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A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes ArchiMer
Aton, Emilie; Renault, Tristan; Gagnaire, Beatrice; Thomas Guyon, Hélène; Cognard, C; Imbert, N.
Bivalve haemocytes are essential in defence mechanisms including phagocytosis. They also produce molecules including hydrolytic enzymes and antimicrobial peptides that contribute to pathogen destruction. Although haemocyte activities have been extensively studied, relatively little is known about the intracellular signalling pathways that are evoked during haemocyte activation and especially the role of calcium. Flow cytometry has been used for the first time to define the effect of cell incubation in haemolymph and artificial sea water (ASW) on Pacific oyster, Crassostrea gigas, haemocytes. Cell viability, enzymatic activities (esterases and amino peptidases), phagocytosis and granulocyte percentage were analysed. Viability and some activities were...
Tipo: Text Palavras-chave: Cellular activities; Fluo 3/AM; Calcium; Flow cytometry; Haemocytes; Crassostrea gigas.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/publication-935.pdf
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A new variety of Bacopa monnieri obtained by in vitro polyploidization Electron. J. Biotechnol.
Salvio Escandón,Alejandro; Hagiwara,Juan Carlos; Alderete,Liliana Marisol.
The commercial value of Bacopa monnieri, a widespread herbaceous plant in Argentina, can be substantially improved increasing its flower size by chromosome doubling with colchicine. MS supplemented with 0.25 mg/L of 6-benzylaminopurine proved to be an appropriate medium for the in vitro multiplication of nodal segments of B. monnieri. Polyploidization was achieved submerging nodal segments during 24 or 48 hrs in colchicine solution 0.001 and 0.01% P/V, in 1% DMSO. Segments submerged in water and in DMSO 1% aqueous solution were used as controls. DNA contents from recovered plants was measured and characterized and their phenotype described. Two tetraploid plants originated from independent events were detected. These plants showed significant differences...
Tipo: Journal article Palavras-chave: Colchicine; Flow cytometry; Ornamental plants; Scrophulariaceae; Tetraploid.
Ano: 2006 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000300002
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A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry BJMBR
Alves,L.P.S.; Almeida,A.T.; Cruz,L.M.; Pedrosa,F.O.; de Souza,E.M.; Chubatsu,L.S.; Müller-Santos,M.; Valdameri,G..
The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps: i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Flow cytometry; Nile red; Poly-3-hydroxybutyrate; Herbaspirillum seropedicae; Azospirillum brasilense.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2017000100606
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A study of autophagy in hemocytes of the Pacific oyster, Crassostrea gigas ArchiMer
Picot, Sandy; Morga, Benjamin; Faury, Nicole; Chollet, Bruno; Dégremont, Lionel; Travers, Marie-agnes; Renault, Tristan; Arzul, Isabelle.
Macroautophagy is a mechanism that is involved in various cellular processes, including cellular homeostasis and innate immunity. This pathway has been described in organisms ranging in complexity from yeasts to mammals, and recent results indicate that it occurs in the mantle of the Pacific oyster, Crassostrea gigas. However, the autophagy pathway has never been explored in the hemocytes of C. gigas, which are the main effectors of its immune system and thus play a key role in the defence of the Pacific oyster against pathogens. To investigate autophagy in oyster hemocytes, tools currently used to monitor this mechanism in mammals, including flow cytometry, fluorescent microscopy and transmission electron microscopy, were adapted and applied to the...
Tipo: Text Palavras-chave: Autophagosome; Autophagy; Crassostrea gigas; Flow cytometry; Fluorescence microscopy; Hemocytes; Transmission electron microscopy.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00487/59905/63087.pdf
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Active caspase-3 expression levels as bioindicator of individual radiosensitivity Anais da ABC (AABC)
SANTOS,NEYLIANE F.G. DOS; SILVA,RAFAEL F.; PINTO,MARCELA M.P.L.; SILVA,EDVANE B. DA; TASAT,DEBORAH R.; AMARAL,ADEMIR.
ABSTRACT Several molecules and events involved in cell response to radiation-induced damage have been investigated towards a personalized radiotherapy. Considering the importance of active caspase-3 in the proteolytic cascade that ensures radiation-induced apoptosis execution, this research was designed to evaluate the expression levels of this protein as a bioindicator of individual radiosensitivity. Peripheral blood samples of 10 healthy individuals were gamma-irradiated (cobalt-60 source) with 1, 2 and 4 Gy (control: non-irradiated samples), and active caspase-3 expression levels were measured in lymphocytes, by flow cytometry, ex vivo and after different times of in vitro incubation (24, 48 and 72 hours). Short-term incubation of 24 h was the most...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Active caspase-3; Apoptosis; Flow cytometry; Ionizing radiation; Lymphocytes.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652017000200649
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Adhesion molecule profiles of B-cell non-Hodgkin's lymphomas in the leukemic phase BJMBR
Matos,D.M.; Rizzatti,E.G.; Garcia,A.B.; Gallo,D.A.P.; Falcão,R.P..
