Nanoplastic toxicity towards freshwater organisms

The fragmentation of plastic litter into smaller fragments, known as microplastics and nanoplastics, as well as their toxicity and environmental distribution have become issues of high concern. Furthermore, the popularization of bioplastics as a greener substitute of conventional plastics represents a challenge for the scientific community in view of the limited information concerning their potential environmental impact. Here, we systematically review the recent knowledge on the environmental fate and toxicity of nanoplastics in freshwater environments, discuss the results obtained thus far, and identify several knowledge gaps. The sources and environmental behaviors of nanoplastics are presented considering in vitro, in vivo, and in silico studies with a focus on real exposure scenarios. Their effects on organisms are classified based on their impact on primary producers, primary consumers, and secondary consumers. This review covers the main results published in the last four years, including all relevant experimental details and highlighting the most sensitive toxicity endpoints assessed in every study. We also include more recent results on the potential environmental impact of biodegradable plastics, a type of material belonging to the category of bioplastics for which there are still scarce data. This review identifies a need to perform studies using secondary nanoplastics rather than synthetic commercial materials as well as to include other polymers apart from polystyrene. There is also an urgent need to assess the possible risk of nanoplastics at environmentally realistic concentrations using sublethal endpoints and long-term assay

Luminescent microbial bioassays and microalgal biosensors as tools for environmental toxicity evaluation

This chapter deals with toxicity bioassays and biosensors based on luminescent microorganisms that report on global toxicity of a sample in such a way that luminescence is reduced or inhibited in the presence of toxic compounds that impair metabolism. Both natural as well as recombinant microorganisms are considered. A detailed description of their main characteristics and environmental applications is reported. A few examples of bioassays for detecting oxidative stress (both bioluminescent and fluorescent bioreporters) are also mentioned and discussed as reactive oxygen species (ROS) formation and subsequent oxidative stress if the antioxidant defenses of the cells are surpassed is one of the main mechanisms of toxicity for most pollutants. There is also a section dedicated to microalgal–based biosensors given their ecological relevance as primary producers, their easiness of culture and immobilization in different matrices, ability to acclimate to low nutrients conditions and ubiquity in aquatic environments. The most used toxicity endpoints for this type of biosensors are the alteration of photosynthetic activity (optical and amperometric biosensors) and the inhibition of enzymes such as alkaline phosphatase (APA) or sterases (mostly conductometric biosensors). The main information is shown in tables that include the microorganisms, their main characteristics (reporter gene systems, transducer types for biosensors) and their main environmental applications as well as relevant references. Although some of these bioassays have already been standardized by different international organizations, there are still many which are also promising tools for environmental global toxicity evaluation and should be fully validated and standardized for regulatory purposesThis research was supported by the Spanish Ministry of Economy and Competitiveness (MINECO), grant CTM2016-74927-C2-2-

Defining an additivity framework for mixture research in inducible whole-cell biosensors

A novel additivity framework for mixture effect modelling in the context of whole cell inducible biosensors has been mathematically developed and implemented in R. The proposed method is a multivariate extension of the effective dose (EDp) concept. Specifically, the extension accounts for differential maximal effects among analytes and response inhibition beyond the máximum permissive concentrations. This allows a multivariate extension of Loewe additivity, enabling direct application in a biphasic dose-response framework. The proposed additivity definition was validated, and its applicability illustrated by studying the response of the cyanobacterial biosensor Synechococcus elongatus PCC 7942 pBG2120 to binary mixtures of Zn, Cu, Cd, Ag, Co and Hg. The novel method allowed by the first time to model complete dose-response profiles of an inducible whole cell biosensor to mixtures. In addition, the approach also allowed identification and quantification of departures from additivity (interactions) among analytes. The biosensor was found to respond in a near additive way to heavy metal mixtures except when Hg, Co and Ag were present, in which case strong interactions occurred. The method is a useful contribution for the whole cell biosensors discipline and related areas allowing to perform appropriate assessment of mixture effects in non-monotonic dose-response frameworksThis research was supported by MINECO grants CGL2010-15675 and CTM2013-45775-C2-2-R

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 environment

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

Título del Post Print: Early and differential bacterial colonization on microplasticsMicrobial 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. β-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 ARGsFinancial support was provided by the Spanish Ministry of Economy and Competitiveness (CTM2016-74927-C2-1/2-R

Effects of petroleum-based and biopolymer-based nanoplastics on aquatic organisms: A case study with mechanically degraded pristine polymers

