Plastic Alters Biofilm Quality as Food Resource of the Freshwater Gastropod Radix balthica.

High amounts of plastic debris enter and accumulate in freshwater systems across the globe. The plastic contamination of benthic habitats in lakes and running waters poses a potential threat to freshwater ecosystems. This study investigates the effects of plastic on two trophic levels of the aquatic food web: primary production, that is, epiplastic biofilm, and primary consumption, that is, a benthic invertebrate grazer. Two plastic types, polymethyl methacrylate (PMMA) and polycarbonate (PC), and glass (control) were used as substrata for natural biofilm establishment. PMMA and PC are, for example, intensively used in the automobile, construction, and electronical industries and in cosmetics (PMMA), CDs, and DVDs (PC). These biofilms were fed to the freshwater gastropod  Radix balthica (Linnaeus 1758) in a laboratory-grazing experiment. Biofilm structure and composition were observed using confocal laser scanning microscopy before the grazing experiment. Sublethal effects on R. balthica were observed measuring consumption of biofilm and growth rates. The biofilm composition on PMMA significantly differed compared to PC and glass. The grazing experiments showed limited biofilm consumption and lower growth rates of R. balthica in both plastic treatments. Concluding, plastic in freshwaters has a direct effect on the primary production and an indirect effect on higher trophic levels.

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.

Floating microplastic debris in a rural river in Germany: Distribution, types and potential sources and sinks.

Microplastic debris affects marine as well as freshwater ecosystems and an increasing number of studies have documented the contamination in aquatic environments worldwide. However, while the research focuses on oceans and larger rivers, little is known about the situation in smaller rivers within rural catchments. Since microplastics pose various risks to ecosystems, wildlife and human health, it is important to identify potential sources, sinks and transport patterns, which are probably different for small rivers. In this study, we investigate the contamination with microplastic debris of the river Ems, representing a smaller river in Northwest Germany with an agricultural catchment. We hypothesised that with increasing river length the plastic concentration increases, especially downstream of towns, waste water treatment plant (WWTP) effluents and major tributaries as they may be important point sources of microplastics. We collected 36 surface water samples at 18 sampling sites within the first 70 km using manual driftnets. We sampled every 7 km and upstream and downstream of three larger towns, four major tributaries and four WWTP effluents. Overall, we found 1.54 ± 1.54 items m-3, which corresponds to the plastic concentrations in larger streams. However, the shape of the detected items differed as we did not find potential primary microplastic. Furthermore, the pattern contradicts our assumption, that the contamination increased with distance to the river's source. Downstream of towns, we found significantly less floating microplastic indicating possible sinks due to sedimentation at sites with slowing flow velocity caused by weirs in towns. Hence, the non-linear distribution pattern of microplastics indicates potential sinks of microplastics due to flow alterations on the river course. This should be considered in future studies modelling microplastic distribution and transport. Furthermore, studies especially in smaller rivers are urgently needed to identify and quantify sources and sinks and to find applicable solutions to reduce microplastic loads.

Environmental conditions affect the food quality of plastic associated biofilms for the benthic grazer Physa fontinalis.

With an ever-increasing amount of plastic pollution in the various aquatic ecosystems around the world, the effects on organisms are still not fully understood. Most studies focus on direct effects posed by plastic intake or entanglement, but plastic debris can also affect primary production of biofilms and have an indirect impact on its consumers. This study investigates the primary production on three common plastic types in freshwater and its food quality for a benthic grazer. We hypothesized that different polymer types affect biofilm composition as well as the life parameters of its consumers. We incubated polyethylene (PE), polyethylene terephthalate (PET) and polystyrene (PS) as well as glass (control) in a productive freshwater creek for natural biofilm establishment. To account for changes in the environmental conditions, the experiment was conducted twice during winter and late spring, respectively. These biofilms were offered to the freshwater gastropod Physa fontinalis as sole food source. Growth and reproduction of the snails were measured to monitor sublethal effects. Additionally, biofilm composition was observed using confocal laser scanning microscopy (CLSM). In winter, snails feeding off PET and PE showed a significantly lower egg production and lower growth rates were observed on PET. No such effects occurred in spring. CLSM data revealed, that algal growth was significantly lower on PE and PET during the winter treatment compared to PS and glass. Since we could only find these effects during the colder and darker months (January-March), the microbial colonization on PE and PET was inhibited by the substrate under less favorable conditions of temperature and light. Hence, benign conditions may mask the adverse effects of microplastic on food webs. Our findings show that future studies on the plastisphere will need to consider such variations to further understand the influence of plastic pollution on primary production and higher trophic levels.

Waves affect predator-prey interactions between fish and benthic invertebrates.

Little is known about the effects of waves on predator-prey interactions in the littoral zones of freshwaters. We conducted a set of mesocosm experiments to study the differential effects of ship- and wind-induced waves on the foraging success of littoral fish on benthic invertebrates. Experiments were conducted in a wave tank with amphipods (Gammarus roeseli) as prey, and age-0 bream (Abramis brama, B0), age-0 and age-1 dace (Leuciscus leuciscus, D0 and D1) as predators. The number of gammarids suspended in the water column was higher in the wave treatments compared to a no-wave control treatment, especially during pulse waves mimicking ship-induced waves in comparison to continuous waves mimicking wind-induced waves. The resulting higher prey accessibility in the water column was differently exploited by the three types of predatory fish. D0 and D1 showed significantly higher foraging success in the pulse wave treatment than in the continuous and control treatments. The foraging success of D0 appears to be achieved more easily, since significantly higher swimming activity and more foraging attempts were recorded only for D1 under the wave treatments. In contrast, B0 consumed significantly fewer gammarids in both wave treatments than in the control. Hence, waves influenced predator-prey interactions differently depending on wave type and fish type. It is expected that regular exposure to ship-induced waves can alter littoral invertebrate and fish assemblages by increasing the predation risk for benthic invertebrates that are suspended in the water column, and by shifting fish community compositions towards species that benefit from waves.

