Assessing the spatiotemporal occurrence and ecological risk of antifouling biocides in a Brazilian estuary.

Diuron and Irgarol are common antifouling biocides used in paints to prevent the attachment and growth of fouling organisms on ship hulls and other submerged structures. Concerns about their toxicity to non-target aquatic organisms have led to various restrictions on their use in antifouling paints worldwide. Previous studies have shown the widespread presence of these substances in port areas along the Brazilian coast, with a concentration primarily in the southern part of the country. In this study, we conducted six sampling campaigns over the course of 1 year to assess the presence and associated risks of Diuron and Irgarol in water collected from areas under the influence of the Maranhão Port Complex in the Brazilian Northeast. Our results revealed the absence of Irgarol in the study area, irrespective of the sampling season and site. In contrast, the mean concentrations of Diuron varied between 2.0 ng L-1 and 34.1 ng L-1 and were detected at least once at each sampling site. We conducted a risk assessment of Diuron levels in this area using the risk quotient (RQ) method. Our findings indicated that Diuron levels at all sampling sites during at least one campaign yielded an RQ greater than 1, with a maximum of 22.7, classifying the risk as "high" based on the proposed risk classification. This study underscores the continued concern regarding the presence of antifouling biocides in significant ports and marinas in Brazilian ports, despite international bans.

Ionic Liquid-Dispersive Micro-Extraction and Detection by High Performance Liquid Chromatography-Mass Spectrometry for Antifouling Biocides in Water.

A simple analytical method was developed and evaluated for the determination of two antifouling biocides using an ionic liquid-dispersive liquid-liquid micro-extraction (IL-DLLME) and a high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) analysis. Irgarol 1051 and Sea-Nine 211 were extracted from deionized water, lake water, and seawater using IL 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIm][PF6]) and ethyl acetate as the extraction solvent and the dispersion solvent. Several factors were considered, including the type and volume of extraction and dispersive solvent, IL amount, sample pH, salt effect, and cooling temperature. The developed method resulted in a recovery range of 78.7-90.3%, with a relative standard deviation (RSD, n = 3) less than 7.5%. The analytes were enriched greater than 40-fold, and the limits of detection (LOD) for two antifouling biocides were 0.01-0.1 µg L-1. The method was effectively applied for the analysis of real samples of freshwater as well as samples of seawater.

Water quality criteria derivation and tiered ecological risk evaluation of antifouling biocides in marine environment.

This study provides a comprehensive compilation of published toxicological and environmental data further used to assess the ecological risks of six antifouling biocides, including tributyltin (TBT), Irgarol 1051, Diuron, Chlorothalonil, 4,5-Dichloro-N-octyl-3(2H)-isothiazolone (DCOIT), and Dichlofluanid. The standard maximum concentration and standard continuous concentration of antifouling biocides were derived by the species susceptibility distribution method. Following that, the ecological risk assessment of antifouling biocides in the aquatic environment was conducted using the hazard quotient, margin of safety, joint probability curve, and Monte Carlo random sampling method. The following is a concise list of the antifouling biocide dangers associated with acute and chronic risks: Irgarol 1051 > TBT > Diuron > DCOIT > Chlorothalonil > Dichlofluanid. It is strongly advised that systematic and ongoing monitoring of these biocides in coastal areas take place, as well as the creation of acceptable and efficient environmental protection measures, to safeguard the coastal environment's services and functions.

Impact of Tralopyril and Triazolyl Glycosylated Chalcone in Human Retinal Cells' Lipidome.

