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- Unveiling the geographical influence on honey's physicochemical profile: a case study from Montesinho Natural ParkPublication . Soares, Sónia; Magalhães, Leandro; Moreira, Manuela M.; Rede, Diana; Fernandes, Virgínia Cruz; Viegas, Olga; Pinto, Edgar; Almeida, Agostinho; Azevedo, Rui; Delerue-Matos, CristinaIn recent years, global honey consumption has significantly increased due to the rising demand for natural products with health benefits. Consumers are particularly drawn to honey recognized for its superior quality, such as Protected Designation of Origin (PDO) honey or honey produced in protected areas, which is associated with distinctive characteristics. Portugal stands among the top producers of PDO honey, with Montesinho Natural Park (MNP) being a notable example of high-quality Portuguese honey. Despite its reputation, environmental challenges threaten both honey production and quality in this region. This study aims to evaluate the physicochemical parameters and nutritional characteristics of honey from MNP and investigate the correlation between these properties and the geographical location of the apiaries. Honey samples (n=13) were collected from local producers and supermarkets. Standard physicochemical parameters, including 5-hydroxymethylfurfural (HMF), diastase activity, moisture and ash content, free acidity, electrical conductivity, and pH, were analysed following honey legislation guidelines. Additional parameters, such as colour, protein content, low-molecular-weight carbohydrates, and mineral composition, were also assessed. The findings indicate that all honey samples complied with legal standards and align with previously published data, showing a high degree of homogeneity across the physicochemical properties analysed. Principal Component Analysis (PCA) revealed that parameters such as colour, HMF, and specific minerals (Mg, Ca, K, Mn, Sr, Cu, Ba) contributed to the clustering of honey samples, suggesting that geographical variations among apiary locations might influence these properties. However, these differences do not affect the overall quality of the PDO honey produced in MNP. This study represents the first comprehensive characterization of honey from different apiaries within MNP. While the results demonstrate the consistency and quality of MNP honey, further research is necessary to explore the relationship between geographic origin and quality under varying environmental conditions. These insights could be valuable for maintaining sustainable beekeeping practices in the region and preserving the unique attributes of MNP honey.
- Effects of cylindrospermopsin and glyphosate at environmentally relevant concentrations on growth and mineral content of beetroot plants (Beta vulgaris)Publication . Priya, V.; Freitas, Marisa; Pinto, Edgar; Almeida, A.; Oliveira, F.; Azevedo, J.; Campos, A.; Sudhakaran, R.; Vasconcelos, V.Natural toxins produced by freshwater cyanobacteria, such as cylindrospermopsin (CYN), have been regarded as an emergent environmental threat. Cyanotoxins can be applied directly to soil by using contaminated water for agricultural irrigation. Despite the risks for food safety, the impact of cyanotoxins in agriculture is not yet fully understood. Furthermore, in soil-plant system the simultaneous occurrence of cyanotoxins and pesticides can be highly expectable. Pesticides, especially residues of glyphosate (GLY), have been frequently detected in soils and have been recognized to contribute to soil toxicity. Beetroot (Beta vulgaris) are root vegetables, extensively consumed worldwide with great importance for human nourishment and economy. It is, therefore, important to evaluate the effects of using water contaminated with CYN and GLY on beetroot cultivation. This study aimed to assess the effects of environmentally relevant concentrations of CYN, GLY and a mixture of both on growth and mineral content of beetroot (roots and leaves) cultivated in soil system. Plants were exposed in controlled conditions to CYN-crude extracts (50μg/kg) (Chrysosporum ovalisporum culture - LEGE X-001), isolated and in mixture with GLY (750 μg/kg) for 4 months. Beetroot growth was assessed by determining its fresh (Fw) and dry weight (Dw). The determination of mineral content was made by inductively coupled plasma-mass spectrometry (ICP-MS), after sample mineralization by microwaveassisted acid digestion. The results denote that Fw and Dw of beetroot (leaves and roots, respectively) were significantly changed (P
