Analysis of Antibiotics in Bivalves by Ultra-High Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry.

The presence of pharmaceuticals in aquatic ecosystems mostly originates from wastewater treatment plants (WWTPs) and such a situation can be responsible for significant negative impacts on natural ecosystems, such as estuarine and coastal areas. Bioaccumulation of pharmaceuticals, namely antibiotics, in exposed organisms is known to have remarkable effects on different trophic levels of non-target organisms such as algae, invertebrates and vertebrates, including the emergence of bacterial resistance. Bivalves are a highly appreciated seafood product, as they are fed by filtering water, and can bioconcentrate chemicals, being ideal for biomonitoring environmental health hazards in coastal and estuarine ecosystems. To use this sentinel species, an analytical strategy was developed to be used in accessing antibiotics, from human and veterinary medicine, and evaluate their occurrence as emerging pollutants in aquatic environments. The optimized analytical method was fully validated according to the European requirements defined by the Commission Implementing Regulation 2021/808. The validation comprised the following parameters: specificity, selectivity, precision, recovery, ruggedness, linearity, and the decision limit CCα, as well as the limit of detection (LoD) and limit of quantification (LoQ). The method was validated for 43 antibiotics to allow their quantification in both contexts, environmental biomonitoring and food safety.

Mycotoxins in Raw Bovine Milk: UHPLC-QTrap-MS/MS Method as a Biosafety Control Tool.

Mycotoxins are compounds produced by several fungi that contaminate agricultural fields and, either directly or by carry-over, final food products. Animal exposure to these compounds through contaminated feed can lead to their excretion into milk, posing threats to public health. Currently, aflatoxin M1 is the sole mycotoxin with a maximum level set in milk by the European Union, as well as the most studied. Nonetheless, animal feed is known to be contaminated by several groups of mycotoxins with relevance from the food safety point of view that can be carried over into milk. To evaluate the multi-mycotoxin occurrence in this highly consumed food product it is crucial to develop precise and robust analytical methodologies towards their determination. In this sense, an analytical method for the simultaneous identification of 23 regulated, non-regulated, and emerging mycotoxins in raw bovine milk using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) was validated. A modified QuEChERS protocol for extraction purposes was used, and further validation was performed by assessing the selectivity and specificity, limits of detection and quantification (LOD and LOQ), linearity, repeatability, reproducibility, and recovery. The performance criteria were compliant with mycotoxin-specific and general European regulations for regulated, non-regulated, and emerging mycotoxins. The LOD and LOQ ranged between 0.001 and 9.88 ng mL-1 and 0.005 and 13.54 ng mL-1, respectively. Recovery values were between 67.5 and 119.8%. The repeatability and reproducibility parameters were below 15 and 25%, respectively. The validated methodology was successfully applied to determine regulated, non-regulated, and emerging mycotoxins in raw bulk milk from Portuguese dairy farms, proving the importance of widening the monitoring scope of mycotoxins in dairy products. Additionality, this method presents itself as a new strategic and integrated biosafety control tool for dairy farms for the analysis of these natural and relevant human risks.

Risk Assessment of Nine Coccidiostats in Commercial and Home-Raised Eggs.

The poultry industry, in order to prevent and control coccidiosis caused by Eimeria spp., widely uses coccidiostats as feed additives. The main objective of this study was to evaluate the presence of nine coccidiostats in 62 egg samples by UHPLC-MS/MS. Overall, detection frequency and average concentration were 90.3% (56/62) and 106.3 µg kg-1, respectively. Only diclazuril and nicarbazin were detected. Diclazuril, only found in home-raised eggs, showed an overall detection frequency of 8.1% (5/62), with average and maximum concentrations of 0.46 ± 1.90 µg kg-1 and 13.6 µg kg-1, respectively. Nicarbazin presented an overall higher frequency, 88.7% (55/62), with levels up to 744.8 µg kg-1. Additionally, four samples (6.5%) presented both nicarbazin and diclazuril. Home-raised egg samples (n = 28) showed a detection frequency of 89.3%, with nicarbazin found in more samples (85.7% vs. 17.9%) and at higher levels (266.3 ± 169.4 µg kg-1 vs. 0.91 ± 2.78 µg kg-1) when compared to diclazuril. In supermarket samples (n = 34), only nicarbazin was detected in 31 samples (91.1%), with an average of 167.6 ± 62.2 µg kg-1. Considering the average contamination scenario, consumers' health should not be adversely affected by egg consumption. In every scenario considered, children were the most vulnerable population group.

