Antifouling activity of portimine, select semisynthetic analogues, and other microalga-derived spirocyclic imines.

A range of natural products from marine invertebrates, bacteria and fungi have been assessed as leads for nature-inspired antifouling (AF) biocides, but little attention has been paid to microalgal-derived compounds. This study assessed the AF activity of the spirocyclic imine portimine (1), which is produced by the benthic mat-forming dinoflagellate Vulcanodinium rugosum. Portimine displayed potent AF activity in a panel of four macrofouling bioassays (EC50 0.06-62.5 ng ml-1), and this activity was distinct from that of the related compounds gymnodimine-A (2), 13-desmethyl spirolide C (3), and pinnatoxin-F (4). The proposed mechanism of action for portimine is induction of apoptosis, based on the observation that portimine inhibited macrofouling organisms at developmental stages known to involve apoptotic processes. Semisynthetic modification of select portions of the portimine molecule was subsequently undertaken. Observed changes in bioactivity of the resulting semisynthetic analogues of portimine were consistent with portimine's unprecedented 5-membered imine ring structure playing a central role in its AF activity.

Nature-Inspired Peptide Antifouling Biocide: Coating Compatibility, Field Validation, and Environmental Stability.

This study reports the development of a class of eco-friendly antifouling biocides based on a cyclic dipeptide scaffold, 2,5-diketopiperazine (2,5-DKP). The lead compound cyclo(N-Bip-l-Arg-N-Bip-l-Arg) (1) was synthesized in gram amounts and used to assess the compatibility with an ablation/hydration coating, efficacy against biofouling, and biodegradation. Leaching of 1 from the coating into seawater was assessed via a rotating drum method, revealing relatively stable and predictable leaching rates under dynamic shear stress conditions (36.1 ± 19.7 to 25.2 ± 9.1 ng-1 cm-2 day-1) but low or no leaching under static conditions. The coatings were further analyzed using time-of-flight secondary ion mass spectrometry (ToF-SIMS), with 1 seen to localize at the surface of the coating in a surfactant-like fashion. When coatings were deployed in the ocean, detectable reductions in biofouling development were measured for up to 11 weeks. After this time, biofouling overwhelmed the performance of the coating, consistent with leaching kinetics. Biodegradation of 1 in seawater was assessed using theoretical oxygen demand and analytical quantification. Masking effects were observed at higher concentrations of 1 due to antimicrobial properties, but half-lives were calculated ranging from 13.4 to 16.2 days. The results can rationally inform future development toward commercial antifouling products.

Characterization of the effects of triclosan on sperm and embryos of Mytilus and Perna mussel species.

The Greenshell™ mussel (GSM), Perna canaliculus, is a culturally and commercially important species in New Zealand. Declines in spat settlement of GSM have been observed in important growing areas and the cause(s) have not been identified. One hypothesis is that chemical contaminants could be a contributing factor. The aim to this study was to investigate the effects of acute exposure on early life stages using the anti-microbial triclosan (TCS) as a benchmark toxicant and the blue mussel (BM), Mytilus galloprovincialis, as a reference species. Sperm and embryos of BM and GSM were exposed to TCS for 1 h and 48 h, respectively. Following exposures, a range of parameters were investigated including spermatozoa cellular characteristics via flow cytometry, fertilization success, larval mortality and size. Exposure to TCS negatively impacted functional parameters of sperm, reduced the fertilization success and larval size, and increased larval mortality in both BM and GSM with LC5048h of 94.3 and 213 µg L-1, respectively. Triclosan increased sperm ROS production in both species, which could cause destabilisation of mitochondrial and other cellular membranes, resulting in reduced mitochondrial membrane potential (BM) and increased sperm size (GSM), leading to apoptosis in both species. Fertilization success of GSM was only affected at the highest TCS concentration tested (391 µg L-1), but development of larvae derived from exposed sperm was affected from the lowest concentrations tested (0.5 and 5.2 µg L-1) in both species. This highlights the importance of assessing the sensitivity of contaminants across developmental stages. Results of this study confirm that TCS causes oxidative stress and has membranotropic effects, and that early life stages of the endemic GSM are suitable to assess ecotoxicity of contaminants such as TCS.

Investigating the Ecotoxicity of Select Emerging Organic Contaminants Toward the Marine Copepod Gladioferens pectinatus.

Estuarine ecosystems are recipients of anthropogenic stressors released from land-based activities. The aim of the present study was to investigate the ecotoxicological hazards of organic contaminants toward the estuarine copepod Gladioferens pectinatus using acute and chronic testing. Most chemicals demonstrated acute toxicity and influenced development of the copepods. Further research should be conducted to investigate these chemicals and their mixtures using long-term, multigenerational testing to characterize mechanisms of toxicity. Environ Toxicol Chem 2022;41:792-799. © 2022 SETAC.

Does plastic type matter? Insights into non-indigenous marine larvae recruitment under controlled conditions.