- Cylindrospermopsin and glyphosate accumulation in lettuce (Lactuca sativa) simultaneously exposed to both toxicants in hydroponic and soil systemsPublication . Sengupta, S.; Freitas, Marisa; Pinto, Edgar; Ferreira, I.; Oliveira, F.; Azevedo, J.; Prieto, A.; Diez-Quijada, L.; Jos, A.; Cameán, A.M.; Campos, A.; Vasconcelos, V.In nature, the simultaneous occurrence of multiple emergent contaminants such as cyanotoxins (e.g., cylindrospermopsin (CYN)) and herbicides (e.g., glyphosate (GLY)), is highly expectable and it can be anticipated, mainly in the aquatic and terrestrial environments. The use of contaminated water for irrigation can be hazardous to the agricultural sector and some studies have reported that, individually, these contaminants can be accumulated in the edible tissues exerting a negative influence on crop plants safety and ultimately in human health. Furthermore, recent studies have suggested that some cyanotoxins (e.g., microcystins) can change the membrane permeability of roots, resulting in changes in the accumulation rates of other contaminants in plants. Since edible plants are exposed to a wide variety of substances through irrigation water, there is increasing concern in the potential adverse effects of the interactions between those substances when present simultaneously, especially when this can have potential public health consequences. The aim of this study was to determine the accumulation of CYN in Lactuca sativa simultaneously exposed to GLY at environmentally relevant concentrations. Lettuce plants were exposed for 15 days to 50 μg/L or kg of CYN-containing crude extract (Chrysosporum ovalisporum culture - LEGE X001) and 750 μg/L or kg of GLY, in hydroponic and soil systems, respectively. The resins (HP20, SP700, and XAD18) were evaluated for MC-LR uptake kinetics, capacities, and extraction efficiencies and simple procedures were developed for determining MC-LR concentration in binding disc extracts by Adda-ELISA (U.S. EPA Method 546). The XAD18-DGT/Adda-ELISA method had a 7-d deployment time detection limit of ≈0.05 μg/L and capacity of > 250 μg/L of MC-LR in water samples which encompass U.S. EPA and WHO advisory concentrations for drinking and recreational waters. The XAD18DGT/Adda-ELISA method determined timeaveraged MC-LR concentrations in waters with wide ranging pH (4.9–8.3) and ionic strength (0.04–0.8 M) under well-stirred and quiescent conditions with 90–101% accuracy. In addition to high sensitivity and accuracy, the method is simple, inexpensive, and applicable for determining MC-LR and related MCs concentrations in waterbodies with wide ranging chemical characteristics and hydrodynamic conditions. 4.11.07 Cylindrospermopsin and Glyphosate Accumulation in Lettuce (Lactuca sativa) Simultaneously Exposed to Both Toxicants in Hydroponic and Soil Systems S. Sengupta, School of Bio Sciences and Technology Vellore Institute of Technology, Vellore - 632 014, Tamil Nadu, India; M.M. Freitas, School of Health, Polytechnic Institute of Porto; E. Pinto, I. Ferreira, LAQV/REQUIMTE, Departament of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua Jorge de Viterbo Ferreira 228, 4050313 Porto, Portugal; F. Oliveira, J. Azevedo, CIIMAR - Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos, s/n, 4450–208 Porto; A. Prieto, L. Diez-Quijada, A. Jos, A.M. Cameán, Area of Toxicology, Faculty of Pharmacy, Universidad de Sevilla, Profesor García González n◦2, 41012 Seville, Spain / Toxicology; A. Campos, Interdisciplinary Centre of Marine and Environmental Sciences, CIIMAR; V. Vasconcelos, CIIMAR - Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos, s/n, 4450–208 Porto In nature, the simultaneous occurrence of multiple emergent contaminants such as cyanotoxins (e.g., cylindrospermopsin (CYN)) and herbicides (e.g., glyphosate (GLY)), is highly expectable and it can be anticipated, mainly in the aquatic and terrestrial environments. The use of contaminated water for irrigation can be hazardous to the agricultural sector and some studies have reported that, individually, these contaminants can be accumulated in the edible tissues exerting a negative influence on crop plants safety and ultimately in human health. Furthermore, recent studies have suggested that some cyanotoxins (e.g., microcystins) can change the membrane permeability of roots, resulting in changes in the accumulation rates of other contaminants in plants. Since edible