Ionan Marigómez - Academia.edu (original) (raw)

Papers by Ionan Marigómez

Integrated Environmental Assessment and Management, 2013

This article describes the outcome and follow-up discussions of an expert group meeting (Amsterda... more This article describes the outcome and follow-up discussions of an expert group meeting (Amsterdam, October 9, 2009) on the applicability of toxicity profiling for diagnostic environmental risk assessment. A toxicity profile was defined as a toxicological "fingerprint" of a sample, ranging from a pure compound to a complex mixture, obtained by testing the sample or its extract for its activity toward a battery of biological endpoints. The expert group concluded that toxicity profiling is an effective first tier tool for screening the integrated hazard of complex environmental mixtures with known and unknown toxicologically active constituents. In addition, toxicity profiles can be used for prioritization of sampling locations, for identification of hot spots, and-in combination with effect-directed analysis (EDA) or toxicity identification and evaluation (TIE) approaches-for establishing cause-effect relationships by identifying emerging pollutants responsible for the observed toxic potency. Small volume in vitro bioassays are especially applicable for these purposes, as they are relatively cheap and fast with costs comparable to chemical analyses, and the results are toxicologically more relevant and more suitable for realistic risk assessment. For regulatory acceptance in the European Union, toxicity profiling terminology should keep as close as possible to the European Water Framework Directive (WFD) terminology, and validation, standardization, statistical analyses, and other quality aspects of toxicity profiling should be further elaborated. Integr Environ Assess Manag 2013;9999:1-7. © 2013 SETAC

In order to obtain sustained conclusions regarding the effects of the Prestige oil spill in the m... more In order to obtain sustained conclusions regarding the effects of the Prestige oil spill in the monitoring campaigns carried out from Galicia to Basque Country since 2003, baseline values for the biomarkers recorded and their variability associated to varying natural factors (water surface temperature, salinity, tidal ranges and food availability) needed to be determined. For this purpose, mussels Mytilus galloprovincialis

Although the overall consensus sets up that mussels in the Northern Iberian Peninsula belong to t... more Although the overall consensus sets up that mussels in the Northern Iberian Peninsula belong to the species Mytilus galloprovincialis, mussels from this geographical area have been repeatedly reported to as M. edulis, even in various contributions assessing the effects of the Prestige oil spill. Several morphologically similar taxa (M. edulis , M. galloprovinci alis, M. californianus and M. trossulus )

Fish and mollusc histopathology provides a useful tool in biological effect monitoring programmes... more Fish and mollusc histopathology provides a useful tool in biological effect monitoring programmes through which environmental stress situations may be diagnosed on the basis of augmented prevalence or intensity of diseases and parasitic infestations. In the present study, the histopathological examination of the digestive gland of wild mussels was performed as a part of the biomonitoring programmes carried out to

The Science of the total environment, Jan 20, 2000

Lysosomal responses are widely accepted cellular effect biomarkers of general stress. Up to now, ... more Lysosomal responses are widely accepted cellular effect biomarkers of general stress. Up to now, these biomarkers have been analysed by means of conventional techniques based on enzyme histochemical methods, where lysosomal enzymes such as acid phosphatase and beta-glucuronidase (beta-GUS) have been employed as markers of lysosomes. The aim of the present work was to develop more advanced and sensitive methods based on the use of polyclonal antibodies to measure lysosomal enzymes in different sentinel organisms. For this purpose, we have studied the cross-reactivity of two commercial polyclonal antibodies against the lysosomal enzymes acid phosphatase and beta-GUS with molluscan digestive gland by means of immunoblotting and immunohistochemistry. The antibody against acid phosphatase cross-reacted specifically with the lysosomal fraction of the digestive gland, while unspecific immunoreaction occurred with digestive gland whole homogenates and tissue sections. The antibody against b...