Marine plastic debris (MPD) are a global threat to marine ecosystems. Among countless ecosystem impacts, MPD can serve as a vector for marine 'hitchhikers' by facilitating transport and subsequent spread of unwanted pests and pathogens. The transport and spread of these non-indigenous species (NIS) can have substantial impacts on native biodiversity, ecosystem services/functions and hence, important economic consequences. Over the past decade, increasing research interest has been directed towards the characterization of biological communities colonizing plastic debris, the so called Plastisphere. Despite remarkable advances in this field, little is known regarding the recruitment patterns of NIS larvae and propagules on MPD, and the factors influencing these patterns. To address this knowledge gap, we used custom-made bioassay chambers and ran four consecutive bioassays to compare the settlement patterns of four distinct model biofouling organisms' larvae, including the three notorious invaders Crassostrea gigas, Ciona savignyi and Mytilus galloprovincialis, along with one sessile macro-invertebrate Spirobranchus cariniferus, on three different types of polymers, namely Low-Linear Density Polyethylene (LLDPE), Polylactic Acid (PLA), Nylon-6, and a glass control. Control bioassay chambers were included to investigate the microbial community composition colonizing the different substrates using 16S rRNA metabarcoding. We observed species-specific settlement patterns, with larvae aggregating on different locations on the substrates. Furthermore, our results revealed that C. savignyi and S. cariniferus generally favoured Nylon and PLA, whereas no specific preferences were observed for C. gigas and M. galloprovincialis. We did not detect significant differences in bacterial community composition between the tested substrates. Taken together, our results highlight the complexity of interactions between NIS larvae and plastic polymers. We conclude that several factors and their potential interactions influenced the results of this investigation, including: (i) species-specific larval biological traits and ecology; (ii) physical and chemical composition of the substrates; and (iii) biological cues emitted by bacterial biofilm and the level of chemosensitivity of the different NIS larvae. To mitigate the biosecurity risks associated with drifting plastic debris, additional research effort is critical to effectively decipher the mechanisms involved in the recruitment of NIS on MPD.

Plastic additives: challenges in ecotox hazard assessment.

The risk of plastic debris, and specifically micro(nano)plastic particles, to ecosystems remains to be fully characterized. One particular issue that warrants further characterization is the hazards associated with chemical additives within micro(nano)plastic as they are not chemically bound within the polymers and can be persistent and biologically active. Most plastics contain additives and are therefore potential vectors for the introduction of these chemicals into the environment as they leach from plastic, a process that can be accelerated through degradation and weathering processes. There are knowledge gaps on the ecotoxicological effects of plastic additives and how they are released from parent plastic materials as they progressively fragment from the meso to micro and nano scale. This review summarizes the current state of knowledge of the ecotoxicity of plastic additives and identifies research needs to characterize the hazard they present to exposed biota. The potential ecological risk of chemical additives is of international concern so key differences in governance between the European Union and New Zealand to appropriately characterize their risk are highlighted.

Trace Metal Residues in Marine Mussels: A Global Survey.

Pressures from anthropogenic activities are causing degradation of estuarine and coastal ecosystems around the world. Trace metals are key pollutants that are released and can partition in a range of environmental compartments, to be ultimately accumulated in exposed biota. The level of pressure varies with locations and the range and intensity of anthropogenic activities. The present study measured residues of trace metals in Mytilus mussel species collected from a range of locations around the world in areas experiencing a gradient of anthropogenic pressures that we classified as low, moderate, or high impact. The data showed no grouping/impact level when sampling sites in all countries were incorporated in the analysis, but there was significant clustering/impact level for most countries. Overall, high-impact areas were characterized by elevated concentrations of zinc, lead, nickel, and arsenic, whereas copper and silver were detected at higher concentrations in medium-impact areas. Finally, whereas most metals were found at lower concentrations in areas classified as low impact, cadmium was typically elevated in these areas. The present study provides a unique snapshot of worldwide levels of coastal metal contamination through the use of Mytilus species, a well-established marine biomonitoring tool. Environ Toxicol Chem 2021;40:3434-3440. © 2021 SETAC.

Assessing the Efficacy of a Sediment Remediation Program Using Benthic and Pelagic Copepod Bioassays.

Tributyltin is an organotin chemical that has been commonly used in ship antifouling paints. Despite the global total prohibition of tributyltin-based paint in 2008, tributyltin continues to be found at toxic levels in areas of high maritime traffic such as ports and harbors. A remediation program was conducted at a New Zealand port to reduce tributyltin and copper concentrations to acceptable values. The present study assessed the efficacy of the program using a combination of chemical analyses and copepod bioassays. Sediment and water samples were collected at 3 locations along a spatial gradient within the port, and concentrations of various organotin compounds and trace metal levels were measured pre- and postremediation. The toxicity of sediment and elutriate samples was estimated by benthic and pelagic copepod bioassays. Although acute toxicity in sediment samples was reduced following remediation, reproductive success was still affected for the benthic copepod. This approach combining chemical analysis and bioassays is promising for assessing the efficacy of remediation processes at contaminated marine sites. Environ Toxicol Chem 2020;39:492-499. © 2019 SETAC.