plants are exposed to a wide variety of substances through irrigation water, there is increasing concern in the potential adverse effects of the interactions between those substances when present simultaneously, especially when this can have potential public health consequences. The aim of this study was to determine the concentration of CYN and GLY in lettuce plants (roots and leaves) was determined by LC/MSMS. The results show that, at the described conditions, CYN was accumulated in roots (0.06-7.62 μg CYN/g Dw) and leaves (0.13-1.1 μg CYN/g Dw) of lettuce, especially when plants were exposed in hydroponic system. However, interestingly, when lettuce plants were exposed simultaneously to both toxicants the concentration of CYN assimilated by lettuce plants (roots and leaves) was respectively, 1.5fold and 1-3-2.2-fold lower than in the exposure to isolated CYN. Conversely, the plants exposed to the mixture in soil system, showed that the concentration of GLY incorporated by lettuce (roots and leaves) was higher than in the exposure to the isolated compound (0.04 - 0.21 µg GLY/g and < LOQ - 0.84 µg GLY/g, respectively). This finding highlights the potential for the enhancement of GLY accumulation in lettuce plants due to their cooccurrence with CYN, and it underlines the importance of further research regarding the mechanism involved.
- Trace element imbalances in hemodialysis patients: new data from a cohort of Portuguese patientsPublication . Duro, Mary; Azevedo, Rui; Costa, Félix; Pinto, Edgar; Almeida, AgostinhoPatients with end-stage renal disease undergoing hemodialysis (HD) therapy are at risk of developing deficiencies of essential trace elements and/or overload of toxic trace elements, both of which may significantly affect their clinical status. Those imbalances may result from the disease itself but also from the therapy, namely the hemodialysis process. This work aimed at evaluate the current trace elements status in a cohort of patients under hemodialysis therapy. Serum (n=88) and whole blood samples (n=108) from HD patients, collected during the routine periodic control of Al levels, were analyzed for a wide panel of trace elements using inductively coupled plasma mass spectrometry (ICP-MS) (HD group). For comparison purposes, both serum (n=16) and whole blood samples (n=59) of individuals attending the same Clinical Laboratory but with no evidence of renal disease according to standard laboratory analytical criteria were also analysed (Control group).
- Low serum bromine levels in chronic hemodialysis patients - is there any clinical impact?Publication . Duro, Mary; Novakova, Gergana; Bonev, Presian; Azevedo, Rui; Couto, Cristina; Pinto, Edgar; Almeida, AgostinhoPatients on chronic hemodialysis therapy are at high risk for disturbed trace element status due to both the underlying disease and the hemodialysis process itself. Data on serum bromine levels in these patients are scarce. Using an ICP-MS analytical procedure, serum bromine levels were determined in a cohort of end-stage renal disease patients on chronic hemodialysis (n=57; 68.9±14.0 years old; male/female ratio: 1.37). The results were compared with those of a control group (individuals without evidence of kidney disease according to standard clinical laboratory criteria who attended the same clinical laboratory for routine analyses: n=59; 57.4±17.9 years old; male/female ratio: 0.90). Hemodialysis patients had much lower serum bromine levels than controls: 1086±244 vs. 4137±770 µg/L; P<0.0001. Bromine (bromide in plasma) showed to be extensively removed from plasma due to the hemodialysis process. The actual impact of this bromine “deficiency” is unknown. Bromine has not generally been considered an “essential” element, however, evidence of possible essentiality has been growing. In particular, bromine has been linked to brain metabolism: it is used as a sedative to induce sleep, increases in animals during hibernation as well as in sleeping humans; and a bromine compound with REM sleep-inducing and anti-choline esterase activities (1-methylheptyl gammabromoacetoacetate) was identified in human cerebrospinal fluid. On the other hand, estimates suggest that 40-85% of hemodialysis patients have sleep disorders, mainly insomnia. Thus, the possibility has been raised that the bromine “deficiency” found in these patients may be associated with sleep the disorders that affect them. This is an interesting hypothesis that requires further clinical investigation.