Assessment of Agricultural Practices for Controlling Fusarium and Mycotoxins Contamination on Maize Grains: Exploratory Study in Maize Farms.

Maize is a significant crop to the global economy and a key component of food and feed, although grains and whole plants can often be contaminated with mycotoxins resulting in a general exposure of the population and animals. To investigate strategies for mycotoxins control at the grain production level, a pilot study and exploratory research were conducted in 2019 and 2020 to compare levels of mycotoxins in grains of plants treated with two fertilizers, F-BAC and Nefusoil, under real agricultural environment. The 1650 grains selected from the 33 samples were assessed for the presence of both Fusarium species and mycotoxins. Only fumonisins and deoxynivalenol were detected. Fumonisin B1 ranged from 0 to 2808.4 µg/Kg, and fumonisin B2 from 0 to 1041.9 µg/Kg, while deoxynivalenol variated from 0 to 465.8 µg/Kg. Nefusoil showed to be promising in regard to fumonisin control. Concerning the control of fungal contamination rate and the diversity of Fusarium species, no significant differences were found between the two treatments in any of the years. However, a tendency for was observed Nefusoil of lower values, probably due to the guaranteed less stressful conditions to the Fusarium spp. present in the soil, which do not stimulate their fumonisins production.

Mycotoxins Contamination in Rice: Analytical Methods, Occurrence and Detoxification Strategies.

The prevalence of mycotoxins in the environment is associated with potential crop contamination, which results in an unavoidable increase in human exposure. Rice, being the second most consumed cereal worldwide, constitutes an important source of potential contamination by mycotoxins. Due to the increasing number of notifications reported, and the occurrence of mycotoxins at levels above the legislated limits, this work intends to compile the most relevant studies and review the main methods used in the detection and quantification of these compounds in rice. The aflatoxins and ochratoxin A are the predominant mycotoxins detected in rice grain and these data reveal the importance of adopting safety storage practices that prevent the growth of producing fungi from the Aspergillus genus along all the rice chain. Immunoaffinity columns (IAC) and QuECHERS are the preferred methods for extraction and purification and HPLC-MS/MS is preferred for quantification purposes. Further investigation is still required to establish the real exposition of these contaminants, as well as the consequences and possible synergistic effects due to the co-occurrence of mycotoxins and also for emergent and masked mycotoxins.

Improving prenatal care during lockdown: Comparing telehealth and in-person care for low-risk pregnant women in the PROTECT pilot study.

OBJECTIVE: to compare telehealth and in-person care during the COVID-19 lockdown in a population of low-risk pregnant women for prenatal care received and perinatal outcome. METHODS: This single-center study began during the first French lockdown in 2020. Women with at least one telehealth (remote) prenatal care visit were compared with those who received care only in person. Data include results from self-administered surveys and perinatal outcomes. The main outcome was the prenatal care experience, assessed by the 5-point Quality of Prenatal Care Questionnaire (QPCQ) score. Exploratory analyses sought to identify connections between perinatal outcomes and any of their levels of QPCQ score, health/eHealth literacy, stress, and social deprivation scores . RESULTS: The experimental group included 55 women and the control group 52. Maternal and neonatal outcomes were similar in both groups. The mean QPCQ scores did not support any difference between the mothers' experience of prenatal care in each group: 4.15±0.52 in the telehealth and 4.26±0.63 in the in-person groups. Similarly, levels of social deprivation, stress, and health and eHealth literacy did not differ between the groups. CONCLUSION: Regardless of social deprivation or literacy level, both telehealth and in-person monitoring appeared to provide equivalent and good-quality prenatal care experiences during the pandemic, ClinicalTrial.gov registration NCT04368832 (30th April 2020).

