Biodegradable microplastics: Uptake by and effects on the rockpool shrimp Palaemon elegans (Crustacea: Decapoda).

Ingestion of microplastics can lead to deleterious consequences for organisms, as documented by numerous laboratory studies. The current knowledge is based on a multitude of effect studies, conducted with conventional fossil-based and non-degradable plastics. However, there is a lack of information about the acceptance and the effects of novel bio-based and biodegradable plastics. Biodegradable plastics are considered an alternative to conventional plastics and are showing rapidly growing production rates. Biodegradable plastics can disperse into the environment in the same way as conventional plastics do, becoming available to marine organisms. This study aims to provide new insights into the uptake and effects of biodegradable microplastics on marine invertebrates. Rockpool shrimp, Palaemon elegans, were fed with algal flakes coated with polylactic acid (PLA), polyhydroxybutyrate-co-valerate (PHBV) and conventional low-density polyethylene (LDPE) microparticles. Live observations showed that all of the different types of microplastics were ingested. After dissection of the shrimp, less LDPE particles were found in the stomachs than PLA and PHBV particles. This indicates a longer retention time of biodegradable microplastics compared to conventional microplastics. Presumably, less LDPE particles were ingested or evacuated from the stomach, probably by regurgitation. The ingestion of microparticles of all types of plastics induced enzymatic activity of short-chain carboxylesterases in the midgut glands of the shrimp. However, only PLA induced enzymatic activity of medium-chain carboxylesterases. Palaemon elegans showed no oxidative stress response after ingestion of microparticles, irrespective of polymer type. From our results we conclude that biodegradable plastics might have different effects than conventional plastics. The longer retention times of biodegradable plastics might enhance exposure to leaching additives and other harmful substances. Our study provides new insights into how biodegradable plastics might affect aquatic fauna and indicate that the use of biodegradable plastics needs to be reconsidered to some extent.

Degradation of a poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) compound in different environments.

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a promising biodegradable bio-based material, which is designed for a vast range of applications, depending on its composite. This study aims to assess the degradability of a PHBV-based compound under different conditions. The research group followed different methodological approaches and assessed visual and mass changes, mechanical and morphological properties, spectroscopic and structural characterisation, along with thermal behaviour. The Ph-Stat (enzymatic degradation) test and total dry solids (TDS)/total volatile solids (TVS) measurements were carried out. Finally, the team experimentally evaluated the amount of methane and carbon dioxide produced, i.e., the degree of biodegradation under aerobic conditions. According to the results, different types of tests have shown differing effects of environmental conditions on material degradation. In conclusion, this paper provides a summary of the investigations regarding the degradation behaviour of the PHBV-based compound under varying environmental factors. The main strengths of the study lie in its multi-faceted approach, combining assessments of PHBV-based compound degradability under different conditions using various analytical tools, such as visual and mass changes, mechanical and morphological properties, spectroscopic and structural characterization, and thermal behavior. These methods collectively contribute to the robustness and reliability of the undertaken work.

Microplastic retention by marine vegetated canopies: Simulations with seagrass meadows in a hydraulic flume.

Marine canopies formed by seagrass and other coastal vegetated ecosystems could act as sinks of microplastics for being efficient particle traps. Here we investigated for the first time the occurrence of microplastic retention by marine canopies in a hydraulic flume under unidirectional flow velocities from 2 to 30 cm s-1. We used as model canopy-forming species the seagrass Zostera marina with four canopy shoot density (0, 50, 100, 200 shoots m-2), and we used as microplastic particles industrial pristine pellets with specific densities from 0.90 to 1.34 g cm-3 (polypropylene PP; polystyrene PS; polyamide 6 PA; and polyethylene terephthalate PET). Overall, microplastics particles transported with the flow were retained in the seagrass canopies but not in bare sand. While seagrass canopies retained floating microplastics (PP) only at low velocities (<12 cm s-1) due to a barrier created by the canopy touching the water surface, the retention of sinking particles (PS, PA, PET) occurred across a wider range of flow velocities. Our simulations revealed that less dense sinking particles (PS) might escape from the canopy at high velocities, while denser sinking particles can be trapped in scouring areas created by erosive processes around the eelgrass shoots. Our results show that marine canopies might act as potential barriers or sinks for microplastics at certain bio-physical conditions, with the probability of retention generally increasing with the seagrass shoot density and polymer specific density and decreasing with the flow velocity. We conclude that seagrass meadows, and other aquatic canopy-forming ecosystems, should be prioritized habitats in assessment of microplastic exposure and impact on coastal areas since they may accumulate high concentration of microplastic particles that could affect associated fauna.

