Morphology and histology of the digestive gland of Oxychilus (Drouetia) atlanticus (Morelet & Drouët) (Gastropoda: Pulmonata).

Little information exists on the histology of Zonitidae digestive system. This study deals with a detailed characterization of the different types of cells comprising epithelium lining the digestive gland of 0xychilus atlanticus. From light and scanning electron microscope (SEM) studies, three types of differentiated cells can be identified in the digestive gland: digestive cells, excretory cells and calcium cells. Digestive cells are the most numerous, and are present in two forms, one believed to be absorbing food material and the other secreting material. Excretory cells are distinguished by having a large central vacuole, containing excretory granules. Calcium cells contain spherules of calcium salts, which have a characteristic birefringence

Freshwater molluscs from volcanic areas as model organisms to assess adaptation to metal chronic pollution

Copyright © 2006 Elsevier B.V. All rights reserved.Cellular biomarkers of exposure and biological effects were measured in digestive gland of snails (Physa acuta) sampled in sites with 12 andwithout active volcanismin SãoMiguel Island (Azores).Metal content in digestive cell lysosomeswas determined by image analysis 13 after autometallography (AMG) as volume density of autometallographed black silver deposits (VvBSD). Lysosomal structural changes 14 (lysosomal volume, surface and numerical densities – VvLYS, SvLYS and NvLYS−, and surface-to-volume ratio – S/VLYS−) were 15 quantified by image analysis, after demonstration of β-glucuronidase activity, on digestive gland cryotome sections. Additional chemical 16 analyses (atomic absorption spectrophotometry) were done in the digestive gland of snails. The highestmetal concentrations were found 17 in snails from the active volcanic site, which agreed with high intralysomal VvBSD. Digestive cell lysosomes in snails inhabiting sites 18 with active volcanismresembled a typical stress situation (enlarged and less numerous lysosomes). In conclusion, the biomarkers used in 19 this work can be applied to detect changes in metal bioavailability due to chronic exposure to metals (volcanism), in combination with 20 chemical analyses.Centro de Investigação de Re- 430 cursos Naturais (CIRN, University of the Azores) and 431 Direcção Regional da Ciência e Tecnologia (Government 432 of the Azores) and by the Basque Government (ETOR- 433 TEK

Bioavailability and cellular effects of metals on Lumbricus terrestris inhabiting volcanic soils

Copyright © 2005 Elsevier Ltd. All rights reserved.Whether the radial thickness (RT) of the chloragogenous tissue and intestinal epithelium of earthworms (Lumbricus terrestris) reflects the bioavailability of metals in soils was investigated in two areas, one with active volcanism (Furnas) and another with no volcanic activity since 3 million years ago (Santa Maria), in the Azores. Metal contents in soil samples and earthworms from the two areas were analyzed. Autometallography and measurements of the RT were performed in the chloragogenous tissue and intestinal epithelium. Earthworms from the active volcanic area demonstrated lower RT of chloragogenous tissue and intestinal epithelium as well as higher levels of bioavailable metals, especially Zn and Cd. Comparison of bioavailable metal contents between both areas suggests a higher risk for uptake of potentially toxic metals in the active volcanic area than in the non-active volcanic area, which is reflected by the lower RT of the chloragogenous tissue and intestinal epithelium in the former.Centro de Investigação de Recursos Naturais (CIRN, University of the Azores), Direcção Regional da Ciência e Tecnologia (Government of the Azores), and the ETORTEK program from the Basque Government, through the BERRILUR project (IE03-110). André Amaral was supported by a PhD grant from Fundação para a Ciência e a Tecnologia (SFRH/BD/8186/2002)

Integrative biomarker assessment of the effects of chemically and mechanically dispersed crude oil in Pacific oysters, Crassostrea gigas

International audienceThe impact of dispersed crude oil and dispersant on adult Pacific oysters, Crassostrea gigas, was evaluated through an integrative biomarker approach including (1) biochemical (plasma catecholase- and laccase-type phenoloxidase and superoxide dismutase), (2) histological (digestive cell lysosomal responses, digestive gland histopathology) and (3) physiological (flesh condition index) endpoints in the haemolymph and digestive gland. Adult oysters were exposed to non-contaminated water (control), chemically-dispersed oil (Brut Arabian Light), mechanically-dispersed oil and dispersant (FINASOL®) alone for 2 days, and further depurated in noncontaminated water for 4 weeks. After exposure to chemically and mechanically dispersed oil oysters exhibited induction of plasma laccase-type phenoloxidase and superoxide dismutase activities, enlargement of digestive cell lysosomes, lipofuscin accumulation, reduced neutral lipid content and atrophy of digestive gland diverticula; more markedly on exposure to chemically dispersed oil. From the studied biomarkers, only lysosomal biomarkers were significantly affected after exposure to the dispersant alone. This included lysosomal enlargement, neutral lipid depletion and lipofuscin accumulation in the digestive gland epithelium. A recovery of plasma enzyme activities was observed after 4 weeks of depuration. The integrative biological response index indicated that chemically dispersed oil caused significantly higher stress to C. gigas than the mechanically-dispersed one or the dispersant alone; nevertheless, the response seems to be reversible after depuration