Vaccine-Induced Thrombotic Thrombocytopenia: A Case Report.

While the number of people who have been vaccinated against coronavirus disease 2019 (COVID-19) in Portugal keeps rising, the risk of complications, although rare, keeps rising too. We report a case of vaccine-induced thrombotic thrombocytopenia (VITT) in a 30-year-old previously healthy male after vaccination with Ad26.COV2.S. The patient presented to the emergency department (ED) with abdominal pain and headache. Laboratory tests revealed thrombocytopenia, high D-dimer levels, and fibrinogen consumption. Thoracoabdominal CT scan showed a thrombus in the portal mesenteric venous axis. A positive PF4 heparin enzyme-linked immunosorbent assay confirmed the VITT diagnosis, and the patient was started on intravenous immunoglobulin. Both clinical complaints and laboratory findings resolved within six days, and he was discharged to follow-up. This case shows that general symptoms after vaccination should not be depreciated, highlights the importance of early diagnosis and treatment, and raises new questions about the follow-up and further study of these patients.

Examination of the Weinberger Adjustment Inventory-Short Form Among Portuguese Young Adults: Psychometrics and Measurement Invariance.

The Weinberger Adjustment Inventory-Short Form (WAI-SF) is a multidimensional measure of behavioral adjustment frequently used with forensic, clinical, and community populations. However, no previous studies have examined the WAI-SF from a more modern psychometric perspective including second-order models, measurement invariance and a better estimation of reliability. The current sample is composed of female and male young adults (N = 610, M = 21.33 years, SD = 3.09, range = 18-37) from a university context in Portugal. Results indicated that both the four-factor intercorrelated and the four-factor second order models of the WAI-SF Distress and Restraint scales showed good fits. The WAI-SF Distress and Restraint scales were negatively and significantly correlated, and the intercorrelations between the subscales of each scale ranged from moderate to high. The WAI-SF scales and subscales mostly showed adequate to good reliability in terms of McDonald's Omega and the more traditional Cronbach's Alpha. Strong cross-gender measurement invariance was demonstrated, with females scoring significantly higher than males on the Anxiety subscale of the Distress scale, and on the Suppression of Aggression, Impulse Control, Consideration of Others, Responsibility subscales, and Restraint scale. The WAI-SF scales and subscales showed distinctive correlates with other measures (e.g., low self-control, psychopathy) and variables (e.g., delinquency seriousness, substance use). Considering our findings, the use of the WAI-SF is recommended among the Portuguese young adult population and its use in criminological research is encouraged.

Roux-en-Y Gastric Bypass Surgery Leading to Postprandial Hypoglycemia: A Case Report.

Postprandial hypoglycemia is a rare complication after Roux-en-Y gastric bypass (RYGB). The underlying pathophysiology remains to be fully understood. We present a case of a 49-year-old woman with a past medical history of mesenteric thrombosis due to prothrombin-related thrombophilia, which culminated in RYGB 10 years prior to presentation. The patient had been given anticoagulation treatment for several years, which she abandoned one year prior to presentation. She presented to our consultation with episodes of postprandial hypoglycemia and severe anemia due to iron and vitamin B12 deficiencies. Dietary adjustments were set in place to prevent hypoglycemia and neuroglycopenic symptoms. Intravenous iron and intramuscular vitamin B12 supplementation led to full recovery of hemoglobin levels, allowing restart of oral anticoagulation to prevent recurrence of thrombotic events.

Improving B1+ parametric estimation in the brain from multispin-echo sequences using a fusion bootstrap moves solver.

