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Ciguatera Poisoning (CP) is caused by consumption of fish or invertebrates contaminated with ciguatoxins (CTXs). Presently CP is a public concern in some temperate regions, such as Macaronesia (North-Eastern Atlantic Ocean). Toxicity analysis was performed to characterize the fish species that can accumulate CTXs and improve understanding of the ciguatera risk in this area. For that, seventeen fish specimens comprising nine species were captured from coastal waters inMadeira and Selvagens Archipelagos. Toxicity was analysed by screening CTX-like toxicity with the neuroblastoma cell-based assay (neuro-2a CBA). Afterwards, the four most toxic samples were analysed with liquid chromatography-high resolution mass spectrometry (LC-HRMS). Thirteen fish specimens presented CTX-like toxicity in their liver, but only four of these in their muscle. The liver of one specimen of Muraena augusti presented the highest CTX-like toxicity (0.270 ± 0.121 µg of CTX1B equiv·kg-1). Moreover, CTX analogues were detected with LC-HRMS, for M. augusti and Gymnothorax unicolor. The presence of three CTX analogues was identified: C-CTX1, which had been previously described in the area; dihydro-CTX2, which is reported in the area for the first time; a putative new CTX m/z 1127.6023 ([M+NH4]+) named as putative C-CTX-1109, and gambieric acid A.
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Ciguatera , Ciguatoxinas , Animais , Cromatografia Líquida , Ciguatoxinas/química , Peixes , Espectrometria de MassasRESUMO
The importance of ciguatoxins (CTXs) in seafood safety and their emerging occurrence in locations far away from tropical areas highlight the need for simple and low-cost methods for the sensitive and rapid detection of these potent marine toxins to protect seafood consumers. Herein, an electrochemical immunosensor for the detection of CTXs is presented. A sandwich configuration is proposed, using magnetic beads (MBs) as immobilization supports for two capture antibodies, with their combination facilitating the detection of CTX1B, CTX3C, 54-deoxyCTX1B, and 51-hydroxyCTX3C. PolyHRP-streptavidin is used for the detection of the biotinylated detector antibody. Experimental conditions are first optimized using colorimetry, and these conditions are subsequently used for electrochemical detection on electrode arrays. Limits of detection at the pg/mL level are achieved for CTX1B and 51-hydroxyCTX3C. The applicability of the immunosensor to the analysis of fish samples is demonstrated, attaining detection of CTX1B at contents as low as 0.01 µg/kg and providing results in correlation with those obtained using mouse bioassay (MBA) and cell-based assay (CBA), and confirmed by liquid chromatography coupled to high-resolution mass spectrometry (LC-ESI-HRMS). This user-friendly bioanalytical tool for the rapid detection of CTXs can be used to mitigate ciguatera risk and contribute to the protection of consumer health.
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Técnicas Biossensoriais , Ciguatoxinas/análise , Técnicas Eletroquímicas , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Cromatografia Líquida , Ciguatoxinas/administração & dosagem , Ciguatoxinas/imunologia , Peixes , Injeções Intraperitoneais , Fenômenos Magnéticos , Masculino , Camundongos , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
Although consumption of Tetraodontidae species is prohibited in the EU, intoxications are still reported. The evaluation of tetrodotoxins (TTXs) by mass spectrometry (LC-MS/MS and LC-HRMS) and a screening immunoassay (mELISA) in tetraodontid fishes caught along the Western Mediterranean Sea revealed high concentrations of TTXs in Lagocephalus sceleratus while no TTXs were identified in L. lagocephalus and Sphoeroides pachygaster individuals. The high TTXs content found in the L. sceleratus analysed herein demonstrate the occurrence of highly toxic puffer fish in the Western Mediterranean Sea. Being L. sceleratus a recent invasive species in the Mediterranean, surveillance, risk assessment and risk management measures are necessary. The strategy used within this research work could be a valuable tool for future food safety monitoring.
