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1
المؤلفون: Magalí Rey-Campos, Rebeca Moreira, Marco Gerdol, Alberto Pallavicini, Beatriz Novoa, Antonio Figueras
المساهمون: Ministerio de Ciencia, Innovación y Universidades (España), Xunta de Galicia, Ministerio de Economía y Competitividad (España), Rey-Campos, Magalí, Moreira, Rebeca, Gerdol, Marco, Pallavicini, Alberto, Novoa, Beatriz, Figueras, Antonio
المصدر: Frontiers in Immunology
Frontiers in Immunology, Vol 10 (2019)
Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: lcsh:Immunologic diseases. Allergy, 0301 basic medicine, mussel, Immunology, Population, Vibrio splendidus, Hemocyte, Apoptosis, Inflammation, Biology, immune memory, Microbiology, Immune tolerance, 03 medical and health sciences, 0302 clinical medicine, Immune system, medicine, Immunology and Allergy, immune challenge, Vibrio, infection, RNA-Seq, education, education.field_of_study, ROS, Mussel, Acquired immune system, biology.organism_classification, Mytilus, 3. Good health, 030104 developmental biology, Mytilus galloprovincialis, medicine.symptom, lcsh:RC581-607, Immune priming, 030215 immunology
الوصف: 15 pages, 8 figures, 3 tables
Mediterranean mussels (Mytilus galloprovincialis) are sessile filter feeders that live in close contact with numerous marine microorganisms. As is the case in all invertebrates, mussels lack an adaptive immune system, but they respond to pathogens, injuries or environmental stress in a very efficient manner. However, it is not known if they are able to modify their immune response when they reencounter the same pathogen. In this work, we studied the transcriptomic response of mussel hemocytes before and after two consecutive sublethal challenges with Vibrio splendidus. The first exposure significantly regulated genes related to inflammation, migration and response to bacteria. However, after the second exposure, the differentially expressed genes were related to the control and inhibition of ROS production and the resolution of the inflammatory response. Our results also show that the second injection with V. splendidus led to changes at the transcriptional (control of the expression of pro-inflammatory transcripts), cellular (shift in the hemocyte population distribution), and functional levels (inhibition of ROS production). These results suggest that a modified immune response after the second challenge allowed the mussels to tolerate rather than fight the infection, which minimized tissue damage.
This work was conducted with the support of the projects AGL2015-65705-R and RTI2018-095997-B-I00 (Ministerio de Ciencia, Innovación y Universidades, Spain), IN607B 2016/12 (Consellería de Economía, Emprego e Industria-GAIN, Xunta de Galicia), and VIVALDI (678589) (EU H2020). MR-C acknowledges additional funding from the Spanish Ministerio de Economía y Competitividad for her predoctoral contract (BES2016-076302).URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7fee4d0af67b1c14ab795cdeebca5b2b
https://doi.org/10.3389/fimmu.2019.01894 -
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المؤلفون: Cristian Gallardo-Escárate, Rebeca Moreira, Antonio Figueras, Valentina Valenzuela-Muñoz, Beatriz Novoa, Magalí Rey-Campos
المساهمون: European Commission, Ministerio de Economía y Competitividad (España), Moreira, Rebeca, Moreira, Rebeca [0000-0001-7797-7221]
المصدر: Scientific Reports, Vol 9, Iss 1, Pp 1-15 (2019)
Scientific Reports
Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: 0301 basic medicine, animal structures, Antimicrobial peptides, lcsh:Medicine, Article, Microbiology, Transcriptome, 03 medical and health sciences, 0302 clinical medicine, Animals, Microbiome, lcsh:Science, Illumina dye sequencing, Vibrio, Mytilus, Multidisciplinary, biology, fungi, lcsh:R, Molecular Sequence Annotation, Mussel, Blood Proteins, biology.organism_classification, Myticin, 030104 developmental biology, lcsh:Q, 030217 neurology & neurosurgery, Antimicrobial Cationic Peptides
الوصف: 15 pages, 4 tables, 9 figures.-- This article is licensed under a Creative Commons Attribution 4.0 International License