Toxicopathic effects of waterborne Cd in sole juveniles, a prospective pollution monitoring sentinel for estuarine ecosystems

peer reviewedEstuarine ecosystems are under high anthropogenic pressure and receive a wide Q7 variety of contaminants, including metals. They can remain trapped in estuarine sediments at high concentrations for long periods but eventually they can be released to the overlying water and become toxic for the resident benthic biota. As a consequence, biomonitoring programs surveying the health status of estuaries and coastal areas count with benthic species for the assessment of health disturbances in these specific ecosystems. The present investigation aims at recognizing toxicopathic effects elicited in estuarine life stages of the benthic fish Solea senegalensis on exposure to waterborne Cd, a model compound for metal contamination. Biological responses to metal exposure were assessed based on the application of a “biomarker + histopathology” approach. Two-year old sole juveniles were exposed to various concentrations of waterborne Cd for 3 and 7 days (Control; Low Cd: 1 µg/l; Mid Cd: 10 µg/l and High Cd: 1000 µg/l). Liver samples were collected for chemical analysis at day 7. Biological samples were collected at days 3 and 7 for biochemical (brain and liver), histochemical (liver) and histopathological analysis (liver, gills and gonad). Brain acetyl cholinesterase was inhibited after 7 days exposure, indicating neurotoxic effects on exposure to 1000 µg Cd/l. Hepatic catalase and glutathione-S transferase were induced at day 3 and inhibited at day 7, which suggests a bell-shaped response. A dose-dependent lysosomal membrane destabilization in hepatocytes was recorded at day 7. In parallel, histopathological lesions in gills, liver and gonad were more frequent at day 7 in soles exposed to high Cd concentrations. Overall, the “biomarker + histopathology” approach revealed that waterborne Cd causes toxicopathic effects in sole juveniles upon exposure to environmentally relevant concentrations (10 µg Cd/l). The effects were clearly time dependent, and thus it is conceivable that more severe biological effects would be elicited by these low concentrations of Cd at longer exposure periods. Therefore, biomarkers and histopathology in combination provide early-warning indications of altered health status after waterborne Cd exposure in sole juveniles, a prospective sentinel for biological effects assessment of metal contamination in pollution monitoring programs in estuaries and coastal areas

Txikispora philomaios n. sp., n. g., a micro-eukaryotic pathogen of amphipods, reveals parasitism and hidden diversity in Class Filasterea

This study provides a morphological, ultrastructural, and phylogenetic characterization of a novel micro-eukaryotic parasite (2.3–2.6 µm) infecting amphipod genera Echinogammarus and Orchestia. Longitudinal studies across two years revealed that infection prevalence peaked in late April and May, reaching 64% in Echinogammarus sp. and 15% in Orchestia sp., but was seldom detected during the rest of the year. The parasite infected predominantly hemolymph, connective tissue, tegument, and gonad, although hepatopancreas and nervous tissue were affected in heavier infections, eliciting melanization and granuloma formation. Cell division occurred inside walled parasitic cysts, often within host hemocytes, resulting in hemolymph congestion. Small subunit (18S) rRNA gene phylogenies including related environmental sequences placed the novel parasite as a highly divergent lineage within Class Filasterea, which together with Choanoflagellatea represent the closest protistan relatives of Metazoa. We describe the new parasite as Txikispora philomaios n. sp. n. g., the first confirmed parasitic filasterean lineage, which otherwise comprises four free-living flagellates and a rarely observed endosymbiont of snails. Lineage-specific PCR probing of other hosts and surrounding environments only detected T. philomaios in the platyhelminth Procerodes sp. We expand the known diversity of Filasterea by targeted searches of metagenomic datasets, resulting in 13 previously unknown lineages from environmental samples.Ministerio de Economía y Competitividad, Grant/Award Number: BFU2017- 90114- P; Horizon 2020 Framework Programme, Grant/Award Number: 747789; Department for Environment, Food and Rural Affairs, UK Government, Grant/Award Number: FB002A and FC1214; Hezkuntza, Hizkuntza Politika Eta Kultura Saila, Eusko Jaurlaritza, Grant/Award Number: PRE_2016_2_0124; Agencia Estatal de Investigación (AEI); Fondo Europeo de Desarrollo Regional (FEDER); Ayuda Juan de la Cierva—Incorporación, Grant/Award Number: IJC2018- 036657-