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Tetraodontiformes/metabolismo , Tetrodotoxina/análise , Animais , Cromatografia Líquida , Ensaio de Imunoadsorção Enzimática , Feminino , Espécies Introduzidas , Masculino , Espectrometria de Massas , Mar Mediterrâneo , Espanha , Especificidade da Espécie , Espectrometria de Massas em Tandem , Distribuição TecidualRESUMO
BACKGROUND: Location-specific patterns of regulated and non-regulated disinfection byproducts (DBPs) were detected in tap water samples of the Barcelona Metropolitan Area. However, it remains unclear if the detected DBPs together with undetected DPBs and organic micropollutants can lead to mixture effects in drinking water. OBJECTIVE: To evaluate the neurotoxicity, oxidative stress response and cytotoxicity of 42 tap water samples, 6 treated with activated carbon filters, 5 with reverse osmosis and 9 bottled waters. To compare the measured effects of the extracts with the mixture effects predicted from the detected concentrations and the relative effect potencies of the detected DBPs using the mixture model of concentration addition. METHODS: Mixtures of organic chemicals in water samples were enriched by solid phase extraction and tested for cytotoxicity and neurite outgrowth inhibition in the neuronal cell line SH-SY5Y and for cytotoxicity and oxidative stress response in the AREc32 assay. RESULTS: Unenriched water did not trigger neurotoxicity or cytotoxicity. After up to 500-fold enrichment, few extracts showed cytotoxicity. Disinfected water showed low neurotoxicity at 20- to 300-fold enrichment and oxidative stress response at 8- to 140-fold enrichment. Non-regulated non-volatile DBPs, particularly (brominated) haloacetonitriles dominated the predicted mixture effects of the detected chemicals and predicted effects agreed with the measured effects. By hierarchical clustering we identified strong geographical patterns in the types of DPBs and their association with effects. Activated carbon filters did not show a consistent reduction of effects but domestic reverse osmosis filters decreased the effect to that of bottled water. IMPACT STATEMENT: Bioassays are an important complement to chemical analysis of disinfection by-products (DBPs) in drinking water. Comparison of the measured oxidative stress response and mixture effects predicted from the detected chemicals and their relative effect potencies allowed the identification of the forcing agents for the mixture effects, which differed by location but were mainly non-regulated DBPs. This study demonstrates the relevance of non-regulated DBPs from a toxicological perspective. In vitro bioassays, in particular reporter gene assays for oxidative stress response that integrate different reactive toxicity pathways including genotoxicity, may therefore serve as sum parameters for drinking water quality assessment.
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Água Potável , Neuroblastoma , Humanos , Carvão Vegetal , Bioensaio , Cromatografia GasosaRESUMO
The Watch List (WL) is a monitoring program under the European Water Framework Directive (WFD) to obtain high-quality Union-wide monitoring data on potential water pollutants for which scarce monitoring data or data of insufficient quality are available. The main purpose of the WL data collection is to determine if the substances pose a risk to the aquatic environment at EU level and subsequently to decide whether a threshold, the Environmental Quality Standards (EQS) should be set for them and, potentially to be listed as priority substance in the WFD. The first WL was established in 2015 and contained 10 individual or groups of substances while the 4th WL was launched in 2022. The results of monitoring the substances of the first WL showed that some countries had difficulties to reach an analytical Limit of Quantification (LOQ) below or equal to the Predicted No-Effect Concentrations (PNEC) or EQS. The Joint Research Centre (JRC) of the European Commission (EC) organised a series of workshops to support the EU Member States (MS) and their activities under the WFD. Sharing the knowledge among the Member States on the analytical methods is important to deliver good data quality. The outcome and the discussion engaged with the experts are described in this paper, and in addition a literature review of the most important publications on the analysis of 17-alpha-ethinylestradiol (EE2), amoxicillin, ciprofloxacin, metaflumizone, fipronil, metformin, and guanylurea from the last years is presented.