Mediterranean mussels (Mytilus galloprovincialis) are sessile filter feeders that live in close contact with numerous marine microorganisms. As all invertebrates, they lack an adaptive immune response and how these animals are able to respond to a bacterial infection and discriminate it from their normal microbiome is difficult to understand. In this work, we conducted Illumina sequencing of the transcriptome of individual mussels before and after being infected with Vibrio splendidus. The control mussels were injected with filtered seawater. We demonstrate that a great variability exists among individual transcriptomes and that each animal showed an exclusive repertoire of genes not shared with other individuals. The regulated genes in both the control and infected mussels were also analyzed and, unexpectedly, the sampling before the injection was considered a stress stimulus strong enough to trigger and modulate the response in hemocytes, promoting cell migration and proliferation. We found a clear response against the injection of filtered seawater, suggesting a reaction against a tissue injury in which the myticins, the most expressed antimicrobial peptides in mussel, appeared significantly up regulated. Functional experiments with flow cytometry confirmed the transcriptomic results since a significant alteration of hemocyte structures and a decrease in the number of hemocytes positive for myticin C were found only after a Vibrio infection and not observed when mussels were bled before, generating a tissue injury. Therefore, we report the involvement of myticins in the response to a danger signal such as a simple injection in the adductor muscle
This work was conducted with the support of the projects AGL2015-65705-R (Ministerio de Economía y Competitividad, Spain), IN607B 2016/12 (Consellería de Economía, Emprego e Industria - GAIN, Xunta de Galicia) and VIVALDI (678589) (EU H2020). MRC thanks additional funding from the Spanish Ministerio de Economía y Competitividad, for her predoctoral contract (BES-2016-076302) -
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المؤلفون: Beatriz Novoa, Magalí Rey-Campos, A. Saco, Antonio Figueras
المصدر: Frontiers in Immunology
Frontiers in Immunology, Vol 11 (2020)
Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: 0301 basic medicine, Gill, Gills, Hemocytes, Aquaculture, immune response, Transcriptome, Hemolymph, Immunology and Allergy, Pathogen, innate immunity, Original Research, Effector, 04 agricultural and veterinary sciences, bath infection, 6. Clean water, Mytilus, 3. Good health, Organ Specificity, Host-Pathogen Interactions, RNA, Long Noncoding, lcsh:Immunologic diseases. Allergy, animal structures, Immunology, mussel, Biology, Real-Time Polymerase Chain Reaction, Microbiology, 03 medical and health sciences, Immune system, Animals, 14. Life underwater, RNA, Messenger, Vibrio, Innate immune system, Tissue Extracts, fungi, gill, Mussel, transcriptomic, biology.organism_classification, Bacterial Load, 030104 developmental biology, Gene Ontology, 040102 fisheries, 0401 agriculture, forestry, and fisheries, RNA-seq, lcsh:RC581-607, transcriptome
الوصف: 18 pages, 10 figures, 3 tables.-- This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY)
Mussels (Mytilus galloprovincialis) are filter feeder bivalves that are constantly in contact with a wide range of microorganisms, some of which are potentially pathogenic. How mussels recognize and respond to pathogens has not been fully elucidated to date; therefore, we investigated the immune mechanisms that these animals employ in response to a bacterial bath infection from the surrounding water, mimicking the response that mussels mount under natural conditions. After the bath infection, mussels were able to remove the bacteria from their bodies and from the water tank. Accordingly, antibacterial activity was detected in gill extracts, demonstrating that this tissue plays a central role in removing and clearing potential pathogens. A transcriptomic study performed after a bath infection with Vibrio splendidus identified a total of 1,156 differentially expressed genes. The expression levels of genes contributing to a number of biological processes, such as immune response activation pathways and their regulation with cytokines, cell recognition, adhesion and apoptosis, were significantly modulated after infection, suggesting that the gills play important roles in pathogen recognition, as well as being activators and regulators of the mussel innate immune response. In addition to RNA-seq analysis, long non-coding RNAs and their neighboring genes were also analyzed and exhibited modulation after the bacterial challenge. The response of gills against bath infection was compared with the findings of a previous transcriptomic study on hemocytes responding to systemic infection, demonstrating the different and specific functions of gills. The results of this study indicate that recognition processes occur in the gill, thereby activating the effector agents of the immune response to overcome bacterial infection