Bacteriostatic suppression in Norway lobster (Nephrops norvegicus) exposed to manganese or hypoxia under pressure of ocean acidification.

Future ocean acidification (OA) and warming following climate change elicit pervasive stressors to the inhabitants of the sea. Previous experimental exposure to OA for 16 weeks at pH levels predicted for 2100 has shown to result in serious immune suppression of the Norway lobster, Nephrops norvegicus. The lobsters are currently affected by stressors such as periodical hypoxia inducing high levels of bioavailable manganese (Mn). Here, we aimed to investigate possible effects of interactions between OA and these stressors on total hemocyte counts (THCs) and on recovery of inoculated bacteria in the lobsters, measured as a proxy for bacteriostatic response. The effects were judged by following numbers of culturable Vibrio parahaemolyticus in hepatopancreas, 4 and 24h post inoculation in lobsters kept in replicate tanks with six different treatments: either ambient (pCO2∼500 µatm/pH∼8.1 U) or CO2-manipulated seawater (OA; pCO2∼1550 µatm/pH∼7.6 U) for 8 weeks. During the last 2 weeks, additional stress of either hypoxia (∼23% oxygen saturation) or Mn (∼9 mg L(-1)) was added except in control treatments. Our results showed clear effect on bacteriostatic response in Norway lobsters exposed to these stressors. In lobsters kept in ambient seawater without additional stressors, the number of culturable bacteria in hepatopancreas was reduced by ∼34%. In combined treatment of ambient seawater and hypoxia, the reduction was ∼23%, while in the Mn-exposed animals, there was no reduction at all. This was also the case in all OA treatments where mean numbers of culturable V. parahaemolyticus tended to increase. In lobsters from ambient seawater with or without hypoxia, the THC was not significantly different as was also the case in OA without additional stressors. However, in OA treatments combined with either hypoxia or Mn, THC was reduced by ∼35%. While the reduction of culturable V. parahaemolyticus in lobsters was clearly affected by these stressors, we found no notable effects on growth, survival or hemolytic properties of the bacteria itself. Thus, we conclude that this predicted stress scenario is beneficial for the pathogen in its interaction with the host. As OA proceeds, it may force the health of the ecologically and economically important N. norvegicus to a tipping point if exposed to more short-term stressors such as the periodical events of hypoxia and Mn. This could impact lobster condition and biomass and may as well increase the risk for bacterial transmission to consumers.

Transition from sea to land: olfactory function and constraints in the terrestrial hermit crab Coenobita clypeatus.

The ability to identify chemical cues in the environment is essential to most animals. Apart from marine larval stages, anomuran land hermit crabs (Coenobita) have evolved different degrees of terrestriality, and thus represent an excellent opportunity to investigate adaptations of the olfactory system needed for a successful transition from aquatic to terrestrial life. Although superb processing capacities of the central olfactory system have been indicated in Coenobita and their olfactory system evidently is functional on land, virtually nothing was known about what type of odourants are detected. Here, we used electroantennogram (EAG) recordings in Coenobita clypeatus and established the olfactory response spectrum. Interestingly, different chemical groups elicited EAG responses of opposite polarity, which also appeared for Coenobita compressus and the closely related marine hermit crab Pagurus bernhardus. Furthermore, in a two-choice bioassay with C. clypeatus, we found that water vapour was critical for natural and synthetic odourants to induce attraction or repulsion. Strikingly, also the physiological response was found much greater at higher humidity in C. clypeatus, whereas no such effect appeared in the terrestrial vinegar fly Drosophila melanogaster. In conclusion, our results reveal that the Coenobita olfactory system is restricted to a limited number of water-soluble odourants, and that high humidity is most critical for its function.

Naphthalene disrupts pheromone induced mate search in the amphipod Corophium volutator (Pallas).

Many crustaceans communicate with pheromones during mating. In the infaunal amphipod Corophium volutator (Pallas), females release gender-specific waterborne pheromones that guide males in their search for receptive burrowed females. Pollutants that affect the pheromone release or impair the ability of males to detect and respond to chemosensory cues, could profoundly affect mating. C. volutator is commonly found on shallow soft sediments in estuaries and coastal waters, and their reproductive season coincides with intense traffic of leisure boats, which discharge high concentrations of fuel into the water and sediment of these areas. This study shows that sublethal exposure to naphthalene, a polycyclic aromatic hydrocarbon abundant in motor fuel, disturbs chemical communication in C. volutator. This was demonstrated in Y-maze bioassays, where males were allowed to follow female pheromones after separately exposing the males or females to naphthalene spiked sediments (0, 0.5, 5 or 50 microg g(-1) dw, nominal concentrations) for 3 days. The results show that exposure to naphthalene at the two lower concentrations significantly affects the males' response to female pheromones (p<0.05 and p<0.01, respectively). Male search activity was reduced by 27-45% and males could no longer find females by the use of the olfactory sense. Analysed sediment samples, however, indicate that the naphthalene concentrations causing this effect were 2-20 times lower than nominal concentrations. At the highest naphthalene treatment, no significant difference from the controls was found, probably explained by a different exposure regime for these amphipods since they seemingly avoided burrowing in sediments. Female C. volutator produce and release pheromones, despite naphthalene exposure. Disrupted chemical communication in C. volutator may affect the reproductive fitness of males and females with possible consequences for populations in contaminated areas.