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In this work, magnetic particles (MPs) are used as supports for the immobilization of biorecognition molecules for the detection of microcystins (MCs). In one approach, a recombinant protein phosphatase 1 (PP1) has been conjugated to MPs via coordination chemistry, and MC-LR detection has been based on the inhibition of the enzyme activity. In the other approach, a monoclonal antibody (mAb) against MC-LR has been conjugated to protein G-coated MPs, and a direct competitive enzyme-linked immunoparticle assay (ELIPA) has been then performed. Conjugation of biomolecules to MPs has been first checked, and after optimization, MC detection has been performed. The colorimetric PPIA with PP1-MP and the best ELIPA strategy have provided limits of detection (LOD) of 7.4 and 3.9 µg/L of MC-LR, respectively. The electrochemical ELIPA has decreased the LOD to 0.4 µg/L, value below the guideline recommended by the World Health Organisation (WHO). The approaches have been applied to the analysis of a cyanobacterial culture and a natural bloom, and MC equivalent contents have been compared to those obtained by conventional assays and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results have demonstrated the viability of the use of MPs as biomolecule immobilization supports in biotechnological tools for MCs monitoring.
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Imunoensaio/métodos , Microcistinas/análise , Anticorpos Monoclonais/imunologia , Colorimetria , Eletroquímica , Iminoácidos/química , Fenômenos Magnéticos , Microcistinas/imunologia , Níquel/química , Proteína Fosfatase 1/química , Proteínas Recombinantes/químicaRESUMO
Microbial biofilms communities in mineral waters and hot springs have a particular composition with species belonging to different groups such as epsilonproteobacteria and gammaproteobacteria or different siderobacteria and other chymoautrophic organisms, in addition to certain bacillaryophytes, chlorophytes and especially cyanobacteria. Balneotherapy can cause adverse reactions to the usual doses of application of treatments, that consists of a non-specific clinical picture, the so-called "thermal crisis" or "balneointoxication". Despite its clinical similarity (gastric discomfort, hepatic congestive outbreaks, cutaneous reactions, etc.) with that observed in acute cyanotoxin poisonings, thermal crisis has never been associated with the abundant growth of potentially toxic cyanobacteria in the mineral water sources. The aim of this work was to verify the hypothetical involvement of cyanotoxins in this clinical picture. Samples from mostly sulphurous water sources, with thermal characteristics ranging from cold to hyperthermal waters were analysed. ELISA (both in solution and in cellular matrix samples), LC-ESI-HRMS (in cellular matrix samples), and analysis of potential toxicity by means of a standardized bioassay were carried out. The toxic effect observed in the toxicity bioassays in one third of the sources may be related to the existence of microcystins and nodularins and even with other cyanobacterial peptides detected. In addition, several responses observed in the toxicity analyses reflect a pattern, probably linked to a type of hormetic response (hormesis is an adaptive response to low levels of stress, characterized by a biphasic dose-response curve).
