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Purpose: Many cancer patients have problems eating which are usually connected to taste and smell alterations due to side effects of cancer treatment. These problems have consequences both in terms of malnutrition and reduced quality of life. In order to explore social and psychological consequences of eating problems in cancer patients, qualitative interviews were conducted with cancer patients, their caregivers and healthcare professionals., Methods: The study was conducted in three European countries (Poland, Spain and the UK) that differed in culture, oncology care approaches and availability of nutritional products targeted to cancer patients in the market., Results: Differences in the social role of eating between the three European countries were observed which subsequently influenced the impact of eating problems for cancer patients in these countries. Furthermore, the study found that problems with food affect not only the quality of life of cancer patients, but can also distress their caregivers, who are often unable to cope with such food-related problems. In addition, the study showed that commercially available nutritional products for cancer patients focus on nutritional value but tend to neglect an important aspect of eating, which is the enjoyment of food, both individually and socially., (© 2024. The Author(s).)

The levels of blood eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are very variable and, in general, low in most of the world population. In this study, the effects of age, sex, COVID-19, and dietary habits on the lipid profile of the erythrocyte membranes were assessed in a sub-cohort of healthy population (N = 203) from a large cohort of individuals from the Basque Country, Spain, (AKRIBEA). Sex did not have an effect on RBC lipid profile. COVID-19 infected participants showed higher levels of DGLA. Oldest participants showed higher oleic acid, EPA and DHA levels. Arachidonic acid in RBC correlated positively with the intake of sunflower oil, butter, eggs, processed and red meat, whereas DHA and EPA correlated positively with oily and lean fish. Basque Country population showed lipid profiles similar to other high fish consuming countries, such as Italy and Japan. Baseline levels of the whole lipidomic profile of the RBC including SFA, MUFA and PUFA should be examined to obtain a better description of the health and nutritional status., Competing Interests: Declaration of Competing Interest C.F. is co-founder and scientific director of the company Lipinutragen srl, born as spin-off recognized officially by the National Council of Research and involved in research and development of the membrane lipidomic analysis. The company had no role in the design of the study; in the collection and interpretation of data; in the writing of the manuscript, or in the decision to publish the results. The rest of the authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Adipokines, as well as the fatty acid profile of red blood cell (RBC) membranes, are known to play important roles in the development and progression of metabolic complications induced by obesity. Thus, the objective of this study is to compare the serum adipokine profile and the RBC membrane fatty acid profile of normal-weight and obese adults, and to analyze their relationship with serum biochemical parameters., Methods: An observational case-control study was performed in 75 normal-weight and obese adult subjects. Biochemical serum parameters, eight serum adipokines and the RBC membrane fatty acid profiles were measured. Associations between parameters were established using regression analysis., Results: Subjects with obesity showed increased levels of leptin, fibroblast growth factor 21 (FGF21) and overexpressed nephroblastoma (NOV/CCN3), decreased adiponectin, and similar levels of vaspin and chemerin compared to normal-weight subjects. Significant positive and negative correlations were found with triglycerides and high-density lipoprotein-cholesterol (HDL-c), respectively. An increase in the total ω-6 fatty acids in the RBC membrane fatty acid profiles in subjects with obesity was observed, because of higher levels of both dihomo-γ-linolenic acid (DGLA) and arachidonic acid (AA), and decreased total ω-3 fatty acids, mainly due to lower levels of docosahexaenoic acid (DHA). The ω-6/ω-3 ratio in the RBCs was significantly higher, suggesting an inflammatory status, as was also suggested by a reduced adiponectin level. A negative association between DGLA and adiponectin, and a positive association between DHA and serum triglycerides, was observed., Conclusions: Important alterations in serum adipokine and RBC fatty acid profiles are found in subjects with obesity.

