Effects of currently used marine antifouling paint biocides on green fluorescent proteins in Anemonia viridis.

Some of the antifouling booster biocides affects the marine ecosystem negatively. The booster biocides that are resistant to degradation are accumulated in the sediment of the oceans. One of the sedentary organisms in the Mediterranean Sea is Anemonia viridis. This study aims at showing the toxicities of common biocides such as irgarol, seanine-211, zinc omadine, and acticide on the fluorescence by GFPs of A. viridis. The decreases in the fluorescence intensities of the GFP were measured within different booster biocide concentrations. The results show that fluorescent intensities of GFP proteins decrease more than 50% when they are exposed to different concentrations of irgarol, zinc omadine, acticide. In conclusion, ecosystem health should be prioritized when new antifouling paint compositions are proposed. From the results, it seems that A. viridis can be considered as a vulnerable organism and it is sensitive to booster biocides within self-polishing antifouling paint formulations.

Environmental risk assessment of using antifouling paints on pleasure crafts in European Union waters.

To ensure sustainable use of antifouling paints, the European Union have developed a new environmental risk assessment tool, which a product must pass prior to its placement on the market. In this new tool, environmental concentrations are predicted based on estimated release rates of biocides to the aquatic environment and risk characterization ratios are calculated in regional spreadsheets. There are currently two methods in use to predict release rates of biocides; a calculation method and a laboratory method. These methods have been believed to overestimate environmental release of biocides and therefore fixed correction factors to reduce the release rate can be applied. An alternative method, known as the XRF method, has recently been developed and used to derive field release rates from antifouling paints. The aim of this study was to review the new environmental risk assessment tool and assess how the choice of release rate method and application of correction factors impact the approval of antifouling paint products. Eight coatings were environmentally risk assessed for usage in four European marine regions; Baltic, Baltic Transition, Atlantic and Mediterranean; by applying release rates of copper and zinc determined with the different methods. The results showed none of the coatings to pass the environmental risk assessment in the Baltic, Baltic Transition and the Mediterranean if field release rates were used. In contrast, most of the coatings passed if the correction factors were applied on the release rates obtained with the calculation or laboratory method. The results demonstrate the importance of release rate method choice on the outcome of antifouling product approval in EU. To reduce the impact of antifouling paints on the marine environment it is recommended that no correction factors should be allowed in the environmental risk assessment or preferably that site-specific field release rates are used. If the regulation in the European Union (and elsewhere) continues to allow correction factors, the pressure of biocides to the environment from leisure boating will result in degradation of marine ecosystems.

Colonisation and succession of marine biofilm-dwelling ciliate assemblages on biocidal antifouling and fouling-release coatings in temperate Australia.

Ciliate assemblages are often overlooked, but ubiquitous components of microbial biofilms which require a better understanding. Ciliate, diatom and bacterial colonisation were evaluated on two fouling-release (FR) coatings, viz. Intersleek 970 and Hempasil X3, and two biocidal antifouling (AF) coatings, viz. Intersmooth 360 and Interspeed 5640, in Port Phillip Bay, Australia. A total of 15 genera were identified during the 10 week deployment. Intersleek 970 displayed the most rapid fouling by ciliates, reaching 63.3(± 5.9) cells cm(-2). After 10 weeks, all four coatings were extensively fouled. However, the toxicity of the AF coatings still significantly inhibited microbial fouling compared to the FR coatings. On all treatments, colonies of sessile peritrichs dominated the ciliate assemblage in the early stage of succession, but as the biofilm matured, vagile ciliates exerted more influence on the assemblage structure. The AF coatings showed selective toxic effects, causing significant differences in the ciliate species assemblages among the treatments.

Metal record of copper-based antifouling paints in sediment core following marina construction and operation.