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Balneologia , Cianobactérias , Cianobactérias/química , Toxinas de CianobactériasRESUMO
BACKGROUND: Knowledge about human exposure and health effects associated with non-routinely monitored disinfection by-products (DBPs) in drinking water is sparse. OBJECTIVE: To provide insights to estimate exposure to regulated and non-regulated DBPs in drinking water. METHODS: We collected tap water from homes (N = 42), bottled water (N = 10), filtered tap water with domestic activated carbon jars (N = 6) and reverse osmosis (N = 5), and urine (N = 39) samples of participants from Barcelona, Spain. We analyzed 11 haloacetic acids (HAAs), 4 trihalomethanes (THMs), 4 haloacetonitriles (HANs), 2 haloketones, chlorate, chlorite, and trichloronitromethane in water and HAAs in urine samples. Personal information on water intake and socio-demographics was ascertained in the study population (N = 39) through questionnaires. Statistical models were developed based on THMs as explanatory variables using multivariate linear regression and machine learning techniques to predict non-regulated DBPs. RESULTS: Chlorate, THMs, HAAs, and HANs were quantified in 98-100% tap water samples with median concentration of 214, 42, 18, and 3.2 µg/L, respectively. Multivariate linear regression models had similar or higher goodness of fit (R2) compared to machine learning models. Multivariate linear models for dichloro-, trichloro-, and bromodichloroacetic acid, dichloroacetonitrile, bromochloroacetonitrile, dibromoacetonitrile, trichloropropnanone, and chlorite showed good predictive ability (R 2 = 0.8-0.9) as 80-90% of total variance could be explained by THM concentrations. Activated carbon filters reduced DBP concentrations to a variable extent (27-80%), and reverse osmosis reduced DBP concentrations ≥98%. Only chlorate was detected in bottled water samples (N = 3), with median = 13.0 µg/L. Creatinine-adjusted trichloroacetic acid was the most frequently detected HAA in urine samples (69.2%), and moderately correlated with estimated drinking water intake (r = 0.48). SIGNIFICANCE: Findings provide valuable insights for DBP exposure assessment in epidemiological studies. Validation of predictive models in a larger number of samples and replication in different settings is warranted. IMPACT STATEMENT: Our study focused on assessing and describing the occurrence of several classes of DBPs in drinking water and developing exposure models of good predictive ability for non-regulated DBPs.
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Six mass spectrometers based on different mass analyzer technologies, such as time-of-flight (TOF), hybrid quadrupole-TOF (Q-TOF), orbitrap, Fourier transform ion cyclotron resonance (FT-ICR), and triple quadrupole (QqQ), installed at independent laboratories have been tested during a single day of work for the analysis of small molecules in negative electrospray ionization (ESI) mode. The uncertainty in the mass measurements obtained from each mass spectrometer has been determined by taking the precision and accuracy of replicate measurements into account. The present study is focused on calibration processes (before, after, and during the mass measurement), the resolving power of the mass spectrometers, and the data processing for obtaining elemental formulae. The mass range between m/z 100 and 600 has been evaluated with a mix of four standards. This mass range includes small molecules usually detected in food and environmental samples. Negative ESI has been tested as there is almost no data on accurate mass (AM) measurements in this mode. Moreover, it has been used because it is the ESI mode for analysis of many compounds, such as pharmaceutical, herbicides, and fluorinated compounds. Natural organic matter has been used to demonstrate the significance of ultrahigh-resolution in complex mixtures. Sub-millidalton accuracy and precision have been obtained with Q-TOF, FT-ICR, and orbitrap achieving equivalent results. Poorer accuracy and precision have been obtained with the QqQ used: 11 mDa root-mean-square error and 6-11 mDa standard deviation. Some advice and requirements for daily AM routine analysis are also discussed here.
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There is increasing awareness that exposure to endocrine disrupters interferes with lipid homeostasis in vertebrates, including fish. Many of these compounds exert their action by binding to nuclear receptors, such as peroxisome proliferator-activated receptors and retinoid X receptor. This work investigates the use of fish liver cells (PLHC-1 and ZFL cells) for the screening of metabolic and lipid disrupters in the aquatic environment by assessing changes in the cell's lipidome after exposure to the model compounds, tributyltin chloride and all-trans retinoic acid. Lipid extracts, analyzed by FIA-ESI (+/-) Orbitrap, evidenced the intracellular accumulation of triglycerides and diglycerides in both cell models after exposure to 100 and 200 nM tributyltin chloride for 24 h. Exposure to 1 µM all-trans retinoic acid led to a significant accumulation of triglycerides in PLHC-1 cells, while few triglycerides were accumulated in ZFL cells. Retinoic acid (cyp26b1, cyp3a65, lrata) and lipid metabolism (fasn, scd, elovl6) related genes were up-regulated by tributyltin chloride and all-trans retinoic acid, while only all-trans retinoic acid down-regulated the expression of dgat1a. The two cell models show sensitivity and responses to tributyltin chloride and all-trans retinoic acid comparable to those previously reported in mammalian cells. These results support the use of fish liver cells as alternative models for the detection of contaminants that act as lipid disrupters in the aquatic environment.
