Controlo sanitário de parasitas de peixes nas pescarias do Atlântico

261 páginas[EN] European fisheries represent one of the leading economic activities in the world. Marine parasites with public health and industrial concern have become a key issue in major European markets, due to three main reasons: (1) the presence of a reported increasing number of allergic and gastrointestinal disorders caused by fish-borne parasitic infections, (2) the commercial impact and high economic losses due to fish rejections, and (3) the applicability of Regulation EC 178/2002, which states that “fish with visible parasites is unfit for human consumption”. Over the last few years, since the entry into force of European and Member States regulations on food and specifically fishery products, co-responsibility for food quality and safety has lain with food industry, which has introduced Hazard Analysis and Critical Control Points (HACCP) programs in all its actions concerning the food chain. Consequently, significant progresses have been achieved regarding the prevention of parasites in seafood products. However, there is a lack of consensus and standardization for parasite inspection at fishing companies, and no efficient and accurate modus operandi exists to be implemented and accepted by the industry as a routine technique. The EU legal framework defined by zoosanitary regulations, scientific opinions from the European Food Safety Authority (EFSA), as well as the European Hygiene Package among others, has provided a basis on which the fishing sector has focussed its activity. Accordingly, this dissertation has been driven by these considerations during the course of its execution. This context led us to carry out a meticulous horizon scanning under a multidisciplinary approach, as an overview tool in proactive risk management. This fundamental practice takes due account of the stringent requirements that new markets are demanding to fishing industry, and the lacks and needs of the fishing sector with regard to the impact of the most relevant parasites with public health and industrial concern. A comprehensive technical evaluation and laboratory testing of the official parasite detection methods evidenced low reliability within the two most commonly used qualitative inspection procedures in fish processing (i.e. candling, gross visual inspection). Consistent parallel research carried out, has given as a result innovative scientific developments for diagnostic purposes and for the optimization of the current detection procedures. These technological improvements have been presented in more accessible and manageable formats for their incorporation into self-control programs at the fishing industry. Furthermore, the huge amount of inspection work carried out in the most relevant fish species, has allowed reaching a deeper knowledge concerning three very important parasite groups that are impacting on fishing industry; microsporidians, anisakids and copepods. Finally, the design and application of two innovating tools for parasite management (a scoring system for predictive assessment of fish lots, and a transfer of knowledge model presented in web format), helpful for seafood producers, policy makers and general public, are good examples of how to contribute stimulating the exchanging of ideas among stakeholders and improving the inspection scheme. They are also the best approach for helping to convert scientific findings and technological advances into industrial and commercial success. Scientific excellence requires investment in R&D&I with regard to acquire and expand a sound scientific basis for policy and regulation on food safety, and also for helping fishing industry to achieve a preventing plan which provides added value products.The high national and international exporting activity carried out daily from the most important fishing ports of Portugal and from the fishing Port of Vigo, requires that strict control measures based on groundbreaking scientific advances, have to be incorporated into proactive selfinspections made by seafood companies. These measures must include effective preventing and corrective actions in the edible part of heavily infected fish species, thus guaranteeing products of the highest safety and quality to final consumers.