Marinas are central hubs of global maritime leisure and transport, yet their operations can deteriorate the environmental quality of sediments. In response, this study investigated the metal contamination history associated with antifouling paint uses in a sediment core collected from Bracuhy marina (Southeast Brazil). Analysis target major and trace elements (Cu, Zn, Pb, Cd and Sn), rare earth elements (REEs), and Pb isotopes. The modification in Pb isotopic ratios and REEs pattern unequivocally revealed sediment provenance disruption following the marina construction. Metal distribution in the sediment core demonstrates that concentrations of Cu and Zn increased by up to 15 and 5 times, respectively, compared to the local background. This severe Cu and Zn contamination coincides with the onset of marina operations and can be attributed to the use of antifouling paints.

Development of methodology for the visual identification of Tributyltin (TBT) in antifouling paint matrices.

Organotin compounds (OTC), mainly tributyltin (TBT), have been used since the 1970s as biocides in the composition of antifouling paints. Due to its physical-chemical characteristics, TBT has high toxicity to the marine environment affecting non-target organisms. The present study aims to develop a method of direct visual identification of TBT in antifouling paints using the cyclopalladate complex, 4- (2-thiazolylazo) resorcinol (TAR-Pd), synthesized in our laboratory. Tests were performed in blank and in the paint matrix with the following OTC: TBT-O; TBT-Cl; TPT-Cl; DBT-Cl (tributyltin oxide, tributyltin chloride, triphenyltin chloride, dibutyltin chloride), in addition to the SnCl4 and SnCl2 compounds (tin IV chloride and tin II chloride), all at a concentration of approximately 20 g/ kg of dry paint). The test was performed by applying paint samples to test bodies and scraping a few tens of milligrams of the dry paint film. The scraped paint samples were submitted to the test, showing a different staining reaction for the TBT-Cl and SnCl4 samples concerning blank and other samples (TBT-O, TPT, DBT-Cl, and SnCl2). Solution tests were performed to characterize reaction products by spectroscopy in the visible band. The method developed has potential for application in real samples, being selective for TBT-Cl and SnCl4 in an acid medium, obtaining a limit of detection, in the range of 1-10 mg/kg dry paint.

Exploring fishery waste potential as antifouling component.

Marine biofouling is a global issue with economic and ecological implications. Existing solutions, such as biocide-based antifouling paints, are toxic for the environment. The search for better antifouling agents remains crucial. Recent research focuses on eco-friendly antifouling paints containing natural compounds like enzymes. This study evaluates enzymatic extracts from fishery residues for antifouling potential. Extracts from Pleoticus muelleri shrimp, Illex argentinus squid, and Lithodes santolla king crab were analyzed. Proteolytic activity and thermal stability were assessed, followed by bioassays on mussel byssus thread formation and barnacle cypris adhesive footprints. All three extracts demonstrated proteolytic activity and 24-h stability at temperate oceanic temperatures, except I. argentinus. P. muelleri extracts hindered cyprid footprint formation and mussel byssus thread generation. Further purification is required for L. santolla extract to assess its antifouling potential activity. This study introduces the use of fishery waste-derived enzyme extracts as a novel antifouling agent, providing a sustainable tool to fight against biofouling formation.

Update on the status of the contamination by organotin compounds in sediment of Nuevo Gulf, Argentina. Insights from field and experimental studies.

Organotin compounds are persistent pollutants and are considered chemicals of high environmental concern. In the present study, the distribution and degradation of tributyltin were evaluated in field sediments and through an ex situ experiment. For this, sediment samples from two locations were analysed: Luis Piedrabuena Harbour, with higher maritime traffic, and Cerro Avanzado, which receives less impact from anthropogenic activities. The results indicated that pollution levels at Luis Piedrabuena Harbour have decreased compared with studies performed 9 years ago for the same area. On the contrary, traces of organotin compounds have been found for the first time at Cerro Avanzado. Moreover, the butyltin degradation index indicated that organotin compounds undergo an advanced degradation process in the collected samples at both sites. Ex situ experiments revealed a limited capacity of sediments to retain tributyltin, and suggested an active role of bioturbation activity in the degradation of these compounds. In addition, visualisation using chemometric techniques (principal components analysis) allowed a simpler analysis of two sediment characteristics: the degree of contamination and the degradation levels of organotin compounds.