Assuntos
Metabolismo dos Lipídeos , Tretinoína/toxicidade , Compostos de Trialquitina/toxicidade , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cyprinidae , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/análise , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores X de Retinoides/metabolismo , Poluentes Químicos da Água/toxicidade , Peixe-ZebraRESUMO
The appearance of a bloom of cyanobacteria in the Sau-Susqueda-El Pasteral system (River Ter, NE Spain) in the autumn of 2015 has been the most recent episode of extensive bloom detected in Catalonia. This system is devoted mainly to urban supply, regulation of the river, irrigation and production of hydroelectric energy. In fact, it is one of the main supply systems for the metropolitan area of cities such as Barcelona and Girona. An assessment and management plan was implemented in order to minimize the risk associated to cyanobacteria. The reservoir was confined and periodic sampling was carried out. Low and high toxicity was detected by cell bioassays with human cell lines. Additionally, analysis studies were performed by enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-high resolution mass spectrometry (LC-HRMS). A microcystin target analysis and suspect screening of microcystins, nodularins, cylindrosperpmopsin and related cyanobacterial peptides by LC-HRMS were applied. The results for the analysis of microcystins were negative (<0.3 µg/L) in all the surface samples. Only traces of microcystin-LR, -RR and -dmRR were detected by LC-HRMS in a few ng/L from both fractions, aqueous and sestonic. In contrast, different anabaenopeptins and oscillamide Y at unusually high concentrations (µg-mg/L) were observed. To our knowledge, no previous studies have detected these bioactive peptides at such high levels. The reliable identification of these cyanobacterial peptides was achieved by HRMS. Although recently these peptides are detected frequently worldwide, these bioactive compounds have received little attention. Therefore, more studies on these substances are recommended, especially on their toxicity, health risk and presence in water resources.
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Cianobactérias/isolamento & purificação , Monitoramento Ambiental/métodos , Água Doce/análise , Água Doce/microbiologia , Espectrometria de Massas/métodos , Cromatografia Líquida/métodos , EspanhaRESUMO
Antibiotics can be uptaken by plants from soil desorption or directly from irrigation water, but their metabolization pathways in plants are largely unknown. In this paper, an analytical workflow based on high-resolution mass spectrometry was applied for the systematic identification of biotransformation products of ofloxacin in lettuce. The targeted metabolites were selected by comparing the mass chromatograms of exposed with control samples using an advanced spectra-processing method (Fragment Ion Search). The innovative methodology presented allowed us to identify a total of 11 metabolites, including 5 ofloxacin metabolites that are being reported for the first time in plants. Accordingly, major transformation pathways were proposed revealing insight into how ofloxacin and related chemicals are metabolized in lettuce. Furthermore, the influence of biotransformation on potential residual antimicrobial activity of identified compounds was discussed. Human exposure to antibiotics at doses below the minimum inhibitory concentrations is crucial in human risk assessment, including food ingestion; however, in the case of ofloxacin presented results reveal that plant metabolites should also be considered so as not to underestimate their risk.
