Volcanic ash in the water column: Cellular, physiological and anatomical implications for the gastropod suspension-feeder Crepipatella peruviana (Lamarck, 1822).

The influx of volcanic ash into seawater alters particle composition with implications for the cellular, physiological and anatomical response of suspension-feeding organisms. Adult females of Crepipatella peruviana were exposed to three diets consisting of a fixed concentration of 50,000 cells ml-1 of the microalga Isochrysis galbana plus different concentrations of ash particles (30, 90 and 150 mg L-1). The objective was to determine the cellular, physiological and anatomical responses. Mortality increased with ash concentrations, while feeding and respiration rates, tissue weight, and condition index decreased. The gills showed severe degradation of cilia and the presence of large mucous aggregates of cilia and ash. An increase in ash resulted in decreased lipid peroxidation and protein carbonyls, but increased total antioxidant capacity and phenols. Thus, volcanic ash particles may exert a high impact at both cellular and physiological levels for C. peruviana, where inhibition of gill function reduces the ability to acquire food.

Imposex Incidence in Gastropod Species from Santa Marta Coastal Zone, Colombian Caribbean Sea.

Imposex is a phenomenon widely associated with environmental exposure to organotin compounds which were quite common components of antifouling paints applied on boats and ship hulls. Here we study the incidence of imposex in neogastropods and its relation with water quality and maritime traffic in the coastal strip of Santa Marta, Colombia. Imposex was determined via specialized indexes and related to the organisms' size, somatic conditions, variables of water quality and maritime traffic, in a space-time assessment. There was evidence of imposex in five species Plicopurpura patula, Vasula deltoidea, Stramonita haemastoma, S. floridana, and Gemophos auritulus. Purpura patula and Vasula deltoidea species were found in all sampling sites. The results have proved that imposex is highly influenced by the maritime traffic variable, with greater prevalence during the dry season, and with P. patula being more sensitive than V. deltoidea.

Trophic transfer of paralytic shellfish toxin (PST): Physiological and reproductive effects in the carnivorous gastropod Acanthina monodon (Pallas, 1774).

Harmful algal blooms can adversely affect different levels of the trophic chain, from primary consumers, such as bivalve molluscs, to higher links such as large fish, birds and mammals, including humans. Among secondary consumers, it has been described that carnivorous gastropods can accumulate these toxins when they prey on bivalves that have been exposed to toxic microalgae; these could also harm human health. In Chile, frequent events of harmful algal blooms caused by the dinoflagellate Alexandrium catenella have been described. This organism produces paralytic shellfish toxin (PST) which has been identified in some carnivorous gastropods. The objective of this research was to identify the physiological and reproductive response of the carnivorous gastropod Acanthina monodon fed on the Mytilid Perumytilus purpuratus, which had previously been maintained on a diet containing PST. Specimens of A. monodon showed a decrease in ingestion and absorption rate when they consumed PST indirectly through their diet. The oxygen consumption rate was also affected by the diet-time interaction. The variations of these parameters were reflected in the scope for growth, since the available energy was lower in gastropods exposed to toxic diet. Consumption of PST had a negative effect on the reproduction of A. monodon, since intoxicated adults presented lower egg-masses and delayed start of oviposition. We observed a delay in the development of the embryos inside the capsules, and a lower number of hatched juveniles, although these few juveniles from intoxicated parents accomplished higher growth rates during the next 6 months. We may therefore suggest that toxin transfer, from harmful microalgae through the trophic chain, can generate deleterious effects on the physiological energetics of the organisms that consume them, affecting their reproductive capacity and early ontogenetic development.

Effect of the earthquake-tsunami (Chile, 2010) on toxic metal content in the Chilean abalone mollusc Concholepas concholepas.