Influence of season-depending ecological variables on biomarker baseline levels in mussels (Mytilus trossulus) from two Baltic Sea subregions

Highlights • Baltic Sea presents regional and seasonal changes that condition responses in mussels. • Food supply dictated reproductive cycle and consequently biomarker response. • Seasonal variability unequally reflected at different biological organization levels. • Combination of biological and ecological data necessary for biomarker approach.For reliable mussel monitoring programmes based on biomarkers, regionally relevant reference values and their natural variability need to be known. The Baltic Sea exhibits high inter-regional and seasonal variability in physical factors such as salinity, temperature and primary production. The aim of this pilot study is to depict the effects of season-related environmental factors in a selected battery of biomarkers in two environmentally different subregions of the Baltic Sea to help establishing reference data for biochemical, cellular and tissue-level biomarkers. In order to achieve that, mussels were collected from reference sites in Kiel (Germany) and Tvärminne (Finland) during three seasons: summer and autumn 2016, and spring 2017. Finally, in order to characterize the ecological situation, analysis of the chemical tissue burden was performed and chlorophyll‑a and particulate organic carbon concentration and temperature changes were analyzed at each sampling locality using satellite remote sensing images. An integrated biomarker response index was performed to summarize the biomarker responses of each locality and season. The biochemical endpoints showed seasonal variability regulated by temperature, food supply and reproductive cycle, while among the cellular endpoints only lipofuscin accumulation and lysosomal structural changes showed slight seasonal variation. Seasonal changes in tissue level biomarkers were observed only at the northern Baltic Sea site Tvärminne, dictated by the demanding energetic trade-off caused by reproduction. In conclusion, the characterization of the ecological variables and physico-chemical conditions at each site, is crucial to perform a reliable assessment of the effects of a hypothetical pollution scenario in the Baltic Sea. Moreover, reference levels of biomarkers and their responses to natural environmental conditions must be established

The EU Horizon 2020 project GRACE : integrated oil spill response actions and environmental effects

This article introduces the EU Horizon 2020 research project GRACE (Integrated oil spill response actions and environmental effects), which focuses on a holistic approach towards investigating and understanding the hazardous impact of oil spills and the environmental impacts and benefits of a suite of marine oil spill response technologies in the cold climate and ice-infested areas of the North Atlantic and the Baltic Sea. The response methods considered include mechanical collection in water and below ice, in situ burning, use of chemical dispersants, natural biodegradation, and combinations of these. The impacts of naturally and chemically dispersed oil, residues resulting from in situ burning, and non-collected oil on fish, invertebrates (e.g. mussels, crustaceans) and macro-algae are assessed by using highly sensitive biomarker methods, and specific methods for the rapid detection of the effects of oil pollution on biota are developed. By observing, monitoring and predicting oil movements in the sea through the use of novel online sensors on vessels, fixed platforms including gliders and the so-called SmartBuoys together with real-time data transfer into operational systems that help to improve the information on the location of the oil spill, situational awareness of oil spill response can be improved. Methods and findings of the project are integrated into a strategic net environmental benefit analysis tool (environment and oil spill response, EOS) for oil spill response strategy decision making in cold climates and ice-infested areas

The NORMAN Association and the European Partnership for Chemicals Risk Assessment (PARC): let’s cooperate! [Commentary]

The Partnership for Chemicals Risk Assessment (PARC) is currently under development as a joint research and innovation programme to strengthen the scientific basis for chemical risk assessment in the EU. The plan is to bring chemical risk assessors and managers together with scientists to accelerate method development and the production of necessary data and knowledge, and to facilitate the transition to next-generation evidence-based risk assessment, a non-toxic environment and the European Green Deal. The NORMAN Network is an independent, well-established and competent network of more than 80 organisations in the field of emerging substances and has enormous potential to contribute to the implementation of the PARC partnership. NORMAN stands ready to provide expert advice to PARC, drawing on its long experience in the development, harmonisation and testing of advanced tools in relation to chemicals of emerging concern and in support of a European Early Warning System to unravel the risks of contaminants of emerging concern (CECs) and close the gap between research and innovation and regulatory processes. In this commentary we highlight the tools developed by NORMAN that we consider most relevant to supporting the PARC initiative: (i) joint data space and cutting-edge research tools for risk assessment of contaminants of emerging concern; (ii) collaborative European framework to improve data quality and comparability; (iii) advanced data analysis tools for a European early warning system and (iv) support to national and European chemical risk assessment thanks to harnessing, combining and sharing evidence and expertise on CECs. By combining the extensive knowledge and experience of the NORMAN network with the financial and policy-related strengths of the PARC initiative, a large step towards the goal of a non-toxic environment can be taken