[ES] La industria pesquera en Europa constituye una de las principales actividades económicas del mundo. Las parasitosis de origen marino con repercusiones comerciales e implicaciones en la salud pública se han convertido en un problema clave en los mercados europeos debido a tres motivos principales: (1) al incremento en el número de notificaciones de alergias y desórdenes gastrointestinales causados por infecciones parasitarias transmitidas tras el consumo de pescado, (2) al impacto comercial y las elevadas pérdidas económicas debidas a los rechazos por la presencia de parásitos visibles (y/o sus lesiones asociadas), y (3) a la aplicación del Reglamento CE 178/2002, el cual establece que “todo pescado visiblemente parasitado es considerado no apto para el consumo humano”. En los últimos años, a partir de la entrada en vigor de reglamentos específicos sobre los productos de la pesca (tanto a nivel europeo como a nivel de los Estados miembros), la corresponsabilidad de la calidad y seguridad alimentaria ha recaído sobre la industria alimentaria, que consecuentemente ha incorporado programas de Análisis de Peligros y Puntos Críticos de Control (APPCC) a todas sus actuaciones entorno a la cadena alimentaria. En consecuencia, todo ello ha comportado el logro de considerables avances concernientes a la prevención de los parásitos en productos marinos. Sin embargo, la ausencia de un modus operandi lo suficientemente eficiente y fiable en la inspección parasitaria como para ser implementado y aceptado por el sector pesquero como técnica de rutina, es fiel reflejo de la falta de consenso y estandarización existente entre las compañías pesqueras. El marco legal de la UE definido por los reglamentos zoosanitarios, las opiniones científicas de la Agencia Europea de Seguridad Alimentaria (AESA), y por el Paquete de Higiene Alimentaria entre otros, ha sentado las bases sobre las que el sector pesquero ha fundamentado su actividad, y en consecuencia, sobre las que el desarrollo de la presente tesis doctoral ha focalizado su atención. Este mismo contexto es el que nos ha llevado a realizar un meticuloso “horizon scanning” bajo un enfoque multidisciplinario y a modo de herramienta “radar”. Este instrumento resulta fundamental para la gestión proactiva de riesgos, y debe tener en cuenta las principales exigencias de los nuevos mercados de la industria pesquera, así como las carencias y necesidades del sector en relación al impacto de los parásitos con mayores implicaciones sanitarias y comerciales. La evaluación técnica y las exhaustivas pruebas de laboratorio realizadas en este trabajo para valorar la fiabilidad de los dos métodos cualitativos de detección oficiales más utilizados durante el procesado de pescado (candling e inspección visual) evidenciaron que estos procedimientos presentan un bajo nivel de fiabilidad. Las investigaciones ejecutadas en paralelo permitieron optimizar los métodos de detección de parásitos en productos de la pesca vigentes, así como desarrollar innovaciones tecnológicas con fines diagnósticos. Algunas de éstas han sido presentadas en un formato más accesible y manejable para facilitar su incorporación en los programas de autocontrol de las industrias pesqueras.Por otra parte, el amplio trabajo de inspección realizado con las especies de pescado de mayor interés comercial, permitió llegar a un conocimiento mucho más detallado de tres importantísimos grupos de parásitos que actualmente tienen un alto impacto sobre el sector; microsporidios, anisákidos y copépodos. Finalmente, el diseño, la creación y la aplicación práctica de dos herramientas innovadoras para la gestión de parasitosis (un sistema de evaluación predictiva en lotes de pescado, y un modelo de transferencia de conocimiento en formato web) útiles para las empresas pesqueras, las autoridades sanitarias, y los consumidores, han demostrado ser buenos ejemplos de cómo contribuir a estimular el intercambio de ideas entre las partes interesadas, a mejorar la eficacia del esquema de inspección, y sobre todo a convertir los hallazgos científicos y los avances tecnológicos en éxito industrial y comercial. La excelencia científica requiere inversión en I+D+i a fin de adquirir y expandir una base científica sólida para la normalización y vigilancia de la seguridad alimentaria, además de para ayudar a la industria pesquera a conseguir un plan de prevención que permita ofrecer productos de alto valor añadido. La intensa actividad diaria de exportación nacional e internacional que tiene lugar en el puerto pesquero de Vigo y en los puertos pesqueros más importantes de Portugal, requiere que las estrictas medidas de control basadas en los avances tecnológicos y científicos más recientes sean integradas en los programas proactivos de autocontrol de las empresas pesqueras. Asimismo, estas medidas deben incluir acciones preventivas y correctivas efectivas sobre la parte comestible de los peces gravemente parasitados, garantizando así, productos con el más alto nivel de calidad y seguridad para el consumidor final.Apoio financeiro da FCT e do FSE no âmbito do III Quadro Comunitário de ApoioPeer reviewe