Toxic metal content were measured in samples of mollusc Concholepas concholepas obtained from the Chilean coast. Samples were collected during two periods, one before and one after the earthquake-tsunami which occurred in the Maule Region, Chile, February 27th, 2010 as a result of an earthquake with a magnitude of 8.8. Quantification of toxic metals in samples of hepatopancreas and muscle tissue was performed using flame atomic absorption spectroscopy. The analytical methodology was validated with certified reference material. The content means measured in muscle tissue for January 2010 were Cu: 10.3; Cr: 0.7; Cd: < 0.1; Mn: 3.6 and Zn: 38.6 µg g-1 (dry weight). For October 2014, the means were Cu: 8; Cr: 2.4; Cd: < 0.1; Mn: 5.6 and Zn: 45.4 µg g-1 (dry weight). In hepatopancreas tissue, the content means were Cu: 14.8; Cr: 2.4; Cd: 246.2; Mn: 4.4 and Zn: 1552.9 µg g-1 (dry weight). For October 2014, the means were Cu: 53.7; Cr: 3.5; Cd: 118; Mn: 13.4 and Zn: 1352.3 µg g-1 (dry weight). Cd, Cr, Cu, Mn and Zn content in the samples of hepatopancreas were generally higher than those recorded in muscle tissue however they were not always statistically significant. Statistical analysis showed that Cu and Mn content in the post-tsunami period increased in the hepatopancreas tissue. The concentrations of Cd, Cr, Cu, Mn, and Zn measured in the muscular tissue (edible part) of the species Conchalepas concholepas, were lower than the maximum contents allowed by the current legislation (FAO/WHO, 2004; EU, 2001) and its consumption is not a risk to human health.

Imposex in Stramonita haemastoma from coastal sites of Cartagena, Colombia / Imposex em Stramonita haemastoma em locais costeiros de Cartagena, Colômbia

Abstract Imposex is the development of male sexual characteristics caused by the toxic effects of some chemicals that acts as an endocrinal disruptor. Antifouling paints contain these chemicals. Cartagena lacks studies to indicate the extent of imposex in its coastal waters. The aim of this study was to determine the prevalence of imposex in the gastropod Stramonita haemastoma in Cartagena, Colombia. Specimens were collected during 2013 from locations of high and low influence of port activity. Morphometric measurements and the frequency of the occurrence of imposex were registered. The comparison among morphometric variables showed statistically significant differences between the two sites studied. Furthermore, the females of the S. haemastoma species presented an imposex frequency of 93.1% in Birds' Island, Cartagena Bay, compared to 31.8% in La Bocana. The relative penis size index or RPLI (10.145 and 3.231) and vas deferens sequence index or VDSI (2.83 and 1.16), showed possible contamination by organotin compounds in both places.

Resumo Imposex é o desenvolvimento de características sexuais masculinas causadas por poluentes tóxicos de alguns produtos químicos que atuam como desreguladores endócrinos. Tintas anti-incrustantes são as que contêm estes produtos químicos. Cartagena carece de estudos para indicar a extensão do imposex nas suas águas costeiras. O objetivo deste estudo foi determinar a prevalência de imposex no gastrópode Stramonita haemastoma em Cartagena, Colômbia. Os espécimes foram coletados durante 2013 de locais de alta e baixa influência da atividade portuária. Foram registradas as medidas morfométricas e a frequência da ocorrência do imposex. A comparação entre as variáveis morfométricas mostrou diferenças estatisticamente significantes entre os dois locais estudados. Além disso, as fêmeas da espécie S. haemastomaapresentaram uma frequência de imposex de 93,1% na Ilha das Aves, Baía das Cartagena, em comparação com 31,8% em La Bocana. O índice do comprimento relativo do pênis ou RPLI (10,145 e 3,231) e o índice da sequência do vaso deferente ou VDSI (2,83 e 1,16), mostraram possível contaminação por compostos organoestânicos em ambos os locais.

Size, season and origin of gastropods matter in imposex assessments.

Through controlled exposure to tributyltin (TBT), the effects of season, size, and population origin were evaluated on imposex incidence in Stramonita brasiliensis. Four experiments were carried out with organisms collected from three different sites on the Brazilian coast (Torres - Rio Grande do Sul, Farol de Santa Marta - Santa Catarina, and Aracruz - Espírito Santo). S. brasiliensis were anesthetized, sexed, measured, classified by size in small (< 30 mm), medium (≥ 30 to ≤ 40 mm), or big (> 40 mm) and injected in the foot muscle with 0.5 µg g-1 of TBT. Organisms were maintained during one month in aquariums with clear marine water under controlled laboratory conditions. One month after injection, the imposex assessments showed that the population from the southeast (tropical) region was more sensitive to TBT than the population from the south of Brazil (the temperate region). A greater sensitivity in the small and medium categories was observed. Females were also more susceptible to TBT when exposed during their reproductive period. Thus, the present study highlighted the need to ensure that intrinsic biological factors related to organisms are considered in such biomonitoring studies to avoid misinterpretation of results.