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Lactuca/metabolismo , Ofloxacino/metabolismo , Antibacterianos/metabolismo , Biotransformação , HumanosRESUMO
BADGE (bisphenol A diglycidyl ether) is a synthesis product of bisphenol A (BPA), which, like other plasticizers, can cross the human placenta and reach the foetus. However, compared to BPA, there is almost no toxicological information. This work investigates the toxicity, endocrine and lipid disruption potential of BADGE and its hydrolysed and chlorinated derivatives (BADGE·H2O and BADGE·2HCl) in human placental JEG-3 cells. The analysis of culture medium by HPLC-ESI(+)-QqQ evidenced a good bioavailability of BADGE·2HCl and BADGE·H2O, but low stability of BADGE. Regardless, BADGE·2HCl and BADGE showed higher cytotoxicity than BADGE·H2O, which was the only compound that significantly inhibited CYP19 activity (IC50 49⯱â¯5⯵M). JEG-3â¯cells lipidome analyzed by FIA-ESI(+/-)-Orbitrap was significantly altered by exposure to BADGE·2HCl and BADGE at concentrations at the low µM range. BADGE·2HCl lead to a strong decrease of diacyl- and triacyl-glycerides (DGs,TGs) together with some membrane lipids, while BADGE lead to an accumulation of TGs. The results evidence the ability of BADGE and derivatives to affect placental lipid handling and to modulate placental CYP19 activity (BADGE·H2O) and highlights the need to monitor human exposure to these compounds, at least as intensely as BPA is monitored.
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Inibidores da Aromatase/toxicidade , Aromatase/metabolismo , Compostos Benzidrílicos/toxicidade , Carcinógenos/toxicidade , Compostos de Epóxi/toxicidade , Fenóis/toxicidade , Placenta/citologia , Plastificantes/toxicidade , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão/métodos , Diglicerídeos/metabolismo , Feminino , Humanos , Lipídeos de Membrana/metabolismo , Placenta/efeitos dos fármacos , Gravidez , Espécies Reativas de Oxigênio/metabolismo , Triglicerídeos/metabolismoRESUMO
Two small Lagocephalus sceleratus juveniles were captured in picarel targeting catches from North Aegean Sea (Greece) in the autumn of 2017. An electrochemical immunosensing tool using magnetic beads as immobilisation support was developed and applied to the rapid screening of tetrodotoxins (TTXs), potent neurotoxins that constitute a food safety hazard when present in seafood. This tool revealed the presence of TTXs in both individuals. Results were compared with those provided by mELISA and LC-HRMS, the latter confirming the presence of TTX. Some of the tissues contained TTX contents close to or above 2â¯mg/kg. L. sceleratus juveniles had been considered as non-toxic and, to our knowledge, this is the first report of high TTX levels in small L. sceleratus individuals. Such specimens can be mistaken with other edible species, posing a threat to consumers. The availability of low-cost and user-friendly tools for TTXs detection will contribute to guarantee seafood safety.
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Técnicas Eletroquímicas/métodos , Tetraodontiformes/metabolismo , Tetrodotoxina/análise , Animais , Bactérias/isolamento & purificação , Grécia , Separação Imunomagnética/métodos , Oceanos e Mares , Alimentos Marinhos/análise , Alimentos Marinhos/microbiologia , Tetraodontiformes/crescimento & desenvolvimento , Tetrodotoxina/isolamento & purificaçãoRESUMO
This study aimed to assess the bioaccessibility of different marine biotoxins in naturally contaminated shellfish and fish gonads using an in vitro digestion methodology. In general, hydrophilic toxins (domoic acid, paralytic shellfish poisoning toxins and tetrodotoxins) showed higher bioaccessibility than lipophilic ones (okadaic acid and azaspiracids). The bioaccessibility of toxins from the okadaic acid group ranged from 69% (raw European razor clams) to 74% (raw donax clams). Regarding azaspiracids, 47% of the initial content was bioaccessible in steamed blue mussel. As for hydrophilic toxins, 100% of the initial content was bioaccessible after digestion in raw shellfish and puffer fish gonads. The total tetrodotoxin bioaccessibility in puffer fish gonads decreased significantly after steaming. The profile of tetrodotoxins changed during the digestion process: TTX and 11-norTTX-6S-ol analogues decreased significantly after digestion, but the 5,6,11-trideoxy TTX analogue increased in both raw and steamed puffer fish gonads. These preliminary findings confirm the need to consider bioaccessibility data in future seafood risk assessment, as such information enables a more accurate and realistic estimation of potential seafood hazards, particularly in what concerns lipophilic toxins, therefore, constituting a crucial tool in the refinement of regulatory limits for the presence of biotoxins in seafood.