Background: An optimal and correctly balanced metabolic status is essential to improve sports performance in athletes. Recent advances in omic tools, such as the lipid profile of the mature erythrocyte membranes (LPMEM), allow to have a comprehensive vision of the nutritional and metabolic status of these individuals to provide personalized recommendations for nutrients, specifically, the essential omega-3 and omega-6 fatty acids, individuating deficiencies/unbalances that can arise from both habitual diet and sportive activity. This work aimed to study the LPMEM in professional female football players during the football season for the first time and compare it with those defined as optimal values for the general population and a control group., Methods: An observational study was carried out on female football players from the Athletic Club (Bilbao) playing in the first division of the Spanish league. Blood samples were collected at three points: at the beginning, mid-season, and end of the season for three consecutive seasons (2019-2020, 2020-2021, and 2021-2022), providing a total of 160 samples from 40 women. The LPMEM analysis was obtained by GC-FID by published method and correlated to other individual data, such as blood biochemical parameters, body composition, and age., Results: We observed a significant increase in docosahexaenoic acid (DHA) ( p 0.048) and total polyunsaturated fatty acid (PUFA) ( p 0.021) in the first season. In the second season, we observed a buildup in the membrane arachidonic acid (AA) ( p  < .001) and PUFA ( p  < .001) contents when high training accumulated. In comparison with the benchmark of average population values, 69% of the football players showed lower levels of omega-6 dihomo-γ-linolenic acid (DGLA), whereas 88%, 44%, and 81% of the participants showed increased values of AA, eicosapentaenoic acid (EPA), and the ratio of saturated and monounsaturated fatty acids (SFA/MUFA), respectively. Regarding relationships between blood biochemical parameters, body composition, and age with LPMEM, we observed some mild negative correlations, such as AA and SFA/MUFA ratio with vitamin D levels (coefficient = -0.34 p = .0019 and coefficient = -.25 p  = .042); DGLA with urea and cortisol (coefficient = -0.27 p  < .006 and coefficient = .28 p  < .0028) and AA with age (coefficient = -0.33 p  < .001)., Conclusion: In conclusion, relevant variations in several fatty acids of the membrane fatty acid profile of elite female football players were observed during the competitive season and, in comparison with the general population, increased PUFA contents were confirmed, as reported in other sportive activities, together with the new aspect of DGLA diminution, an omega-6 involved in immune and anti-inflammatory responses. Our results highlight membrane lipidomics as a tool to ascertain the molecular profile of elite female football players with a potential application for future personalized nutritional strategies (diet and supplementation) to address unbalances created during the competitive season.

The social, environmental and health concerns associated with the massive consumption of meat products has resulted in calls for a reduction in meat consumption. A simplex lattice design was used for studying the effect of combining broccoli, upcycled brewer’s spent grain (BSG) and insect flours from Tenebrio molitor (IF) as alternative sources of protein and micronutrients, in hybrid sausages formulation. The techno-functional properties of the ingredients and the nutritional and textural properties of nine hybrid sausages were analysed. The effect of adding these ingredients (constituting 35% of a turkey-based sausage) on protein, fat, fibre, iron and zinc content, and textural properties (Texture Profile Analysis (TPA) and Warner−Bratzler parameters) were modelled employing linear regression (0.72 < R2 < 1). The “desirability” function was used for multi-response optimisation of the samples for the highest protein content, optimum chewiness and a* value (closeness to red). The analysis of sensory data for the three optimised samples showed no significant differences in juiciness and odour between the hybrid meat sausage with 22% broccoli, 3% BSG, and 10% IF and the commercial Bratwurst sausage elaborated exclusively with animal protein. Colour, appearance, chewiness and pastiness were rated higher than for the reference. The instrumental chewiness highly correlated with sensorial chewiness (R2 = 0.98). Thus, a strategy introducing less refined and more sustainable sources of protein and micronutrients was successfully employed to model and statistically optimise a meat product formulation with reduced animal protein content., Competing Interests: The authors declare no conflict of interest.

Fatty acids have an important place in both biological and nutritional contexts and, from a clinical point of view, they have known consequences for diseases' onset and development, including cancer. The use of fatty acid-based food and nutraceuticals to support cancer therapy is a multidisciplinary subject, involving molecular and clinical research. Knowledge regarding polyunsaturated fatty acids essentiality/oxidizability and the role of lipogenesis-desaturase pathways for cell growth, as well as oxidative reactivity in cancer cells, are discussed, since they can drive the choice of fatty acids using their multiple roles to support antitumoral drug activity. The central role of membrane fatty acid composition is highlighted for the application of membrane lipid therapy. As fatty acids are also known as biomarkers of cancer onset and progression, the personalization of the fatty acid-based therapy is also possible, taking into account other important factors such as formulation, bioavailability and the distribution of the supplementation. A holistic approach emerges combining nutra- and pharma-strategies in an appropriate manner, to develop further knowledge and applications in cancer therapy.