PURPOSE: To simultaneously estimate the B1+ field (along with the T2 ) in the brain with multispin-echo (MSE) sequences and dictionary matching. METHODS: T2 mapping provides clinically relevant information such as in the assessment of brain degenerative diseases. It is commonly obtained with MSE sequences, and accuracy can be further improved by matching the MSE signal to a precomputed dictionary of echo-modulation curves. For additional T1 quantification, transmit B1+ field knowledge is also required. Preliminary work has shown that although simultaneous brain B1+ estimation along with T2 is possible, it presents a bimodal distribution with the main peak coinciding with the true value. By taking advantage of this, the B1+ maps are expected to be spatially smooth by applying an iterative method that takes into account each pixel neighborhood known as the fusion bootstrap moves solver (FBMS). The effect of the FBMS on B1+ accuracy and piecewise smoothness is investigated and different spatial regularization levels are compared. Total variation regularization was used for both B1+ and T2 simultaneous estimation because of its simplicity as an initial proof-of-concept; future work could explore non edge-preserving regularization independently for B1+ . RESULTS: Improvements in B1+ accuracy (up to 45.37% and 16.81% B1+ error decrease) and recovery of spatially homogeneous maps are shown in simulations and in vivo 3.0T brain data, respectively. CONCLUSION: Accurate B1+ estimated values can be obtained from widely available MSE sequences while jointly estimating T2 maps with the use of echo-modulation curve matching and FBMS at no further cost.

Environmental risk assessment and bioaccumulation of pharmaceuticals in a large urbanized estuary.

We screened for the presence of 66 different pharmaceutical residues in surface waters and in multiple invertebrate and fish species of the Tejo estuary to produce an environmental risk assessment of individual pharmaceuticals and their mixtures, as well as evaluate the bioaccumulation of pharmaceuticals in one of Europe's largest estuarine systems. Sixteen pharmaceutical residues, from seven therapeutic classes, were detected in estuarine waters, with environmental mixture concentrations ranging from 42 to 1762 ng/L. Environmental risk assessment via the determination of risk quotients, demonstrated high ecological risk for the antibiotic amoxicillin and angiotensin II receptor blockers irbesartan and losartan. Moderate risk was estimated for antidepressants, antiepileptics, anxiolytics and beta-blockers, but the risk quotient of the accumulated mixture of compounds was over 380-fold higher than the no risk threshold, driven by antibiotics and angiotensin II receptor blockers. In biota, higher risk therapeutic groups were found in higher concentrations, with nine pharmaceutical residues detected, including six antibiotics and two neuroactive compounds, and maximum tissue concentrations up to 250 µg/kg. Bioaccumulation was species- and compound-specific, with only two compounds found simultaneously in water and biota, likely a result of the complex dynamics and fate of pharmaceuticals in estuarine waters. Nonetheless, higher detection frequencies were observed in species living directly on or just above the substrate (i.e. benthic and demersal species), underpinning the importance of habitat use, as well the potential role of sediment and diet based routes for pharmaceutical uptake. Ultimately, results support urgent action on managing the impact of pharmaceuticals in coastal environments, striving for improved monitoring schemes tailored to the dynamic nature and ecological diversity of estuaries and coastal ecosystems.

Study on the efficiency of a covalent organic framework as adsorbent for the screening of pharmaceuticals in estuary waters.

Herein, we demonstrate, for the first time, that covalent organic frameworks (COFs) can be efficient adsorbents for the screening of pharmaceuticals in real water samples, obtaining highly representative data on their occurrence and avoiding the cost of carrying high volume samples and tedious and costly clean-up and preconcentration steps. Of the 23 pharmaceuticals found present in the water samples from the Tagus river estuary using state-of-the-art solid-phase extraction (SPE), 22 were also detected (adsorbed and recovered for analysis) using a COF as the adsorbent material with adsorption efficiency of over 80% for nearly all compounds. In specific cases, acidification of the water samples was identified to lead to a dramatic loss of adsorption efficiency, underlining the effect of sample pre-treatment on the results. The COF efficiently adsorbed (>80%) 19 pharmaceuticals without acid treatment of the sample, highlighting the potential of this class of materials for representative in situ passive adsorption of pharmaceuticals, making this material suitable for being used in water monitoring programs as a simple and cost-efficient sample preparation procedure. In the case of α-hydroxyalprazolam and diclofenac, the COF outperformed the SPE procedure in the recovery efficiency. Although further efforts should be made in tailoring the desorption of the pharmaceuticals from the COF by using different solvents or solvent mixtures, we propose COFs as convenient adsorbent for broad-scope screening and as an efficient adsorbent material to target specific classes of pharmaceuticals. To the best of our knowledge, this is the first study on the use of COFs for contaminant screening in real, naturally contaminated water samples.

