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Sweet cherry pits, date seeds, and grape seeds are abundant fruit by-products in the Mediterranean region. Assessing their antioxidant capacity is crucial for their valorization. Grape and date seeds exhibited higher concentrations of total phenolic and flavonoid contents, and significant antioxidant capacity. Epicatechin was the main flavonoid in sweet cherry pits and date seeds (29–85 mg/g), while vanillic acid was the predominant phenolic acid across all by-products (5–23 mg/g). However, some sweet cherry pit varieties exceeded Maximum Residue Levels (MRL) for five pesticides, while grape seeds contained thirteen fungicide residues, all below MRL. Ochratoxin A was detected in one date seed but below the limit of quantification. Additionally, grape seeds showed an Al content of approximately 130 mg/kg, along with levels of As, Cd, and Pb. Date seeds exhibited high potential for food and pharmaceutical applications, pending evaluation for chemical contaminants.

The European population is ageing. Food Supplements (FSs) are foods with particular characteristics, consumed by elderly people for various purposes, including combating nutritional deficits. Their consumption in this age group, associated with a high prevalence of polypharmacy, can enhance interactions. Potential drug-food (or food supplements), drug-drug interactions and polypharmacy are common health issues among older adults. The prevalence of polypharmacy is high, and preliminary data also indicate that there is significant FS use, increasing the risk of the duplication of therapies and various adverse reactions as well as drug–FS and FS-FS interactions. Therefore, the intervention of health professionals in mitigating these risks is essential. This review highlights and discusses the association between FSs, polypharmacy, and adverse reactions due to the risk of potential interactions between these products. Moreover, it also provides current scientific evidence regarding the use of FSs by the elderly. A review of the challenges, advantages, and risks of using FSs in elderly people who are malnourished and/or polymedicated, focusing on the good practises needed to support healthy ageing, is presented. In this regard, this paper aims to help health professionals better deal with the issue of the use of multiple FSs and polypharmacy, overcome the malnutrition problem, and improve the health and well-being of older adults.

In recent years, contaminants of emerging concern have been reported in several environmental matrices due to advances in analytical methodologies. These anthropogenic micropollutants are detected at residual levels, representing an ecotoxicological threat to aquatic ecosystems. In particular, the pharmacotherapeutic group of non-steroidal anti-inflammatories (NSAIDs) is one of the most prescribed and used, as well as one of the most frequently detected in the aquatic environment. Bivalves have several benefits as a foodstuff, and also as an environment bioindicator species. Therefore, they are regarded as an ideal tool to assess this issue from both ecotoxicological and food safety perspectives. Thus, the control of these residues in bivalves is extremely important to safeguard environmental health, also ensuring food safety and public health. This paper aims to review NSAIDs in bivalves, observing their consumption, physicochemical characteristics, and mechanisms of action; their environmental occurrence in the aquatic environment and aquatic biota; and their effects on the ecosystem and the existent legal framework. A review of the analytical methodologies for the determination of NSAIDs in bivalves is also presented.

Urban wastewaters contain pharmaceuticals that are not appropriately removed in conventional wastewater treatments, limiting treated water reuse. Microalgae have been shown to remove pharmaceuticals from urban wastewater in laboratory trials, but few studies have been conducted under natural conditions. In this work, pharmaceutical removal was assessed in a pilot-scale microalgal tertiary wastewater treatment in real conditions. Even after secondary treatment, the water contained measurable amounts of pharmaceuticals (an average of 218.4 ng/L) that significantly decreased to 39.83 ng/L at the exit of the microalgal system. Pharmaceuticals’ average removal rates were slightly higher in the summer (79.1%) than in autumn (71.1%). Antibiotics and antipsychotics were better removed (88.8 and 86.4%, respectively) than antihypertensives (75.3%) and others (Bezafibrate and Diclofenac; 64.0%). Physicochemical characteristics of the wastewater influenced pharmaceutical removal; significant positive correlations were observed between anti-hypertensive drug removal and ammonium concentration (r = 0.53; p < 0.05), total nitrogen and total pharmaceutical removal (r = 0.46; p < 0.05), and total nitrogen and antipsychotic drug removal (r = 0.47; p < 0.05). The results demonstrate the effectiveness of microalgal tertiary treatment in the removal of pharmaceuticals.