This work intends to describe the physical properties of red blood cell (RBC) membranes in obese adults. The hypothesis driving this research is that obesity, in addition to increasing the amount of body fat, will also modify the lipid composition of membranes in cells other than adipocytes. Forty-nine control volunteers (16 male, 33 female, BMI 21.8 ± 5.6 and 21.5 ± 4.2 kg/m 2 , respectively) and 52 obese subjects (16 male and 36 female, BMI 38.2± 11.0 and 40.7 ± 8.7 kg/m 2 , respectively) were examined. The two physical techniques applied were atomic force microscopy (AFM) in the force spectroscopy mode, which allows the micromechanical measurement of penetration forces, and fluorescence anisotropy of trimethylammonium diphenylhexatriene (TMA-DPH), which provides information on lipid order at the membrane polar-nonpolar interface. These techniques, in combination with lipidomic studies, revealed a decreased rigidity in the interfacial region of the RBC membranes of obese as compared to control patients, related to parallel changes in lipid composition. Lipidomic data show an increase in the cholesterol/phospholipid mole ratio and a decrease in sphingomyelin contents in obese membranes. ω-3 fatty acids (e.g., docosahexaenoic acid) appear to be less prevalent in obese patient RBCs, and this is the case for both the global fatty acid distribution and for the individual major lipids in the membrane phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS). Moreover, some ω-6 fatty acids (e.g., arachidonic acid) are increased in obese patient RBCs. The switch from ω-3 to ω-6 lipids in obese subjects could be a major factor explaining the higher interfacial fluidity in obese patient RBC membranes.

Metabolically healthy obesity (MHO) has been described as BMI ≥ 30 kg/m 2 , without metabolic disorders traditionally associated with obesity. Beyond this definition, a standardized criterion, for adults and children, has not been established yet to explain the absence of those metabolic disorders. In this context, biomarkers of inflammation have been proposed as suitable candidates to describe MHO. The use of mature red blood cell fatty acid (RBC FA) profile is here proposed since its membrane lipidome includes biomarkers of pro- and anti-inflammatory conditions with a strict relationship with metabolic and nutritional status. An observational study was carried out in 194 children (76 children with obesity and 118 children with normal weight) between 6 and 16 years old. RBC FAs were analyzed by gas chromatography-flame ionization detector (GC-FID). An unsupervised hierarchical clustering method was conducted on children with obesity, based on the RBC FA profile, to isolate the MHO cluster. The MHO cluster showed FA levels similar to children with normal weight, characterized by lower values of arachidonic acid, (total ω-6 FA, ω6/ω3 FA ratios and higher values for EPA, DHA, and total ω-3 FA) (for all of them p ≤ 0.01) compared to the rest of the children with obesity (obese cluster). The MHO cluster also presented lipid indexes for higher desaturase enzymatic activity and lower SFA/MUFA ratio compared to the obese cluster. These differences are relevant for the follow-up of patients, also in view of personalized protocols providing tailored nutritional recommendations for the essential fatty acid intakes.

As the obesity epidemic continues to grow inexorably worldwide, the need to develop effective strategies to prevent and control obesity seems crucial. The use of molecular tools can be useful to characterize different obesity phenotypes to provide more precise nutritional recommendations. This study aimed to determine the fatty acid (FA) profile of red blood cell (RBC) membranes, together with the evaluation of their dietary intake and biochemical parameters, of children and adults with obesity. An observational study was carried out on 196 children (113 with normal weight and 83 with obesity) and 91 adults (30 with normal weight and 61 with obesity). Mature RBC membrane phospholipids were analyzed for FA composition by gas chromatography-mass spectrometry (GC-MS). Dietary habits were evaluated using validated food frequency questionnaires (FFQ). Children with obesity presented higher levels of ω-6 polyunsaturated FAs (mainly linoleic acid, p = 0.01) and lower values of ω-3 FAs (mainly DHA, p < 0.001) compared with adults. Regarding blood biochemical parameters, children with obesity presented lower levels of glucose, LDL cholesterol, and alanine aminotransferase compared with adults with obesity. These lipidomic differences could be considered to provide specific nutritional recommendations for different age groups, based on an adequate fat intake.