This research was funded by the Spanish Ministerio de Ciencia, Innovación y Universidades (AEI/EU-FEDER RTI2018-095997-B-I00) and the EU-H2020 VIVALDI (678589). Our laboratory is funded by Interreg VA Spain-Portugal cooperation programme (POCTEP) 2014-2020, 0474_BLUEBIOLAB project, co-funded by the European Regional Development Fund (FEDER) and IN607B 2019/01 from Consellería de Economía, Emprego e Industria (GAIN), Xunta de Galicia. MR-C was supported by a Spanish AEI/EU-FEDER predoctoral contract BES-2016-076302. AS was supported by a Spanish AEI/EU-FSE predoctoral contract PRE2019-090760URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7038c77a4f646eb35f238795d430cad6
http://europepmc.org/articles/PMC7772429 -
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المصدر: Italian Journal of Food Safety
Italian Journal of Food Safety, Vol 5, Iss 1 (2016)مصطلحات موضوعية: 0301 basic medicine, 030106 microbiology, Article, law.invention, Microbiology, 03 medical and health sciences, law, Pathogen, Polymerase chain reaction, Detection limit, Chromatography, biology, Aeromonas sp, lcsh:TP368-456, Inoculation, Water, GCAT, biology.organism_classification, DNA extraction, Vibrio, lcsh:Food processing and manufacture, Detection, PCR, Aeromonas, Food Science
الوصف: The genus Aeromonas is present in a wide variety of water environments and is recognized as potentially pathogenic to humans and animals. Members of this genus are often confused with Vibrio when using automated, commercial identification systems that are culture-dependent. This study describes a PCR detection method for Aeromonas that is cultureindependent and that targets the gcat (Glycerophospholopid-Cholesterol Acyltransferase) gene, which is specific for this genus. The GCAT-PCR was 100% specific in artificially inoculated water samples, with a detection limit that ranged from 2.5 to 25 cfu/mL. The success at detecting this pathogen in 86 water samples using the GCAT-PCR method was identical to the conventional culturing method when a pre-enrichment step was carried out, yielding 83.7% positive samples. On the other hand, without a pre-enrichment step, only 77.9% of the samples were positive by culturing and only 15.1% with the GCAT-PCR. However, 83.7% positive samples were obtained for the GCAT-PCR when the water volume for the DNA extraction was increased from 400 μL to 4 mL. The proposed molecular method is much faster (5 or 29 h) than the culturing method (24 or 48 h) whether performed directly or after a pre-enrichment step and it will enable the fast detection of Aeromonas in water samples helping to prevent a possible transmission to humans.
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a918fa36a6ef2fadec4a30f086ba24d1
http://europepmc.org/articles/PMC5076704 -
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المؤلفون: Antonio Figueras, María Prado-Álvarez, Alejandro Romero, Pablo Balseiro, B. Novoa, S. Dios
المصدر: Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: Lipopolysaccharides, Programmed cell death, Hemocytes, Lipopolysaccharide, Cell Survival, Phagocytosis, Stimulation, Aquatic Science, Flow cytometry, Microbiology, chemistry.chemical_compound, Immune system, Adjuvants, Immunologic, Ruditapes decussatus, medicine, Animals, Environmental Chemistry, Viability assay, Vibrio, biology, medicine.diagnostic_test, Haemocyte, General Medicine, biology.organism_classification, Bivalvia, Gene Expression Regulation, chemistry, Bacteria
الوصف: The morphology and functionality of Ruditapes decussatus haemocytes have been characterized by light microscopy and flow cytometry, leading to the identification of three different cellular subpopulations. Granulocytes were the largest cells, the hyalinocytes were smaller and contained fewer granules and the intermediate cells showed a size similar to hyalinocytes and a higher number of granules. The phagocytosis of different particles and the associated production of oxygen radicals were measured by flow cytometric methods. Granulocytes were the most active cells, followed by the intermediate cells and hyalinocytes. The effect of stimulation of haemocytes with lipopolysaccharide (LPS), with a heat inactivated bacterial mixture or with the infection of Vibrio splendidus on the cell viability and the expression of selected immune-related genes were studied. While significant low levels of damaged cells were registered in LPS-stimulated cells, the treatment with dead bacteria or V. splendidus reduced cell viability 1 h, 3 h and 6 h after treatment. The stimulation of haemocytes with LPS and dead bacteria induced changes in the expression of defender against cell death (DAD-1), thrombin, prosaposin, inhibitor of apoptosis (IAP), factor B and C3 complement component.