The Histochemical journal, 1999

The distribution of proliferating cells in the digestive gland of the common marine mussel, Mytil... more The distribution of proliferating cells in the digestive gland of the common marine mussel, Mytilus galloprovincialis Lmk, was investigated by means of immunochemical techniques employing PC10, a commercial monoclonal antibody to the proliferating cell nuclear antigen (PCNA). Immunoblot analysis of digestive gland whole homogenates revealed a single crossreactive band of 36-37 kDa, identical to the corresponding protein of rat liver and murine melanoma cells. A band of slightly higher electrophoretic mobility (34-35 kDa) was found in fish liver. In mussel digestive gland, the samples obtained from young specimens presented a more intense signal for PCNA than in those obtained from old mussels, suggesting that the digestive gland cells of young mussels exhibit a higher proliferative activity. In paraffin sections, PC10 specifically labelled nuclei of all cell types, but only a smaller number of cells lining the different digestive epithelia. PCNA expression was more intense in digest...

Tissue & cell, 1996

The localization of metals in selected tissues of metal-exposed mussels was investigated by means... more The localization of metals in selected tissues of metal-exposed mussels was investigated by means of autometallography. Mussels collected from a Zn-polluted site were (a) depurated or, alternatively, (b) exposed to either Cu, Zn or Cd for 41 d. Mussels collected from a clean site were used as experimental reference. Autometallographically demonstrated black silver deposits (BSD), indicating the presence of metals, were observed in gills, (a) in frontal cells and haemocytes of Cu-exposed mussels, (b) in secretory postlateral and abfrontal cells and in endothelial cells of Zn-exposed mussels, and (c) in frontal, postlateral and endothelial cells but mainly in abfrontal cells and haemocytes of Cd-exposed mussels. Autometallography also revealed the presence of BSD in connective tissue brown cells underlying the mantle. Additionally, adipogranular cells of the connective tissue surrounding the gonad follicles were positively stained but no BSD was found in gonad tissue. Scarce BSD were ...

Aquatic Toxicology, 1998

The amount of black silver deposits (BSDs) revealed by autometallography in several tissues of ma... more The amount of black silver deposits (BSDs) revealed by autometallography in several tissues of marine winkles (Littorina littorea) exposed to sublethal concentrations of copper (Cu) and zinc (Zn) was semi-quantified using a light microscope. No significant change in the amount of BSDs was recorded in the cell types comprising the gill and mantle tissues. Changes in the amount of BSDs

Archives of Environmental Contamination and Toxicology, 2014

Bioavailability is affected by soil physicochemical characteristics such as pH and organic matter... more Bioavailability is affected by soil physicochemical characteristics such as pH and organic matter (OM) content. In addition, OM constitutes the energy source of Eisenia fetida, a well established model species for soil toxicity assessment. The present work aimed at assessing the effects of changes in OM content on the toxicity of Cd in E. fetida through the measurement of neutral red uptake (NRU) and mortality, growth, and reproduction (Organisation for Economic Co-operation and Development [OECD] Nos. 207 and 222). Complementarily, metallothionein (MT) and catalase transcription levels were measured. To decrease variability inherent to natural soils, artificial soils (Organization for Economic Cooperation and Development 1984) with different OM content (6, 10, and 14%) and spiked with Cd solutions at increasing concentrations were used. Low OM in soil decreased soil ingestion and Cd bioaccumulation but also increased Cd toxicity causing lower NRU of coelomocytes, 100 % mortality, and stronger reproduction impairment, probably due to the lack of energy to maintain protection mechanisms (production of MT).Cd bioaccumulation did not reflect toxicity, and OM played a pivotal role in Cd toxicity. Thus, OM content should be taken into account when using E. fetida in in vivo exposures for soil health assessment.

