Contribution to the risk assessment of ciguatera poisoning in europe (the balearic and Canary Islands and Madeira archipelago)

Actualment, la intoxicació alimentaria de la ciguatera (CP) es considera una malaltia emergent a les zones temperades d'Europa. La present tesi contribueix a l'avaluació del risc d'intoxicació per ciguatera a les Illes Balears (Mediterrània Occidental), Illes Canàries i arxipèlag de Madeira (Macaronèsia, Atlàntic Est). El present estudi caracteritza la biodiversitat i la distribució geogràfica dels gèneres Gambierdiscus i Fukuyoa a les illes més importants de Balears i Canàries. A més, s'avalua la producció potencial de CTX de les espècies Gambierdiscus i Fukuyoa. El risc de CP també s'avalua mitjançant la detecció de ciguatoxines en peixos de les Illes Balears i de l'Arxipèlag de Madeira. Tot i que la CP no és present a les Illes Balears, com es de-mostrarà aquí, la ciguatera es troba en l'etapa en la qual les poblacions de microalgues estan produint toxines similars a CTX i ho estan fent des de fa alguns anys. A la Macaronèsia, les troballes recents sobre les espècies de Gambierdiscus i les ciguatoxines en els mariscs han demostrat que encara hi ha poc coneixement sobre el fenomen de la CP i els seus agents causants.Actualmente, la intoxicación alimentaria de la ciguatera (CP) se considera una enfermedad emergente en las zonas tem-pladas de Europa. La presente tesis contribuye a la evaluación del riesgo de intoxicación por ciguatera en las Islas Baleares (Mediterráneo Occidental), Islas Canarias y Archipiélago de Madeira (Macaronensia, Atlántico Este). El presente estudio caracteriza la biodiversidad y la distribución geográfica de los gé-neros Gambierdiscus y Fukuyoa en las islas más importantes de Baleares y Canarias. Además, se evalúa la producción potencial de CTX de las especies Gambierdiscus y Fukuyoa. El riesgo de CP también se evalúa mediante la detección de ciguatoxinas en peces de las Islas Baleares y del Archipiélago de Madeira. Aun-que la CP no está presente en las Islas Baleares, como se demostrará aquí, la ciguatera se encuentra en la etapa en la que las poblaciones de microalgas están produciendo toxinas similares a CTX y lo están haciendo desde hace algunos años. En la Macaronesia, los hallazgos recientes sobre las especies de Gam-bierdiscus y las ciguatoxinas en los mariscos han demostrado que todavía hay poco conocimiento sobre el fenómeno de la CP y sus agentes causantes.Currently, ciguatera poisoning (CP) is now considered to be an emerging disease in temperate areas of Europe. The present thesis is a contribution to the risk assessment of ciguatera poisoning in the Balearic Islands (West Mediterranean Sea), the Canary Islands and the Madeira Archipelago (Macaronesia, East Atlantic). The current study characterizes the biodiversity and the geographical distribution of the Gam-bierdiscus and Fukuyoa genera in the most important islands of the Balearic and Canary Islands. In addi-tion, the potential CTX production of Gambierdiscus and Fukuyoa species is evaluated. The risk of CP is also evaluated by the detection of the ciguatoxins in fish from the Balearic Islands and Madeira Archipe-lago. Even though CP is not present in the Balearic Islands, as will be demonstrated here, ciguatera is at the stage that populations of microalgae are producing CTX-like toxins and have been doing so for some years. In the Macaronesia, recent findings about Gambierdiscus species and ciguatoxins in seafood have shown that it is still little knowledge about CP phenomenon and their causative agents

Diversity of introduced terrestrial flatworms in the Iberian Peninsula: a cautionary tale