Antifouling (AF) coatings containing booster biocides are used worldwide as one of the most cost-effective ways to prevent the attachment of marine organisms to submerged structures. Nevertheless, many of the commercial biocides, such as Econea® (tralopyril), are toxic in marine environments. For that reason, it is of extreme importance that new efficient AF compounds that do not cause any harm to non-target organisms and humans are designed. In this study, we measured the half-maximal inhibitory concentration (IC50) of a promising nature-inspired AF compound, a triazolyl glycosylated chalcone (compound 1), in an immortalized human retinal pigment epithelial cell line (hTERT-RPE-1) and compared the results with the commercial biocide Econea®. We also investigated the effects of these biocides on the cellular lipidome following an acute (24 h) exposure using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS). Our results showed that compound 1 did not affect viability in hTERT-RPE-1 cells at low concentrations (1 µM), in contrast to Econea®, which caused a 40% reduction in cell viability. In total, 71 lipids were found to be regulated upon exposure to 10 µM of both compounds. Interestingly, both compounds induced changes in lipids involved in cell death, membrane modeling, lipid storage, and oxidative stress, but often in opposing directions. In general, Econea® exposure was associated with an increase in lipid concentrations, while compound 1 exposure resulted in lipid depletion. Our study showed that exposure to human cells at sublethal Econea® concentrations results in the modulation of several lipids that are linked to cell death and survival.

Does pH variation influence the toxicity of organic contaminants in estuarine sediments? Effects of Irgarol on nematode assemblages.

Natural pH values in coastal waters vary largely among locations, ecosystems, and time periods; still, there is an ongoing acidification trend. In this scenario, more acidic pH values can alter bioavailability of organic contaminants, to organisms. Despite this, interactive effects between pH and chemical substances are not usually considered in Ecological Risk Assessment protocols. This study investigated the effects of pH on the toxicity of a hydrophobic organic compound on a benthic community using a microcosm experiment setup to assess the response of nematode assemblages exposed to environmentally relevant concentrations of Irgarol at two natural pH conditions. Estuarine nematode assemblages were exposed to two concentrations of Irgarol at pH 7.0 and 8.0 for periods of 7 and 35 days. Lower diversity of nematode genera was observed at the highest tested Irgarol concentration (1281 ± 65 ng.g-1). The results showed that the effects of Irgarol contamination were independent of pH variation, indicating no influence of acidification within this range on the toxicity of Irgarol to benthic meiofauna. However, the results showed that estuarine nematode assemblages are impacted by long-term exposure to low (but naturally occurring) pHs. This indicates that estuarine organisms may be under naturally high physiological pressure and that permanent changes in the ecosystem's environmental factors, such as future coastal ocean acidification, may drive organisms closer to the edges of their tolerance windows.

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.

Antifouling paint biocides (Irgarol 1051 and diuron) in the selected ports of Peninsular Malaysia: occurrence, seasonal variation, and ecological risk assessment.

Irgarol 1051 and diuron are photosystem II inhibitors in agricultural activities and antifouling paints in the shipping sector. This study focused on three major ports (western, southern, and eastern) surrounding Peninsular Malaysia to construct the distribution of both biocides on the basis of the seasonal and geographical changes. Surface seawater samples were collected from November 2011 to April 2012 and pretreated using the solid-phase extraction technique followed by quantification with GC-MS and LC-MS-MS for Irgarol 1051 and diuron, respectively. Generally, the distribution of Irgarol 1051 was lowest during November 2011 and highest during April 2012, and similar patterns were observed at all ports, whereas the distribution of diuron was rather vague. The increasing pattern of Irgarol 1051 from time to time is probably related to its accumulation in the seawater as a result of its half-life and consistent utilization. On the basis of the discriminant analysis, the temporal distribution of Irgarol 1051 varied at Klang North Port, Klang South Port, and Pasir Gudang Port, whereas diuron was temporally varied only at Kemaman Port. Furthermore, Irgarol 1051 was spatially varied during November 2011, whereas diuron did not show any significant changes throughout all sampling periods. Ecological risk assessment exhibited a high risk for diuron and Irgarol 1051, but Irgarol 1051 should be of greater concern because of its higher risk compared to that of diuron. Thus, it is recommended that the current Malaysian guidelines and regulations of biocide application should be reevaluated and improved to protect the ecosystem, as well as to prevent ecological risks to the aquatic environment.

Photochemical fate of medetomidine in coastal and marine environments.