Antifouling booster biocides in Latin America and the Caribbean: A 20-year review.

This review summarized booster biocides studies from Latin America and the Caribbean during the last two decades. Studies were focused on six countries, with most of them in Brazil. In water and sediment, diuron and Irgarol were the most abundant and frequent biocides, probably due to their former intense use. Antifouling paint particles were also reported and had mainly DCOIT, which is currently the most used booster biocide. Toxicity of individual booster biocides was tested in laboratory, and most effects were related to chlorothalonil, DCOIT, dichlofluanid, and Irgarol, including, but not limited to DNA damage, fertility decrease, and mortality at different trophic levels. This review highlighted the need for further studies on environmental occurrence of booster biocides in Latin America and Caribbean associated to ecotoxicological studies. Such information is essential to determine the potential ecological risks and to create directives regarding safe limits of booster biocides in aquatic systems.

Potential disruptive effects of copper-based antifouling paints on the biodiversity of coastal macrofouling communities.

The expanded use of copper(I)-based antifouling paints (AF) has increased copper leaching into coastal environments, requiring attention and legislative restrictions for potential long-term effects on benthic populations. The ecological succession of macrofouling communities was analysed on wooden and stainless steel panels coated with four copper(I)-based AF (Paints A-D) immersed for 10 months in the Lagoon of Venice. With the exception of Paint B, which contained only copper(I) compounds and was based on hard-matrix technology, the other paints were based on self-polishing matrices and various booster biocides. The booster content was a mix of TBT compounds for Paint A, dichlofluanid for Paint C, Irgarol 1051, and chlorothalonil for Paint D. The macrofouling communities appeared dissimilar to those on the reference uncoated panels as regard the species richness, the coverage areas, and the biocoenosis structure. Generally, green algae, bryozoans, and barnacles were the most tolerant taxa and a negative species selection occurred for sponges, serpulids, and ascidians. Paints A and D showed the highest performance, and Paint D also prevented molluscs on wood panels. Paints B and C rapidly decreased their efficiency, the first probably due to the insoluble matrix with the highest biocidal leaching rate, and the second due to the presence of a booster with low toxicity. Paint B also inhibited red algae and molluscs, but Paint C did not reveal significant differences in types of species settlements with reference panels.

Immunotoxic effects of exposure to the antifouling copper(I) biocide on target and nontarget bivalve species: a comparative in vitro study between Mytilus galloprovincialis and Ruditapes philippinarum.

Edible bivalves constitute an important bioresource from an economic point of view, and studies on their immune responses to environmental pollutants are crucial for both the preservation of biodiversity and economic reasons. The worldwide diffusion of copper(I)-based antifouling paints has increased copper leaching into coastal environments and its potential impact on both target and nontarget organisms. In this study, immunotoxicity assays were carried out with short-term (60 min) cultures of hemocytes from the bivalves Mytilus galloprovincialis-a mussel dominant in the macrofouling community-and Ruditapes philippinarum-a clam dominant in the soft-sediment community-exposed to CuCl to compare the toxic effects on their immune responses. The LC50 values were similar, 40 µM (3.94 mg L-1) for the mussel and 44 µM (4.33 mg L-1) for the clam. In both species, apoptosis occurred after exposure to 1 µM (98.9 µg L-1) CuCl, the concentration able to significantly increase the intracellular Ca2+ content. Biomarkers of cell morphology and motility revealed microfilament disruption, a significant decrease in yeast phagocytosis and lysosome hydrolase (ß-glucuronidase) inhibition beginning from 0.5 µM (49.5 µg L-1) CuCl in both the mussel and clam. The same concentration of CuCl affected biomarkers of oxidative stress, as a significant decrease in reduced glutathione content in the cytoplasm and inhibition of mitochondrial cytochrome-c oxidase (COX) were detected in both species. Comparison of the biomarkers showed that clam is more sensitive than the mussel regarding alterations to the lysosomal membrane and reactive oxygen species (ROS) production, which supports the potential harmful effects of antifouling biocides on the survival of nontarget pivotal species in the coastal community.