Mass ingestion of gastroliths and other foreign bodies in three juvenile hooded seals (Cystophora cristata) stranded in north-western Iberian Peninsula

We present here three clinical cases involving mass ingestion of foreign bodies in Hooded seals (Cystophora cristata) stranded in north-western Iberian Peninsula. Although the presence of gastroliths is considered to be normal in pinnipeds, the cases presented here highlight how an excessive presence of them as well as other foreign bodies could result in rapid onset of a potentially lethal gastrointestinal stasis syndrome, which has to be quickly resolved, medically or surgically. Ultrasound examinations and posterior X-ray confirmation have demonstrated their utility to a rapid detection of gastric bodies, and have to be taken into account in Hooded seal routine clinical protocols. Finally, we conclude that it is particularly important to avoid the use of loose stones or sand over resting areas and to take extreme precautions with small items near the rehabilitation pools when dealing with this seal species.JM Alonso-Farré and M. Llarena-Reino are currently funded by Fundação para a Ciência e a Tecnologia, (Portugal), under post-doctoral fellowship SFRH/BPD/47251/2008, and pre-doctoral grant SFRH/BD/45398/2008, respectively.Peer reviewe

A scoring system approach for the parasite predictive assessment of fish lots: a proof of concept with anisakids

8 páginas, 2 figuras, 2 tablasA total of 982 individuals distributed in 11 lots belonging to 10 fish species from three Atlantic FAO fishing areas were sampled and examined to detect the presence of anisakid larvae in fish muscle. After hazard identification by genetic sequencing and exposure assessment by anatomic extent and demographic characterization of infection, all data were fitted for each fish species to a new proposed scoring schema of parasite prediction. In the absence of a criterion standard method for inspection and precise definition of the quantum satis for parasites in contaminated fish lots, the inspection rating scheme called SADE (Site of infection, Assurance of quality, Demography, Epidemiology) may help fish industries to precisely handle and to evaluate the likely outcome of infected fish lots after being diagnosed. For this purpose, a supporting flow diagram for decision was defined and suggested. This new performance assessment tool has the aim of staging fish lots, thus helping in planning manufacture, commercial, and research decisions during self-management programs. This novel scoring system provides an improved inspection format by implementing the occurrence stratification for parasites to guide Hazard Analysis and Critical Control Points (HACCP) programs for the uniform exchange of information among fish industries, administration and researchers, thus facilitating standardization and communication. In the future, this scoring version could be validated (in terms of classification and wording) for similar overall predictive purposes in other muscular parasites infecting seafood products.Xunta de Galicia, Projects INCITE-07MMA015CT, IN841C, 10TAL001CT, and 10TAL033EFundaçao para a Ciencia e a Tecnologia, Grant SFRH/BD/45398/2008Peer reviewe

Horizon scanning for management of emerging parasitic infections in fishery products

10 páginas, 3 figuras, 1 tablaPublic organizations operating in health and food-safety sectors are increasingly realizing the advantages of the long-term view of risk uncertainties associated to biological hazards, served-up in the short-term to anticipate the problem and its handling. Thus, the horizon scanning is becoming a major strand in proactive risk management and patient-consumer protection continuity. This approach was recently explained in the scientific opinion on risk assessment of parasites in fishery products by the European Food Safety Authority, EFSA (2010), followed by the launching of a funding scheme for a specific EU Framework Program Project under the Knowledge Based Bio-Economy concept, KBBE (FP7-KBBE-2012-6), which drives the new EU 2020 strategy. The aim of this paper is to examine horizon scanning issues in relation to public health and industrial concern on the presence of parasites in fishery products recorded in the Rapid Alert System for Food and Feed (RASFF) System. We focus on specific threats, targets, methods and challenges as a means of acquiring management goals and future objectives. The proposed horizon scanning identifies emerging ideas/technologies for an early handling of parasitized fish stocks/products for priority setting to inform strategic planning of stakeholders, policy-makers and health services. In order to accomplish this, a set of risk GIS maps illustrating the state of art about the presence of the zoonotic Anisakis spp. on commercial fish stocks of the last 65 years was firstly developed. Secondly, a program of 108 surveys among fish sellers of Galicia (NW Spain) were carried out with the main objective of getting information about hazard recognition, fish product management practices, quality self-controls and corrective and preventive measures in use. Additionally, during the “I International Symposium on strategies for management of parasitized seafood products” (Vigo, Spain), groups of researchers, technologists, official inspectors and industries participated in round tables with 3 different perspectives: market-industry, inspection and academia. All scanners agreed that the status quo to manage fish parasites in the production-to-consumption food pathway is unsatisfactory. The central message proposed a stable network performance based on collaborative software to provide multi-level information for industrial management of parasite contaminants in fish products. The discussion group also proposed to invigorate collaborative translational research and professional training as key drivers to fuel technological innovations and tech transfer, which may help to minimize/eliminate the risk of parasites that have public health and economic impacts in fish products.We thank Xunta de Galicia for financial support under Projects Parcode (10TAL033E), Anitech (10TAL001CT) and IN841C. Authors would like to thanks to Miguel Bao (ECOBIOMAR) the excellent bibliographic work, and to Carlos Vello/Luis Outeiriño (Comercial Hospitalaria Grupo 3 S.L.) and Rosa Fernández (CETMAR) for their help in the organization of the I International Symposium on strategies for management of parasitized seafood products. M. Llarena-Reino and M. Regueira thanks Fundação para a Ciência e a Tecnologia and European Social Fund for financial support under grants SFRH/BD/45398/2008 and SFRH/BD/51038/2010 respectively.Peer reviewe