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Alimentos Marinhos/análise , Água do Mar/química , Toxinas Biológicas/farmacocinética , Animais , Disponibilidade Biológica , Humanos , Interações Hidrofóbicas e HidrofílicasRESUMO
This data article presents a set of non-wood native biomass sources identified for bioenergy production in isolated communities living in Amazon׳s extractive reserves. The data were inventoried using management plan documentation, which provides technical information on Amazon׳s protected areas. The sample was collected from a virtual database published by the Ministry of Environment, the federal body responsible for managing protected areas in Brazil. Five variables were extracted from the management plans to produce the inventory, which includes data on biomass typology, nomenclature and occurrence, as well as mode of access to communities and availability of energy sources in the protected areas.
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This data article focuses on sustainability indicators for bioenergy generation from Brazilian Amazon׳s non-woody native biomass sources, considered to be modern forms of biomass. In the construction of the indicators, the Indicator-based Framework for Evaluation of Natural Resource Management Systems (MESMIS, from the original Spanish) method was used, with the application of the seven sustainability attributes to identify critical points and limiting and favorable factors for sustainability. The data yielded a list of 29 indicators distributed across 27 critical points, selected from three system evaluation areas: 11 environmental indicators, 11 social indicators, and 7 economic indicators.
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In August 2014, a puffer fish poisoning incidence resulting in one fatality was reported in New Caledonia. Although tetrodotoxin (TTX) intoxication was established from the patients' signs and symptoms, the determination of TTX in the patient's urine, serum or plasma is essential to confirm the clinical diagnosis. To provide a simple cost-effective rapid screening tool for clinical analysis, a maleimide-based enzyme-linked immunosorbent assay (mELISA) adapted for the determination of TTX contents in human body fluids was assessed. The mELISA was applied to the analysis of urine samples from two patients and a response for the presence of TTX and/or structurally similar analogues was detected in all samples. The analysis by LC-MS/MS confirmed the presence of TTX but also TTX analogues (4-epiTTX, 4,9-anhydroTTX and 5,6,11-trideoxyTTX) in the urine. A change in the multi-toxin profile in the urine based on time following consumption was observed. LC-MS/MS analysis of serum and plasma samples also revealed the presence of TTX (32.9â¯ng/mL) and 5,6,11-trideoxyTTX (374.6â¯ng/mL) in the post-mortem plasma. The results provide for the first time the TTX multi-toxin profile of human samples from a puffer fish intoxication and clearly demonstrate the implication of TTX as the causative agent of the reported intoxication case.
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Ensaio de Imunoadsorção Enzimática/métodos , Doenças Transmitidas por Alimentos/diagnóstico , Toxinas Marinhas/química , Alimentos Marinhos/intoxicação , Tetraodontiformes , Tetrodotoxina/química , Animais , Cromatografia Líquida de Alta Pressão , Contaminação de Alimentos/análise , Doenças Transmitidas por Alimentos/sangue , Doenças Transmitidas por Alimentos/urina , Humanos , Toxinas Marinhas/sangue , Toxinas Marinhas/urina , Nova Caledônia , Espectrometria de Massas em Tandem , Tetrodotoxina/análogos & derivados , Tetrodotoxina/sangue , Tetrodotoxina/urinaRESUMO
ABSTRACT Objective: to investigate situations that interfere with health professionals' performance, in the identification and maintenance of potential brain-dead donors in an Adult Emergency Care Unit, and to indicate actions, from the health team's perception, that can promote care for these patients. Method: this is a qualitative, convergent care research, carried out with health professionals from an adult emergency unit, a large public teaching hospital located in southern Brazil. Data were collected between February 2020 and January 2021 through structured participant observation, semi-structured interview and convergence group. Data analysis was performed according to the stages of gathering, synthesis, theorization and transfer. Results: based on data analysis, the difficulties in caring for brain-dead persons, lack of information, inadequate structure and difficulties in the work process were identified as situations that interfere with health professionals' performance in relation to brain-dead potential donors. Actions were signaled to promote care for these people through continuing education activities on the subject, dissemination of information, construction of technologies/protocols, organization of infrastructure and the work process with support for professionals who assist these people. Conclusion: it was evidenced that professional education, the organization of the donation process and systematized actions to improve the work are fundamental factors for the effective care for brain-dead potential organ donors.