This work has been funded by the Spanish Ministerio de Ciencia e Innovación (MICINN) (AGL2008-05111). -
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المؤلفون: M.M. Costa, María Prado-Álvarez, Philippe Roch, Beatriz Novoa, Camino Gestal, Antonio Figueras, Hui Li
المصدر: Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: Vibrio anguillarum, PAMPs, Hemocytes, Gram-negative bacteria, Aquatic Science, Nitric Oxide, NO, Micrococcus, Microbiology, chemistry.chemical_compound, Phagocytosis, Animals, Environmental Chemistry, Respiratory Burst, Vibrio, Mytilus, biology, Gene Expression Profiling, Pathogen-associated molecular pattern, Mytilin, Zymosan, AMPs, ROS, General Medicine, biology.organism_classification, Myticin, Respiratory burst, Mytilus galloprovincialis, Gene Expression Regulation, chemistry, Mussel, Lysozyme
الوصف: 9 páginas, 6 figuras, 1 tabla
The effect of live bacteria (Micrococcus lysodeikticus and Vibrio anguillarum), and PAMPs (poly I:C, zymosan, LPS, LTA and CpG) on the production of intermediate toxic radicals (respiratory burst activity and production of nitric oxide) and mytilin B, myticin C and lysozyme gene expression was studied in vivo and in vitro. In vitro, bacteria were able to modulate the haemocytes’ respiratory burst activity, being significantly increased after 6 h of incubation. The effect of pathogen-associated molecular patterns (PAMPs) was also studied. Zymosan produced an increase of the PMA-mediated response but an inhibition of the zymosan-mediated response. A significant increase of nitric oxide production was found at all the sampled time points (1, 3 and 6 h) in comparison with controls on both, the Gram-positive and Gram-negative bacteria. The in vivo responses measured on haemocytes after M. lysodeikticus injection were faster than those induced by V. anguillarum. However, V. anguillarum induced stronger in vitro effects. Mytilin B, myticin C and lysozyme in vitro gene expression, occurred at short times after infection. The maximum in vitro expression was detected 3 h post-infection. The differences between M. lysodeikticus and V. anguillarum in different measured parameters may suggest that different signalling pathways might be involved. Moreover, among all assayed PAMPs, LPS elicited the highest response.