Morphological and genetic identification of Pennella instructa (Copepoda: Pennellidae) on Atlantic swordfish (Xiphias gladius, L. 1758)

8 pages, 6 figures, 1 tableDuring the last years the presence of parasites in commercial fish species has increased becoming an emergent major sanitary problem. For seafood companies the control of this biological hazard is turning into a priority issue, as quality of the products is now being seriously compromised. The swordfish Xiphias gladius, as one of the most important commercial species among European markets, has been inspected for the presence of pennellids. A total of 214 mesoparasitic copepods were sampled for epidemiology and genetic analysis, 5 pennellid cephalothoraxes’ for morphological and genetic studies and 50 slices of parasitized swordfishes were examined for internal lesions and diagnostics on the basis of morphological characters. All hosts were captured in Atlantic waters. Pennella instructa was morphologically identified as the only metazoan species in the 5 cephalothoraxes. Morphological characters and internal and external lesions caused by P. instructa were acutely described. Prevalence and mean Intensity of infection were determined for the total fishes parasitized. Copepods were molecularly characterized and the phylogenetic relationships were explored based on 18S and 28S rDNA sequences. Phylogenetic analyses using maximum likelihood models were performed including pennellid external portions and cephalothoraxes (morphologically identified as P. instructa). All pennellid samples were clustered into the same clade with bootstrap values of 100% for both genes sequenced. The prevention of rejections due to these parasites must be the first step to ensure safer and high quality standard products to final consumers. Monitoring actions and proactive self-inspections including preventive and corrective measures should be more intensively integrated into HACCP systems of seafood companiesXunta de Galicia has funded this work under Projects INCITE‐07MMA015CT, IN841C, 10TAL001CT and 10TAL033E. M. Llarena‐Reino thanks Fundação para a Ciência e a Tecnologia for its financial support under Grant SFRH/BD/45398/2008Peer reviewe

Microsporidians xenomas of anglerfish from NE Atlantic waters

12 pages, 9 figures, 2 tablesThe presence of emergent visible parasites at commercial valuable fish species is increasingly causing problems at fisheries and seafood industries. Microsporidians have been previously reported to appear forming apparent xenomas complexes in anglerfish species, but no effort has been carried out to simultaneously integrate epidemiological data, phenotypic, genotypic and fine structural characterizations in the same parasite sample. In this work, specimens of Lophius budegassa and Lophius piscatorius from NE Atlantic waters were sampled and examined to provide information about specific site of infection and demographic data of two groups of different sizes of xenomas present at both fish species. Histological descriptions and scanning and transmission electron microscopy were carried out on fresh spores of Lophius budegassa for ultrastructural studies. In both types of xenomas, it was observed simultaneously the microsporidian genus Spraguea in the form of two different types of spores. Molecular analyses of both xenomas from the two fish species, based on the small subunit ribosomal DNA gene, were also performed to genetically support the morphological diagnostic provided.Xunta de Galicia has funded this work under Projects INCITE-07MMA015CT, IN841C, 10TAL 001CT and 10TAL033E. M. Llarena-Reino thanks Fundação para a Ciência e a Tecnologia for its financial support (Grant SFRH/BD/45398/2008).Peer reviewe