TBT is still a matter of concern in Peru.

Tributyltin (TBT) environmental concentrations as well as imposex levels have been declining in several coastal areas worldwide. However, recent studies have shown that TBT contamination is still an important issue along some Latin American coastal areas. Thus, the incidence of imposex and butyltin compounds (BTs) in sediments and gastropod tissues was spatially and temporally (2009 and 2012) appraised in Lima (Peru) along potential TBT sources (harbors and marinas). Despite the general pattern of reduction close to Callao harbor, a clear increment in the levels of imposex (RPLI = 0.0 to 8.4 and 0.0 to 28.4/VDSI = 0.0 to 3.3 and 0.0 to 3.5) and TBT in tissues (25 to 112 and 62 to 146 ng Sn g-1) of Thaisella chocolata were seen from 2009 to 2012 in two sampling sites, respectively, located nearby a newly established marina. Thus, despite the international restrictions on the use of TBT-based antifouling paints are apparently reducing the inputs at international harbors, the present study clearly shows that marinas are still acting as important sources of TBT to the study area due possibly to the lack of legal restrictions on production, trading and/or use of TBT. The present findings reinforce what has been seen along many other Latin American coastal areas.

Toxicokinetic and toxicodynamic studies of carbaryl alone or in binary mixtures with azinphos methyl in the freshwater gastropod Planorbarius corneus.

Carbamate insecticides such as carbaryl and organophosphates such as azinphos-methyl share the ability to inhibit the activity of B-esterases. This study aimed to (1) assess the inhibitory effects of carbaryl on B-esterase activity in soft tissues and hemolymph of Planorbarius corneus; (2) establish whether binary mixtures of carbaryl and azinphos-methyl depart or not from a model of concentration addition on the inhibition of cholinesterase activity; (3) determine the bioconcentration and elimination of the pesticides. The results showed that exposure of gastropods to increasing concentrations of carbaryl (0.1-5 mg L-1) for 48 h inhibited cholinesterase activity in a concentration-dependent manner, with an EC50 of 1.4 ±â€¯0.3 mg L-1 and 1.2 ±â€¯0.1 mg L-1 for soft tissue and hemolymph, respectively. Carboxylesterase activity, measured with the substrates p-nitrophenyl butyrate and p-nitrophenyl acetate, was between 2.3 and 25 times more sensitive to carbaryl inhibition than cholinesterase activity. Binary mixtures corresponding to 0.5 EC50 carbaryl + 0.5 EC50 azinphos-methyl and 0.75 EC50 carbaryl + 0.75 EC50 azinphos-methyl produced inhibitions of cholinesterase activity similar to those of individual pesticides, following a model of concentration addition. Bioconcentration was analyzed using a one-compartment model. The absorption kinetics (k1) for both pesticides alone (1.4 mg L-1 of carbaryl or 1.8 mg L-1 of azinphos-methyl) or mixed (1.4 mg L-1 of carbaryl + 1.8 mg L-1 of azinphos-methyl) were similar. The elimination kinetics ratio (k2) estimated for the pesticides alone or in the mixtures showed that carbaryl was eliminated 3.5 times faster than azinphos-methyl. These results suggest that exposure of Planorbarius corneus to binary mixtures of carbaryl and azinphos-methyl for 48 h follow a concentration addition model on inhibition of cholinesterase activity and that the pesticide mixtures do not change the toxicokinetic parameters of the parent compounds.

Imposex in Stramonita haemastoma from coastal sites of Cartagena, Colombia.

Imposex is the development of male sexual characteristics caused by the toxic effects of some chemicals that acts as an endocrinal disruptor. Antifouling paints contain these chemicals. Cartagena lacks studies to indicate the extent of imposex in its coastal waters. The aim of this study was to determine the prevalence of imposex in the gastropod Stramonita haemastoma in Cartagena, Colombia. Specimens were collected during 2013 from locations of high and low influence of port activity. Morphometric measurements and the frequency of the occurrence of imposex were registered. The comparison among morphometric variables showed statistically significant differences between the two sites studied. Furthermore, the females of the S. haemastoma species presented an imposex frequency of 93.1% in Birds' Island, Cartagena Bay, compared to 31.8% in La Bocana. The relative penis size index or RPLI (10.145 and 3.231) and vas deferens sequence index or VDSI (2.83 and 1.16), showed possible contamination by organotin compounds in both places.