Transfer and effects of PET microfibers in Chironomus riparius.

Multiple studies in freshwater environments have verified that microplastic particles are present in water columns, sediment, and aquatic organisms. These studies indicated that certain freshwater ecosystems may act as temporary sinks of microplastic particles, leading to accumulation in the sediment and the ingestion by benthic organisms. Polyethylene terephthalate (PET) is one of the non-buoyant polymers that has been frequently found in aquatic sediments. This study aims to investigate a possible transfer of PET microfibers from aquatic to the terrestrial habitats and addressed selected effects (i.e. survival, general stress response, and growth) of PET microfibers using Chironomus riparius, a frequently applied model organism in ecotoxicological research. To assess the growth and development of C. riparius, a modified 28-day sediment chronic toxicity test was conducted, in which the main endpoint is time until emergence of the larvae. In this assay, C. riparius were exposed to artificial sediments spiked with PET microfibers. In addition, weight and head capsule lengths of the larvae were also measured. As a general stress response marker on the molecular level, Heat Shock Protein 70 (HSP70) levels were measured in two involved life stages, i.e. larvae and adults. Using staining method, ingestion of PET microfibers was verified in the adult sample. Our results clearly demonstrated that ingested microfibers by C. riparius larvae can be carried through subsequent life stages and end up in the adults. Accordingly, this is the first proof of aquatic-terrestrial transfer of PET microfibers for C. riparius. However, toxicity test results showed that there was no significant effect on the time until emergence, weight or head capsule lengths in the organisms exposed to PET microfibers compared to control organisms. HSP70 measurements showed no significant effects between control and exposure groups in the same life stage. The result suggests that PET microfibers in the applied concentration do not exert adverse effects both on organism and subcellular level in one generation.

Thermal limits in native and alien freshwater peracarid Crustacea: The role of habitat use and oxygen limitation.

In order to predict which species can successfully cope with global warming and how other environmental stressors modulate their vulnerability to climate-related environmental factors, an understanding of the ecophysiology underpinning thermal limits is essential for both conservation biology and invasion biology.Heat tolerance and the extent to which heat tolerance differed with oxygen availability were examined for four native and four alien freshwater peracarid crustacean species, with differences in habitat use across species. Three hypotheses were tested: (1) Heat and lack of oxygen synergistically reduce survival of species; (2) patterns in heat tolerance and the modulation thereof by oxygen differ between alien and native species and between species with different habitat use; (3) small animals can better tolerate heat than large animals, and this difference is more pronounced under hypoxia.To assess heat tolerances under different oxygen levels, animal survival was monitored in experimental chambers in which the water temperature was ramped up (0.25°C min-1). Heat tolerance (CTmax) was scored as the cessation of all pleopod movement, and heating trials were performed under hypoxia (5 kPa oxygen), normoxia (20 kPa) and hyperoxia (60 kPa).Heat tolerance differed across species as did the extent by which heat tolerance was affected by oxygen conditions. Heat-tolerant species, for example, Asellus aquaticus and Crangonyx pseudogracilis, showed little response to oxygen conditions in their CTmax, whereas the CTmax of heat-sensitive species, for example, Dikerogammarus villosus and Gammarus fossarum, was more plastic, being increased by hyperoxia and reduced by hypoxia.In contrast to other studies on crustaceans, alien species were not more heat-tolerant than native species. Instead, differences in heat tolerance were best explained by habitat use, with species from standing waters being heat tolerant and species from running waters being heat sensitive. In addition, larger animals displayed lower critical maximum temperature, but only under hypoxia. An analysis of data available in the literature on metabolic responses of the study species to temperature and oxygen conditions suggests that oxygen conformers and species whose oxygen demand rapidly increases with temperature (low activation energy) may be more heat sensitive.The alien species D. villosus appeared most susceptible to hypoxia and heat stress. This may explain why this species is very successful in colonizing new areas in littoral zones with rocky substrate which are well aerated due to continuous wave action generated by passing ships or prevailing winds. This species is less capable of spreading to other waters which are poorly oxygenated and where C. pseudogracilis is the more likely dominant alien species. A http://onlinelibrary.wiley.com/doi/10.1111/1365-2435.13050/suppinfo is available for this article.

Effects of ship-induced waves on aquatic ecosystems.

Most larger water bodies worldwide are used for navigation, and the intensity of commercial and recreational navigation is expected to further increase. Navigation profoundly affects aquatic ecosystems. To facilitate navigation, rivers are trained and developed, and the direct effects of navigation include chemical and biological impacts (e.g., inputs of toxic substances and dispersal of non-native species, respectively). Furthermore, propagating ships create hydrodynamic alterations, often simply summarized as waves. Although ship-induced waves are recognized as influential stressors, knowledge on their effects is poorly synthesized. We present here a review on the effects of ship-induced waves on the structure, function and services of aquatic ecosystems based on more than 200 peer reviewed publications and technical reports. Ship-induced waves act at multiple organizational levels and different spatial and temporal scales. All the abiotic and biotic components of aquatic ecosystems are affected, from the sediment and nutrient budget to the planktonic, benthic and fish communities. We highlight how the effects of ship-induced waves cascade through ecosystems and how different effects interact and feed back into the ecosystem finally leading to altered ecosystem services and human health effects. Based on this synthesis of wave effects, we discuss strategies for mitigation. This may help to develop scientifically based and target-oriented management plans for navigational waters that optimize abiotic and biotic integrity and their ecosystem services and uses.