Red blood cell (RBC) membrane can reflect fatty acid (FA) contribution from diet and biosynthesis. In cancer, membrane FAs are involved in tumorigenesis and invasiveness, and are indicated as biomarkers to monitor the disease evolution as well as potential targets for therapies and nutritional strategies. The present study provides RBC membrane FA profiles in recently diagnosed breast cancer patients before starting chemotherapy treatment. Patients and controls were recruited, and their dietary habits were collected. FA lipidomic analysis of mature erythrocyte membrane phospholipids in blood samples was performed. Data were adjusted to correct for the effects of diet, body mass index (BMI), and age, revealing that patients showed lower levels of saturated fatty acids (SFA) and higher levels of monounsaturated fatty acid, cis-vaccenic (25%) than controls, with consequent differences in desaturase enzymatic index (∆9 desaturase, -13.1%). In the case of polyunsaturated fatty acids (PUFA), patients had higher values of ω-6 FA (C18:2 (+11.1%); C20:4 (+7.4%)). RBC membrane lipidomic analysis in breast cancer revealed that ω-6 pathways are favored. These results suggest new potential targets for treatments and better nutritional guidelines.

Obesity is a chronic metabolic disease of high complexity and of multifactorial origin. Understanding the effects of nutrition on childhood obesity metabolism remains a challenge. The aim of this study was to determine the fatty acid (FA) profile of red blood cell (RBC) membranes as a comprehensive biomarker of children's obesity metabolism, together with the evaluation of their dietary intake. An observational study was carried out on 209 children (107 healthy controls, 41 who were overweight and 61 with obesity) between 6 and 16 years of age. Mature RBC membrane phospholipids were analyzed for FA composition by gas chromatography-mass spectrometry (GC-MS). Dietary habits were evaluated using validated food frequency questionnaires (FFQ) and the Mediterranean Diet Quality Index for children (KIDMED) test. Compared to children with normal weight, children with obesity showed an inflammatory profile in mature RBC FAs, evidenced by higher levels of ω-6 polyunsaturated FAs (mainly arachidonic acid, p < 0.001). Children who were overweight or obese presented lower levels of monounsaturated FA (MUFA) compared to children with normal weight ( p = 0.001 and p = 0.03, respectively), resulting in an increased saturated fatty acid (SFA)/MUFA ratio. A lower intake of nuts was observed for children with obesity. A comprehensive membrane lipidomic profile approach in children with obesity will contribute to a better understanding of the metabolic differences present in these individuals.

Fatty acids are closely involved in lipid synthesis and metabolism in cancer. Their amount and composition are dependent on dietary supply and tumor microenviroment. Research in this subject highlighted the crucial event of membrane formation, which is regulated by the fatty acids' molecular properties. The growing understanding of the pathways that create the fatty acid pool needed for cell replication is the result of lipidomics studies, also envisaging novel fatty acid biosynthesis and fatty acid-mediated signaling. Fatty acid-driven mechanisms and biological effects in cancer onset, growth and metastasis have been elucidated, recognizing the importance of polyunsaturated molecules and the balance between omega-6 and omega-3 families. Saturated and monounsaturated fatty acids are biomarkers in several types of cancer, and their characterization in cell membranes and exosomes is under development for diagnostic purposes. Desaturase enzymatic activity with unprecedented de novo polyunsaturated fatty acid (PUFA) synthesis is considered the recent breakthrough in this scenario. Together with the link between obesity and cancer, fatty acids open interesting perspectives for biomarker discovery and nutritional strategies to control cancer, also in combination with therapies. All these subjects are described using an integrated approach taking into account biochemical, biological and analytical aspects, delineating innovations in cancer prevention, diagnostics and treatments.

Purpose: Taste and smell changes are common side effects in cancer patients undergoing chemotherapy treatments (CT). This can lead to a reduced food enjoyment and an inadequate nutrient intake with a high impact on nutritional status and quality of life. The aim of this study was to evaluate the self-reported chemosensory alterations of patients undergoing chemotherapy according to CT type., Methods: An observational study was conducted with 151 patients undergoing CT at Oncology Outpatient Unit from Onkologikoa Foundation. An interviewer-assisted questionnaire was designed to investigate chemosensory changes in patients undergoing CT., Results: Seventy-six percent patients reported taste disorders and 45% smell changes. Xerostomia is the most frequent symptom reported by patients receiving chemotherapy in our study (63.6%), and it is strongly associated to bad taste in mouth (OR = 5.96; CI = 2.37-14.94; p value = 0.000) and taste loss (OR = 5.96; CI = 2.37-14.94; p value = 0.000). Anthracyclines, paclitaxel, carboplatin, and docetaxel were the CT agents producing the highest taste disturbance rates. Cisplatin and 5-Fluorouracil are the CT resulting in the lowest complaints. Logistic regression revealed statistically significant associations between taste loss and carboplatin and docetaxel (OR = 3.50; CI = 1.12-10.90; p value = 0.031) and cold hypersensitivity and oxaliplatin (OR = 12.14; CI = 4.18-35.25; p value = 0.000). Not only platin-based CT such as carboplatin produced dysgeusia, but also anthracyclines and paclitaxel treatments., Conclusions: The better knowledge of taste and smell alterations according to CT type may provide valuable information for the design of new strategies to tackle CT side effects. It is important to take into account taste and smell dysfunctions and other alterations such as xerostomia together.