Marine Environmental Research, 2015

Science of The Total Environment, 1993

ABSTRACT The present work investigates the relationship between cellular responses and organismic... more ABSTRACT The present work investigates the relationship between cellular responses and organismic responses to the water accommodated fraction (WAF) of different petroleum hydrocarbons in mussels, Mytilus galloprovincialis. The results show that effects of WAF-induced environmental stress measured at the cellular and organismic levels exhibit a high degree of concordance. Positive correlation coefficients between indices of good digestive cell condition (high values of digestive epithelial thickness and high percentages of holding and absorpting tubules) and indices of good individual condition are found, while negative correlation coefficients explain the associations of parameters indicating cell or tissue damage with the same organismic parameters. There are few significant correlations between the cellular and the organismic responses when both are recorded at long exposure times but many significant correlations arise when cellular responses are recorded at short exposure times and the organismic responses at long exposure times. Then, selected cellular responses could be useful as ‘early warning biomarkers’ to predict responses at the individual level previous to their display. On the other hand, correlation values between catalase activity (involved in the detoxication of excess free oxygen radicals produced during hydrocarbon metabolism) in the digestive epithelium and organismic parameters show higher significance after long exposure times than at short exposure times. It is suggested that changes in catalase levels are not indicative of cell damage; conversely, they may represent a cellular adaptative mechanism against hydrocarbon toxicity. Nevertheless, development of adaptation mechanisms leads to significant deterioration of individual condition. Finally, the degree of gonad development is found to be a very important variable affecting both cellular and organismic responses to pollutants, and changes in parameters defining the structure of oocyte lysosomes are suggested to be useful as indicators of gamete spawning and also of the general health condition of mussels.

Science of The Total Environment, 2006

Cellular biomarkers of exposure and biological effects were measured in digestive gland of snails... more Cellular biomarkers of exposure and biological effects were measured in digestive gland of snails (Physa acuta) sampled in sites with and without active volcanism in São Miguel Island (Azores). Metal content in digestive cell lysosomes was determined by image analysis after autometallography (AMG) as volume density of autometallographed black silver deposits (Vv(BSD)). Lysosomal structural changes (lysosomal volume, surface and numerical densities - Vv(LYS,) Sv(LYS) and Nv(LYS-), and surface-to-volume ratio - S/V(LYS)-) were quantified by image analysis, after demonstration of beta-glucuronidase activity, on digestive gland cryotome sections. Additional chemical analyses (atomic absorption spectrophotometry) were done in the digestive gland of snails. The highest metal concentrations were found in snails from the active volcanic site, which agreed with high intralysomal Vv(BSD). Digestive cell lysosomes in snails inhabiting sites with active volcanism resembled a typical stress situation (enlarged and less numerous lysosomes). In conclusion, the biomarkers used in this work can be applied to detect changes in metal bioavailability due to chronic exposure to metals (volcanism), in combination with chemical analyses.

Science of The Total Environment, 2000

The occurrence of changes in flesh and shell weights and in other biometric parameters of mussels... more The occurrence of changes in flesh and shell weights and in other biometric parameters of mussels, Mytilus gallopro¨incialis, has been related to different metal levels found in their soft tissues. The effects of clean and Zn-polluted environments and laboratory experiments where Zn-polluted mussels were exposed to sublethal concentrations of Zn, Cu and Cd were investigated. Zinc-polluted mussel shell weights increased significantly after a 51-day depuration period. Exposure of Zn-polluted mussels to Zn or Cd, however, caused a slightly reduced shell growth in comparison with depurating mussel Cu-exposures not causing any reduction in growth. Apart from metal concentrations, metalrshell weight indices have been used to assess metal bioavailability. Metal concentrations recorded in the soft tissues of depurating mussels increased without a source of 'extra' metals, while the Znrshell-weight index was reduced, as expected from a depuration process, Cu and Cdrshell-weight indices remaining constant. Experimental exposure to Zn, Cu and Cd caused augmented values of Zn, Cu and Cdrshell-Ž weight indices, respectively. These different findings were attributed to changes in flesh weight related with gamete . spawning that would produce inconsistent estimates of whole metal concentration in soft tissues. Since changes in the tissue composition and in growth rates do not affect ZnrCu ratios and metalrshell-weight indices, these parameters are proposed as reliable indices of metal bioavailability for 'Mussel-Watch' monitoring programmes. The most sensitive parameter is the metalrshell-weight index, which is, therefore, highly recommended to be used in 'Mussel-Watch' monitoring programmes in order to determine metal bioavailability in seawaters. ᮊ M. Soto 0048-9697r00r$ -see front matter ᮊ 2000 Elsevier Science B.V. All rights reserved. Ž . PII: S 0 0 4 8 -9 6 9 7 9 9 0 0 4 8 9 -1 ( ) M. Soto et al. r The Science of the Total En¨ironment 247 2000 175᎐187 176