RESUMEN Objetivo: investigar situaciones que interfieren en la actuación de los profesionales de la salud, en la identificación y mantenimiento de potenciales donantes en muerte encefálica en una unidad de emergencia de adultos y señalar acciones, en la percepción del equipo de salud, que puedan promover el cuidado de estos pacientes. Método: Investigación cualitativa, de asistencia convergente, realizada con profesionales de la salud de una unidad de emergencia de adultos, de un gran hospital público de enseñanza ubicado en el sur de Brasil. Los datos fueron recolectados entre febrero de 2020 y enero de 2021 a través de observación participante estructurada, entrevista semiestructurada y grupo de convergencia. El análisis de los datos se realizó según las etapas: aprehensión, síntesis, teorización y transferencia. Resultados: con base en el análisis de los datos, las dificultades en la asistencia a la persona con muerte encefálica, falta de información, estructura inadecuada y dificultades en el proceso de trabajo fueron identificadas como situaciones que interfieren en la actuación de los profesionales de la salud en relación al potencial donante en cerebro muerte. Se señalaron acciones para promover la asistencia a estas personas a través de actividades de educación continua en el tema, difusión de información, construcción de tecnologías/protocolos, organización de la infraestructura y del proceso de trabajo con apoyo a los profesionales que asisten a estas personas. Conclusión: se evidenció que la educación profesional, la organización del proceso de donación y acciones sistematizadas para mejorar el trabajo son factores fundamentales para la asistencia eficaz al potencial donante de órganos en muerte encefálica.
RESUMO Objetivo: investigar situações que interferem na atuação dos profissionais da saúde, na identificação e manutenção do potencial doador em morte encefálica em uma unidade de pronto-socorro adulto e sinalizar ações, na percepção da equipe de saúde, que possam promover a assistência a esses pacientes. Método: pesquisa qualitativa, convergente-assistencial, realizada com profissionais de saúde de uma unidade de pronto-socorro adulto, de um hospital-escola público e de grande porte localizado no Sul do Brasil. Os dados foram coletados entre fevereiro de 2020 e janeiro de 2021 por meio de observação estruturada participante, entrevista semiestruturada e grupo de convergência. A análise dos dados foi realizada de acordo com as etapas: apreensão, síntese, teorização e transferência. Resultados: a partir da análise dos dados identificaram-se como situações que interferem na atuação dos profissionais da saúde frente ao potencial doador em morte encefálica as dificuldades na assistência à pessoa em morte encefálica, a falta de informação, estrutura inadequada e as dificuldades no processo de trabalho. Houve sinalização de ações para promover a assistência a essas pessoas por meio de atividades de educação permanente sobre o tema, divulgação de informações, construção de tecnologias/protocolos, organização da infraestrutura e do processo de trabalho com suporte aos profissionais que atendem essas pessoas. Conclusão: evidenciou-se que a educação profissional, a organização do processo de doação e ações sistematizadas para aperfeiçoamento do trabalho são fatores fundamentais para a efetiva assistência ao potencial doador de órgãos em morte encefálica.