This research has been supported by the Ministerio de Ciencia e Innovación (project AGL2003-02454) and by European Community (project IMAQUANIM (FOOD-CT-2005-007103)). -
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المؤلفون: Antonio Figueras, M.M. Costa, B. Novoa
المصدر: Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: Mediterranean mussel, Hemocytes, beta-Glucans, Chemiluminescence, animal structures, Hemocyte, macromolecular substances, Bactericidal activity, Aquatic Science, Nitric Oxide, Microbiology, chemistry.chemical_compound, Immune system, Hemolymph, Escherichia coli, Animals, Environmental Chemistry, Carpet shell clam, Vibrio alginolyticus, Respiratory Burst, Vibrio, Mytilus, Respiratory burs, biology, Ecology, B-glucans, fungi, Zymosan, Nitric oxide, General Medicine, Mussel, biology.organism_classification, Bivalvia, Respiratory burst, chemistry, Luminescent Measurements, Carcinogens, Tetradecanoylphorbol Acetate
الوصف: 8 páginas, 5 figuras
The effects of b-glucans on several immune functions of carpet shell clam (Ruditapes decussatus) and Mediterranean mussel (Mytilus galloprovincialis) hemocytes were determined. Nitric oxide (NO) production increased significantly in b-glucan treated mussels and clams. In mussels, b-glucans increased by themselves the release of free oxygen radicals and also were able to enhance the phorbol 12-myristate 13-acetate (PMA) mediated effect on this hemocyte activity. However, high doses of b-glucans when combined with zymosan decreased this respiratory burst. In clams, hemolymph treated with several doses of b-glucans limited the growth of the three bacteria, Vibrio algynolyticus (strain TA15), Vibrio splendidus (strain TA2) and Escherichia coli (strain ATCC 13706). This modulation on the antibacterial activity, however, was not observed when mussel hemolymph was incubated with b-glucans. These results suggest that the immune responses of these animals can be up and down modulated by external stimuli and, although clams and mussels are both relatively closely related species, their behaviour concerning immune responses can be different
This research has been supported by the Ministerio de Educacio´n y Ciencia (project AGL2003-02454), the European Community (project IMAQUANIM (FOOD-CT-2005-007103)) and the Xunta de Galicia (project PGIDIT06PXIC402124PM). Ms Costa thanks the Ministerio de Educacio´n y Ciencia for her predoctoral fellowship. -
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المؤلفون: Beatriz Novoa, Massimiliano Babbucci, Pablo Balseiro, Luca Bargelloni, Rebeca Moreira, Massimo Milan, Alejandro Romero, Antonio Figueras
المساهمون: Ministerio de Ciencia e Innovación (España), European Commission
المصدر: Digital.CSIC. Repositorio Institucional del CSIC
instname
BMC Genomicsمصطلحات موضوعية: 0106 biological sciences, Time Factors, Hemocytes, Microarray, 01 natural sciences, Ruditapes philippinarum, Transcription (biology), Gene expression, Regulation of gene expression, 0303 health sciences, biology, Blast2GO, General Medicine, Cell biology, 3. Good health, DNA microarray, Databases, Nucleic Acid, Research Article, Biotechnology, animal structures, Aquatic Science, Microbiology, 03 medical and health sciences, Immune system, Genetics, Animals, Environmental Chemistry, Immune response, Gene, Vibrio alginolyticus, 030304 developmental biology, Analysis of Variance, Innate immune system, Gene Expression Profiling, Computational Biology, Reproducibility of Results, Molecular Sequence Annotation, Oligo-microarray, biology.organism_classification, Vibrio, Bivalvia, Fishery, Gene expression profiling, 010602 entomology, Gene Expression Regulation, 13. Climate action, Vibrio Infections, Gene ontology, Transcriptome
الوصف: [Background] The Manila clam (Ruditapes philippinarum) is a cultured bivalve with worldwide commercial importance, and diseases cause high economic losses. For this reason, interest in the immune genes in this species has recently increased. The present work describes the construction of the first R. philippinarum microarray containing immune-related hemocyte sequences and its application to study the gene transcription profiles of hemocytes from clams infected with V. alginolyticus through a time course.
[Results] The complete set of sequences from R. philippinarum available in the public databases and the hemocyte sequences enriched in immune transcripts were assembled successfully. A total of 12,156 annotated sequences were used to construct the 8 ×15 k oligo-microarray. The microarray experiments yielded a total of 579 differentially expressed transcripts. Using the gene expression results, the associated Gene Ontology terms and the enrichment analysis, we found different response mechanisms throughout the experiment. Genes related to signaling, transcription and apoptosis, such as IL-17D, NF-κB or calmodulin, were typically expressed as early as 3 hours post-challenge (hpc), while characteristic immune genes, such as PGRPs, FREPs and defense proteins appeared later at 8 hpc. This immune-triggering response could have affected a high number of processes that seemed to be activated 24 hpc to overcome the Vibrio challenge, including the expression of many cytoskeleton molecules, which is indicative of the active movement of hemocytes. In fact functional studies showed an increment in apoptosis, necrosis or cell migration after the infection. Finally, 72 hpc, activity returned to normal levels, and more than 50% of the genes were downregulated in a negative feedback of all of the previously active processes.