Science of The Total Environment, 1997

The tissue distribution of Zn in mussels collected from a Zn-polluted site and the changes in the... more The tissue distribution of Zn in mussels collected from a Zn-polluted site and the changes in the pattern of distribution for Zn and Cu in response to experimental exposures to Cu, Zn and Cd, as well as the tissue distribution of Cd have been studied. The organs of mussels were accurately separated avoiding mixtures except the kidney area that was included within the remainder tissues. Mussels exposed to intermediate dose (ID)-Cd showed the highest Cd burden fist in gills and then more or less equally distributed between the various tissues examined with a dominance of the digestive gland. Mussels exposed to high dose (HD)-Cd, showed Cd equally distributed through all the organs at both exposure times. Below 0.8 kg Cd/l seawater, the gills of Zn-polluted mussels were able to either protect against Cd entry or mobilise uptaken Cd towards other tissues. On the other hand, Zn-and Cu-burdens did not rise in gill tissue, although in depurating mussels Zn-levels were higher than in any other exposure group. Similar conclusions were obtained for the digestive gland. However, the accumulative capacity of the digestive gland tissue for Cd was not exhausted and Cu burdens were depressed on exposure to Cd. Interactions between metals in the various organs studied could be explained by redistribution of haemocytes or brown cells and by enhanced release of metal-bearing digestive lysosomes in response to metal exposure. It is suggested that (a) the pattern of metal accumulation and tissue distribution as well as a part of the interactions between Cu, Zn and Cd cannot be explained solely by the availability of metallothioneins in specific organs; and (b) the specific cell type in which these ligands occur as well as the relative abundance and distribution of these cells are relevant factors controlling metal burdens in organs of mussels. Therefore, in addition to the chemical and the biochemical approaches, investigations on tissue sections are required to better understand these phenomena. Meanwhile, the digestive gland of mussels could be used as the target organ to monitor metal pollution, including essential and non-essential metals, on the basis of metal/shell-weight indices. 0 1997 Elsevier Science B.V.

Microscopy Research and Technique, 2002

Marine Pollution Bulletin, 2011

The biological impact of a treated produced water (PW) was investigated under controlled laborato... more The biological impact of a treated produced water (PW) was investigated under controlled laboratory conditions in the blue mussel, Mytilus edulis. Mussel health status was assessed using an integrated biomarker approach in combination with chemical analysis of both water (with SPMDs), and mussel tissues. Acyl-CoA oxidase activity, neutral lipid accumulation, catalase activity, micronuclei formation, lysosomal membrane stability in digestive cells and haemocytes, cell-type composition in digestive gland epithelium, and the integrity of the digestive gland tissue were measured after 5 week exposure to 0%, 0.01%, 0.1%, 0.5% and 1% PW. The suite of biomarkers employed were sensitive to treated PW exposure with significant sublethal responses found at 0.01-0.5% PW, even though individual chemical compounds of PW were at extremely low concentrations in both water and mussel tissues. The study highlights the benefits of an integrated biomarker approach for determining the potential effects of exposure to complex mixtures at low concentrations. Biomarkers were integrated in the Integrative Biological Response (IBR/n) index.

Marine Pollution Bulletin, 2006

In order to assess the biological effects of the Prestige oil spill (POS), mussels (Mytilus gallo... more In order to assess the biological effects of the Prestige oil spill (POS), mussels (Mytilus galloprovincialis), European hake (Merluccius merluccius) and European anchovy (Engraulis encrasicolus) were sampled between April and September 2003 in various geographical areas of Bay of Biscay: Galicia, Central Cantabrian and East Cantabrian. In mussels, several cell and tissue biomarkers were measured: peroxisome proliferation as induction of acyl-CoA oxidase (AOX) activity, lysosomal responses as changes in the structure (lysosomal volume density, V(V(L)), surface-to-volume ratio, S/V(L), and numerical density, N(V(L))) and in membrane stability (labilization period, LP), cell type replacement as relative proportion of basophilic cells (volume density of basophilic cells, V(V(BAS))) in digestive gland epithelium, and changes in the morphology of digestive alveoli as mean luminal radius to mean epithelial thickness (MLR/MET). Additionally, flesh condition index (FCI) and gonad index (GI) were measured as supporting parameters. In hake and anchovy, liver histopathology was examined to determine the prevalence of parasites, melanomacrophage centers, non-specific lesions (inflammatory changes, atrophy, necrosis, apoptosis), early non-neoplastic toxicopathic lesions (i.e., hepatocellular nuclear polymorphism), foci of cellular alteration, benign and malignant neoplasms. In mussels, AOX induction was noticeable in April except in Galicia. LP values were low in all the geographical areas studied, indicating disturbed health, especially in Galicia. Alike, lysosomal enlargement was observed in most stations as shown by the extremely low S/V(L) values. V(V(BAS)) and MLR/MET values were markedly high. Overall, employed biomarkers detected exposure to toxic chemicals and disturbed health in mussels from Bay of Biscay, with impact decreasing from April to September. Although hepatocellular nuclear polymorphism and nematode parasitization in fish liver were remarkably prominent in some areas, they cannot be hitherto related to POS, since we lack historical data to determine whether the prevalences found were normal or significantly raised after POS.

