Reduced pH increases mortality and genotoxicity in an Arctic coastal copepod, Acartia longiremis.

This study investigates DNA damage and mortality in an Arctic marine copepod after long-term exposure to lowered pH. Acartia longiremis were collected from northern Norway and incubated in ambient pH 8.1, and reduced pH 7.6 and 7.2 over 3-4 weeks. Cumulative mortality was significantly elevated in the lowered pH treatments in all exposures. The fluorescence-based fast micromethod for analysis of DNA strand breaks and alkali-labile sites was modified for use on crustaceous zooplankton. DNA damage initially increased in the lowered pH treatments, decreasing after >14 days, and DNA damage was significantly higher in lowered pH conditions. This method is ideal for investigating oxidative stress and genotoxicity response to low pH in Arctic marine copepods exposed to future ocean acidification conditions.

Moving forward in microplastic research: A Norwegian perspective.

Given the increasing attention on the occurrence of microplastics in the environment, and the potential environmental threats they pose, there is a need for researchers to move quickly from basic understanding to applied science that supports decision makers in finding feasible mitigation measures and solutions. At the same time, they must provide sufficient, accurate and clear information to the media, public and other relevant groups (e.g., NGOs). Key requirements include systematic and coordinated research efforts to enable evidence-based decision making and to develop efficient policy measures on all scales (national, regional and global). To achieve this, collaboration between key actors is essential and should include researchers from multiple disciplines, policymakers, authorities, civil and industry organizations, and the public. This further requires clear and informative communication processes, and open and continuous dialogues between all actors. Cross-discipline dialogues between researchers should focus on scientific quality and harmonization, defining and accurately communicating the state of knowledge, and prioritization of topics that are critical for both research and policy, with the common goal to establish and update action plans for holistic benefit. In Norway, cross-sectoral collaboration has been fundamental in supporting the national strategy to address plastic pollution. Researchers, stakeholders and the environmental authorities have come together to exchange knowledge, identify knowledge gaps, and set targeted and feasible measures to tackle one of the most challenging aspects of plastic pollution: microplastic. In this article, we present a Norwegian perspective on the state of knowledge on microplastic research efforts. Norway's involvement in international efforts to combat plastic pollution aims at serving as an example of how key actors can collaborate synergistically to share knowledge, address shortcomings, and outline ways forward to address environmental challenges.

Aging of microplastics promotes their ingestion by marine zooplankton.

Microplastics (<5 mm) are ubiquitous in the marine environment and are ingested by zooplankton with possible negative effects on survival, feeding, and fecundity. The majority of laboratory studies has used new and pristine microplastics to test their impacts, while aging processes such as weathering and biofouling alter the characteristics of plastic particles in the marine environment. We investigated zooplankton ingestion of polystyrene beads (15 and 30 µm) and fragments (≤30 µm), and tested the hypothesis that microplastics previously exposed to marine conditions (aged) are ingested at higher rates than pristine microplastics. Polystyrene beads were aged by soaking in natural local seawater for three weeks. Three zooplankton taxa ingested microplastics, excluding the copepod Pseudocalanus spp., but the proportions of individuals ingesting plastic and the number of particles ingested were taxon and life stage specific and dependent on plastic size. All stages of Calanus finmarchicus ingested polystyrene fragments. Aged microbeads were preferred over pristine ones by females of Acartia longiremis as well as juvenile copepodites CV and adults of Calanus finmarchicus. The preference for aged microplastics may be attributed to the formation of a biofilm. Such a coating, made up of natural microbes, may contain similar prey as the copepods feed on in the water column and secrete chemical exudates that aid chemodetection and thus increase the attractiveness of the particles as food items. Much of the ingested plastic was, however, egested within a short time period (2-4 h) and the survival of adult Calanus females was not affected in an 11-day exposure. Negative effects of microplastics ingestion were thus limited. Our findings emphasize, however, that aging plays an important role in the transformation of microplastics at sea and ingestion by grazers, and should thus be considered in future microplastics ingestion studies and estimates of microplastics transfer into the marine food web.

Seawater pH Predicted for the Year 2100 Affects the Metabolic Response to Feeding in Copepodites of the Arctic Copepod Calanus glacialis.

Widespread ocean acidification (OA) is transforming the chemistry of the global ocean, and the Arctic is recognised as a region where the earliest and strongest impacts of OA are expected. In the present study, metabolic effects of OA and its interaction with food availability was investigated in Calanus glacialis from the Kongsfjord, West Spitsbergen. We measured metabolic rates and RNA/DNA ratios (an indicator of biosynthesis) concurrently in fed and unfed individuals of copepodite stages CII-CIII and CV subjected to two different pH levels representative of present day and the "business as usual" IPCC scenario (RCP8.5) prediction for the year 2100. The copepods responded more strongly to changes in food level than to decreasing pH, both with respect to metabolic rate and RNA/DNA ratio. However, significant interactions between effects of pH and food level showed that effects of pH and food level act in synergy in copepodites of C. glacialis. While metabolic rates in copepodites stage CII-CIII increased by 78% as a response to food under present day conditions (high pH), the increase was 195% in CII-CIIIs kept at low pH-a 2.5 times greater increase. This interaction was absent for RNA/DNA, so the increase in metabolic rates were clearly not a reaction to changing biosynthesis at low pH per se but rather a reaction to increased metabolic costs per unit of biosynthesis. Interestingly, we did not observe this difference in costs of growth in stage CV. A 2.5 times increase in metabolic costs of growth will leave the copepodites with much less energy for growth. This may infer significant changes to the C. glacialis population during future OA.