The fatty acid (FA) composition of red blood cell (RBC) membrane phospholipids of cancer patients can reflect tumor status, dietary intakes, and cancer type or therapy. However, the characteristic membrane profiles have so far not yet defined as a potential biomarker to monitor disease evolution. The present work provides the first evidence of cancer metabolic signatures affecting cell membranes that are independent of nutritional habits. From the Oncology Outpatient Unit of the Onkologikoa hospital, two groups of cancer patients ( n = 54) and healthy controls ( n = 37) were recruited, and mature RBCs membrane phospholipids were analyzed for FA profiling (GC-MS). Dietary habits were evaluated using a validated food frequency questionnaire. The adjusted Analysis of Covariance Test (ANCOVA) model revealed cancer patients to have a lower relative percentage of saturated fatty acids (SFA) (C16:0 (5.7%); C18:0 (15.9%)), and higher monounsaturated fatty acids (MUFA) (9c-C18:1 (12.9%) and 11c-C18:1 (54.5%)), compared to controls. In line with this, we observe that the desaturase enzymatic index (delta-9 desaturase (Δ9D), +28.3%) and the membrane saturation index (SI = SFA/MUFA; -27.3%) were similarly modulated. Polyunsaturated fatty acids (PUFA) families showed an increase of n -6 C18:2 and C20:3 (15.7% and 22.2% respectively), with no differences in n -6 C20:4 and n -3 PUFA (docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA)). Importantly, these changes were found independent of foods and fat intakes from the diet. The membrane lipid profile in RBC was useful to ascertain the presence of two main metabolic signatures of increased desaturation activity and omega-6 in cancer patients, statistically independent from dietary habits.

Docosahexaenoic acid (DHA) is a semiessential polyunsaturated fatty acid (PUFA) for eukaryotic cells that is found in natural sources such as fish and algal oils and widely used as an ingredient for omega-3 containing foods or supplements. DHA effects are connected to its natural structure with six cis double bonds, but geometrical monotrans isomers can be formed during distillation or deodorization processes, as an unwanted event that alters molecular characteristics and annihilates health benefits. The characterization of the six monotrans DHA regioisomers is an open issue to address for analytical, biological, and nutraceutical applications. Here we report the preparation, separation, and first identification of each isomer by a dual approach consisting of the following: (i) the direct thiyl radical-catalyzed isomerization of cis-DHA methyl ester and (ii) the two-step synthesis from cis-DHA methyl ester via monoepoxides as intermediates, which are separated and identified by nuclear magnetic resonance spectroscopy, followed by elimination for the unequivocal assignment of the double bond position. This monotrans DHA isomer library with NMR and GC analytical characterization was also used to examine the products of thiyl-radical-catalyzed isomerization of a fish oil sample and to evaluate the trans isomer content in omega-3 containing supplements commercially available in Italy and Spain.

