be toxic to the surrounding biota; both PCL-NPs and PCL oligomers, but largely the nanoparticulate fraction, were harmful for the two aquatic primary producers. Efforts should be made to thoroughly understand the fragmentation of bioplastics and the toxicity of the smallest fractions resulting from that degradation.

Novel cyanobacterial bioreporters of phosphorus bioavailability based on alkaline phosphatase and phosphate transporter genes of Anabaena sp. PCC 7120.

There is heterogeneity in the way cyanobacteria respond to P starvation and subsequently how they adapt to environments with low or fluctuating P concentrations. In this study, we have fused the promoterless lux operon luxCDABE to the promoter regions of Anabaena sp. PCC 7120 phoA genes putatively encoding alkaline phosphatases, phoA (all2843) and phoA-like (alr5291) and to the promoter region of one operon putatively encoding a high affinity phosphate transporter pst1 (all4575-4572). The self-bioluminescent strains constructed in this way, Anabaena AP (phoA promoter), Anabaena AP-L (phoA-like promoter), and Anabaena PST (pst1 promoter) have been used to study the expression of these genes in response to P starvation and P re-feeding with inorganic and organic phosphate sources. Our data showed that the pst1 promoter was activated at much higher level than the phoA-like promoter following P starvation; however, we did not observe activation of the phoA promoter. The P re-feeding experiments revealed that both strains, Anabaena (A.) PST and A. AP-L could be used as novel bioreporters of P availability in environmental samples. Both strains were used to estimate bioavailable P in environmental samples (fresh- and wastewaters) with a wide range of soluble P concentrations. The results indicated that most of the P in the water samples was in chemical forms available to the cyanobacterium; however there were some differences in the estimates given by both strains as A. PST appeared to be more adequate for the samples with the lowest P load while A. AP-L gave similar or even higher values of P concentrations than those chemically measured in samples with higher P load.

Free Ca2+ as an early intracellular biomarker of exposure of cyanobacteria to environmental pollution.

Calcium functions as a versatile messenger in a wide variety of eukaryotic and prokaryotic cells. Cyanobacteria are photoautotrophs which have a great ecological impact as primary producers. Our research group has presented solid evidence of a role of calcium in the perception of environmental changes by cyanobacteria and their acclimation to these changes. We constructed a recombinant strain of the freshwater cyanobacterium Anabaena sp. PCC 7120 that constitutively expresses the calcium-binding photoprotein apoaequorin, enabling in-vivo monitoring of any fluctuation in the intracellular free calcium concentration of the cyanobacterium in response to any environmental stimulus. The "Ca(2+) signature" is the combination of changes in all Ca(2+) signal properties (magnitude, duration, frequency, source of the signal) produced by a specific stimulus. We recorded and analyzed the Ca(2+) signatures generated by exposure of the cyanobacterium to different groups of environmental pollutants, for example cations, anions, organic solvents, naphthalene, and pharmaceuticals. We found that, in general, each group of tested chemicals triggered a specific calcium signature in a reproducible and dose-dependent manner. We hypothesize that these Ca(2+) signals may be related to the cellular mechanisms of pollutant perception and ultimately to their toxic mode of action. We recorded Ca(2+) signals triggered by binary mixtures of pollutants and a signal induced by a real wastewater sample which could be mimicked by mixing its main constituents. Because Ca(2+) signatures were induced before toxicity was evident, we propose that intracellular free Ca(2+) may serve as an early biomarker of exposure to environmental pollution.

Microplastics can act as vector of the biocide triclosan exerting damage to freshwater microalgae.