Many tropical terrestrial planarians (Platyhelminthes, Geoplanidae) have been introduced around the globe. One of these species is known to cause significant decline in earthworm populations, resulting in a reduction of ecological functions that earthworms provide. Flatworms, additionally, are a potential risk to other species that have the same dietary needs. Hence, the planarian invasion might cause significant economic losses in agriculture and damage to the ecosystem. In the Iberian Peninsula only Bipalium kewense Moseley, 1878 had been cited till 2007. From that year on, four more species have been cited, and several reports of the presence of these animals in particular gardens have been received. In the present study we have: (1) analyzed the animals sent by non-specialists and also the presence of terrestrial planarians in plant nurseries and garden centers; (2) identified their species through morphological and phylogenetic molecular analyses, including representatives of their areas of origin; (3) revised their dietary sources and (4) used Species Distribution Modeling (SDM) for one species to evaluate the risk of its introduction to natural areas. The results have shown the presence of at least ten species of alien terrestrial planarians, from all its phylogenetic range. International plant trade is the source of these animals, and many garden centers are acting as reservoirs. Also, landscape restoration to reintroduce autochthonous plants has facilitated their introduction close to natural forests and agricultural fields. In conclusion, there is a need to take measures on plant trade and to have special care in the treatment of restored habitats

First report of Gambierdiscus in the Western Mediterranean Sea (Balearic Islands)

Gambierdiscus (Dinophyceae) species are benthic dinoflagellates living in marine littoral zones of circumtropical areas and have recently been described in temperate waters [1]. Some species are producers of potent neurotoxins: ciguatoxins (CTXs) and maitotoxins (MTXs). Ciguatoxins are linked to Ciguatera Fish Poisoning (CFP). Ciguatera used to be restricted to tropical and subtropical areas, but since the last decade, it appears to be expanding to more temperate latitudes. For example, outbreaks of ciguatera have been reported in the Canary Islands and Madeira (eastern Atlantic Ocean), where several species of the genus Gambierdiscus have been identified.info:eu-repo/semantics/publishedVersio

Further Advance of Gambierdiscus Species in the Canary Islands, with the First Report of Gambierdiscus belizeanus

Ciguatera Poisoning (CP) is a human food-borne poisoning that has been known since ancient times to be found mainly in tropical and subtropical areas, which occurs when fish or very rarely invertebrates contaminated with ciguatoxins (CTXs) are consumed. The genus of marine benthic dinoflagellates Gambierdiscus produces CTX precursors. The presence of Gambierdiscus species in a region is one indicator of CP risk. The Canary Islands (North Eastern Atlantic Ocean) is an area where CP cases have been reported since 2004. In the present study, samplings for Gambierdiscus cells were conducted in this area during 2016 and 2017. Gambierdiscus cells were isolated and identified as G. australes, G. excentricus, G. caribaeus, and G. belizeanus by molecular analysis. In this study, G. belizeanus is reported for the first time in the Canary Islands. Gambierdiscus isolates were cultured, and the CTX-like toxicity of forty-one strains was evaluated with the neuroblastoma cell-based assay (neuro-2a CBA). G. excentricus exhibited the highest CTX-like toxicity (9.5–2566.7 fg CTX1B equiv. cell−1) followed by G. australes (1.7–452.6.2 fg CTX1B equiv. cell−1). By contrast, the toxicity of G. belizeanus was low (5.6 fg CTX1B equiv. cell−1), and G. caribaeus did not exhibit CTX-like toxicity. In addition, for the G. belizeanus strain, the production of CTXs was evaluated with a colorimetric immunoassay and an electrochemical immunosensor resulting in G. belizeanus producing two types of CTX congeners (CTX1B and CTX3C series congeners) and can contribute to CP in the Canary Islands.info:eu-repo/semantics/publishedVersio

Chronic intestinal pseudoobstruction and ophthalmoplegia in a patient with mitochondrial myopathy

A 38 year old woman having chronic intestinal pseudoobstruction associated with mitochondrial myopathy is reported. The clinical and radiographic features suggested the diagnosis of chronic intestinal pseudoobstruction. Muscular atrophy and ophthalmoplegia led to muscle biopsy, which disclosed accumulation of normal and abnormal mitochondria ('ragged red fibres'), characteristic of mitochondrial myopathy

Tuberculosis recurrence after completion treatment in a European city: reinfection or relapse?