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.

Rice is the second most important cereal crop and is vital for the diet of billions of people. However, its consumption can increase human exposure to chemical contaminants, namely mycotoxins and metalloids. Our goal was to evaluate the occurrence and human exposure of aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEN), and inorganic arsenic (InAs) in 36 rice samples produced and commercialized in Portugal and evaluate their correlation. The analysis of mycotoxins involved ELISA, with limits of detection (LODs) of 0.8, 1 and 1.75 μg kg−1 for OTA, AFB1, and ZEN, respectively. InAs analysis was carried out by inductively coupled plasma mass spectrometry (ICP-MS; LOD = 3.3 μg kg−1). No sample showed contamination by OTA. AFB1 was present in 2 (4.8%) samples (1.96 and 2.20 μg kg−1), doubling the European maximum permitted level (MPL). Concerning ZEN, 88.89% of the rice samples presented levels above the LOD up to 14.25 µg kg−1 (average of 2.75 µg kg−1). Regarding InAs, every sample presented concentration values above the LOD up to 100.0 µg kg−1 (average of 35.3 µg kg−1), although none surpassed the MPL (200 µg kg−1). No correlation was observed between mycotoxins and InAs contamination. As for human exposure, only AFB1 surpassed the provisional maximum tolerable daily intake. Children were recognized as the most susceptible group.

Herbal infusions are highly popular beverages consumed daily due to their health benefits and antioxidant properties. However, the presence of plant toxins, such as tropane alkaloids, constitutes a recent health concern for herbal infusions. This work presents an optimized and validated methodology based on the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction procedure followed by Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC-ToF-MS) for the determination of tropane alkaloids (atropine, scopolamine, anisodamine, and homatropine) in herbal infusions, in accordance with criteria established by Commission Recommendation EU No. 2015/976. One of the seventeen samples was contaminated with atropine, exceeding the current European regulation regarding tropane alkaloids. In addition, this study evaluated the antioxidant capacity of common herbal infusions available on Portuguese markets, indicating the high antioxidant capacity of yerba mate (Ilex paraguariensis), lemon balm (Melissa officinalis), and peppermint (Mentha x piperita).

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.

Coccidiosis remains one of the major problems of the poultry industry. Caused by Eimeria species, Coccidiosis is a contagious parasitic disease affecting poultry with great economic significance. Currently, in order to prevent health problems caused by this disease, broiler farmers make extensive use of coccidiostats in poultry feed, maintaining animal health and, in some cases, enhancing feed conversion. The presence of unauthorized substances, residues of veterinary products and chemical contaminants in the food industry is of concern, since they may pose a risk to public health. As the use of coccidiostats has been increasing without any requirements for veterinary prescription, research and surveillance of coccidiostat residues in poultry meat is becoming imperative. This review presents an up-to-date comprehensive discussion of the state of the art regarding coccidiosis, the most used anticoccidials in poultry production, their mode of action, their prophylactic use, occurrence and the European Union (EU) applicable legislation.

Breast milk is considered the ideal form of nutrition for newborns and infants. However, it can carry over contaminants, namely mycotoxins, with biological effects to which this population is particularly vulnerable. Human biomonitoring and surveillance programs are particularly scarce in low-income countries, where food security is a more urgent priority in comparison with food safety. This pilot survey aims to assess exposure of breastfed infants to aflatoxin M1 (AFM1), zearalenone (ZEN), and ochratoxin A (OTA) in Angola, and to evaluate the main socio-demographical and food consumption determinants of lactating mothers. All 37 breast milk samples analyzed are found to be contaminated with ZEN and OTA, although none are found contaminated with AFM1. Contamination levels are lower than previously reported for ZEN but higher in the case of OTA. A significant association between ZEN levels in breast milk and the consumption of cookies by the lactating mothers is found. As for OTA, higher levels are observed in the milk from mothers with younger infants, for which high estimated daily intake (EDI) is determined. As far as the authors are aware, this is the first survey of the occurrence of mycotoxins in breast milk in Angola, so further human biomonitoring works should follow, given that mycotoxins are a global health issue that directly impact the health of populations.