Portuguese honeys as antimicrobial agents against Candida species.

BACKGROUND AND AIM: Honey has been recognized worldwide for its antioxidant, anti-tumor, anti-inflammatory and antimicrobial properties. Among them, the antifungal properties associated to honey make it an attractive alternative treatment for Candida-associated infections, particularly for topical application to the mucous membranes and skin. In this sense, the main purpose of this work was to evaluate physicochemical properties of five Portuguese honeys and Manuka honey (an Australian honey with well recognized medical proprieties, used as control) and to evaluate the antifungal activity in Candida species planktonic and biofilm assays. EXPERIMENTAL PROCEDURE: Pollen analysis, pH determination, color, concentration of protein and methylglyoxal, conductivity, total phenolics and flavonoids, hydrogen peroxide concentration, and characterization by differential scanning calorimetry in honey samples were determined. Additionally, the effect of honeys on planktonic growth of Candida was initially evaluated by determination of the minimum inhibitory concentrations. Then, the same effect of those honeys was evaluated in biofilms, by Colony Forming Units enumeration. RESULTS AND CONCLUSION: It has been shown that Portuguese heather (Erica cinereal) honey presented the most similar physicochemical properties to manuka honey (specially phenolic and flavonoids contents). The five Portuguese honeys under study, presented in general a potent activity against planktonic multi-resistant yeast pathogens (several clinical isolates and reference strains of Candida species) and S. aureus and P. aeruginosa bacteria cultures. Additionally, it was also concluded that Portuguese heather honey (50% and 75% (w/v)) can also act as a good Candida species biofilm reducer, namely for C. tropicalis.

Occurrence of Fusarium spp. in Maize Grain Harvested in Portugal and Accumulation of Related Mycotoxins during Storage.

Maize is an important worldwide commodity susceptible to fungal contamination in the field, at harvest, and during storage. This work aimed to determine the occurrence of Fusarium spp. in maize grains produced in the Tagus Valley region of Portugal and the levels of related mycotoxins in the 2018 harvest and during their storage for six months in barrels, mimicking silos conditions. Continuous monitoring of temperature, CO2, and relative humidity levels were done, as well as the concentration of mycotoxins were evaluated and correlated with the presence of Fusarium spp. F. verticillioides was identified as the predominant Fusarium species. Zearalenone, deoxynivalenol and toxin T2 were not found at harvest and after storage. Maize grains showed some variability in the levels of fumonisins (Fum B1 and Fum B2). At the harvest, fumonisin B1 ranged from 1297 to 2037 µg/kg, and fumonisin B2 ranged from 411 to 618 µg/kg. Fumonisins showed a tendency to increase (20 to 40%) during six months of storage. Although a correlation between the levels of fumonisins and the monitoring parameters was not established, CO2 levels may be used to predict fungal activity during storage. The composition of the fungal population during storage may predict the incidence of mycotoxins.

Analysis Protocols for MRI Mapping of the Blood Oxygenation-Sensitive Parameters T2* and T2 in the Kidney.