Microplastics Alter the Properties and Sinking Rates of Zooplankton Faecal Pellets.

Plastic debris is a widespread contaminant, prevalent in aquatic ecosystems across the globe. Zooplankton readily ingest microscopic plastic (microplastic, < 1 mm), which are later egested within their faecal pellets. These pellets are a source of food for marine organisms, and contribute to the oceanic vertical flux of particulate organic matter as part of the biological pump. The effects of microplastics on faecal pellet properties are currently unknown. Here we test the hypotheses that (1) faecal pellets are a vector for transport of microplastics, (2) polystyrene microplastics can alter the properties and sinking rates of zooplankton egests and, (3) faecal pellets can facilitate the transfer of plastics to coprophagous biota. Following exposure to 20.6 µm polystyrene microplastics (1000 microplastics mL(-1)) and natural prey (∼1650 algae mL(-1)) the copepod Calanus helgolandicus egested faecal pellets with significantly (P < 0.001) reduced densities, a 2.25-fold reduction in sinking rates, and a higher propensity for fragmentation. We further show that microplastics, encapsulated within egests of the copepod Centropages typicus, could be transferred to C. helgolandicus via coprophagy. Our results support the proposal that sinking faecal matter represents a mechanism by which floating plastics can be vertically transported away from surface waters.

The impact of polystyrene microplastics on feeding, function and fecundity in the marine copepod Calanus helgolandicus.

Microscopic plastic debris, termed "microplastics", are of increasing environmental concern. Recent studies have demonstrated that a range of zooplankton, including copepods, can ingest microplastics. Copepods are a globally abundant class of zooplankton that form a key trophic link between primary producers and higher trophic marine organisms. Here we demonstrate that ingestion of microplastics can significantly alter the feeding capacity of the pelagic copepod Calanus helgolandicus. Exposed to 20 µm polystyrene beads (75 microplastics mL(­1)) and cultured algae ([250 µg C L(­1)) for 24 h, C. helgolandicus ingested 11% fewer algal cells (P = 0.33) and 40% less carbon biomass (P < 0.01). There was a net downward shift in the mean size of algal prey consumed (P < 0.001), with a 3.6 fold increase in ingestion rate for the smallest size class of algal prey (11.6­12.6 µm), suggestive of postcapture or postingestion rejection. Prolonged exposure to polystyrene microplastics significantly decreased reproductive output, but there were no significant differences in egg production rates, respiration or survival. We constructed a conceptual energetic (carbon) budget showing that microplastic-exposed copepods suffer energetic depletion over time. We conclude that microplastics impede feeding in copepods, which over time could lead to sustained reductions in ingested carbon biomass.

Isolation of microplastics in biota-rich seawater samples and marine organisms.

Microplastic litter is a pervasive pollutant present in aquatic systems across the globe. A range of marine organisms have the capacity to ingest microplastics, resulting in adverse health effects. Developing methods to accurately quantify microplastics in productive marine waters, and those internalized by marine organisms, is of growing importance. Here we investigate the efficacy of using acid, alkaline and enzymatic digestion techniques in mineralizing biological material from marine surface trawls to reveal any microplastics present. Our optimized enzymatic protocol can digest >97% (by weight) of the material present in plankton-rich seawater samples without destroying any microplastic debris present. In applying the method to replicate marine samples from the western English Channel, we identified 0.27 microplastics m(-3). The protocol was further used to extract microplastics ingested by marine zooplankton under laboratory conditions. Our findings illustrate that enzymatic digestion can aid the detection of microplastic debris within seawater samples and marine biota.

Potential acidification impacts on zooplankton in CCS leakage scenarios.

Carbon capture and storage (CCS) technologies involve localized acidification of significant volumes of seawater, inhabited mainly by planktonic species. Knowledge on potential impacts of these techniques on the survival and physiology of zooplankton, and subsequent consequences for ecosystem health in targeted areas, is scarce. The recent literature has a focus on anthropogenic greenhouse gas emissions into the atmosphere, leading to enhanced absorption of CO2 by the oceans and a lowered seawater pH, termed ocean acidification. These studies explore the effects of changes in seawater chemistry, as predicted by climate models for the end of this century, on marine biota. Early studies have used unrealistically severe CO2/pH values in this context, but are relevant for CCS leakage scenarios. Little studied meso- and bathypelagic species of the deep sea may be especially vulnerable, as well as vertically migrating zooplankton, which require significant residence times at great depths as part of their life cycle.