From TBT to booster biocides: Levels and impacts of antifouling along coastal areas of Panama.

Antifouling biocides in surface sediments and gastropod tissues were assessed for the first time along coastal areas of Panama under the influence of maritime activities, including one of the world's busiest shipping zones: the Panama Canal. Imposex incidence was also evaluated in five muricid species distributed along six coastal areas of Panama. This TBT-related biological alteration was detected in three species, including the first report in Purpura panama. Levels of organotins (TBT, DBT, and MBT) in gastropod tissues and surficial sediments ranged from <5 to 104 ng Sn g-1 and <1-149 ng Sn g-1, respectively. In addition, fresh TBT inputs were observed in areas considered as moderate to highly contaminated mainly by inputs from fishing and leisure boats. Regarding booster biocides, TCMTB and dichlofluanid were not detected in any sample, while irgarol 1051, diuron and DCOIT levels ranged from <0.08 to 2.8 ng g-1, <0.75-14.1 ng g-1, and <0.38-81.6 ng g-1, respectively. The highest level of TBT (149 ng Sn g-1) and irgarol 1051 (2.8 ng g-1), as well as relevant level of DCOIT (5.7 ng g-1), were detected in a marina used by recreational boats. Additionally, relatively high diuron values (14.1 ng g-1) were also detected in the Panama Canal associate to a commercial port. DCOIT concentrations were associated with the presence of antifouling paint particles in sediments obtained nearby shipyard or boat maintenance sites. The highest levels of TBT, irgarol 1051, and diuron exceeded international sediment quality guidelines indicating that toxic effects could be expected in coastal areas of Panama. Thus, the simultaneous impacts produced by new and old generations of antifouling paints highlight a serious environmental issue in Panamanian coastal areas.

Butyltin contamination in Northern Chilean coast: Is there a potential risk for consumers?

Imposex is the superimposition of non-functional male sex organs in gastropod females. This syndrome is a hormonal imbalance induced by tributyltin (TBT) which have been used in antifouling paints formulation. The present study aimed to perform an integrated environmental assessment of imposex and butyltin (BT) contamination using surface sediments and tissues of Thaisella chocolata (an edible gastropod) from northern Chile. The results showed imposex incidence in 11 out of 12 sites. In the most contaminated sites, which are areas under the influence of maritime activities, and also used for fishing and aquaculture, RPLI were over 60 and VDSI over 4 (high incidence of sterile females). Exceptionally high contamination levels and evidences of fresh inputs of tributyltin (TBT) were detected along the studied area. TBT levels above 300 and 90ngSng-1, respectively, were recorded in sediments and edible gastropod tissues of 6 sites. Thus, a daily ingestion of 90 to 173g of T. chocolata foot (4 to 8 organisms) from the most contaminated sites will certainly lead to the consumption of BT exceeding the tolerable daily intake recommended by European Food Safety Authority. It is reasonable to consider that human risk is even higher if daily consumption of additional seafood is considered. Moreover, some contaminated sites were located within the marine reserve "Isla Grande Atacama", indicating that even marine protected areas are under the influence of TBT contamination. These findings suggest that current levels of TBT in the studied area are sufficient to induce harmful effects on the environment and constitutes a potential threat to seafood consumers. Thus, national regulatory actions toward environmental protection and food safety of local populations are still mandatory, even after 8years of the TBT global ban by IMO.

Evaluation of the toxic effect of endocrine disruptor Bisphenol A (BPA) in the acute and chronic toxicity tests with Pomacea lineata gastropod.