Effects of chronic exposure to benzophenone and diclofenac on DNA methylation levels and reproductive success in a marine copepod.

The UV-filter benzophenone and the anti-inflammatory diclofenac are commonly detected in the environment. The aim of this study was to assess the multigenerational effects of chronic exposure to low concentrations of these chemicals on toxicity and DNA methylation levels in the copepod Gladioferens pectinatus. Acute toxicity tests were conducted to determine the sensitivity of G. pectinatus to the chemicals. All chemicals impacted breeding, hatching and egg viability. Diclofenac (1 mg.L-1) reduced the number of eggs per gravid female. Benzophenone (0.5 mg.L-1) decreased egg hatching success. Exposure to the reference toxicant copper (0.02 mg.L-1) led to unsuccessful hatching. Effects on DNA methylation was estimated by the percentage of 5- methylcytosine. The treatments resulted in strong differences in DNA methylation with increased methylation in the exposed animals. The two chemicals impacted both egg viability and the induction of differential DNA methylation, suggesting potential intra- and trans-generational evolutionary effects.

Effects of tributyltin and 17ß-estradiol on immune and lysosomal systems of the Asian clam Corbicula fluminea (M.).

Freshwater clams Corbicula fluminea were experimentally exposed to a range of tributyltin (TBT) (50, 250 and 500ng Sn/L) and 17ß-estradiol (20, 200, 2000ng/L) for 30 days. After 15 and 30 days, phagocytosis activity of haemocytes and lysosomal structural changes in the digestive cells were assayed. 17ß-Estradiol exerted a higher inhibition on phagocytosis than tributyltin. This would suggest the existence of estrogen receptors, influencing the immune function. The stereological parameters measured for lysosomal structural changes in animals exposed to tributyltin varied as observed in other studies. Tributyltin is then depurated as other contaminants via digestive cell lysosomal compartment. This experiment emphasized a possible approach on the influence of endocrine disrupting compounds on a hermaphroditic species for environmental surveys.

Ecological impacts of long-term application of biosolids to a radiata pine plantation.

Assessment of the ecological impact of applying biosolids is important for determining both the risks and benefits. This study investigated the impact on soil physical, chemical and biological properties, tree nutrition and growth of long-term biosolids applications to a radiata pine (Pinus radiata D. Don) plantation growing on a Sandy Raw Soil in New Zealand. Biosolids were applied to the trial site every 3 years from tree age 6 to 19 years at three application rates: 0 (Control), 300 (Standard) and 600 (High) kg nitrogen (N) ha(-1), equivalent to 0, 3 and 6 Mg ha(-1) of dry biosolids, respectively. Tree nutrition status and growth have been monitored annually. Soil samples were collected 13 years after the first biosolids application to assess the soil properties and functioning. Both the Standard and High biosolids treatments significantly increased soil (0-50 cm depth) total carbon (C), N, and phosphorus (P), Olsen P and cation exchange capacity (CEC), reduced soil pH, but had no significant effects on soil (0-20 cm depth) physical properties including bulk density, total porosity and unsaturated hydraulic conductivity. The High biosolids treatment also increased concentrations of soil total cadmium (Cd), chromium (Cr), copper (Cu) and lead (Pb) at 25-50 cm depth, but these concentrations were still considered very low for a soil. Ecotoxicological assessment showed no significant adverse effects of biosolids application on either the reproduction of springtails (Folsomia candida) or substrate utilisation ability of the soil microbial community, indicating no negative ecological impact of bisolids-derived heavy metals or triclosan. This study demonstrated that repeated application of biosolids to a plantation forest on a poor sandy soil could significantly improve soil fertility, tree nutrition and pine productivity. However, the long-term fate of biosolids-derived N, P and litter-retained heavy metals needs to be further monitored in the receiving environment.

Scoring approach based on fish biomarkers applied to French river monitoring.

The aim was to apply a multimarker scoring approach as complementary to freshwater monitoring programmes carried out by the Water Agency Adour-Garonne. Fish (chub, barbel and trout) were collected in 11 sites in rivers in south-west France. Five biomarkers of response were measured either in muscle or brain for acetylcholinesterase (AChE) and in liver for glutathione S-transferase, catalase and 7-ethoxyresorufine O-deethylase. As a result of multivariate analysis, sites were clearly discriminated mainly by 7-ethoxyresorufine O-deethylase and acetylcholinesterase activities. According to the scoring approach, a multimarker pollution index was calculated for each sampling site as the sum of the response index of the five measured biomarkers (pollution index). Sorting was established by ranging the sites from lightly to highly contaminated locations.