Medetomidine has been authorized in ship hull paints as an antifouling biocide under the biocidal product regulation in Europe since 2016. Its release into marine systems causes concerns over persistence and toxicity. However, the environmental fate of medetomidine has not been fully investigated. In this study, the photodegradation of medetomidine under natural sunlight conditions was investigated using collected coastal and sea waters. In addition, the phototransformation of medetomidine with reactive species (i.e., singlet oxygen, excited triplet state organic matter, and hydroxyl radicals) under UVA light was examined. Photoproducts were isolated by high-performance liquid chromatography (HPLC), identified by a combination of nuclear magnetic resonance (NMR) spectroscopy and time-of-flight mass spectrometry (qTOF), and reaction mechanisms were proposed. The results show that medetomidine is a neutral base (pKa of protonated form = 7.2) that leads to two different protonation states in the aquatic environment. Photodegradation of neutral medetomidine was dominated by reaction with singlet oxygen, while protonated medetomidine was relatively photostable. The contribution of reactive species to the overall photodegradation of neutral medetomidine was calculated to provide an assessment of phototransformation of medetomidine. The half-live of medetomidine was < 1.5 days in natural waters (pHcoastal = 8.3; pHsea = 8.1) under sunlit near-surface conditions, suggesting that it is not persistent in the aquatic environment. Because medetomidine has a relatively short half-life in sunlit aquatic ecosystems, a number of products, such as 2-(2,3-dimethylphenyl)propanamide, can be formed by photochemical reactions of medetomidine, with unknown consequences for marine and coastal waters.

Abiotic fate of tolylfluanid and dichlofluanid in natural waters.

To prevent the growth of unwanted organisms on ship hulls, antifouling paints, containing biocides such as tolylfluanid (N-[dichlor(fluor)methyl]sulfanyl-N-(dimethylsulfamoyl)-4-methylaniline) and dichlofluanid (N-(dichlorfluormethylthio)-N',N'-dimethyl-N-phenylsulfamid), are applied. There are concerns over their occurrence and fate in the marine environment due to long-term immersion in water. In the present study, the hydrolysis and photolysis of these compounds were investigated. Results showed that tolylfluanid and dichlofluanid hydrolyzed completely to their respective hydrolysis products DMST (N,N-dimethyl-N'-p-tolylsulfamide) and DMSA (N,N-dimethyl-N'-phenylsulfamide) in coastal water within 24 h. Furthermore, the transformation of tolylfluanid and dichlofluanid under natural sunlight was determined in selected marine waters (coastal water and sea water) in comparison to deionized water. The experiments revealed that photodegradation rates of DMST and DMSA in coastal water were higher than in sea water or deionized water. The indirect phototransformation of the hydrolysis products with selected reactive species (triplet state organic matter, singlet oxygen, and hydroxyl radicals) showed that DMST and DMSA mainly display triplet reactivity. The measured half-lives of the hydrolysis products in natural waters were 2.7 and 23 days, with DMST being considerably faster transformed than DMSA. However, several direct and indirect photoproducts have been newly identified and measured. DMS (N,N-dimethylsulfamide), was identified as the major phototransformation product in natural waters. It is generated by indirect photodegradation processes and exhibits potential persistence in the environment.

Biodegradation of third-generation organic antifouling biocides and their hydrolysis products in marine model systems.