Marine Pollution Bulletin, 2008

Fisheries have been vital to coastal communities around the North Sea for centuries, but this sem... more Fisheries have been vital to coastal communities around the North Sea for centuries, but this semi-enclosed sea also receives large amounts of waste. It is therefore important to monitor and control inputs of contaminants into the North Sea. Inputs of effluents from offshore oil and gas production platforms (produced water) in the Norwegian sector have been monitored through an integrated chemical and biological effects programme since 2001. The programme has used caged Atlantic cod and blue mussels. PAH tissue residues in blue mussels and PAH bile metabolites in cod have confirmed exposure to effluents, but there was variation between years. Results for a range of biological effects methods reflected exposure gradients and indicated that exposure levels were low and caused minor environmental impact at the deployment locations. There is a need to develop methods that are sufficiently sensitive to components in produced water at levels found in marine ecosystems.

Marine Environmental Research, 2006

To assess the biological effects of the Prestige oil spill (November 2002) on coastal ecosystems,... more To assess the biological effects of the Prestige oil spill (November 2002) on coastal ecosystems, mussels (Mytilus galloprovincialis) were sampled in 22 locations along the coast of Galicia and the Bay of Biscay in April, July and September 2003 and in February, April, July and October 2004. Several cell and tissue-level biomarkers were measured in digestive gland. Flesh condition index and gamete development stages were assessed as supporting parameters. AOX activity, a marker of exposure to peroxisome proliferating compounds including PAHs, was particularly low in Galicia in 2003 and further was markedly increased in several localities in 2004. Values of the labilization period (LP) of the lysosomal membrane were low in all the studied localities, especially in Galicia in 2003. In 2004, LP values raised, evidencing a certain recovery in mussel&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s health. In agreement, the volume density of basophilic cells was markedly high in 2003 and showed a decreasing trend throughout 2004. Parameters defining the structure of digestive alveoli showed few variations between 2003 and 2004. Significant correlations between several biomarkers and total polycyclic aromatic hydrocarbons (PAHs) were found. In conclusion, employed biomarkers detected highest degree of disturbance in areas most impacted by the oil spill (Galicia) and were able to evidence a recovery trend during 2004, related to a decrease in total PAH concentrations in mussels.

Marine Environmental Research, 2000

The aim of this study was to investigate the cellular and biochemical response of mussels, Mytilu... more The aim of this study was to investigate the cellular and biochemical response of mussels, Mytilus galloprovincialis, transplanted from a relatively pristine site to a polluted one and vice versa in the Lagoon of Venice (northeast of Italy) and to apply auto-metallography, a rapid and sensitive histochemical technique, to determine the bio-available fraction of heavy metals accumulated in the body tissues of organisms. Animal digestive glands have been used for morphological analyses (lysosomal volume, surface and numerical density, mean epithelial thickness, mean diverticular and luminar radius), autometallographical black silver deposits quantification and biochemical assays (superoxide dismutase and catalase activity). Furthermore, heavy metal content was determined by atomic absorption spectrophotometry (AAS) using standard procedures. The overall results indicate a direct influence of the environment in the thinning of the digestive cells and in the increasing number of lysosomes in mussels from the more polluted site. These data are in agreement with the metal content in digestive cell lysosomes as determined by autometallography, whereas AAS measurements show less significant differences.