[Conclusions] Using a new version of the R. philippinarum oligo-microarray, a putative timing for the response against a Vibrio infection was established. The key point to overcome the challenge seemed to be 8 hours after the challenge, when we detected immune functions that could lead to the destruction of the pathogen and the activation of a wide variety of processes related to homeostasis and defense. These results highlight the importance of a fast response in bivalves and the effectiveness of their innate immune system.
This work has been partially funded by the EU Project REPROSEED (245119). RM wishes to acknowledge the Spanish MICINN for her FPI Spanish research grant (BES-2009-029765). -
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المؤلفون: Juan J. Borrego, Dolores Castro, Antonio Figueras, A. Luque, Beatriz Novoa
المصدر: Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: Vibrio tapetis, Infectivity, animal structures, Virulence, biology, Clams, biology.organism_classification, Bivalvia, Vibrio, Brown Ring Disease, Microbiology, Agglutination (biology), Vibrionaceae, Hemolymph, Pathogenicity, Bivalve mollusc, Ecology, Evolution, Behavior and Systematics, Bacteria
الوصف: 8 páginas, 2 figuras, 3 tablas
Four bacterial strains isolated from carpet-shell clams (Ruditapes decussatus) and manila clams (R. philippinarum) exhibiting Brown Ring Disease (BRD) symptoms in Galicia (Northwest of Spain) were characterized using biochemical and serological techniques. Moreover their pathogenicity for clams and resistance against clam hemolymph were determined. All of the Vibrio isolates exhibited biochemical characteristics typical of the genus. Strains GR1703RP and GR6301RP, isolated from R. philippinarum showing signs of BRD, shared phenotypical and antigenic characteristics and bacterial extracellular products enzymatic activities with the Vibrio tapetis. Strains GR0705RD and GR0202RD, isolated from R. decussatus showing BRD signs, although biochemically similar to V. tapetis, did not show positive agglutination with V. tapetis antiserum. The virulence of the bacteria varied depending on the bacterial strain and clam species assayed. Mortalities were recorded for a 30-day period, reaching values ranging from 6 to 80%. Strain GR0202RD was the most virulent for R. philippinarum with a cumulative mortality rate of 45%, while the strain GR0705RD induced the highest prevalence of BRD signs, affecting 75% of the survivors. In the case of R. decussatus, the four bacterial strains produced a cumulative mortality rate higher than 45%. In general, bacterial strains, showed resistance to clams hemolymph.
This work was financed by the FAR Project AQ 3.763 of the EEC -
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المؤلفون: L Villamil, Antonio Figueras, B. Novoa, Miquel Planas, Alicia E. Toranzo
المصدر: Digital.CSIC. Repositorio Institucional del CSIC
instnameمصطلحات موضوعية: Gill, Gills, animal structures, Veterinary (miscellaneous), Histopathology, Aquatic Science, medicine.disease_cause, Kidney, Bacterial Adhesion, Microbiology, Cell Line, Lethal Dose 50, Fish Diseases, Scophthalmus maximus, Intestinal mucosa, Vibrio Infections, medicine, Animals, Seawater, Intestinal Mucosa, Vibrio, biology, Virulence, Myocardium, fungi, Pathogenic bacteria, Turbot, biology.organism_classification, Scophthalmus, Phenotype, Larva, Flatfishes, Vibrio pelagius, Bacteria
الوصف: 11 páginas, 7 figuras, 2 tablas
A bacterial strain, characterized as Vibrio pelagius (Hq 222), was isolated from a turbot, Scophthalmus maximus (L.), larvae mass mortality in a commercial fish farm in Spain. Turbot larvae, post-larvae (0.2 g) and juveniles (5 and 15 g) were experimentally infected. The bacterium appeared to be very virulent for larvae and post-larvae, LD50 being
This work was funded by Project 1FD97-044-C03- 03 from FEDER funds.
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