Cu isotope records of Cu-based antifouling paints in sediment core profiles from the largest European Marina, The Port Camargue.

The intensive use of copper (Cu) compounds as an alternative biocide in antifouling paints (APs) has resulted in wide Cu contamination into the marine environment, especially near marina harbor activities. In this work, the applicability of Cu isotopes to discriminate Cu origins related to the use of Cu-based APs in marine environments was tested. To this, Cu isotopes in APs, shipyard sludges, and sediment cores sampled in the Cu-contaminated Mediterranean marina of Port Camargue were determined. APs represent an important dominant anthropogenic source for metals in this site, making it ideal to test Cu isotopes as tracers. The overall isotope composition of four sediment cores and a surface sample varied between -0.13 and 0.44 ‰ (δ65Cu relative to NIST-976). Selected APs brands show a similar Cu concentration ~0.15 % and δ65Cu average of 0.54 ± 0.05 ‰. The plot of δ65Cu vs concentration for all datasets allowed dissociating natural and APs end-members. However, sample isotope systematics were not consistent with a conservative mixing binary source process. Heavily Cu-contaminated sediments show isotope signatures lighter than APs brands. However, the most Cu-contaminated sample, located directly above the careening area, shows a δ65Cu slightly lighter than APs (0.44 ‰ vs 0.54 ‰, respectively). Results suggest the preferential releasing of a heavy isotope pool by APs when these compounds are solubilized in seawater. The isotope fractionation was attributed to potential chemical Cu coordination changes during its elemental partition between paint and marina seawater and the fractionation induced by the organic ligands in the water column, before deposition. Further laboratory experiments are recommended to model the isotope fractionation mechanisms related to Cu release by APs. Because the APs' isotope signature is modified in marine environments, the use of Cu isotopes as tracers of AP in marine environments is challenging and needs more investigation.

Biocides in antifouling paint formulations currently registered for use.

Antifouling paints incorporate biocides in their composition seeking to avoid or minimize the settlement and growing of undesirable fouling organisms. Therefore, biocides are released into the aquatic environments also affecting several nontarget organisms and, thus, compromising ecosystems. Despite global efforts to investigate the environmental occurrence and toxicity of biocides currently used in antifouling paints, the specific active ingredients that have been used in commercial products are poorly known. Thus, the present study assessed the frequencies of occurrence and relative concentrations of biocides in antifouling paint formulations registered for marketing worldwide. The main data were obtained from databases of governmental agencies, business associations, and safety data sheets from paint manufacturers around the world. The results pointed out for 25 active ingredients currently used as biocides, where up to six biocides have been simultaneously used in the examined formulations. Cuprous oxide, copper pyrithione, zinc pyrithione, zineb, DCOIT, and cuprous thiocyanate were the most frequent ones, with mean relative concentrations of 35.9 ± 12.8%, 2.9 ± 1.6%, 4.0 ± 5.3%, 5.4 ± 2.0%, 1.9 ± 1.9%, and 18.1 ± 8.0% (w/w) of respective biocide present in the antifouling paint formulations. Surprisingly, antifouling paints containing TBT as an active ingredient are still being registered for commercialization nowadays. These results can be applied as a proxy of biocides that are possibly being used by antifouling systems and, consequently, released into the aquatic environment, which can help to prioritize the active ingredients that should be addressed in future studies.

An absurd scenario in 2021: Banned TBT-based antifouling products still available on the market.

Despite the efforts of the International Marine Organization (IMO), through the International Convention on the Control of Harmful Antifouling Systems on Ships to ban the use of TBT-based antifouling paints (September 2008), and the Rotterdam Convention (RC) to forbid the trade of tributyltin (TBT), the situation does not seem to have significantly changed since 2014 when TBT-based paints were shown to be still being manufactured in the United States and offered for sale in stores throughout the Caribbean and Central America. In fact, this study shows that seven years later the same situation not only persists, but may have an even more global distribution than previously thought.