Bisphenol A (BPA) is a plasticizer and a risk when it interacts with organisms, and can cause changes in the development and reproduction of them. This study aimed to evaluate the effects of BPA, by acute and chronic toxicity tests with neonates and adults of Pomacea lineata. Adults and neonates were divided into groups exposed to BPA (1-20mg/L), or 17ß-estradiol (1mg/L) and control in the acute and chronic toxicity tests. Behavior, heart rate, reproduction and hemolymph biochemical analysis were measured. In the acute toxicity test, the 96-h LC50 with adults was 11.09 and with neonates was 3.14mg/L. In this test, it was observed lethargic behavior and an increase of 77.6% of aspartate aminotransferase in the adults' hemolymph (p<0.05); and neonates' heart rate decreased 72.7% (p<0.05). In the chronic toxicity test, it was observed behaviors associated with reproduction, as Copulate, in the groups exposed to BPA. The results that were found in this study proved that BPA is a potentially toxic agent to Pomacea lineata according to biological parameters evaluated. These data contribute to the understanding of BPA toxic effects' in the aquatic invertebrates.

Assessment of tributyltin contamination based on imposex in Stramonita rustica (Mollusca: Gastropoda) along southern Bahia coast, northeastern Brazil / Avaliação da contaminação por tributil-estanho baseada em imposex em Stramonita rustica (Mollusca: Gastropoda) ao longo do Litoral Sul da Bahia

Abstract This study investigated the effects of tributyltin (TBT) on the morphology of the genital system of the gastropod Stramonita rustica in southern Bahia, Brazil. For this, 330 specimens were collected during the summer of 2014 at eight sampling points to ascertain whether male sex organs had developed in addition to the complete female genital tract in females (= imposex). The analyses were made under a stereoscopic microscope. Imposex and their associated indexes, and the sterile females, exhibited the highest rates in harbors and shipyards areas. Despite the total ban of TBT in anti-fouling paints on a global scale since 2003, the results of this and other studies indicate the continued use of those paints on the Brazilian coast. This shows the inefficiency of existing legislation and the need to strengthen enforcement of the ban.

Resumo Este estudo investigou os efeitos de tributil-estanho (TBT) sobre a morfologia do sistema genital do gastrópode Stramonita rustica (Lamarck, 1822) no Litoral Sul da Bahia, Brasil. Para isso, 330 espécimens foram coletados durante o verão de 2014 em oito pontos de amostragem, para verificar se os órgãos sexuais masculinos tinham se desenvolvido para além do trato genital feminino completo (= imposex). As análises foram feitas sob um microscópio estereoscópico. O imposex e seus índices associados, assim como as fêmeas estéreis, apresentaram as maiores taxas em áreas portuárias e estaleiros. Apesar da proibição total do TBT em tintas anti-incrustantes em escala global desde 2003, os resultados deste e de outros estudos indicam a continuidade de uso dessas tintas na costa brasileira. Isso mostra a ineficácia da legislação vigente e reforça a necessidade de se cumprir integralmente o banimento.

Shell shape as a biomarker of marine pollution historic increase.

Buccinanops globulosus is a TBT sensitive marine gastropod, classified as a good indicator of imposex incidence and used as a model to study adverse contamination effects. Population and maritime industries has incremented pollution in Nuevo gulf harbor since 1970s, promoting morphological changes in B. globulosus shell shape. We study the shell shape of the species comparing present day's specimens from the harbor zone with those collected in the same zone before the increasing of maritime activity and pre-Hispanic archaeological Middens. We demonstrated that harbor pollution produces globular shell shape in B. globulosus, an effect that probably allows gastropods to isolate themselves from the external adverse environment. On the contrary, shells from pre-Hispanic periods, unpolluted sites and those collected before the expansion of maritime activities, presented an elongated shell shape. Our study confirms that shell shape variation in marine gastropods can be used as a biomarker of harbor pollution.

Assessment of tributyltin contamination based on imposex in Stramonita rustica (Mollusca: Gastropoda) along southern Bahia coast, northeastern Brazil.

This study investigated the effects of tributyltin (TBT) on the morphology of the genital system of the gastropod Stramonita rustica in southern Bahia, Brazil. For this, 330 specimens were collected during the summer of 2014 at eight sampling points to ascertain whether male sex organs had developed in addition to the complete female genital tract in females (= imposex). The analyses were made under a stereoscopic microscope. Imposex and their associated indexes, and the sterile females, exhibited the highest rates in harbors and shipyards areas. Despite the total ban of TBT in anti-fouling paints on a global scale since 2003, the results of this and other studies indicate the continued use of those paints on the Brazilian coast. This shows the inefficiency of existing legislation and the need to strengthen enforcement of the ban.

Evidence of metabolic microevolution of the limpet Nacella concinna to naturally high heavy metal levels in Antarctica.