Microplastic ingestion by zooplankton.

Small plastic detritus, termed "microplastics", are a widespread and ubiquitous contaminant of marine ecosystems across the globe. Ingestion of microplastics by marine biota, including mussels, worms, fish, and seabirds, has been widely reported, but despite their vital ecological role in marine food-webs, the impact of microplastics on zooplankton remains under-researched. Here, we show that microplastics are ingested by, and may impact upon, zooplankton. We used bioimaging techniques to document ingestion, egestion, and adherence of microplastics in a range of zooplankton common to the northeast Atlantic, and employed feeding rate studies to determine the impact of plastic detritus on algal ingestion rates in copepods. Using fluorescence and coherent anti-Stokes Raman scattering (CARS) microscopy we identified that thirteen zooplankton taxa had the capacity to ingest 1.7-30.6 µm polystyrene beads, with uptake varying by taxa, life-stage and bead-size. Post-ingestion, copepods egested faecal pellets laden with microplastics. We further observed microplastics adhered to the external carapace and appendages of exposed zooplankton. Exposure of the copepod Centropages typicus to natural assemblages of algae with and without microplastics showed that 7.3 µm microplastics (>4000 mL(-1)) significantly decreased algal feeding. Our findings imply that marine microplastic debris can negatively impact upon zooplankton function and health.

Effects of elevated CO2 on the reproduction of two calanoid copepods.

Some planktonic groups suffer negative effects from ocean acidification (OA), although copepods might be less sensitive. We investigated the effect of predicted CO2 levels (range 480-750ppm), on egg production and hatching success of two copepod species, Centropages typicus and Temora longicornis. In these short-term incubations there was no significant effect of high CO2 on these parameters. Additionally a very high CO2 treatment, (CO2=9830ppm), representative of carbon capture and storage scenarios, resulted in a reduction of egg production rate and hatching success of C. typicus, but not T. longicornis. In conclusion, reproduction of C. typicus was more sensitive to acute elevated seawater CO2 than that of T. longicornis, but neither species was affected by exposure to CO2 levels predicted for the year 2100. The duration and seasonal timing of exposures to high pCO2, however, might have a significant effect on the reproduction success of calanoid copepods.

The relevance of marine chemical ecology to plankton and ecosystem function: an emerging field.

Marine chemical ecology comprises the study of the production and interaction of bioactive molecules affecting organism behavior and function. Here we focus on bioactive compounds and interactions associated with phytoplankton, particularly bloom-forming diatoms, prymnesiophytes and dinoflagellates. Planktonic bioactive metabolites are structurally and functionally diverse and some may have multiple simultaneous functions including roles in chemical defense (antipredator, allelopathic and antibacterial compounds), and/or cell-to-cell signaling (e.g., polyunsaturated aldehydes (PUAs) of diatoms). Among inducible chemical defenses in response to grazing, there is high species-specific variability in the effects on grazers, ranging from severe physical incapacitation and/or death to no apparent physiological response, depending on predator susceptibility and detoxification capability. Most bioactive compounds are present in very low concentrations, in both the producing organism and the surrounding aqueous medium. Furthermore, bioactivity may be subject to synergistic interactions with other natural and anthropogenic environmental toxicants. Most, if not all phycotoxins are classic secondary metabolites, but many other bioactive metabolites are simple molecules derived from primary metabolism (e.g., PUAs in diatoms, dimethylsulfoniopropionate (DMSP) in prymnesiophytes). Producing cells do not seem to suffer physiological impact due to their synthesis. Functional genome sequence data and gene expression analysis will provide insights into regulatory and metabolic pathways in producer organisms, as well as identification of mechanisms of action in target organisms. Understanding chemical ecological responses to environmental triggers and chemically-mediated species interactions will help define crucial chemical and molecular processes that help maintain biodiversity and ecosystem functionality.

Microplastics as contaminants in the marine environment: a review.

Since the mass production of plastics began in the 1940s, microplastic contamination of the marine environment has been a growing problem. Here, a review of the literature has been conducted with the following objectives: (1) to summarise the properties, nomenclature and sources of microplastics; (2) to discuss the routes by which microplastics enter the marine environment; (3) to evaluate the methods by which microplastics are detected in the marine environment; (4) to assess spatial and temporal trends of microplastic abundance; and (5) to discuss the environmental impact of microplastics. Microplastics are both abundant and widespread within the marine environment, found in their highest concentrations along coastlines and within mid-ocean gyres. Ingestion of microplastics has been demonstrated in a range of marine organisms, a process which may facilitate the transfer of chemical additives or hydrophobic waterborne pollutants to biota. We conclude by highlighting key future research areas for scientists and policymakers.