Mismanaged plastic litter submitted to environmental conditions may breakdown into smaller fragments, eventually reaching nano-scale particles (nanoplastics, NPLs). In this study, pristine beads of four different types of polymers, three oil-based (polypropylene, PP; polystyrene, PS; and low-density polyethylene, LDPE) and one bio-based (polylactic acid, PLA) were mechanically broken down to obtain more environmentally realistic NPLs and its toxicity to two freshwater secondary consumers was assessed. Thus, effects on the cnidarian Hydra viridissima (mortality, morphology, regeneration ability, and feeding behavior) and the fish Danio rerio (mortality, morphological alterations, and swimming behavior) were tested at NPLs concentrations in the 0.001 to 100 mg/L range. Mortality and several morphological alterations were observed on hydras exposed to 10 and 100 mg/L PP and 100 mg/L LDPE, whilst regeneration capacity was overall accelerated. The locomotory activity of D. rerio larvae was affected by NPLs (decreased swimming time, distance or turning frequency) at environmentally realistic concentrations (as low as 0.001 mg/L). Overall, petroleum- and bio-based NPLs elicited pernicious effects on tested model organisms, especially PP, LDPE and PLA. Data allowed the estimation of NPLs effective concentrations and showed that biopolymers may also induce relevant toxic effectsThe authors acknowledge the financial support provided by the Spanish Government (Ministerio de Ciencia e Innovación): PID2020-113769RBC21/22, PLEC2021-007693 and the Thematic Network of Micro- and Nanoplastics in the Environment (RED2018-102345-T, EnviroPlaNet Network) and by the Portuguese Government: Project NanoPlanet (2022.02340.PTDC) and financial support to CESAM (UIDP/ 50017/ 2020+UIDB/50017/2020+LA/P/0094/2020), through FCT/MCTES. MTB is the recipient of a FPU (FPU17/01789) pre-doctoral contract by the Spanish Ministerio de Ciencia, Innovación y Universidade

Honeybees as active samplers for microplastics

Microplastics are ubiquitous and their sampling is a difficult task. Honeybees interact with the environment inside their foraging range and take pollutants with them. In this work, we demonstrated for the first time that worker bees can act as active samplers of microplastics. We collected honeybees from apiaries located in the centre of Copenhagen and from nearby semiurban and rural areas. We showed the presence of microplastics in all sampled locations mostly in the form of fragments (52%) and fibres (38%) with average equivalent diameter of 64 ± 39 μm for fibres and 234 ± 156 μm for fragments. The highest load corresponded to urban apiaries, but comparable number of microplastics was found in hives from suburban and rural areas, which can be explained by the presence of urban settlements inside the foraging range of worker bees and to the easy dispersion of small microplastics by wind. Micro-FTIR analysis confirmed the presence of thirteen synthetic polymers, the most frequently of which was polyester followed by polyethylene and polyvinyl chloride. Our results demonstrated the presence of microplastics attached to the body of the honeybees and opens a new research pathway to their use as active biosamplers for anthropogenic pollutionThe authors acknowledge the financial support provided by the Spanish Government: CTM2016-74927-C2-1-R/2-R, and the Thematic Network of Micro- and Nanoplastics in the Environment (RED2018-102345-T, EnviroPlaNet Network). CE thanks the Spanish Ministry of Science, Innovation and Universities for the award of a pre-doctoral grant (FPI

Construction of a self-luminescent cyanobacterial bioreporter that detects a broad range of bioavailable heavy metals in aquatic environments

Frontiers in Microbiology 6 (2015): 186 This Document is Protected by copyright and was first published by Frontiers. All rights reserved. It is reproduced with permissionA self-luminescent bioreporter strain of the unicellular cyanobacterium Synechococcus sp. PCC 7942 was constructed by fusing the promoter region of the smt locus (encoding the transcriptional repressor SmtB and the metallothionein SmtA) to luxCDABE from Photorhabdus luminescens; the sensor smtB gene controlling the expression of smtA was cloned in the same vector. The bioreporter performance was tested with a range of heavy metals and was shown to respond linearly to divalent Zn, Cd, Cu, Co, Hg, and monovalent Ag. Chemical modeling was used to link bioreporter response with metal speciation and bioavailability. Limits of Detection (LODs), Maximum Permissive Concentrations (MPCs) and dynamic ranges for each metal were calculated in terms of free ion concentrations. The ranges of detection varied from 11 to 72 pM for Hg2+ (the ion to which the bioreporter was most sensitive) to 1.54-5.35 μM for Cd2+ with an order of decreasing sensitivity as follows: Hg2+ >> Cu2+ >> Ag+ > Co2+ = Zn2+ > Cd2+. However, the maximum induction factor reached 75-fold in the case of Zn2+ and 56-fold in the case of Cd2+, implying that Zn2+ is the preferred metal in vivo for the SmtB sensor, followed by Cd2+, Ag+ and Cu2+ (around 45-50-fold induction), Hg2+ (30-fold) and finally Co2+ (20-fold). The bioreporter performance was tested in real environmental samples with different water matrix complexity artificially contaminated with increasing concentrations of Zn, Cd, Ag, and Cu, confirming its validity as a sensor of free heavy metal cations bioavailability in aquatic environmentsThis study was funded by MINECO grants CGL2010-15675 and CTM2013-45775-C2-2-

