Alonso M.,Applied Proteomics |
Lago F.C.,Applied Proteomics |
Gomez-Reino M.,ANFACO CECOPESCA |
Fernandez J.,INSUINA S.L. |
And 3 more authors.
Parasitology | Year: 2013
Global aquaculture production of turbot has rapidly increased worldwide in the last decade and it is expected to have even bigger growth in the next years due to new farms operating. The losses caused by pathogen infections have grown at the same time as the production of this species. Parasitological infections are among the main relevant pathologies associated with its culture and produce serious losses in aquaculture, reduce the growth rate in fish and may lead to unmarketable fish due to skeletal muscle abnormalities in cases with high intensity of infection. The microsporidian parasite Tetramicra brevifilum causes severe infections and generates major losses in farmed turbot. Infections are difficult to control due to spore longevity and its direct transmission. To facilitate the infection management, an effective tool for fast detection and identification of T. brevifilum is needed. This study provides a molecular methodology of fast Real-Time PCR for T. brevifilum detection to the aquaculture industry, useful for routine control of T. brevifilum at turbot farms. The method is characterized by its high specificity and sensitivity, and it can be applied to cultured turbot for parasite detection regardless of the life-cycle stage of the pathogen or the infection intensity. © Cambridge University Press 2012.
Otero P.A.Z.,University of Santiago de Compostela |
Alfonso A.,University of Santiago de Compostela |
Vieytes M.R.,University of Santiago de Compostela |
Cabado A.G.,ANFACO CECOPESCA |
And 2 more authors.
Environmental Toxicology and Chemistry | Year: 2010
Environmental conditions are key factors in the development of marine toxic phytoplankton. Spirolides are marine toxins with a heptacyclic imine ring responsible for the toxicity in mice. Alexandrium ostenfeldii (A. ostenfeldii) is the main producer of these toxins, although this dinoflagellate often produces toxins belonging to the paralytic shellfish poisoning (PSP) group. The present study shows the first evidence that external environmental factors can influence the toxin profile produced by the dinoflagellate A. ostenfeldii. The species investigated is indigenous to the North Atlantic coast, and their cells grew under several environmental parameters. Toxin production was measured by means of liquid chromatography-mass spectrometry (LC-MS) and the chromatograms reflect the presence of two spirolides in all cultures; one in the region m/z 692.5, corresponding to 13-desmethyl spirolide C (13-desMeC) and the other in the region m/z 678.5, which corresponds to 13,19-didesmethyl spirolide C (13,19-didesMeC). The physical parameters studied were salinity, culture media, and photoperiod. The highest amount of toxin per cell was obtained when dinoflagellates grew in F/2 and Walne medium, 28‰ salinity, and 24 h of light. However, the highest proportion of 13,19-didesMeC with respect to 13-desMeC was achieved in L1 medium, 33‰ salinity, and 14:10 h light:dark. On the contrary, the highest proportion of 13-desMeC in cells was obtained when A. ostenfeldii was cultured in F/2 medium, 28‰ salinity, and the same photoperiod. Therefore, from these data the optimum conditions to culture A. ostenfeldii and to obtain the highest amount of spirolide per cell are shown. In addition, these environmental conditions can be considered a tool to predict and avoid A. ostenfeldii blooms. © 2009 SETAC.
Mena-Bueno S.,University of Santiago de Compostela |
Atanasova M.,ANFACO CECOPESCA |
Fernandez-Trasancos A.,University of Santiago de Compostela |
Paradela-Dobarro B.,University of Santiago de Compostela |
And 6 more authors.
Food and Function | Year: 2016
Objective: epicardial adipose tissue (EAT) from patients with coronary artery disease (CAD) contains higher levels of inflammatory proteins and lower adiponectin levels than subcutaneous adipose tissue (SAT), enhancing the progression of atherosclerosis. Since products from sea cucumber have anti-inflammatory properties, we investigated its effect on EAT, SAT and endothelial cells. Methods: stromal cells or explants from EAT and SAT were obtained from patients with cardiovascular disease. Extracts were obtained after hydrolysis by food-grade enzymes at different times. Proteins were identified by LC-MALDI mass spectrometry. Adipogenesis and adiponectin induction were determined on stromal cells in the presence/absence of extracts. The bioavailability of the extracts was tested on a Caco-2 cell culture model in vitro. The bioavailable fraction was probed on endothelial cells and EAT or SAT explants. Vascular cell adhesion protein (VCAM-1), intercellular adhesion molecule (ICAM-1), IL-6 and adiponectin were determined by real time polymerase chain reaction (RT-PCR). Results: our results showed that H. forskali and P. tremulus extracts contained compounds with anti-oxidant and anti-inflammatory properties. The bioavailable fraction of P. tremulus reduced VCAM-1 (p < 0.01) and IL-6 (p < 0.05) expression levels in endothelial cells while bioavailable compounds from H. forskali decreased ICAM-1 expression in SAT (p < 0.05). No effect was observed on EAT. Conclusion: these results suggest that sea cucumber extracts might be used for the prevention of endothelial cells and SAT inflammation. © The Royal Society of Chemistry 2015.