Multivariate optimization of microwave-assisted digestion methods for Cu and Sn determination in antifouling paints using inductively coupled plasma optical emission spectrometry.

Antifouling paints containing Cu, Zn, organotins, and many organic booster biocides may be found in ships and watercraft hulls to avoid the fouling of marine organisms. This type of paint can be harmful to the environment, therefore, the monitoring of toxic elements and compounds in antifouling paints are of great importance to access its quality and potential toxicity to the environment. Hence, this work describes the development of microwave-assisted digestion methods for the determination of Cu and Sn in antifouling paints by inductively coupled plasma optical emission spectrometry (ICP OES). The factors: sample mass and solutions of HNO3, HF, and HCl were optimized using the central composite design (CCD). Dry ashing with a muffle furnace and laser ablation-ICP-MS were used for methodological comparison with the microwave digestion-assisted ICP-OES methods. All the mixtures of acids allowed efficient extraction of the analytes; however, the one that stood out was the use of HF, HNO3, and H2O2.

Assessment of anthropogenic metals in shipyard sediment in the Amazon delta estuary in northern Brazil.

Shipyard activities have contributed to the release of anthropogenic metals in sediment in the Amazon delta estuary, but no studies of the issue have been carried out in northern Brazil. This study evaluated the sediment that is under the influence of shipyard activities in the Guajará Bay and in the channel of the Maguari River, in Belém, Pará (PA) state, northern Brazil. Sediment samples were collected in the vicinity of the shipyards, while samples of paint and metal fragments were collected from hulls of abandoned vessels. Metals under analysis were Cu, Zn, Pb, Ni, Cr, Ba, V, Li, Fe and Al. Mean Cu concentrations found in the sediment in two shipyards - 28.3 mg kg-1 and 41.0 mg kg-1 - were above the threshold effect level (TEL) for the amphipod Hyalella azteca. The highest concentrations of metals found in paint fragments from abandoned vessels were 29,588 mg kg-1 Ba, 9,350 mg kg-1 Zn, 1,097 mg kg-1 Pb and 548 mg kg-1 Cr. This fact suggests that vessel abandonment is a major source of contamination in shipyard areas. The principal component analysis (PCA) showed that most metals under study are closely related to sediment contamination in the shipyards. Geoaccumulation index and screening concentrations of inorganic contaminants for metals in freshwater ecosystems confirmed that a shipyard was contaminated by copper. Results may support further studies of contamination and application of waste management to shipyards and vessel graveyards around the world.

Impacts of copper contamination on a rocky intertidal predator-prey interaction.

Metal contamination can change ecological interactions with potential effects on community dynamics. However, understanding real effects of metals on biota relies on studies undertaken in natural conditions. Through a field experiment, we investigated the effects of copper contamination on the responses of a barnacle prey and its predator, the dogwhelk, and explicitly their interaction. Contamination increased barnacle mortality and reduced predation with no effects on interaction strength. This was because the higher mortality of the prey compensated for the lower consumption of the predator. Despite not affecting the interaction strength, these results suggest a decrease in energy flow in the trophic chain that may lead to important changes in community structure and ecosystem functioning. This study shows the importance of manipulative experiments designed to provide mechanistic insights into ecological interactions to better clarify the effect of stressors on the structure and dynamic of communities.

Biological Effects Assessment of Antibiofouling EDCs: Gaeta Harbor (South Italy) Benthic Communities' Analysis by Biodiversity Indices and Quantitative gpx4 Expression.