The gastropod Nacella concinna is the most conspicuous macroinvertebrate of the intertidal zone of the Antarctic Peninsula and adjacent islands. Naturally high levels of copper and cadmium in coastal marine ecosystems are accumulated in N. concinna tissues. We aimed to study the effects of metal cations on N. concinna arginase in the context of possible adaptive microevolution. Gills and muscle had the highest argininolytic activity, which was concentrated in the cytosol in both tissues. Gills had the highest levels of arginase and may be involved in the systemic control of l-arginine levels. The relatively high argininolytic activity of the N. concinna muscular foot, with KM=25.3±3.4mmolL-1, may be involved in the control of l-arginine levels during phosphagen breakdown. N. concinna arginases showed the following preferences for metal cations: Ni2+>Mn2+>Co2+>Cu2+ in muscle and Mn2+>Cu2+ in gills. Cu2+ activation is a unique characteristic of N. concinna arginases, as copper is a potent arginase inhibitor. Cu2+ partly neutralized N. concinna arginase inhibition by Cd2+, worked synergistically in muscle arginase activation by Co2+ and neutralized muscle arginase activation by Ni2+. Mn2+ was able to activate muscle arginase in the presence of Fe3+ and Pb2+. The selection of arginases that are activated by Cu2+ and resistant to inhibition by Cd2+ in the presence of Cu2+ over evolutionary timescales may have favored N. concinna occupation of copper- and cadmium-rich niches.

Plant extracts, metaldehyde and saline solutions on the population control of Bradybaena similaris / Extratos vegetais, metaldeído e soluções salinas para o controle populacional de Bradybaena similaris

This study aimed to test the efficiency of plant extracts, metaldehyde and saline solutions, as alternatives to the population control of the snail Bradybaena similaris , and to investigate the effect of the plant extracts in reducing the damage of the snail on Brassica oleracea . The experiments were performed at the Entomology Laboratory of the Universidade Comunitária da Região de Chapecó (Unochapecó), using a random experimental design with nine treatments in triplicate. Five adult individuals of B. similaris were subjected to each trial, totaling 135 snails. The following treatments were tested: cinnamon ( Melia azedarach ), timbó ( Ateleia glazioveana ), rosemary ( Rosmarinus officinalis ), mate herb ( Ilex paraguariensis ), two concentrations of metaldehyde (3% and 5%), two concentrations of salt solution (5% and 10 %), and a control treatment (distilled water). To evaluate the survival of B. similaris it was checked the treatments every 24 hours, over four consecutive days. The results revealed that the two concentrations of metaldehyde were fully efficient, that the saline solution (10%) had and intermediate efficiency, and that all other treatments were not effective. The treatment with the M. azedarach extract induced a higher consumption of B. oleracea , while the saline solution at 10% and the extracts of R. officinalis and I. paraguariensis inhibited leaf consumption.(AU)

O objetivo deste trabalho foi avaliar a eficiência de extratos vegetais, metaldeído e soluções salinas como alternativas para o controle populacional de Bradybaena similaris e o efeito dos extratos para a redução dos danos sobre Brassica oleracea , em condições de laboratório (Laboratório de Entomologia da Unochapecó, Chapecó, Santa Catarina). O experimento foi conduzido utilizando cinco indivíduos adultos de B. similares para cada um dos nove tratamentos em triplicatas, totalizando 135 caracóis. Os seguintes tratamentos foram testados: extrato de cinamomo ( Melia azedarach ), timbó ( Ateleia glazioveana ), alecrim ( Rosmarinus officinalis ), erva-mate ( Ilex paraguariensis ), metaldeído (3% e 5%), soluções salinas (5% e 10%) e a testemunha (água destilada). Para avaliação dos dados, foi determinada a taxa de sobrevivência de B. similaris a cada 24 horas em um período de quatro dias. Os resultados obtidos indicam que as duas concentrações de metaldeído foram eficientes, a solução salina (10%) apresentou eficiência intermediária, enquanto os demais tratamentos não foram eficazes. Observou-se que o tratamento com extrato de M. azedarach induziu a um consumo maior de B. oleracea , enquanto a solução salina a 10% e os extratos de R. officinalis e I. paraguariensis inibiram o consumo.(AU)

Antioxidant responses in estuarine invertebrates exposed to repeated oil spills: Effects of frequency and dosage in a field manipulative experiment.