Arsenic is a metalloid with natural and anthropogenic sources and its inorganic form is toxic to humans. Rice is highly consumed worldwide and is prone to arsenic contamination; therefore, this study evaluated the inorganic arsenic content of 70 Portuguese rice samples. These were analysed through inductively coupled plasma mass spectrometry (ICP-MS) with a detection limit of 3.3 µg kg−1. The average contamination was of 29.3 µg kg−1, with brown and short rice presenting higher values than white and long rice. The highest concentration, 100 µg kg−1, equalled the maximum residue limit (MRL) for rice destined for infants’ consumption. The estimated daily intake (EDI) surpassed the benchmark dose (lower confidence limit 10%) (BMDL10) of 0.3 µg kg−1 of bw/day considering children in the 95th percentile of rice consumption and the worst-case scenario concentration. However, other sources also contribute to the EDI and some population groups can exceed the BMDL10.

Human biomonitoring (HBM) data provide information on total exposure regardless of the route and sources of exposure. HBM studies have been applied to quantify human exposure to contaminants and environmental/occupational pollutants by means of determining the parent compounds, their metabolites, or even their reaction products in biological matrices. HBM studies performed among the Portuguese population are dispersed and limited. Thus, to overcome this knowledge gap, this work reviews the published Portuguese HBM information concerning mycotoxins detected in the urine, serum, milk, hair, and nails of different groups of the Portuguese population. This integrative approach to the available HBM data allows us to analyze the main determinants and patterns of exposure of the Portuguese population to the selected hazardous compounds, as well as to assess the potential health risks. We also aimed to identify the main difficulties and challenges of HBM through the analysis of the enrolled studies. Ultimately, this study aims to support national and European policies in promoting human health by summarizing the most important outcomes and lessons learned through the HBM studies carried out in Portugal.

Human biomonitoring (HBM) data provide information on total exposure regardless of the route and sources of exposure. HBM studies have been applied to quantify human exposure to contaminants and environmental/occupational pollutants by determining the parent compounds, their metabolites or even their reaction products in biological matrices. HBM studies performed among the Portuguese population are disperse and limited. To overcome this knowledge gap, this review gathers, for the first time, the published Portuguese HBM information concerning polycyclic aromatic hydrocarbons (PAHs), metals, metalloids, and pesticides concentrations detected in the urine, serum, milk, hair, and nails of different groups of the Portuguese population. This integrative insight of available HBM data allows the analysis of the main determinants and patterns of exposure of the Portuguese population to these selected hazardous compounds, as well as assessment of the potential health risks. Identification of the main difficulties and challenges of HBM through analysis of the enrolled studies was also an aim. Ultimately, this study aimed to support national and European policies promoting human health and summarizes the most important outcomes and lessons learned through the HBM studies carried out in Portugal.

Glyphosate is a systemic, broad-spectrum and post-emergent herbicide. The use of glyphosate has grown in the last decades, and it is currently the most used herbicide worldwide. The rise of glyphosate consumption over the years also brought an increased concern about its possible toxicity and consequences for human health. However, a scientific community consensus does not exist at the present time, and glyphosate’s safety and health consequences are controversial. Since glyphosate is mainly applied in fields and can persist several months in the soil, concerns have been raised about the impact that its presence in food can cause in humans. Therefore, this work aims to review the glyphosate use, toxicity and occurrence in diverse food samples, which, in certain cases, occurs at violative levels. The incidence of glyphosate at levels above those legally allowed and the suspected toxic effects of this compound raise awareness regarding public health.

Pistachios are one of the types of tree nut fruits with the highest mycotoxin contamination, especially of aflatoxins, worldwide. This study developed a Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method that was followed by Ultra-High Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC–ToF-MS) for the determination of mycotoxins in pistachios. Different approaches to dispersive solid phase extraction as a clean-up method for high lipid matrices were evaluated. For this, classic sorbents such as C18 (octadecyl-modified silica) and PSA (primary secondary amine), and new classes of sorbents, namely EMR-Lipid (enhanced matrix removal-lipid) and Z-Sep (modified silica gel with zirconium oxide), were used. The QuEChERS method, followed by Z-Sep d-SPE clean-up, provided the best analytical performance for aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), zearalenone (ZEA), toxin T2 (T2) and toxin HT-2 (HT2) in pistachios. The method was validated in terms of linearity, sensitivity, repeatability, interday precision and recovery; it achieved good results according to criteria imposed by Commission Regulation (EC) no. 401/2006. The method was applied to real samples and the results show that pistachios that are available in Portuguese markets are safe from mycotoxins that are of concern to human health.