The most representative organisms of the Harbor of Gaeta Gulf in South Italy were analyzed for biofouling by visual census and confirmed later by molecular approach on an artificial Conatex panel dipped 3 m into a eutrophic area during the Covid-19 pandemic. Mitochondrial Cytochrome C oxidase subunit 1(COI) gene was sequenced from 20 different marine species (flora: 2 families, 2 orders; fauna: 16 families, 11 orders) to test whether the morphology-based assignment of the most common biofouling member was supported by DNA-based species identification. Twelve months of submersion resulted in generation of sufficient data to obtain a facies climax represented mainly by the bivalve mollusk, Mytilus galloprovincialis. Specific diversity and variations of the biofouling biomasses were analyzed using two different anti-biofouling paints: an endocrine disrupting chemical (EDC)-containing metal biocide, and a biocide-free paint. Also, their effects on detoxification and reproductive health of M. galloprovincialis were evaluated using glutathione S-transferase enzymatic activity and RTqPCR expression of the fertility antioxidant gene glutathione peroxidase 4 (gpx4). The obtained data provide useful indications on which future investigations may be focused and may become a potential management tool for a harbor biofouling database to assist local administrations in EDCs protection of autochthonous benthic communities and their fertility using innovative antifouling paints.

Unusual tin organics, DDX and PAHs as specific pollutants from dockyard work in an industrialized port area in China.

In order to recognize organic contaminants responsible for ecological stresses from intensive shipping traffic and dockyard works, this study aimed at characterizing the sediment contamination of a large industrialized port located in Hainan Island, China. Surface sediment samples were collected from 17 stations including the main docks, the dockyards and the major industrial wastewater outlets. Organotin compounds, the pesticide DDT (bis(chlorophenyl)trichloroethane) and its metabolites and polycyclic aromatic compounds were identified as main pollutant groups by GC/MS applying a non-target screening approach. The pesticide DDT and its metabolites were found in the same samples as the organotin derivatives pointing to similar emission sources. The concurrent presence of these compounds in the dockyard samples suggests a combined usage of organotin compounds and DDT as active ingredients in antifouling paints in Yangpu. As highly specific molecular indicators for dockyard activities, butyltin and phenyltin compounds were identified. Noteworthy, also tributylmethyltin and triphenylmethyltin were detected, likely resulting from microbial assisted biomethylation of synthetic organotin compounds in the sediments. The concentrations of PAHs, DDX and TBT in sediments from dockyards exceeded global sediment quality guidelines and the toxicity thresholds, and potentially have adverse biological effects on marine organisms.

A review on the environmental impacts of shipping on aquatic and nearshore ecosystems.

There are several environmental and ecological effects of shipping. However, these are rarely assessed in total in the scientific literature. Thus, the aim of this study was to summarize the different impacts of water-based transport on aquatic and nearshore ecosystems and to identify knowledge gaps and areas for future research. The review identified several environmental and ecological consequences within the main impact categories of water discharges, physical impacts, and air emissions. However, although quantitative data on these consequences are generally scarce the shipping contribution to acidification by SOx- and NOx-emissions has been quantified to some extent. There are several knowledge gaps regarding the ecological consequences of, for example, the increasing amount of chemicals transported on water, the spread of non-indigenous species coupled with climate change, and physical impacts such as shipping noise and artificial light. The whole plethora of environmental consequences, as well as potential synergistic effects, should be seriously considered in transport planning.

Nickel, vanadium, and lead as indicators of sediment contamination of marina, refinery, and shipyard areas.

Metallic elements found in the aquatic environment may originate in areas where petroleum is refined and vessels are maintained and repaired. This study aims to assess contamination caused by nickel (Ni), lead (Pb), and vanadium (V) in sediment of the Lagoa dos Patos estuary (RS, Brazil) and to evaluate them as indicators of areas under the influence of petroleum products and antifouling paints. Surface sediments were collected in summer and in winter in areas of marinas, shipyards, refinery, and a control station. High Pb and V concentrations in shipyards and at the Yacht Club showed that some organisms may be affected by toxicity. High Pb results of the index of geoaccumulation (Igeo) were found at the Yacht Club and shipyards. Al, Ni, and V had similar distribution in the sediment in both seasons. Ni and V had high relation in winter at the Yacht Club and at the Santos Shipyard, thus suggesting that these elements come mainly from petroleum products. The same happened to the relations between Pb and V, as well as Pb and Ni at the Santos Shipyard. These elements are employed as useful tools as indicators to identify places with moderate to high localized anthropogenic inputs of petroleum derivatives and antifouling paints.