The environmental fate for some selected antifouling biocides, dichlofluanid, tolylfluanid, tralopyril, and medetomidine, is relatively poorly understood with nearly all data derived from the assessment reports. Water/sediment systems and biofilms were used to determine biodegradation of the antifouling biocides. Dichlofluanid and tolylfluanid are known to hydrolyze to form DMSA (N,N-dimethyl-N'-phenylsulfamide) and DMST (N,N-Dimethyl-N'-(4-methylphenyl)sulfamide), respectively. DMSA did not show biodegradation, but it was shown to transform abiotically into N,N-dimethylsulfamide (N,N-DMS). In contrast, the structurally similar DMST did show biodegradation with a half-life of 5.78 days. The resulting transformation product of the biodegradation of DMST is also N,N-DMS. N,N-DMS accounted for the majority of the mass balance after 27 days in the water/sediment systems. Moreover, the biofilm systems also degraded both DMSA and DMST to N,N-DMS. The hydrolysis product of tralopyril, called BCCPCA (3-bromo-5-(4-chlorophenyl)-4-cyano-1 H-pyrrole-3-carboxylic acid), was not metabolized in the experiments and remained persistent. For this compound, a new log Kow of 2.47 was determined since the previously reported Kow value seemed to overestimate sediment partitioning. Medetomidine was removed from the water/sediment system, though, not significantly more than the control. However, a transformation product (medetomidine-acid) was detected in the incubation but not in the control, pointing to limited biodegradation. These results show that tolylfluanid can be rapidly removed by biodegradation in the marine environment, while dichlofluanid, tralopyril, and medetomidine remained in the system for a longer period of time. The prolonged stability of these biocides could mean that there is potential for accumulation in the environment. This potential is also there for the DMSA (dichlofluanid) and DMST (tolylfluanid) derived transformation product N,N-DMS, which was recalcitrant.

Are antifouling residues a matter of concern in the largest South American port?

In the present study, levels of booster biocides (diuron, Irgarol, chlorothalonil, dichlofluanid and DCOIT), butyltin compounds (TBT, DBT and MBT) and antifouling paint particles (APPs) were assessed in sediments of areas under the influence of the largest Latin American port, marinas, boat traffic and ship/boat maintenance facilities located within Santos-São Vicente Estuarine System (SSES). Contamination profile was directly related to local maritime activities, where sediments from the main navigation channel (MNC) presented low levels of antifouling residues while adjacent areas (AA), characterized by the presence of boats and boatyards, showed higher contamination considering all analyzed residues. Moreover, areas under the influence of fishing boats/yards presented relevant levels of butyltins (ΣBTs > 300 ng g-1) and APPs (>100 µg g-1), while marinas dominated by recreational boats showed higher booster biocides occurrence. Sites located nearby shipyards in the MNC and boatyards in the AA presented expressive amounts of APPs (>200 µg g-1). These APPs represent an important long-term source of biocides to the SSES. Thus, the profile of maritime activities in association to local oceanographic conditions drive the spatial distribution of antifouling residues within SESS, which in some case presented levels above sediment guidelines for TBT, DCOIT and diuron.

Rapid toxicity assessment of six antifouling booster biocides using a microplate-based chlorophyll fluorescence in Undaria pinnatifida gametophytes.

Biocides of antifouling agents can cause problems in marine ecosystems by damaging to non-target algal species. Aquatic bioassays are important means of assessing the quality of water containing mixtures of contaminants and of providing a safety standard for water management in an ecological context. In this study, a rapid, sensitive and inexpensive test method was developed using free-living male and female gametophytes of the brown macroalga Undaria pinnatifida. A conventional fluorometer was employed to evaluate the acute (48 h) toxic effects of six antifouling biocides: 4,5-Dichloro-2-octyl-isothiazolone (DCOIT), diuron, irgarol, medetomidine, tolylfluanid, zinc pyrithione (ZnPT). The decreasing toxicity in male and female gametophytes as estimated by EC50 (effective concentration at which 50% inhibition occurs) values was: diuron (0.037 and 0.128 mg l-1, respectively) > irgarol (0.096 and 0.172 mg l-1, respectively) > tolylfluanid (0.238 and 1.028 mg l-1, respectively) > DCOIT (1.015 and 0.890 mg l-1, respectively) > medetomidine (12.032 and 12.763 mg l-1, respectively). For ZnPT, 50% fluorescence inhibition of U. pinnatifida gametophytes occurred at concentrations above 0.4 mg l-1. The Undaria method is rapid, simple, practical, and cost-effective for the detection of photosynthesis-inhibiting biocides, thus making a useful tool for testing the toxicity of antifouling agents in marine environments.

Effects of tributyltin chloride on cell structures of epithelial layer in different stages of Artemia salina (Linnaeus, 1758).