Renal hypoxia is generally accepted as a key pathophysiologic event in acute kidney injury of various origins and has also been suggested to play a role in the development of chronic kidney disease. Here we describe step-by-step data analysis protocols for MRI monitoring of renal oxygenation in rodents via the deoxyhemoglobin concentration sensitive MR parameters T2* and T2-a contrast mechanism known as the blood oxygenation level dependent (BOLD) effect.This chapter describes how to use the analysis tools provided by vendors of animal and clinical MR systems, as well as how to develop an analysis software. Aspects covered are: data quality checks, data exclusion, model fitting, fitting algorithm, starting values, effects of multiecho imaging, and result validation.This chapter is based upon work from the PARENCHIMA COST Action, a community-driven network funded by the European Cooperation in Science and Technology (COST) program of the European Union, which aims to improve the reproducibility and standardization of renal MRI biomarkers. This experimental protocol chapter is complemented by two separate chapters describing the basic concept and data analysis.

Pleural Effusion Secondary to Obstructive Uropathy: A Case of Urinothorax.

Urinothorax is a rare type of pleural effusion and usually the result of genitourinary tract disease. An accurate and early diagnosis is crucial as resolution of the underlying pathology is the mainstay of treatment. We report the case of a 69-year-old man who was admitted to the Internal Medicine ward due to obstructive acute kidney injury of unknown origin. The patient was submitted to urinary catheterization and to right percutaneous nephrostomy. Two weeks after admission he developed a large left pleural effusion; a left urinoma was also visible on computed tomography. After thoracentesis, pleural fluid analysis demonstrated a paucicellular transudate with pH <7.40 and pleural fluid/serum creatinine ratio >1.0. The diagnosis of urinothorax was made and further study allowed the diagnosis of prostate cancer as the aetiology of the obstruction. When bilateral percutaneous nephrostomy was performed, resolution of the urinothorax and normalization of renal function occurred. LEARNING POINTS: Urinothorax is a rare and possibly under-recognized cause of transudative pleural effusion; over the last five decades, fewer than 60 cases have been reported in the English literature worldwide.Diagnosis of urinothorax depends upon a high index of clinical suspicion and on pleural fluid analysis, but imaging can also support the diagnosis, for example by showing the presence of a urinoma.Clinicians should recognize urinothorax early in its clinical course as favourable outcomes are often achieved after correction of the underlying genitourinary tract pathology.

Tambaqui females (Colossoma macropomum) spawn after hormonal induction with buserelin acetate.

The aim of this study was to evaluate induced reproduction in tambaqui females using buserelin acetate as compared with the traditional treatment regimen with carp pituitary extract (CPE). Reproductive traits of females with a body weight (BW) of 8.47 ±â€¯1.52 kg were evaluated in ten females treated with buserelin acetate at the dose of 0.5 mL/kg BW, in a single application, and in ten females treated with CPE at the dose of 5.5 mg/kg BW, in two applications (10 % and 90 %, with a 12-h interval between applications). Spawning rate did not differ between the females treated with buserelin acetate (40 %) and CPE (40 %). Weight, fertilization rate and hatching rate did not differ between the two treatment groups. Degree-hours (determined as the average temperature multiplied by time, in hours, for spawning after the treatment with the second dose of CPE and after the single treatment with buserelin acetate) for spawning and number of oocytes per gram of gametes collected were greater (P < 0.05) in the females treated with buserelin acetate than in the females treated with CPE. Production index, absolute fecundity and relative fecundity were greater (P < 0.05) in the females treated with CPE. The hormone buserelin acetate promotes reproduction in tambaqui females with there being a similar spawning rate and oocyte quality, however, lesser production indices and fecundity than when there is the conventional treatment regimen imposed with carp pituitary extract.

The use of ultra-high-pressure-liquid-chromatography tandem time-of-flight mass spectrometry as a confirmatory method in drug residue analysis: Application to the determination of antibiotics in piglet liver.