We have experimentally investigated the effects of repeated diesel spills on the bivalve Anomalocardia brasiliana, the gastropod Neritina virginea and the polychaete Laeonereis culveri, by monitoring the responses of oxidative stress biomarkers in a subtropical estuary. Three frequencies of exposure events were compared against two dosages of oil in a factorial experiment with asymmetrical controls. Hypotheses were tested to distinguish between (i) the overall effect of oil spills, (ii) the effect of diesel dosage via different exposure regimes, and (iii) the effect of time since last spill. Antioxidant defense responses and oxidative damage in the bivalve A. brasiliana and the polychaete L. culveri were overall significantly affected by frequent oil spills compared to undisturbed controls. The main effects of diesel spills on both species were the induction of SOD and GST activities, a significant increase in LPO levels and a decrease in GSH concentration. N. virginea was particularly tolerant to oil exposure, with the exception of a significant GSH depletion. Overall, enzymatic activities and oxidative damage in A. brasiliana and L. culveri were induced by frequent low-dosage spills compared to infrequent high-dosage spills, although the opposite pattern was observed for N. virginea antioxidant responses. Antioxidant responses in A. brasiliana and L. culveri were not affected by timing of exposure events. However, our results revealed that N. virginea might have a delayed response to acute high-dosage exposure. Experimental in situ simulations of oil exposure events with varying frequencies and intensities provide a useful tool for detecting and quantifying environmental impacts. In general, antioxidant biomarkers were induced by frequent low-dosage exposures compared to infrequent high-dosage ones. The bivalve A. brasiliana and the polychaete L. culveri are more suitable sentinels due to their greater responsiveness to oil and also to their wider geographical distribution.

Fe, oxidative and nitrosative metabolism in the Antarctic limpet Nacella concinna.

The hypothesis of this work was that oxidative and nitrosative metabolism in the digestive gland (DG) of two limpet populations (intertidal and subtidal) of the Antarctic species Nacella concinna show different behavior when they were exposed to either intermittent (intertidal) or constant (subtidal) natural Fe. Total Fe content and labile Fe pool were higher in the DG of the subtidal compared to the intertidal population. However, no significant differences between populations were seen on the Fe atoms content of the isolated ferritin. Ascorbyl radical content was 2.0±0.4 and 6.5±0.8pmol/mg FW in the DG of the intertidal and subtidal animals, respectively. Lipid damage, assessed as content of thiobarbituric reactive substances, was different between the tissues of intertidal and subtidal samples, 491±102 and 1242±367pmol/mg FW, respectively. Catalase and superoxide dismutase activities showed no differences between the limpets. Nitric oxide (NO) content was 25±3 and 22±2pmol/mg FW in DG from intertidal and subtidal animals, respectively. NO synthase-like (NOS-like) activity was evaluated supplementing the samples with the enzyme co-factors, and the inhibitory effect of Nω-nitro-L-arginine methyl ester hydrochloride was tested. NO generation rate was 3.4±0.3 and 4.7±0.6pmol/minmg FW in DG from the intertidal and subtidal population, respectively. These results showed that the oxidative condition of the limpet population constantly covered by the Fe enriched water is more affected than the intertidal population. However, the nitrosative metabolism seems to be independent of the environmental high Fe content since similar NO steady state concentration and NOS-like activity were measured in both populations.

Sex steroid imbalances in the muricid Stramonita haemastoma from TBT contaminated sites.

Imposex incidence, organotin tissue levels, and sex steroid (free and esterified testosterone and estradiol) levels were assessed in Stramonita haemastoma from Babitonga Bay (Santa Catarina State, Southern Brazil). The imposex levels showed a reduction when compared to a previous evaluation performed in the same area. In spite of that, the detected imposex incidence indicated the occurrence of tributyltin (TBT) inputs that were still able to produce endocrine disruption in local gastropods. In addition, a high level of organotins was observed in tissues of imposexed females. These females also showed a hormonal imbalance, especially in the total testosterone/total estradiol ratio. These findings obtained under realistic field conditions suggest that the steroid pathway could be responsible by the imposex induction after exposure to TBT. In this case, measurements of sex steroid levels can be an additional evidence for monitoring sites and impose affected gastropod populations.