One of the main challenges for cancer patients under treatment is to prevent and tackle malnutrition. The current clinical nutrition market offers different food supplements or oral nutritional support products (mainly milkshakes or modified texture products) for cancer patients under risk of malnutrition. However, it is worth mentioning that these products do not address the pleasure of eating, since they do not meet sensory requirements, such as taste and smell alterations, nor patients' food preferences, leading to a big impact on their quality of life (QOL). Still, controversy remains regarding the specific nutritional requirements for cancer patients during the disease. Several randomized controlled clinical trials yield opposite results when using different bioactive compounds such as omega-3 fatty acids or antioxidants in order to prevent malnutrition or improve QOL. The use of 'omics' technologies in oncology, such as membrane lipidomics, as a powerful tool to provide new insights for the understanding of diet and cancer and their interacting metabolic pathways, will be discussed. The better knowledge of specific requirements (nutrients, sensory parameters and food preferences) for cancer patients provides valuable information for the food industry in the design of customized food products capable of preventing malnutrition, alleviating symptoms and improving QOL. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Resveratrol, a polyphenolic phytoalexin found in many plants, has been reported to have antiobesogenic effects in several animal and in vitro models. Zebrafish present several technical advantages that place them at an interesting, halfway point between in vitro and rodent models. The aim of the present work was to evaluate the metabolization of resveratrol and its glucoside (piceid) in zebrafish and their ability to induce the consumption of fat reserve in zebrafish larvae. Resveratrol and piceid were both able to reduce yolk sac fat content depending on the dose tested. Furthermore, resveratrol showed a potent and rapid action, whereas piceid needed more time and higher doses to be as effective as resveratrol. In accordance with other animal models and humans, the principal metabolites found in zebrafish larvae were monoglucoronide and monosulfate forms of resveratrol. In conclusion, zebrafish are a potentially excellent animal model for polyphenol research as they present several advantageous characteristics for efficacy screening and metabolomic studies before rodents.

By-products generated every year by the European fruit and cereal processing industry currently exceed several million tons. They are disposed of mainly through landfills and thus are largely unexploited sources of several valuable biobased compounds potentially profitable in the formulation of novel food products. The opportunity to design novel strategies to turn them into added value products and food ingredients via novel and sustainable processes is the main target of recently EC-funded FP7 project NAMASTE-EU. NAMASTE-EU aims at developing new laboratory-scale protocols and processes for the exploitation of citrus processing by-products and wheat bran surpluses via the production of ingredients useful for the formulation of new beverage and food products. Among the main results achieved in the first two years of the project, there are the development and assessment of procedures for the selection, stabilization and the physical/biological treatment of citrus and wheat processing by-products, the obtainment and recovery of some bioactive molecules and ingredients and the development of procedures for assessing the quality of the obtained ingredients and for their exploitation in the preparation of new food products., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The European Water Framework Directive (WFD) establishes a framework for the protection and improvement of estuarine (transitional) and coastal waters, attempting to achieve good water status by 2015; this includes, within the assessment, biological and chemical elements. The European Commission has proposed a list of priority dangerous substances (including metals such as Cd, Hg, Ni and Pb), with the corresponding list of environmental quality standards (EQS), to assess chemical status, but only for waters. In this contribution, a long-term (1995-2007) dataset of transitional and coastal water and sediment trace elements concentrations, from the Basque Country (northern Spain), has been used to investigate the response of these systems to water treatment programmes. Moreover, the approach proposed in the WFD, for assessing water chemical status (the 'one out, all out' approach), is compared with the integration of water and sediment data, into a unique assessment. For this exercise, background levels are used as reference conditions, identifying the boundary between high and good chemical status. EQS are used as the boundary between good and moderate chemical status. This contribution reveals that the first approach can lead to misclassification, with the second approach representing the pattern shown by the long-term data trends. Finally, the management implications, using each approach are discussed.

The European Water Framework Directive (WFD) establishes a framework for the protection of estuarine and coastal waters, with the most important objective being to achieve 'good ecological status' for all waters, by 2015. Hence, Member States are establishing programmes for the monitoring of water quality status, through the assessment of ecological and chemical elements. These monitoring programmes can be of three types: surveillance monitoring; operational monitoring (both undertaken on a routine basis); and investigative monitoring (carried out where the reason of any exceedance for ecological and chemical status is unknown). Until now, nothing has been developed in relation to investigative monitoring and no clear guidance exists for this type of monitoring, as it must be tackled on a 'case-by-case' basis. Consequently, the present study uses slag disposal from a blast furnace, into a coastal area, as a case-study in the implementation of investigative monitoring, according to the WFD. In order to investigate the potential threat of such slags, this contribution includes: a geophysical study, to determine the extent of the disposal area; sediment analysis; a chemical metal analysis; and an ecotoxicological study (including a Microtox test and an amphipod bioassay). The results show that metal concentrations are several times above the background concentration. However, only one of the stations showed toxicity after acute toxicological tests, with the benthic communities being in a good status. The approaches used here show that contaminants are not bioavailable and that no management actions are required with the slags.