Consumption of food containing antibiotic residues can cause direct toxic effects, such as allergic reactions, or indirect effects, like the possibility to contribute to the development of antimicrobial resistance. It is essential for consumer food safety, as well as for public health, to monitor antibiotics in foods of animal origin intended for human consumption, particularly in piglets, a highly consumed meat in Portugal. Concerning the lack of regulations regarding the validation criteria for the ToF-MS (time-of-flight mass spectrometry) drug residues determination, this study presents an approach for the use of UHPLC (ultra-high-pressure-liquid-chromatography)-ToF-MS not only for screening but also for confirmatory purposes of 45 antibiotics from 7 different classes. Thus, a method was optimized and validated in accordance with Decision 2002/657/EC by evaluating CCα, CCß, specificity, selectivity, recovery, ruggedness, repeatability and reproducibility. For antibiotics with no MRL set, the lowest values achieved for CCα and CCß were, respectively, of 0.1 µg kg-1 and 0.2 µg kg-1 for cinoxacin. Regarding precision, the CV values obtained ranged from 5% to 21% for repeatability and from 6% to 22% for reproducibility. Recoveries were calculated falling between 82% and 120%. After validation, the method was used to analyse piglet liver samples. Antibiotic residues were determined in 47% of the 21 analysed samples. Nevertheless, all the antibiotics were detected at concentrations below the MRLs.

Evaluation of the mycotoxins content of Salicornia spp .: a gourmet plant alternative to salt.

SALICORNIA: spp. are traditionally used in Asia as food, forage and medicine. In the West, its popularity has been increasing as an alternative to salt. Nevertheless, the potential presence of contaminants needs to be considered. Hence this study, which aimed to investigate the presence of potentially mycotoxigenic fungi and the occurrence of aflatoxins and ochratoxin A in Salicornia sampled in Portugal. Fungi isolation was performed by the direct plating technique in selective media. The mycotoxins contamination levels were determined by an Ultra-High Performance Liquid Chromatography coupled with Time-of-Flight Mass Spectrometry (UHPLC-ToF-MS). The results suggest that Salicornia is highly susceptible to fungi infestation, e.g. by Aspergillus and Penicillium species. Relevant levels of AFB1 > 5 µg/Kg and total aflatoxins (sum of AFB1, AFB2, AFG1 and AFG2) > 10 µg/Kg were found in various samples.

Depressed, hypertense and sore: Long-term effects of fluoxetine, propranolol and diclofenac exposure in a top predator fish.

Pharmaceutical compounds are continuously released into the aquatic environment, resulting in their ubiquitous presence in many estuarine and coastal systems. As pharmaceuticals are designed to produce effects at very low concentrations and target specific evolutionary conserved pathways, there are growing concerns over their potential deleterious effects to the environment and specifically to aquatic organisms, namely in early life-stages. In this context, the long-term effects of exposure of juvenile meagre Argyrosomus regius to three different pharmaceuticals were investigated. Fish were exposed to environmental concentrations of one of three major used pharmaceuticals: the antidepressant fluoxetine (0.3 and 3 µg/L for 15 days), the anti-hypertensive propranolol and the non-steroidal anti-inflammatory agent diclofenac (0.3 and 15 µg/L for 30 days). Pharmaceuticals bioconcentration in fish muscle was examined, along with biomarkers in different tissues related with antioxidant and biotransformation responses (catalase, superoxide dismutase, ethoxyresorufin-O-deethylase and glutathione S-transferase), energetic metabolism (lactate dehydrogenase, isocitrate dehydrogenase and electron transport system activities), neurotransmission (acetylcholinesterase activity) and oxidative damage (DNA damage and lipid peroxidation levels). Overall, each pharmaceutical had different potential for bioconcentration in the muscle (FLX > PROP > DCF) and induced different biological responses: fluoxetine was the most toxic compound to juvenile meagre, affecting fish growth, triggering antioxidant defense responses, inhibiting detoxification mechanisms and increasing lipid peroxidation and DNA damage in the liver; propranolol exposure increased DNA damage and decreased aerobic metabolism in fish muscle; and diclofenac showed no potential to bioconcentrate, yet it affected fish metabolism by increasing cellular energy consumption in the muscle and consequently reducing fish net energy budget. The diverse response patterns evidence the need for future research focused on pharmaceuticals with different modes of action and their exposure effects on organismal physiological mechanisms and homeostatic status. Ultimately, the combination of sub-individual and individual responses is key for ecologically relevant assessments of pharmaceutical toxicity.