Effects of a candidate antifouling compound (medetomidine) on pheromone induced mate search in the amphipod Corophium volutator.

Environmental hazards associated with traditional, toxic antifouling coatings based on heavy metals calls for the development of alternative, environmentally acceptable antifouling compounds. Medetomidine ((+/-)-4-[1-(2,3-dimethylphenyl)ethyl]-1H-imidazole) is a candidate antifouling biocide which impedes settlement of barnacles in the nanomolar range. Prior to introducing novel biocides it is of great importance to consider potential effects on non-target organisms. This study is the first to investigate the effects of medetomidine on the amphipod Corophium volutator, specifically effects on male mate search behaviour. In a laboratory, Y-maze bioassay, C. volutator males were allowed to follow female pheromones after 24 h exposure to 0 (control), 0.01 and 0.1 microg mL(-1) medetomidine. We found that exposure to medetomidine at both concentrations significantly reduced pheromone induced mate search (by 42-71%), with fewer males crawling towards female odour. The results obtained indicate that medetomidine may impair the reproductive fitness of non-target crustaceans, an aspect that needs to be considered before further commercialisation.

Copper reduced mating behaviour in male shore crabs (Carcinus maenas (L.)).

Many crustaceans use pheromones to find mates and induce mating behaviours. If pollutants impair the ability to detect chemosensory cues and respond to pheromone signals, they could profoundly affect mating. In a series of laboratory experiments, the effect of copper (0, 0.1 or 0.5 mg Cu(II) per litre for 5 days) on specific components of the mating behaviour of male shore crab Carcinus maenas was investigated, as well as differences in sensitivity between red and green colour morphs. The results show that copper exposure clearly altered the response of C. maenas males to a pheromone stimulus (pre-moult female urine) presented alone, together with a dummy female (a sponge injected with pre-moult female urine) or with a real female. Crabs exposed to the highest copper treatment took more than twice as long to initiate search activity after pheromone introduction and their search behaviour was less directed. When offered a dummy female, male crabs showed decreased pheromone discrimination in both copper treatments. Stroking was the only mating behaviour significantly affected, with a 90% reduction in red crabs in the highest copper treatment. Additionally, crabs of the highest copper treatment more often pinched the dummy female (non-mating behaviour). Finally, male crabs exposed to copper more often pinched pre-moult females and it took about three times longer to establish cradle-carrying. Thus, copper affects the ability of males to detect female pheromones, perform specific mating behaviours and to form pairs.

Effects of manganese on chemically induced food search behaviour of the Norway lobster, Nephrops norvegicus (L.).

The decapod Norway lobster, Nephrops norvegicus (L.), lives on muddy sediments rich in manganese (Mn). In hypoxic conditions, manganese is reduced and released from the sediment, so increased concentrations of dissolved Mn(2+) become bioavailable. In excess, manganese acts as a neurotoxin and may inhibit vital functions of benthic organisms, such as muscle contraction. We investigated in a laboratory flume experiment, the effect of environmentally realistic concentrations of manganese (0.1 and 0.2mM for 12 days) on the food search behaviour of N. norvegicus. We found that lobsters exposed to manganese had a more than doubled reaction time to food odour stimuli compared to the controls (p<0.05). In addition, manganese exposure reduced the number of N. norvegicus reaching the food stimuli source. Compared to the controls where 86% reached the stimuli source, only about half of the lobsters exposed to 0.1mM Mn and one-third of the lobsters exposed to 0.2mM Mn reached the stimuli source (p<0.05 and 0.001, respectively). There was no significant difference between treatments in the number of lobsters leaving their shelter or in the time from reaction until leaving the shelter and there was no difference in search time for those animals that eventually did locate the stimuli source. This study shows that environmentally realistic manganese concentrations affect parts of the food search behaviour of N. norvegicus, likely due to impaired chemosensory ability or reduced motivation for feeding. Thus, the ability of N. norvegicus to detect and find food can be reduced in areas with high manganese concentrations, with possible consequences on individual and population levels.