Optimization of the pepsin digestion method for anisakis inspection in the fish industry

8 páginas, 2 figuras, 3 tablasDuring the last 50 years human anisakiasis has been rising while parasites have increased their prevalence at determined fisheries becoming an emergent major public health problem. Although artificial enzymatic digestion procedure by CODEX (STAN 244-2004: standard for salted Atlantic herring and salted sprat) is the recommended protocol for anisakids inspection, no international agreement has been achieved in veterinary and scientific digestion protocols to regulate this growing source of biological hazard in fish products. The aim of this work was to optimize the current artificial digestion protocol by CODEX with the purpose of offering a faster, more useful and safer procedure for factories workers, than the current one for anisakids detection. To achieve these objectives, the existing pepsin chemicals and the conditions of the digestion method were evaluated and assayed in fresh and frozen samples, both in lean and fatty fish species. Results showed that the new digestion procedure considerably reduces the assay time, and it is more handy and efficient (the quantity of the resulting residue was considerably lower after less time) than the widely used CODEX procedure. In conclusion, the new digestion method herein proposed based on liquid pepsin format is an accurate reproducible and user-friendly off-site tool, that can be useful in the implementation of screening programs for the prevention of human anisakiasis (and associated gastroallergic disorders) due to the consumption of raw or undercooked contaminated seafood productsXunta de Galicia has provided financial support under Projects INCITE-07MMA015CT, IN841C, 10TAL001CT and 10TAL033E. María Llarena-Reino thanks Fundação para a Ciência e a Tecnologia and European Social Fund for financial support under Grant SFRH/BD/45398/2008.Peer reviewe

The accuracy of visual inspection for preventing risk of Anisakis spp. infection in unprocessed fish

The importance of the zoonoses caused by L3 Anisakidae larvae lies in the repercussion that this parasite exerts on food safety and quality. EU legislation recommends fish operators to do visual inspection of the whole fish abdominal cavity and gut to control the risk of visible parasites, thus ensuring that no contaminated fish reach the consumers. The accuracy of the above visual inspection method should fall on a well-tested statistical significance between the number of observable parasites in the abdominal cavity and the number of parasites in the edible part of the fish (i.e., musculature). The aim of this study was to analyze this statistical significance, and the efficacy of the washing practice to remove Anisakis spp. from gut. To carry out this work, 322 fresh individuals of Micromesistius poutassou and 230 of Scomber scombrus were necropsied within 12 h and 48 h post-capture. Then, descriptive statistics, correlation and regression analyses were used to evaluate the significant statistical relationship between the number of anisakid larvae found in the gut and musculature of both fish species. Additionally, livers and gonads of 25 fresh specimens of Merluccius merluccius were vigorously washed under tap water, and examined under stereomicroscope looking for Anisakis spp. larvae. Results evidenced the low efficiency of visual inspection of gut parasites as a commonly recommended method for predicting nematode larvae in the flesh of fish. Therefore, a direct-invasive inspection of musculature is stressed as the only criteria with scientific merit for accurately detecting contaminated fishes by anisakids. Moreover, fresh European hake liver and gonads showed at least one larva remained inside the tissue after washing vigorously under tap water. Results suggested that critical control points at Hazard Analysis Critical Control Point (HACCP) programmes should be reviewed to improve the risk of anisakid-induced allergies and gastrointestinal anisakiasis among consumers.publishe

Ultrasound and magnetic resonance imaging of a tumorous growth in the urinary bladder of an adult male common dolphin (Delphinus delphis)

23rd Annual Conference of the European Cetacean Society, “Climate Change and Marine Mammals”, Istanbul, Turkey, 2-4 March 2009The diagnostic imaging techniques: ultrasound, computed tomography (CT) and magnetic resonance imaging (MRI) are increasingly being used with clinical and anatomical purposes in marine speciesN

Seals on the Galician coast: from sporadic to regular records

23rd Annual Conference of the European Cetacean Society, “Climate Change and Marine Mammals”, Istanbul, Turkey, 2-4 March 2009Since 1991, the NGO CEMMA has been attending strandings and surveying marine mammal populations along the Galician coastN