Combined toxicity of graphite-diamond nanoparticles and thiabendazole to Daphnia magna

Carbon-based nanomaterials exhibit unique properties that make them suitable for a wide variety of industrial and biomedical applications. In this work, we studied the acute toxicity of graphite-diamond nanoparticles (GDN) combined with the fungicide thiabendazole (TBZ) to the immobilization of the cladoceran Daphnia magna in the presence and absence of the micro green algae Raphidocelis subcapitata, supplied as food source. The toxicity of GDN to D. magna decreased in the presence of R. subcapitata, while that of TBZ increased, the latter suggesting a carrier effect to TBZ. GDN-TBZ mixtures were fitted to the most common conceptual models applied to mixture toxicity: Concentration Addition (CA), Independent Action (IA) and Combination Index (CI). For GDN-TBZ mixtures in the absence of food the best fit was obtained with dose ratio deviation from CA model, while in the presence of food, dose level deviation from CA gave a better fit. The binary mixtures of GDN and TBZ showed synergistic toxic interactions at low concentrations, which could be attributed to the increased bioavailability of TBZ adsorbed on GDN. For higher concentrations of GDN, the binary mixtures turned antagonistic due to particle agglomeration. Our study provides evidence that deviations from additivity are dose dependent and relevant for the risk assessment of mixtures of nanoparticles with other chemical pollutants.publishe

Physicochemical and biological interactions between cerium oxide nanoparticles and a 1,8-naphthalimide derivative

This Accepted Manuscript is available for reuse under a CC BY-NC-ND licence after the 24 month embargo period provided that all the terms of the licence are adhered toCerium (Ce) oxide nanoparticles (CNPs) have attracted attention due to their high bioactivity and unique redox-chemistry. The oxygen vacancies at the surface of the nanoparticle explain the autocatalytic properties of CNPs in which the Ce3+ atoms occupy the center of the oxygen vacancies surrounded by Ce4+ atoms. Until now, CNPs have been associated with organic molecules at the synthesis stage to extend their applications or improve their stability. However, there is a lack of information regarding the post-synthesis interaction of CNPs and organic molecules that could enhance or induce new properties. Due to their unique optical properties and their many uses in different areas such as supramolecular chemistry or biomedicine, we have chosen a derivative from the family of naphthalimides (the 4-amino-1,8-naphthalimide-N-substituted; ANN) to study the interaction with different CNPs (CNP1-4) and their joint bioactivity compared to that of the same compounds alone. ANN-CNP complexes were formed as revealed by spectroscopic studies, but, the interaction was markedly different depending on the physicochemical properties of CNPs and their surface content of Ce3+ sites. The ANN adsorption on all CNPs involved the amino group in the naphthalene moiety as shown by NMR spectroscopy, while the pyrrolidine ring was mainly involved in the specific interaction between ANN and CNP1. The biological effect of each CNP and ANN individually and forming complexes was assessed using a bioluminescent model bacterium. The results showed that ANN and CNP with the higher content of surface Ce3+ (CNP1) when combined acted additively towards the used model organism. In the opposite, ANN-CNP2, ANN-CNP3 and ANN-CNP4 complexes were antagonistic when the nanoparticles dominated the mixture. The results of this study contribute to expand the knowledge of the interaction between nanoparticles and organic molecules which may be useful for understanding the behavior of nanoparticles in complex matricesThis research was supported by CTM2013-45775-C2-1-R and CTM2013-45775-C2-2-R grants from MINECO. Cerium oxide nanoparticles were kindly provided by S. Seal and S. Das (University of Central Florida, U.S.A). Gerardo Pulido-Reyes thanks the Spanish Ministry of Education for the award of an FPU grant (FPU12/01796