The consumption of pistachios (Pistacia vera L.) has been increasing, given their important benefit to human health. In addition to being an excellent nutritional source, they have been associated with chemical hazards, such as mycotoxins, resulting in fungal contamination and its secondary metabolism. Aflatoxins (AFs) are the most common mycotoxins in pistachio and the most toxic to humans, with hepatotoxic effects. More mycotoxins such as ochratoxin A (OTA), fumonisins (FBs), zearalenone (ZEA) and trichothecenes (T2, HT2 and DON) and emerging mycotoxins have been involved in nuts. Because of the low levels of concentration and the complexity of the matrix, the determination techniques must be very sensitive. The present paper carries out an extensive review of the state of the art of the determination of mycotoxins in pistachios, concerning the trends in analytical methodologies for their determination and the levels detected as a result of its contamination. Screening methods based on immunoassays are useful due to their simplicity and rapid response. Liquid chromatography (LC) is the gold standard with new improvements to enhance accuracy, precision and sensitivity and a lower detection limit. The reduction of Aspergillus’ and aflatoxins’ contamination is important to minimize the public health risks. While prevention, mostly in pre-harvest, is the most effective and preferable measure to avoid mycotoxin contamination, there is an increased number of decontamination processes which will also be addressed in this review.

The occurrence of pharmaceuticals in water catchments and drinking waters raises potential risks to public health. Therefore, after addressing the major aquatic contamination pathway, the wastewater treatment plants (WWTPs), and, subsequently, surface waters, 18 human pharmaceuticals from 6 therapeutic groups (antibiotics, lipid regulators, selective serotonin reuptake inhibitors, non-steroidal anti-inflammatory drugs (NSAIDs) and hormones) were analyzed in drinking water catchments, tap and drinking fountain waters. This was performed by solid phase extraction (SPE) and liquid chromatography coupled with tandem mass detection (LC-MS/MS). The 97 samples analyzed were collected from 31 different sites in the center of Portugal. All samples presented concentrations below the method detection limits (MDLs) that ranged between 1.13 to 5.45 ng L−1. The achieved results contributed to a better knowledge on the Portuguese and European context of drinking water, since there is a knowledge gap regarding this matrix. Comparing our data with other studies, published worldwide, we can observe that median concentrations of pharmaceuticals were reported in the low ng L−1 levels, values close to our MDLs. Consequently, it is unlikely that, in light of the current knowledge, the presence of pharmaceuticals in drinking water presents a threat to human health.

Citrinin (CIT) deserves attention due to its known toxic effects in mammalian species and its widespread occurrence in food commodities, often along with ochratoxin A, another nephrotoxic mycotoxin. Human exposure, a key element in assessing risk related to food contaminants, depends upon mycotoxin contamination levels in food and on food consumption. Commercial supplements, commonly designated as red rice, usually used in daily diets in Asiatic countries due to their medicinal properties, may pose a health problem as a result of high CIT levels. In addition to the worldwide occurrence of CIT in foods and supplements, a wide range of several analytical and detection techniques with high sensitivity, used for evaluation of CIT, are reviewed and discussed in this manuscript. This review addresses the scientific literature regarding the presence of CIT in foods of either vegetable or animal origin, as well as in supplements. On what concerns analytical methodologies, sample extraction methods, such as shaking extraction and ultrasonic assisted extraction (UAE), clean-up methods, such as liquid-liquid extraction (LLE), solid phase extraction (SPE) and Quick, Easy, Cheap, Effective, Rugged and Safe (QuECHERS), and detection and quantification methods, such as thin layer chromatography (TLC), high performance liquid chromatography (HPLC), capillary electrophoresis (CE), biosensors, and ELISA, are also reviewed.