Background Tuberculosis (TB) recurrence can be due to reinfection or relapse. The contribution of each to TB incidence and the factors associated with recurrence are not well known. Effectiveness of TB control programs is assessed in part by recurrence rates. The aim of this study was to establish the recurrence rate of TB in Barcelona, the associated risk factors and the role of reinfection. Methods A population-based retrospective longitudinal study was performed in Barcelona, Spain. TB patients with positive culture results who completed treatment between Jan 1st, 2003 and Dec 31st, 2006 were followed-up until December 31st, 2009 by the TB Control Program. The incidence rate of recurrence was calculated per person-year of follow-up (py). Kaplan-Meier and Cox regression methods were used for the survival analysis by calculating the hazard ratio (HR) with 95% confidence intervals (CI). Results Of the 1,823 TB cases identified, 971 fulfilled the inclusion criteria and 13 (1.3%) had recurrent TB. The recurrence rate was 341 cases per 100,000 py, 13 times higher than the TB incidence of the general population. Likelihood of TB recurrence at the 1st, 3rd and 5th year of follow-up was 0.1%, 1.4% and 1.6%, respectively. Factors associated with recurrence were HIV infection (HR: 4.7, CI: 1.4-15.7), living in the inner city district (HR: 3.9, CI: 1.3-11.8) and history of TB treatment (HR: 5.2, CI: 1.7-16.2). Genotyping results of recurrent cases were available for 6 patients (3 reinfections and 3 relapses). Conclusion The rate of TB recurrence in Barcelona is low and most episodes occur within the first three years. Patients at higher risk of recurrence are co-infected with HIV, living in neighborhoods with high TB incidence or with a history of TB treatment. When available, genotyping results help determine whether the recurrence is due to reinfection or relapse

Identification of New CTX Analogues in Fish from the Madeira and Selvagens Archipelagos by Neuro-2a CBA and LC-HRMS

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.The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) under the ECsafeSEAFOOD project (grant agreement n° 311820). À.T. acknowledges IRTA-URV-Banco Santander for the Ph.D. grant (2016 PMF-PIPF-74).Peer reviewe

Genetic study and geographical modelling distribution of the Ciguatera-Causing dinoflagellates, Gambierdiscus and Fukuyoa genera

Gambierdiscus y Fukuyoa son dos géneros de dinoflagelados que se encuentran principalmente en zonas tropicales, pero en las últimas décadas se han detectado en zonas temperadas o más frías. Parece ser que hay una expansión de estas microalgas mediada por el cambio climático. Con este trabajo se quiere hacer una aproximación para examinar la diversidad genética de este género, ver si hay una relación genética y geográfica. Para ello se han utilizado herramientas clásicas de análisis genético. También se ha querido modelizar la presencia o ausencia de especies o de cada género, mediante modelos logísticos con un gran número de variables Como resultado se han creado largo datasets de secuencias asociadas a coordenadas. Se ha podido ver la diversidad de ambos géneros y se ha podido calcular modelos logísticos para determinar una presencia o ausencia de las microalgas.Los trópicos albergan una gran diversidad de especies de estos dinolfagellados, pero podría haber índices de que se están expandiendo las especies. Por ahora con nuestros resultados, no se pueden concluir que haya una expansión, pero este trabajo es una primera aproximación para ver este tipo de expansiones de las microalgas. También hay un primer análisis con modelos logísticos basado en la presencia y ausencia de las microalgas para ver comprender qué variables determinan la distribución geográfica de las especies, estos análisis se pueden perfeccionar posteriormente con modelos más potentes.Gambierdiscus i Fukuyoa són dos gèneres de dinoflagel·lats que es troben principalment en zones tropicals, però en les últimes dècades s'han detectat en zones temperades o més fredes. Sembla ser que hi ha una expansió d'aquestes microalgues intervinguda pel canvi climàtic. Amb aquest treball es vol fer una aproximació per examinar la diversitat genètica d'aquest gènere, veure si hi ha una relació genètica i geogràfica. Per a això s'han utilitzat eines clàssiques d'anàlisi genètica. També s'ha volgut modelitzar la presència o absència d'espècies o de cada gènere, mitjançant models logístics amb un gran nombre de variables Com a resultat s'han creat llarg datasets de seqüències associades a coordenades. S'ha pogut veure la diversitat de tots dos gèneres i s'ha pogut calcular models logístics per determinar una presència o absència de les microalgas.Los tròpics alberguen una gran diversitat d'espècies d'aquests dinolfagellados, però podria haver índexs que s'estan expandint les espècies. Per ara amb els nostres resultats, no es poden concloure que hi hagi una expansió, però aquest treball és una primera aproximació per veure aquest tipus d'expansions de les microalgues. També hi ha una primera anàlisi amb models logístics basat en la presència i absència de les microalgues per veure comprendre quines variables determinen la distribució geogràfica de les espècies, aquestes anàlisis es poden perfeccionar posteriorment amb models més potents.Gambierdiscus and Fukuyoa are two genera of dinoflagellates found mainly in tropical zones, but in recent decades these species have been detected in cooler-temperate zones. It seems that there is an expansion of these microalgae mediated by climate change. Aims of this work are study the genetic diversity of these genera, see if there is a genetic and geographical relationships and analyse the possible expansion. To this end, classical genetic analysis tools have been used. In addition, efforts have been done to model the presence or absence of species for each genera through logistic models with a large number of environmental variables. As a result, long datasets of sequences associated with coordinates have been created. Throughout the created dataset has been analysed the diversity of both genera. Also, logistic models have been calculated to determine the presence or absence of microalgae. Tropical zones are hotspots of these dinoflagellates, but genetic indices of expansion might exist. For now, with our results, it is not possible to conclude that there is an expansion to cool areas, but this work is a first approach to observe this type of expansions in dinoflagellate. In addition, first analysis with logistic models based on the presence and absence of microalgae are studied, these analyses can be further completed in the future with more powerful models

