A domesticated photoautotrophic microbial community as a biofilm model system for analyzing the influence of plastic surfaces on invertebrate grazers in limnic environments

The environmental fate of plastic particles in water bodies is influenced by microbial biofilm formation. Invertebrate grazers may be affected when foraging biofilms on plastics compared to biofilms on natural substrata but the mechanistic basis for these effects is unknown. For analyzing these effects in ecotoxicological assays stable and reproducible biofilm communities are required that are related to the environmental site of interest. Here, a defined biofilm community was established and used to perform grazing experiments with a freshwater snail. For this, snippets of different plastic materials were incubated in the photic zone of three different freshwater sites. Amplicon sequencing of biofilms formed on these snippets showed that the site of incubation and not the plastic material dominated the microbial community composition. From these biofilms, individual microbial strains as well as photoautotrophic consortia were isolated; these consortia consisted of heterotrophic bacteria that were apparently nourished by microalga. While biofilms formed by defined dual cultures of a microalga and an Alphaproteobacterium were not accepted by the snail P. fontinalis, a photoautotrophic consortium (Co_3) sustained growth and metabolism of this grazer. Amplicon sequencing revealed that consortium Co_3, which could be stably maintained on solid medium under photoautotrophic conditions, reproducibly formed biofilms of a defined composition on three different plastic materials and on glass surfaces. In conclusion, our study shows that the generation of domesticated photoautotrophic microbial communities is a valid novel approach for establishing laboratory ecotoxicological assays with higher environmental relevance than those based on defined microbiota

Uptake and Transfer of Polyamide Microplastics in a Freshwater Mesocosm Study

Steadily increasing inputs of microplastics pose a growing threat to aquatic fauna, but laboratory studies potentially lack realism to properly investigate its effects on populations and ecosystems. Our study investigates the trophic and ontogenetic transfer of microplastics in a near-natural exposure scenario. The controlled outdoor freshwater mesocosms were exposed to polyamide (PA) 5–50 µm in size in concentrations of 15 and 150 mg L−1 and a control without microplastic addition. To verify the uptake of particles via the food chain, larvae and imagines of the midges Chaoborus crystallinus and C. obscuripes were examined, which feed on zooplankton during their larval stage. Larvae were captured after 117 days and imagines were caught in emergence traps that were emptied weekly. To detect the microparticles within the organisms, 200 larvae and 100 imagines per application were macerated and treated with fluorescent dye before investigation under a fluorescent microscope. We could detect up to 12 PA particles per individual larvae, while nearly no plastic was found in the imagines. This shows that, while Chaoborus sp. takes up microplastics via predation, most of the pollutant is egested through regurgitation and remains in the water, where it can further accumulate and potentially harm other organisms

Uptake and Transfer of Polyamide Microplastics in a Freshwater Mesocosm Study

Steadily increasing inputs of microplastics pose a growing threat to aquatic fauna, but laboratory studies potentially lack realism to properly investigate its effects on populations and ecosystems. Our study investigates the trophic and ontogenetic transfer of microplastics in a near-natural exposure scenario. The controlled outdoor freshwater mesocosms were exposed to polyamide (PA) 5–50 µm in size in concentrations of 15 and 150 mg L−1 and a control without microplastic addition. To verify the uptake of particles via the food chain, larvae and imagines of the midges Chaoborus crystallinus and C. obscuripes were examined, which feed on zooplankton during their larval stage. Larvae were captured after 117 days and imagines were caught in emergence traps that were emptied weekly. To detect the microparticles within the organisms, 200 larvae and 100 imagines per application were macerated and treated with fluorescent dye before investigation under a fluorescent microscope. We could detect up to 12 PA particles per individual larvae, while nearly no plastic was found in the imagines. This shows that, while Chaoborus sp. takes up microplastics via predation, most of the pollutant is egested through regurgitation and remains in the water, where it can further accumulate and potentially harm other organisms