Tiago Simões

Fatty acid profile of the sea snail Gibbula umbilicalis as a biomarker for coastal metal pollution

Metals are among the most common environmental pollutants with natural or anthropogenic origin that can be easily transferred through the food chain. Marine gastropods are known to accumulate high concentrations of these metals in their tissues. Gibbula umbilicalis ecological importance and abundant soft tissues, which enables extent biochemical assessments, makes this particular organism a potentially suitable species for marine ecotoxicological studies. Fatty acids are carbon-rich compounds that are ubiquitous in all organisms and easy to metabolize. Their biological specificity, relatively well-studied functions and importance, and the fact that they may alter when stress is induced, make fatty acids prospect biomarkers. This work aimed to assess fatty acid profile changes in the gastropod G. umbilicalis exposed to three metal contaminants. After a 168h exposure to cadmium, mercury, and nickel, the following lipid related endpoints were measured: total lipid content; lipid peroxidation; and fatty acid profile (FAP). The analysis of the FAP suggested an alteration in the fatty acid metabolism and indicated a link between metals exposure and homeoviscous adaptation and immune response. In particular, five fatty acids (palmitic, eicosatrienoic, arachidonic, eicosapentaenoic, and docosahexaenoic acids), demonstrated to be especially good indicators of G. umbilicalis responses to the array of metals used, having thus the potential to be used as biomarkers for metal contamination in this species. This work represents a first approach for the use of FAP signature as a sensitive and informative parameter and novel tool in environmental risk assessment (ERA) of coastal environments, using G. umbilicalis as model species.

An integrative omics approach to unravel toxicity mechanisms of environmental chemicals: effects of a formulated herbicide

The use of integrative molecular approaches can aid in a comprehensive understanding of the effects of toxicants at different levels of biological organization, also supporting risk assessment. The present study aims to unravel the toxicity mechanisms of a widely used herbicide to the arthropod Folsomia candida exposed in a natural soil, by linking effects on reproduction, proteomics and genome-wide gene expression. The EC50 effects on reproduction over 4 weeks was 4.63 mg glyphosate/kg of soil. The formulation included a polyethoxylated tallowamine as an adjuvant, which at 50% effect on reproduction had an estimated concentration of 0.87–1.49 mg/kg of soil. No effects were observed on survival and reproduction when using the isolated active substance, pointing the toxicity of the formulated product to the co-formulant instead of the active ingredient, glyphosate. RNA sequencing and shotgun proteomics were applied to assess differential transcript and protein expressions between exposed and control organisms in time, respectively. Specific functional categories at protein and transcriptome levels were concordant with each other, despite overall limited correlations between datasets. The exposure to this formulation affected normal cellular respiration and lipid metabolism, inducing oxidative stress and leading to impairment in biological life cycle mechanisms such as molting and reproduction.

Seasonal Variation in Proximate Composition and Fatty Acid Profile of Grey Triggerfish (Balistes capriscus) Captured Along the Coast of Portugal

Proximate composition (ash, moisture, total protein, and fat contents) and fatty acid profile of Balistes capriscus (grey triggerfish) were assessed over a 6-months period (April to September, 2011) in animals captured along the coast of Peniche (Portugal). High protein (18.9% to 21.4%) and low lipid (0.5% to 0.8%) contents were determined. The highest lipid level was found in June-captured animals. High proportions of n-3 polyunsaturated fatty acids (n-3 PUFA) were observed (53.87 ± 2.00%) together with substantial quantities of oleic acid (27.58 ± 1.09%) and palmitic acid (16.52 ± 0.93%). Docosahexaenoic acid accounted for 77% of the total PUFA and was 7 times more abundant than eicosapentaenoic acid, and the n-3/n-6 ratio was 7.2. Gender did not influence lipid levels and the fatty acid profile, but seasonal variations were observed for PUFA and monounsaturated fatty acid contents. Overall, the nutritional properties of this underexploited species may be comparable to those of other lean fish species with higher economic value.

Sex-steroids and hypolipidemic chemicals impacts on brown trout lipid and peroxisome signaling — Molecular, biochemical and morphological insights

Lipid metabolism involves complex pathways, which are regulated in a similar way across vertebrates. Hormonal and hypolipidemic deregulations cause lipid imbalance from fish to humans, but the underlying mechanisms are far from understood. This study explores the potential of using juvenile brown trout to evaluate the in vivo interferences caused by estrogenic (17α-ethinylestradiol - EE2), androgenic (testosterone - T), and hypolipidemic (clofibrate - CLF) compounds in lipidic and/or peroxisomal pathways. Studied endpoints were from blood/plasma biochemistry, plasma fatty acid profile, ultrastructure of hepatocytes and abundance of their peroxisomes to mRNA expression in the liver. Both T and CLF caused minimal effects when compared to EE2. Estrogenized fish had significantly higher hepatosomatic indexes, increased triglycerides and very-low density lipoproteins (VLDL) in plasma, compared with solvent control. Morphologically, EE2 fish showed increased lipid droplets in hepatocytes, and EE2 and T reduced volume density of peroxisomes in relation to the hepatic parenchyma. Polyunsaturated fatty acids (PUFA) in plasma, namely n-3 PUFA, increased with EE2. EE2 animals had increased mRNA levels of vitellogenin A (VtgA), estrogen receptor alpha (ERα), peroxisome proliferator-activated receptor alpha (PPARα), PPARαBa and acyl-CoA long chain synthetase 1 (Acsl1), while ERβ-1, acyl-CoA oxidase 1-3I (Acox1-3I), Acox3, PPARγ, catalase (Cat), urate oxidase (Uox), fatty acid binding protein 1 (Fabp1) and apolipoprotein AI (ApoAI) were down-regulated. In summary, in vivo EE2 exposure altered lipid metabolism and peroxisome dynamics in brown trout, namely by changing the mRNA levels of several genes. Our model can be used to study possible organism-level impacts, viz. in gonadogenesis.