Automated Patch Clamp for the Detection of Tetrodotoxin in Pufferfish Samples

Tetrodotoxin (TTX) is a marine toxin responsible for many intoxications around the world. Its presence in some pufferfish species and, as recently reported, in shellfish, poses a serious health concern. Although TTX is not routinely monitored, there is a need for fast, sensitive, reliable, and simple methods for its detection and quantification. In this work, we describe the use of an automated patch clamp (APC) system with Neuro-2a cells for the determination of TTX contents in pufferfish samples. The cells showed an IC50 of 6.4 nM for TTX and were not affected by the presence of muscle, skin, liver, and gonad tissues of a Sphoeroides pachygaster specimen (TTX-free) when analysed at 10 mg/mL. The LOD achieved with this technique was 0.05 mg TTX equiv./kg, which is far below the Japanese regulatory limit of 2 mg TTX equiv./kg. The APC system was applied to the analysis of extracts of a Lagocephalus sceleratus specimen, showing TTX contents that followed the trend of gonads > liver > skin > muscle. The APC system, providing an in vitro toxicological approach, offers the advantages of being sensitive, rapid, and reliable for the detection of TTX-like compounds in seafood.This research was funded by the Ministerio de Ciencia e Innovación (MICIN) and the Agencia Estatal de Investigación (AEI) (Spain) through the CELLECTRA (PID2020-112976RB-C21 and PID2020-112976RB-C22) project. J.R. acknowledges IRTA for his Ph.D. grant (CPI0422). A.T. acknowledges Ministerio de Universidades (Spain) for her Margarita Salas postdoc grant (2021URV-MS-24). M.A. acknowledges MICIN and AEI for her Ph.D. grant (PRE2019-088181). Authors also acknowledge the support from the CERCA Program/Generalitat de Catalunya. The article processing fees were funded by MDPI.info:eu-repo/semantics/publishedVersio

New information about the toxicological profile of Prorocentrum panamense (Prorocentrales, Dinophyceae) and its global distribution

Dinoflagellates of the genera Prorocentrum and Dinophysis are known producers of toxic compounds belonging to the okadaic acid (OA) group. The ingestion of shellfish contaminated with these toxins causes a human disease named diarrheic shellfish poisoning (DSP). In this study, the first record of Prorocentrum panamense, a potential toxin-producer species, was reported in the Canary Islands together with its toxicological characterization. Prorocentrum panamense cells were collected during April 2017 from natural pools located in the Northeastern part of Gran Canaria. This new record represents an expansion of P. panamense distribution area, previously restricted to the Pacific Ocean, Indian Ocean, Arabian Gulf and the Caribbean, and its introduction mechanisms are discussed. Laboratory cultures of P. panamense were settled and toxin production was assessed in both cell pellets and culture media at four different growth phases (latency, exponential, early stationary and late stationary) implementing LC-MS/MS and neuro-2a cell-based assay (CBA). LC-MS/MS allowed the identification of OA in the fraction corresponding to the late stationary phase, and tests performed on neuro-2a cells showed, for most of the fractions, OA-like activity observable by both cell morphology changes and cell mortality. This information is fundamental for a better understanding of the genus Prorocentrum global distribution, its ecology, and risks associated to toxic producing species.info:eu-repo/semantics/acceptedVersio