Background: Tributylin chloride (TBTCl) has been demonstrated to be acutely toxic to aquatic organisms. Aim: This study was conducted to investigate the effects of TBTCl on epithelial cell of gut Artemia salina in different stages (Nauplii, Juveniles, and Adults). Methods: Samples of A. salina used were cultivated in incubators for hatching. Nauplii were harvested at 24 hours of age, while the juveniles and adults were harvested at 21 and 35 days of age, respectively. These three stages of A. salina were exposed to different concentrations of TBTCl (1 ng.L-1 to 500 ng.L-1) for 24 hours. For nauplii, juveniles, and adults, 100 individuals were exposed, and those that survived in the exposure test were harvested for histological analysis. Results: The histological examinations revealed significant differences (p < 0.05) in type of lesions associated with different TBTCl concentrations and at different stages. The predominant lesions associated with different stages and different concentrations of TBTCl were epithelial cell necroasis, degeneration, cell loss, disruption, piknosis, and submucosal necrosis. Cell scoring was a significant difference (p < 0.05) between the groups of different TBTCl concentrations and different life stages. Conclusion: Overall, in this study, the generality of the lesion scores showed that the adults are relatively more susceptible to the effects of TBTCl compared to the juvenile and the nauplii.

Effects of chlorothalonil on the antioxidant defense system of mussels Perna perna.

Chlorothalonil is an effective fungicide used in agriculture and formulations of antifouling paints, which use and possible toxicity has been generating great concern. Thus, the present study investigated the effects of chlorothalonil on the antioxidant defense system (ADS) of the mussel Perna perna. The ADS was evaluated in gills and digestive gland after 24 h and 96 h of exposure to environmental relevant levels of chlorothalonil (0.1 and 10 µg/L). The activity of the enzymes superoxide dismutase (SOD), catalase (CAT), glutamate cysteine-ligase (GCL) and glutathione S-transferase (GST), levels of non-enzymatic defenses, represented by glutathione (GSH), and lipoperoxidation (LPO) and protein carbonyls (PCO) were evaluated. Results indicated that exposure to chlorothalonil is affecting the ADS in both tissues. While the activity of SOD increased and GST and GSH were not altered in gills, they decreased in digestive gland after 24 h of exposure to 10 µg/L of chlorothalonil. The contrasting results indicate that gills and digestive gland presented different patterns of responses after exposure to chlorothalonil. Moreover, a tissue-specific response to chlorothalonil was observed. Gills could be acting as the first line of defense, presenting higher enzymatic levels with minor effects on the parameters analyzed. On the other hand, digestive gland, with lower levels of antioxidant defenses, was the most affect organ by chlorothalonil. It also should be highlighted that the fungicide reduced the glutathione metabolism in the digestive gland, which can lead to an imbalance of the redox state within the cells of animals.

Development of complementary CE-MS methods for speciation analysis of pyrithione-based antifouling agents.

In the recent decade, metal pyrithione complexes have become important biocides for antifouling purposes in shipping. The analysis of metal pyrithione complexes and their degradation products/species in environmental samples is challenging because they exhibit fast UV degradation, transmetalation, and ligand substitution and are known to be prone to spontaneous species transformation within a chromatographic system. The environmental properties of the pyrithione species, e.g., toxicity to target and non-target organisms, are differing strongly, and it is therefore inevitable to identify as well as quantify all species separately. To cope with the separation of metal pyrithione species with minimum species transformation during analysis, a capillary electrophoresis (CE)-based method was developed. The hyphenation of CE with selective electrospray ionization- and inductively coupled plasma-mass spectrometry (ESI-, ICP-MS) provided complementary molecular and elemental information for the identification and quantification of pyrithione species. To study speciation of pyrithiones, a leaching experiment of several commercial antifouling paints containing zinc pyrithione in ultrapure and river water was conducted. Only the two species pyrithione (HPT) and dipyrithione ((PT)2) were found in the leaching media, in concentrations between 0.086 and 2.4 µM (HPT) and between 0.062 and 0.59 µM ((PT)2), depending on the paint and leaching medium. The limits of detection were 20 nM (HPT) and 10 nM ((PT)2). The results show that complementary CE-MS is a suitable tool for mechanistical studies concerning species transformation (e.g., degradation) and the identification of target species of metal pyrithione complexes in real surface water matrices, laying the ground for future environmental studies. Graphical abstract Hyphenation of CE with ESI- and ICP-MS provided complementary molecular and elemental information. Metal pyrithione species released from commercial antifouling paints could be identified and quantified in ultrapure and river water matrices.