Despite the large number of recent studies on microplastics (MPs) and their ability to act as carriers of pollutants, the knowledge about the biological effects of MPs loaded with chemicals is scarce. The aim of this study was to evaluate the potential of MPs as vectors for the antimicrobial triclosan (TCS). For it, we tested low-density polyethylene (LDPE), polyamide (PA), polyethylene terephthalate (PET), polyoxymethylene (POM), polypropylene (PP), polystyrene (PS) and the biodegradable polylactic acid (PLA). Thus, chemical analysis of sorption and desorption of TCS by these MPs was evaluated. The effect of TCS-loaded MPs to Anabaena sp. PCC7120, a cyanobacterium model of primary producers in freshwater ecosystems, was investigated. Chemical analyses showed different capacity of sorption depending on the MP type, which was related to some of their physicochemical properties. PA (104.7 µg/g), POM (57.4 µg/g) and LDPE (18.3 µg/g) were the polymers that sorbed the highest amounts of TCS. Glass transition temperature of polymers and their physicochemical interaction with TCS explained the extent of sorption. Significant decreases were found in growth, 22.3%, 94.6% and 81.0%, and chlorophyll a content, 58.4%, 95.0% and 89.6%, of Anabaena when exposed to TCS-loaded LDPE, PA and POM beads, respectively, which were the only MPs displaying significant sorption-desorption of TCS, implying that these MPs could act as vectors of TCS towards freshwater microalgae. This finding is of fundamental relevance as microalgae are at the base of the aquatic trophic chain and support growth of upper organisms.

Early and differential bacterial colonization on microplastics deployed into the effluents of wastewater treatment plants.

Microbial colonization of microplastics (MPs) in aquatic ecosystems is a well-known phenomenon; however, there is insufficient knowledge of the early colonization phase. Wastewater treatment plant (WWTP) effluents have been proposed as important pathways for MPs entry and transport in aquatic environments and are hotspots of bacterial pathogens and antibiotic resistance genes (ARGs). This study aimed at characterizing bacterial communities in the early stage of biofilm formation on seven different types of MPs deployed in two different WWTPs effluents as well as measuring the relative abundance of two ARGs (sulI and tetM) on the tested MPs. Illumina Miseq sequencing of the 16S rRNA showed significant higher diversity of bacteria on MPs in comparison with free-living bacteria in the WWTP effluents. ß-diversity analysis showed that the in situ environment (sampling site) and hydrophobicity, to a lesser extent, had a role in the early bacterial colonization phase. An early colonization phase MPs-core microbiome could be identified. Furthermore, specific core microbiomes for each type of polymer suggested that each type might select early attachment of bacteria. Although the tested WWTP effluent waters contained antibiotic resistant bacteria (ARBs) harboring the sulI and tetM ARGs, MPs concentrated ARBs harboring the sulI gene but not tetM. These results highlight the relevance of the early attachment phase in the development of bacterial biofilms on different types of MP polymers and the role that different types of polymers might have facilitating the attachment of specific bacteria, some of which might carry ARGs.

Generation of nanoplastics during the photoageing of low-density polyethylene.

In this work, we studied the hydrolytic and photochemical degradation of three low-density polyethylene (LDPE) materials, within the size range of microplastics (MP). The MPs were exposed to mechanical agitation and UV irradiation equivalent to one year of solar UVB + UVA in a stirred photoreactor. Flow cytometry was used to track the formation of small (1-25 µm) MPs by applying Mie's theory to derive the size of MP particles from scattering intensity readings. The calculation was based on a calibration with polystyrene (PS) beads. The results showed that the generation of 1-5 µm MP reached 104-105 MPs in the 1-25 µm range per gram of LDPE. ATR-FTIR and micro-FTIR measurements evidenced the formation of oxygenated moieties, namely hydroxyl, carbonyl, and carbon-oxygen bonds, which increased with irradiation time. We also found evidence of the production of a high number of nanoplastics (<1 µm, NPs). The Dynamic Light Scattering size of secondary NPs was in the hundreds of nm range and might represent up to 1010 NPs per gram of LDPE. Our results allowed the unambiguous spectroscopic assessment of the generation of NPs from LDPE under conditions simulating environmental exposure to UV irradiation and used flow cytometry for the first-time to track the formation of secondary MPs.

Microbial colonizers of microplastics in an Arctic freshwater lake.