Until now, the available data regarding citrinin (CIT) levels in food and the consumption of contaminated foods are insufficient to allow a reliable estimate of intake. Therefore, biomonitoring configuring analysis of parent compound and/or metabolites in biological fluids, such as urine or blood, is being increasingly applied in the assessment of human exposure to CIT and its metabolite, dihydrocitrinone (DH-CIT). Most studies report urinary levels lower for the parent compound when compared with DH-CIT. A high variability either in the mean levels or in the inter-individual ratios of CIT/DH-CIT between the reported studies has been found. Levels of DH-CIT in urine were reported as being comprised between three to seventeen times higher than the parent mycotoxin. In order to comply with this objective, sensitive analytical methodologies for determining biomarkers of exposure are required. Recent development of powerful analytical techniques, namely liquid chromatography coupled to mass spectrometry (LC-MS/MS) and ultra-high-performance liquid chromatography (UHPLC-MS/MS) have facilitated biomonitoring studies, mainly in urine samples. In the present work, evidence on human exposure to CIT through its occurrence and its metabolite, in biological fluids, urine and blood/plasma, in different countries, is reviewed. The analytical methodologies usually employed to evaluate trace quantities of these two molecules, are also presented. In this sense, relevant data on sampling (size and pre-treatment), extraction, cleanup and detection and quantification techniques and respective chromatographic conditions, as well as the analytical performance, are evidenced.

Potential risks associated with releases of human pharmaceuticals into the environment have become an increasingly important issue in environmental health. This concern has been driven by the widespread detection of pharmaceuticals in all aquatic compartments. Therefore, 22 pharmaceuticals, 6 metabolites and transformation products, belonging to 7 therapeutic groups, were selected to perform a review on their toxicity and environmental risk assessment (ERA) in different aquatic compartments, important issues to tackle the water framework directive (WFD). The toxicity data collected reported, with the exception of anxiolytics, at least one toxicity value for concentrations below 1 µg L−1. The results obtained for the ERA revealed risk quotients (RQs) higher than 1 in all the aquatic bodies and for the three trophic levels, algae, invertebrates and fish, posing ecotoxicological pressure in all of these compartments. The therapeutic groups with higher RQs were hormones, antiepileptics, anti-inflammatories and antibiotics. Unsurprisingly, RQs values were highest in wastewaters, however, less contaminated water bodies such as groundwaters still presented maximum values up to 91,150 regarding 17α-ethinylestradiol in fish. Overall, these results present an important input for setting prioritizing measures and sustainable strategies, minimizing their impact in the aquatic environment.

Ochratoxin A (OTA) is produced by fungi present in several agricultural products with much relevance to food safety. Since this mycotoxin is widely found in cereals, beer has a potential contamination risk. Therefore, it was deemed essential to quantify, for the first time, the levels of OTA in beer, a cereal-based product that is marketed in Portugal, as well as to calculate the human estimated weekly intake (EWI) and risk assessment. A total of 85 samples were analyzed through immunoaffinity clean-up, followed by liquid chromatography-fluorescence detection (LC-FD). This analytical methodology allowed a limit of quantification (LOQ) of 0.43 µg/L. The results showed that 10.6% were contaminated at levels ranging between

Potential risks associated with releases of human pharmaceuticals into the environment have become an increasingly important issue in environmental health. This concern has been driven by the widespread detection of pharmaceuticals in all aquatic compartments. Therefore, 22 pharmaceuticals, 6 metabolites and transformation products, belonging to 7 therapeutic groups, were selected to perform a systematic review on their source, fate and occurrence in different aquatic compartments, important issues to tackle the Water Framework Directive (WFD). The results obtained evidence that concentrations of pharmaceuticals are present, in decreasing order, in wastewater influents (WWIs), wastewater effluents (WWEs) and surface waters, with values up to 14 mg L−1 for ibuprofen in WWIs. The therapeutic groups which presented higher detection frequencies and concentrations were anti-inflammatories, antiepileptics, antibiotics and lipid regulators. These results present a broad and specialized background, enabling a complete overview on the occurrence of pharmaceuticals in the aquatic compartments.

Antibiotics represent a pharmacotherapeutic group widely used in both human and veterinary medicine for which ecosurveillance has been continually recommended. It is urgent to rank the antibiotics and highlight those that may pose potential risk to the environment, a key step for the risk management. The absence of this type of contributions applied to the Portuguese reality supported the idea of compiling the data presented herein. With such purpose the most recent and representative data is used to draw a comparative contribution of each antimicrobial classes according to their intended use, i.e., in human versus veterinary medicine. The aim was to assess: (1) the amount and patterns of antimicrobials usage between human and animals; (2) the qualitative comparison between the antimicrobial classes used in each practice (human and veterinary) or specific use; (3) the potential to enter the environment, metabolism, mode of action and environmental occurrences. This manuscript will, thus, identify priorities for the environmental risk assessment, considering the ranking of the antimicrobials by their usage and potential environmental exposure. Ultimately, this study will serve as a basis for future monitoring programs, guiding the policy of regulatory agencies.