Toxicity of emerging antifouling biocides to non-target freshwater organisms from three trophic levels.

Antifouling (AF) systems provide the most cost-effective protection against biofouling. Several AF biocides have, however, caused deleterious effects in the environment. Subsequently, new compounds have emerged that claim to be more environment-friendly, but studies on their toxicity and environmental risk are necessary in order to ensure safety. This work aimed to assess the toxicity of three emerging AF biocides, tralopyril, triphenylborane pyridine (TPBP) and capsaicin, towards non-target freshwater organisms representing three trophic levels: algae (Chlamydomonas reinhardtii), crustacean (Daphnia magna) and fish (Danio rerio). From the three tested biocides, tralopyril had the strongest inhibitory effect on C. reinhardtii growth, effective quantum yield and adenosine triphosphate (ATP) content. TPBP caused sub-lethal effects at high concentrations (100 and 250µgL-1), and capsaicin had no significant effects on algae. In the D. magna acute immobilisation test, the most toxic compound was TPBP. However, tralopyril has a short half-life and quickly degrades in water. With exposure solution renewals, tralopyril's toxicity was similar to TPBP. Capsaicin did not cause any effects on daphnids. In the zebrafish embryo toxicity test (zFET) the most toxic compound was tralopyril with a 120h - LC50 of 5µgL-1. TPBP's 120h - LC50 was 447.5µgL-1. Capsaicin did not cause mortality in zebrafish up to 1mgL-1. Sub-lethal effects on the proteome of zebrafish embryos were analysed for tralopyril and TPBP. Both general stress-related and compound-specific protein changes were observed. Five proteins involved in energy metabolism, eye structure and cell differentiation were commonly regulated by both compounds. Tralopyril specifically induced the upregulation of 6 proteins implicated in energy metabolism, cytoskeleton, cell division and mRNA splicing whilst TPBP lead to the upregulation of 3 proteins involved in cytoskeleton, cell growth and protein folding. An ecological risk characterization was performed for a hypothetical freshwater marina. This analysis identified capsaicin as an environment-friendly compound while tralopyril and TPBP seem to pose a risk to freshwater ecosystems. Noneless, more studies on the characterization of the toxicity, behaviour and fate of these AF biocides in the environment are necessary since this information directly affects the outcome of the risk assessment.

Comparative toxicological effects of two antifouling biocides on the marine diatom Chaetoceros lorenzianus: Damage and post-exposure recovery.

Antifouling biocides are commonly used in coastal electric power stations to prevent biofouling in their condenser cooling systems. However, the environmental impact of the chemical biocides is less understood than the thermal stress effects caused by the condenser effluents. In this study, Chaetoceros lorenzianus, a representative marine diatom, was used to analyse the toxicity of two antifouling biocides, chlorine and chlorine dioxide. The diatom cells were subjected to a range of concentrations of the biocides (from 0.05 to 2mg/L, as total residual oxidants, TRO) for contact time of 30min. They were analysed for viability, genotoxicity, chlorophyll a and cell density endpoints. The cells were affected at all concentrations of the biocides (0.05-2mg/L), showing dose-dependent decrease in viability and increase in DNA damage. The treated cells were later incubated in filtered seawater devoid of biocide to check for recovery. The cells were able to recover in terms of overall viability and DNA damage, when they had been initially treated with low concentrations of the biocides (0.5mg/L of Cl2 or 0.2mg/L of ClO2). Chlorophyll a analysis showed irreparable damage at all concentrations, while cell density showed increasing trend of reduction, if treated above 0.5mg/L of Cl2 and 0.2mg/L of ClO2. The data indicated that in C. lorenzianus, cumulative toxic effects and recovery potential of ClO2 up to 0.2mg/L were comparable with those of Cl2, up to 0.5mg/L concentration in terms of the studied endpoints. The results indicate that at the biocide levels currently being used at power stations, recovery of the organism is feasible upon return to ambient environment. Similar studies should be carried out on other planktonic and benthic organisms, which will be helpful in the formulation of future guidelines for discharge of upcoming antifouling biocides such as chlorine dioxide.