Microplastics (MPs) have been found everywhere as they are easily transported between environmental compartments. Through their transport, MPs are quickly colonized by microorganisms; this microbial community is known as the plastisphere. Here, we characterized the plastisphere of three MPs, one biodegradable (PHB) and two non-biodegradables (HDPE and LDPE), deployed in an Arctic freshwater lake for eleven days. The plastisphere was found to be complex, confirming that about a third of microbial colonizers were viable. Plastisphere was compared to microbial communities on the surrounding water and microbial mats on rocks at the bottom of the lake. Microbial mats followed by MPs showed the highest diversity regarding both prokaryotes and eukaryotes as compared to water samples; however, for fungi, MPs showed the highest diversity of the tested substrates. Significant differences on microbial assemblages on the three tested substrates were found; regarding microbial assemblages on MPs, bacterial genera found in polar environments such as Mycoplana, Erythromicrobium and Rhodoferax with species able to metabolize recalcitrant chemicals were abundant. Eukaryotic communities on MPs were characterized by the presence of ciliates of the genera Stentor, Vorticella and Uroleptus and the algae Cryptomonas, Chlamydomonas, Tetraselmis and Epipyxis. These ciliates normally feed on algae so that the complexity of these assemblages may serve to unravel trophic relationships between co-existing taxa. Regarding fungal communities on MPs, the most abundant genera were Betamyces, Cryptococcus, Arrhenia and Paranamyces. MPs, particularly HDPE, were enriched in the sulI and ermB antibiotic resistance genes (ARGs) which may raise concerns about human health-related issues as ARGs may be transferred horizontally between bacteria. This study highlights the importance of proper waste management and clean-up protocols to protect the environmental health of pristine environments such as polar regions in a context of global dissemination of MPs which may co-transport microorganisms, some of them including ARGs.

Microplastics as vectors of the antibiotics azithromycin and clarithromycin: Effects towards freshwater microalgae.

Water pollution due to microplastics (MPs) is recognized as a major anthropogenic impact. Once MPs reach the ecosystems, they are exposed to a variety of other pollutants, which can be sorbed on them, transported and eventually desorbed. In this work, we tested the hypothesis that MPs can behave as conveyors for delivering chemicals toxic to aquatic microorganisms by investigating the vector role of MPs of polyethylene terephthalate (PET), polylactic acid (PLA), polyoxymethylene (POM) and polystyrene (PS) to the macrolide antibiotics azithromycin (AZI) and clarithromycin (CLA). AZI and CLA were chosen, as they are included in the Watch List for EU monitoring concerning water policy by Decision (EU) 2018/840. MPs were loaded in contact with 500 µg/L of AZI or 1000 µg/L of CLA. Results showed that both antibiotics were sorbed on all tested MPs. The more hydrophobic AZI was sorbed in higher proportion than CLA. Both antibiotics were desorbed from MPs upon contact with water with percentages between 14.6 ± 2.6% for AZI and 1.9 ± 1.4% for CLA of the concentrations to which the MPs were initially exposed. Virgin MPs were not toxic to the cyanobacterium Anabaena sp. PCC7120. However, antibiotic-loaded MPs significantly inhibited the growth and chlorophyll content of the cyanobacterium. Most of the sorbed antibiotics became released upon contact with cyanobacterial cultures, which was the cause for the observed toxicity. Therefore, MPs can play a role as vectors of antibiotics in freshwaters systems affecting the basic trophic level of photosynthetic microorganisms.

Occurrence and transport of microplastics sampled within and above the planetary boundary layer.

Nowadays, there is no direct evidence about the presence of microplastics (MPs) in the atmosphere above ground level. Here, we investigated the occurrence, chemical composition, shape, and size of MPs in aircraft sampling campaigns flying within and above the planetary boundary layer (PBL). The results showed that MPs were present with concentrations ranging from 1.5 MPs m-3 above rural areas to 13.9 MPs m-3 above urban areas. MPs represented up to almost one third of the total amount of microparticles collected. Fourier Transform Infrared Spectroscopy allowed identifying seven types of MPs with the highest diversity corresponding to urban areas. Atmospheric transport and deposition simulations were performed using the HYbrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) model. Air mass trajectory analyses showed that MPs could be transported more than 1000 km before being deposited. This pioneer study is the first evidence of the microplastic presence above PBL and their potential long-range transport from their point of release even crossing distant borders.