In Portugal, the climate, dietary habits, and food contamination levels present the characteristics for higher population susceptibility to ochratoxin A (OTA), one of the known mycotoxins with the greatest public health and agro-economic importance. In this review, following a brief historical insight on OTA research, a summary of the available data on OTA occurrence in food (cereals, bread, wine, meat) and biological fluids (blood, urine) is made. With this data, an estimation of intake is made to ascertain and update the risk exposure estimation of the Portuguese population, in comparison to previous studies and other populations.

La unidad de manejo más utilizada en pesquerías está limitada a las especies objetivo y sus especies asociadas. La finalidad de este estudio fue adecuar una metodología para regionalizar y caracterizar los sistemas socio-ecológicos (SSE) para la evaluación y el manejo de las pesquerías de pequeña escala del estado de Campeche, México, debido a que los componentes espaciales pueden facilitar el entendimiento de la relación humano-ambiente. Se identificaron ocho regiones del SSE pesquero con una combinación de características únicas a partir del contexto biofísico, el volumen de captura y esfuerzo pesquero, así como la dinámica de la flota.

Ventricular contraction is roughly proportional to the amount of calcium released from the Sarcoplasmic Reticulum (SR) during systole. While it is rather straightforward to measure calcium levels and contractibility under different physiological conditions, the complexity of calcium handling during systole and diastole has made the prediction of its release at steady state impossible. Here we approach the problem analyzing the evolution of intracellular and extracellular calcium fluxes during a single beat which is away from homeostatic balance. Using an in-silico subcellular model of rabbit ventricular myocyte, we show that the high dimensional nonlinear problem of finding the steady state can be reduced to a two-variable general equilibrium condition where pre-systolic calcium level in the cytosol and in the SR must fulfill simultaneously two different equalities. This renders calcium homeostasis as a problem that can be studied in terms of its equilibrium structure, leading to precise predictions of steady state from single-beat measurements. We show how changes in ion channels modify the general equilibrium, as shocks would do in general equilibrium macroeconomic models. This allows us to predict when an enhanced entrance of calcium in the cell reduces its contractibility and explain why SERCA gene therapy, a change in calcium handling to treat heart failure, might fail to improve contraction even when it successfully increases SERCA expression.

BACKGROUND: Rapid pacing rates induce alternations in the cytosolic calcium concentration caused by fluctuations in calcium released from the sarcoplasmic reticulum (SR). However, the relationship between calcium alternans and refractoriness of the SR calcium release channel (RyR2) remains elusive. METHODOLOGY/PRINCIPAL FINDINGS: To investigate how ryanodine receptor (RyR2) refractoriness modulates calcium handling on a beat-to-beat basis using a numerical rabbit cardiomyocyte model. We used a mathematical rabbit cardiomyocyte model to study the beat-to-beat calcium response as a function of RyR2 activation and inactivation. Bi-dimensional maps were constructed depicting the beat-to-beat response. When alternans was observed, a novel numerical clamping protocol was used to determine whether alternans was caused by oscillations in SR calcium loading or by RyR2 refractoriness. Using this protocol, we identified regions of RyR2 gating parameters where SR calcium loading or RyR2 refractoriness underlie the induction of calcium alternans, and we found that at the onset of alternans both mechanisms contribute. At low inactivation rates of the RyR2, calcium alternans was caused by alternation in SR calcium loading, while at low activation rates it was caused by alternation in the level of available RyR2s. CONCLUSIONS/SIGNIFICANCE: We have mapped cardiomyocyte beat-to-beat responses as a function of RyR2 activation and inactivation, identifying domains where SR calcium load or RyR2 refractoriness underlie the induction of calcium alternans. A corollary of this work is that RyR2 refractoriness due to slow recovery from inactivation can be the cause of calcium alternans even when alternation in SR calcium load is present.