- A detailed and comparative elemental analysis (Poland vs. Portugal) of espresso coffeePublication . Almeida, Agostinho; Cunha, Ana; Pinto, Edgar; Azevedo, Rui; Seroczynska, Kinga; Konieczynski, Pawel; Wesolowski, MarekCoffee is one of the most popular beverages in the world. There are many different types of coffee, but “espresso” has become the favourite in many countries. In Portugal, 80% of people drink coffee daily and prefer to make it as an espresso. Annual per capita coffee consumption (dry beans) is estimated at 5 kg, being higher in many other Western European countries, namely the Nordic countries. In Poland, also more than 80% of adults consume coffee every day, but instant coffee still takes up the largest share. National per capita consumption (2021) is estimated at between 0.72.5 kg. Espresso coffee is prepared by passing water at high temperature (88-92 ºC) and under high pressure (8-10 bar) through ground coffee (6-8 g) for 20-30 seconds to produce around 25-40 ml of beverage. The extraction of substances existing in coffee powder is very efficient, so their concentration in coffee (beverage) is also very high. In this work, we carried out a comparative study of the elemental composition (n=34 macrominerals and trace elements) of espresso coffee from two different European origins: Northern Poland (n=60, areas of Gdańsk, Gdynia and Elbląg) and Northern Portugal (n=22, Porto Metropolitan Area). Samples (5-10 ml) were taken from the beverage as served in cafes, restaurants and gas stations. In order to better document the contribution of coffee (powder) to the final composition of the beverage, water samples were taken in parallel at the same locations. The analysis was performed by ICP-MS. In both cases, espresso coffee proved to be a concentrated source of many elements (Table 1), with levels much higher than those of water from the same location. Polish waters were globally much more mineralized than Portuguese waters, and generally Polish coffee also showed significantly higher levels of macrominerals and trace elements than the Portuguese ones.
- Association between aluminium levels and cognitive decline in environmentally exposed eldersPublication . Gerardo, Bianca; Pinto, Marina Cabral; Nogueira, Joana; Almeida, Agostinho; Pinto, Edgar; Simões, Mário Rodrigues; Freitas, SandraNormal brain functioning depends on metals homeostasis, as imbalances may increase oxidative stress and cause other detrimental intracellular events that lead to cell loss. Furthermore, environmental exposure to toxic metals is hypothesized to induce neuroinflammation and neuropathology, paving the way to neurodegeneration. The aim of this study was to examine whether such Potentially Toxic Elements (PTE) are associated with cognitive performance over time in a sample of elderly individuals of environmentally contaminated regions. A total of 146 elders permanently residing in environmental risk areas were recruited. Participants completed the Mini Mental State Examination (MMSE) for cognitive assessment and were biomonitored for several PTE through hair and urine analysis. After 5 years, the cognitive assessment was repeated. Cognitive performance significantly declined from baseline to follow-up (p < 0.001), with participants scoring on average 4.41 less points on MMSE (Baseline: M = 25.79, SD = 3.520, Md = 27.00; Follow-up: M = 21.38, SD = 5.343, Md = 21.00). Regression analysis on the relationship between PTE and cognitive performance revealed that Aluminium (β = -0.322, p = 0.32) and Zinc (β = -0.372, p = 0.15) in hair and Aluminium in urine (β = -0.368, p = 0.010) significantly predicted MMSE scores past 5 years of biomonitoring (p = 0.002). Higher contents of these PTE were associated with worse cognitive performance at follow-up, after controlling the effects of age, education and supplement intake. Higher levels of Aluminium significantly predict worse general cognitive performance 5 years after biomonitoring across different biological samples. Environmental exposure to this metal may constitute a risk factor for cognitive decline.