Herrero B.,ANFACO CECOPESCA |
Lago F.C.,ANFACO CECOPESCA |
Vieites J.M.,ANFACO CECOPESCA |
Espineira M.,ANFACO CECOPESCA
European Food Research and Technology | Year: 2011
Sardines are a small pelagic species belonging to the Clupleidae family and have high commercial value because of the properties of their meat. One of the most valuable fish in this family is Sardina pilchardus, which in addition to its high commercial value is the one that should be labeled on canned sardines, the other species being labeled as sardines X. In this study, a fast real-time polymerase chain reaction (PCR) assay based on TaqMan probe technology, which amplifies a fragment of the internal transcribed spacers (ITS1 and ITS2) and the 5.8 S rRNA region, was developed for the authentication of two commercially important species of sardines, S. pilchardus and S. aurita. This methodology can be applied to a variety of products, including those that have been subjected to intensive processing treatments such as canning. The present methodology was validated and subsequently applied to 50 commercial samples in order to determine whether correct labeling is being carried out in the market. This assay combines the high specificity, sensitivity, and rapidity of fast real-time PCR with the advantages of using two different probes in the same reaction, making it a powerful tool for verifying the correct application of food labeling regulations. © 2011 Springer-Verlag.
Garrido-Maestu A.,Microbiology and Bioassays Laboratory |
Chapela M.-J.,ANFACO CECOPESCA |
Penaranda E.,Microbiology and Bioassays Laboratory |
Cabado A.G.,ANFACO CECOPESCA
Food Biotechnology | Year: 2015
The current study was conducted in order to reduce the overall time needed in a previously validated qPCR method, and extend the procedure for the simultaneous detection of an additional pathogen (Shigella spp.). The original method was modified by extending the primary enrichment and eliminating the secondary enrichment. Additionally, a rapid commercial DNA extraction kit was evaluated against our reference protocol taking into consideration DNA concentration obtained, purity of the DNA extracts evaluating two absorbance ratios (260/280 and 260/230), and Cq and final fluorescence after qPCR analysis. Comparable results were obtained with both DNA extraction methods, but the commercial kit performed worse with low bacterial concentrations. A total of 84 spiked and blind samples were analyzed with the rapid protocol without finding significant differences with respect to the original method regarding qPCR efficiency (90-103%), and all the method’s parameters that were previously analyzed (above 91%). Additionally, a very low limit of detection was still obtained after the method optimization (below 10 cfu/25 g). The modified method represents a significant advance in detection of foodborne pathogens because it can provide accurate and reliable results for producers and health authorities in less than 27 h including the enrichment step. This protocol implementation resulted in an overall 5 h reduction, with less hands-on work, and without any loss in the quality of the method. In addition the inclusion of an additional pathogen extends the method’s applicability to 4 pathogens of interest. © Taylor & Francis Group, LLC.
Fajardo P.,ANFACO CECOPESCA |
Atanassova M.,ANFACO CECOPESCA |
Cotterill J.,FERA |
Wontner-Smith T.,FERA |
And 2 more authors.
WIT Transactions on the Built Environment | Year: 2013
The EU FP7 funded project "Bio-engineered micro Encapsulation of Active agents Delivered to Shellfish (BEADS)" is focused on mitigating the impact of marine biotoxins (ASP/DSP), microbial contamination (bacteria/norovirus) and the parasitic protozoan Bonamia ostreae on shellfish aquaculture. Purpose: to develop probiotic diets and a microencapsulated delivery system in the digestive tract of shellfish to improve depuration. Feeding experiments were performed to identify the optimum size of alginate microcapsules, testing three different sizes and colours containing nondegradable fluorescent dye microbeads. Oysters and mussels were placed in tanks containing filtered seawater. Three sizes of microcapsules were mixed and shellfish were fed for 3 hours. Shellfish were removed during the feeding period at 0.5, 1, 2 and 3 hours and dissected. Any capsules remaining in the water and in the digestive organs of shellfish were extracted and measured using a fluoroskan analyzer. A higher concentration of the smaller capsules was found in the digestive gland, indicating that the smaller capsules were preferentially ingested. The ingestion increased during the three-hour period. Fluorescent beads of different colours embedded in alginate capsules were observed bound to the mucus string. Mussels were more efficient than oysters in incorporating alginate beads that were observed by light microscopy as intact in the digestive gland of mussels and oysters during feeding period. Broken alginate beads were found in faeces. During passage through the intestine, active agents embedded by alginate capsules are released into the digestive gland providing a useful tool to transport active agents. © 2013 WIT Press.