Long-term 2007-2013 monitoring of reproductive disturbance in the dun sentinel Assiminea grayana with regard to polymeric materials pollution at the coast of Lower Saxony, North Sea, Germany.

During biological effect monitoring studies of endocrine active compounds with the snail Assiminea grayana in 2007-2013, reproductive disorders including atresia, transformation of capsule/albumen glands into prostates in females and ovotestis, transformation of prostates to capsule/albumen glands, disruption of spermatogenesis, and calcification of tubules in males, were encountered in several years. The search of sources of endocrine active substances was first directed to antifouling biocides from paint particles and extended to leaching compounds from polymeric materials. In contrast to the reference sites, most of the observed disorders occurred at a station near harbors and dockyards polluted with residues from antifouling paints and polymeric materials. Beside of investigations about the potential ingestion of polymer particles by the snails, further investigations of compounds of polymeric materials with endocrine potential should follow.

Tralopyril bioconcentration and effects on the gill proteome of the Mediterranean mussel Mytilus galloprovincialis.

Antifouling (AF) systems are used worldwide as one of the most cost-effective ways of protecting submerged structures against heavy biofouling. The emergence of environmentally friendly AF biocides requires knowledge on their environmental fate and toxicity. In this study we measured the bioconcentration of the emerging AF biocide tralopyril (TP) in the Mediterranean mussel Mytilus galloprovincialis and investigated the effects of TP on the mussel gill proteome following acute (2days) and chronic (30days) exposure, as well as after a 10-day depuration period. The experiments were carried out with 1µg/L TP; blank and solvent (5×10(-5)% DMSO) controls were also included. Proteomics analysis was performed by mass spectrometry-based multidimensional protein identification technology (MudPIT). Differentially expressed proteins were identified using a label-free approach based on spectral counts and G-test. Our results show that TP is rapidly accumulated by mussels at concentrations up to 362ng/g dw (whole tissues), reaching steady-state condition within 13days. Ten days of depuration resulted in 80% elimination of accumulated TP from the organism, suggesting that a complete elimination could be reached with longer depuration times. In total, 46 proteins were found to be regulated in the different exposure scenarios. Interestingly, not only TP but also DMSO alone significantly modulated the protein expression in mussel gills following acute and chronic exposure. Both compounds regulated proteins involved in bioenergetics, immune system, active efflux and oxidative stress, often in the opposite way. Alterations of several proteins, notably several cytoskeletal ones, were still observed after the depuration period. These may reflect either the continuing chemical effect due to incomplete elimination or an onset of recovery processes in the mussel gills. Our study shows that exposure of adult mussels to sublethal TP concentration results in the bioconcentration of this biocide in the tissues and modulates the expression of several proteins that may intervene in important metabolic pathways.

Affinity states of biocides determine bioavailability and release rates in marine paints.

A challenge for the next generation marine antifouling (AF) paints is to deliver minimum amounts of biocides to the environment. The candidate AF compound medetomidine is here shown to be released at very low concentrations, ie ng ml(-1) day(-1). Moreover, the release rate of medetomidine differs substantially depending on the formulation of the paint, while inhibition of barnacle settlement is independent of release to the ambient water, ie the paint with the lowest release rate was the most effective in impeding barnacle colonisation. This highlights the critical role of chemical interactions between biocide, paint carrier and the solid/aqueous interface for release rate and AF performance. The results are discussed in the light of differential affinity states of the biocide, predicting AF activity in terms of a high surface affinity and preserved bioavailability. This may offer a general framework for the design of low-release paint systems using biocides for protection against biofouling on marine surfaces.