- Bioaccessibility and changes on cylindrospermopsin concentration in edible mussels over storage and processing time.Publication . Freitas, Marisa; Azevedo, Joana; Carvalho, António Paulo; Mendes, Vera; Manadas, Bruno; Campos, Alexandre; Vasconcelos, VítorThe cyanotoxin cylindrospermopsin has been recognized of increased concern due to the global expansion of its main producer, Cylindrospermopsis raciborskii. Previous studies have shown that aquatic organisms, especially bivalves, can accumulate high levels of cylindrospermopsin. Based on the potential for human health risks, a provisional tolerable daily intake of 0.03 µg/kg body-weight has been recommended. However, human exposure assessment has been based on the cylindrospermopsin concentration in raw food items. This study aimed to assess the changes on cylindrospermopsin concentration in edible mussels over storage and processing time as well as cylindrospermopsin bioaccessibility. Mussels, (Mytilus galloprovincialis) fed cylindrospermopsin-producing C. raciborskii, were subjected to the treatments and then analyzed by LC-MS/MS. Mussels stored frozen allowed a significantly higher recovery of cylindrospermopsin (52.5%/48 h and 57.7%/one week). The cooking treatments did not produce significant differences in cylindrospermopsin concentration in mussel matrices (flesh), however, cylindrospermopsin was found in the cooking water, suggesting that heat processing can be used to reduce the availability of cylindrospermopsin in this food item. The in vitro digestion with salivary and gastrointestinal juices considerably decreased the cylindrospermopsin availability in uncooked and steamed mussels, highlighting the importance in integrating the bioaccessibility in the human health risk assessment.
- Raw chicken meat is a vehicle of Ampicillin-multidrug-resistant Enterococcus faecium: is it a on health problem?Publication . Novais, Carla; Freitas, Ana R.; Rebelo, Andreia; Ferreira, Carolina; Duarte, Bárbara; Ribeiro, Sofia; Mourão, Joana; Scholes, Christopher; Antunes, Patrícia; Peixe, LuísaResistance to ampicillin-AmpR is associated with Enterococcus faecium-Efm from hospitalized-humans (clade-A1) and at lesser extent with community-based isolates (clade-A2: human and animal strains; clade-B: human-commensal strains). Recently, AmpR combined with specific putative virulence factors were proposed as molecular markers of Efm linked to human infections, which can have an impact in Efm risk assessment in different public health contexts (PMID:29519512). Here we evaluated the occurrence of AmpR-Efm with potential public health impact in chicken-meat samples.
- Toxic cyanobacteria and cyanotoxins monitoring studies in the framework of TOXICROP ProjectPublication . Uro, V. H.; Ranilla, C. A.; Oliveira, F.; Freitas, Marisa; Azevedo, J.; Ferreira, J.; Vasconcelos, Vítor; Campos, AlexandreFresh water resources in many countries are vulnerable due to their biogeographical and climatic characteristics. Moreover higher water consumption and higher human impacts in the downstream water bodies is leading to a higher eutrophication with increased incidence and intensity of cyanobacteria blooms and their toxins. The scarcity of clean water resources leads to the compulsory use of water containing cyanobacteria and their toxins in agriculture. TOXICROP is a European project funded by the Marie Skłodowska-Curie Research and Innovation Staff Exchange (RISE) programme aimed, among other objectives, to map agricultural risk areas of cyanotoxin occurrence, in countries that participate in the Project. In 2019 two monitoring campaigns were accomplished, in Pisão reservoir, located in the South region of Portugal, in September 2019, and in El Pañe an Aguada-Blanca reservoirs, located in the Andean Plateau, in the region of Arequipa, Peru, in October 2019. Sampling points were identified in these lakes and 1L samples were collected with assistance of a boat. Other water infrastructures relevant for this study, i.e., waterways, irrigation canals, irrigation hidrants and small irrigation reservoirs, were identified and sampled as well. Moreover, soil and plant samples were collected from an important agricultural area in the region of Arequipa city, which will be also subject to chemical analyses for detection and quantification of cyanotoxins. Here we present the monitoring work performed in the field, in the framework of TOXICROP project.