Blanco L.,ANFACO CECOPESCA |
Martinez A.,ANFACO CECOPESCA |
Ferreira M.,ANFACO CECOPESCA |
Vieites J.,ANFACO CECOPESCA |
Cabado A.,ANFACO CECOPESCA
Food Additives and Contaminants: Part B Surveillance | Year: 2013
A total of 84 samples of wild and farmed fish, cephalopods and fish oils for animal feeding, traded in Spain, were analysed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in 2009-2012, by gas chromatography-ion trap tandem mass spectrometry (GC-MS-MS). The method was optimised for screening at moderate costs, allowing PCDD/Fs determination at 1 pg World Health Organization-toxic equivalent quantities (WHO-TEQ) g-1 wet weight (w w) and dl-PCBs at 0.02 pg WHO-TEQ g-1 w w. Concentrations in fish and cephalopods ranged from values below the limit of detection to 1.7 pg g-1 WHO-TEQ sum PCDD/Fs and dl-PCBs, considered as safe with regard to EU legislation. Higher levels were found in cod livers (5.4-54.2) and fish oils (3.3-30.7), with one noncompliant sample in each group. © 2013 Copyright Copyright Taylor & Francis.
PubMed | National Taiwan Ocean University, ANFACO CECOPESCA and Alagappa University
Type: | Journal: Results in immunology | Year: 2013
Penaeidins are members of a special family of antimicrobial peptide existing in penaeid shrimp and play an important role in the immunological defense of shrimp. Here, we report a penaeidin sequence cloned from the Indian white shrimp Fenneropenaus indicus (Fein-Penaeidin). The Fein-Penaeidin open reading frame encodes a 77 amino acid peptide including a 19 amino acid signal peptide. The deduced amino acid sequences of Fein-Penaeidin include a proline rich N-terminal domain and a carboxyl-domain that contains six cysteine residues. Structural analysis revealed an alpha-helix in its secondary structure and the predicted 3D structure indicated two-disulphide bridges in the alpha-helix. Phylogenetic analysis and sequence comparison with other known peaneidin suggest the gene shows high similarity to that of penaeidin from Peneaus monodon (95%), F. indicus (80%) and Fenneropenaeus chinensis (74%). Fein-Penaeidin was examined in normal and microbial challenged shrimp and was found to be constitutively expressed in haemocytes, Heart, gills, muscles, intestine, hepatopancreas and eyestalk. Bacterial challenge resulted in mRNA up-regulation, inducing expression at 6h post injection indicating the penaeidin involved in the innate immunity.
PubMed | Ebiotec EuroEspes Biotecnologia, ANFACO CECOPESCA, University of Santiago de Compostela and Proteomic Unit
Type: Journal Article | Journal: Food & function | Year: 2016
epicardial adipose tissue (EAT) from patients with coronary artery disease (CAD) contains higher levels of inflammatory proteins and lower adiponectin levels than subcutaneous adipose tissue (SAT), enhancing the progression of atherosclerosis. Since products from sea cucumber have anti-inflammatory properties, we investigated its effect on EAT, SAT and endothelial cells.stromal cells or explants from EAT and SAT were obtained from patients with cardiovascular disease. Extracts were obtained after hydrolysis by food-grade enzymes at different times. Proteins were identified by LC-MALDI mass spectrometry. Adipogenesis and adiponectin induction were determined on stromal cells in the presence/absence of extracts. The bioavailability of the extracts was tested on a Caco-2 cell culture model in vitro. The bioavailable fraction was probed on endothelial cells and EAT or SAT explants. Vascular cell adhesion protein (VCAM-1), intercellular adhesion molecule (ICAM-1), IL-6 and adiponectin were determined by real time polymerase chain reaction (RT-PCR).our results showed that H. forskali and P. tremulus extracts contained compounds with anti-oxidant and anti-inflammatory properties. The bioavailable fraction of P. tremulus reduced VCAM-1 (p < 0.01) and IL-6 (p < 0.05) expression levels in endothelial cells while bioavailable compounds from H. forskali decreased ICAM-1 expression in SAT (p < 0.05). No effect was observed on EAT.these results suggest that sea cucumber extracts might be used for the prevention of endothelial cells and SAT inflammation.