We evaluated the expression of 10 adhesion molecules on peripheral blood tumor cells of 17 patients with chronic lymphocytic leukemia, 17 with mantle-cell lymphoma, and 13 with nodal or splenic marginal B-cell lymphoma, all in the leukemic phase and before the beginning of any therapy. The diagnosis of B-cell non-Hodgkin's lymphomas was based on cytological, histological, immunophenotypic, and molecular biology methods. The mean fluorescence intensity of the adhesion molecules in tumor cells was measured by flow cytometry of CD19-positive cells and differed amongst the types of lymphomas. Comparison of chronic lymphocytic leukemia and mantle-cell lymphoma showed that the former presented a higher expression of CD11c and CD49c, and a lower expression of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Non-Hodgkin's lymphoma; Adhesion molecules; Flow cytometry; CD49c/CD49d.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006001000011
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Age-related changes of the multidrug resistance P-glycoprotein function in normal human peripheral blood T lymphocytes BJMBR
Machado,C.G.; Calado,R.T.; Garcia,A.B.; Falcão,R.P..
The multidrug resistance P-glycoprotein is a transmembrane efflux pump expressed by lymphocytes and is involved in their cytolytic activity. In the present study, we investigated the age-related changes of P-glycoprotein function in normal peripheral blood lymphocytes. Blood samples from 90 normal volunteers (age range, 0 to 86 years) were analyzed. P-glycoprotein function was assessed by the flow cytometric rhodamine 123 assay. P-glycoprotein function was highest in cord blood and progressively declined with age in peripheral blood T CD4+ and CD8+ cells. In contrast, P-glycoprotein function did not vary with age in CD19+ B or CD16+CD56+ natural killer cells. These data suggest that the decline in P-glycoprotein function in T CD4+ and CD8+ lymphocytes as a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: P-glycoprotein; Multidrug resistance; Lymphocytes; Rhodamine 123; Flow cytometry.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003001200006
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Amélioration des productions phytoplanctoniques en écloserie de mollusques : caractérisation des microalgues fourrage ArchiMer
Robert, Rene; Chretiennot-dinet, Marie-josèphe; Kaas, Raymond; Martin-jezequel, Véronique; Moal, Jeanne; Le Coz, Jean-rene; Nicolas, Jean-louis; Bernard, Eudes; Connan, Jean-paul; Le Dean, Loic; Le Gourrierec, Gaetane; Leroy, Bertrand; Quere, Claudie.
in mollusc hatcheries. Consequently, we assessed themost commonly used species to establish their growth and size profiles, their cytomorphological, biochemical, biomolecular identification as well as their cytometer signs, and also their ability to grow under extreme conditions of temperature and salinity. This work aimed at matching the conformity of the strains used in Argenton (dispatched to most of the French commercial hatcheries) to those of a referenced culture collection. A previous screening preceded this study, because of the 45 species and/or clones held originally, Argenton kept no more than 15 species. As most of the latter are also represented in referenced collection, our strains are therefore correctly identified. On the other hand, the...
Tipo: Text Palavras-chave: Ecological limits; Biomolecular profile; Flow cytometry; Biochemical composition; Cytomorphology; Phytoplankton; Mollusc; Hatchery; Ecotolérance; Profil biomoléculaire; Cytométrie en flux; Composition biochimique; Cytomorphologie; Phytoplancton; Mollusques; Ecloserie.
Ano: 2004 URL: http://archimer.ifremer.fr/doc/2004/rapport-1546.pdf
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An approach on the in vitro maintenance of sugarcane with views for conservation and monitoring of plant nuclear DNA contents via flow cytometry. Repositório Alice
NOGUEIRA, G. F.; PIO, L. A. S.; PASQUAL, M.; AMARAL, A. L. do; CHERWINSKI-PEREIRA, J. E..
In vitro conservation techniques can be utilized for germplasm maintenance. However, few reports on the in vitro conservation of sugarcane species are present in the literature. The objective of this study was to subject sugarcane plants to in vitro under minimal growth conditions and to evaluate the survival, regeneration, and the monitoring of nuclear DNA content levels of the plants. Shoots from 10 sugarcane varie-ties (Saccharum spp.) were introduced into two media: MC1, consisting of half-strength Murashige and Skoog (MS) salts and 3% sorbitol, or MC2, similar to the first formulation, but additionally supplemented with 3.8 ?Mabscisicacid(ABA).
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Cana de açúcar; Flow cytometry; Sugarcane.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1023802
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Analysis of hemocyte parameters in Pacific oysters, Crassostrea gigas, reared in the field — Comparison of hatchery diploids and diploids from natural beds ArchiMer
Gagnaire, Beatrice; Soletchnik, Patrick; Faury, Nicole; Kerdudou, Nolwenn; Le Moine, Olivier; Renault, Tristan.
Internal defence mechanisms of bivalves include the hemocytes which assume several functions. Relationships exist between environmental factors, physiological state of oysters, defence system and pathogens. Hatchery oysters are more and more produced and reared in the field. The characteristics and particularly the defence system of these animals are poorly documented. In this context, two field trials were carried out from May to September 2002 and from April to July 2003 in order to monitor hemocyte parameters of diploid oysters from natural beds and hatchery-produced diploid oysters using flow cytometry. Several hemocyte parameters (cell mortality, granulocyte percentage, phagocytosis, esterase and peroxidase activities) were monitored. For both oyster...
Tipo: Text Palavras-chave: Hatchery; Natural beds; Diploids; Flow cytometry; Peroxidase activity; Esterase activity; Phagocytosis; Hemocytes; Crassostrea gigas; Pacific oyster.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-3119.pdf
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Analysis of larval oyster grazing by flow cytometry ArchiMer
Chretiennot-dinet, Marie-josèphe; Vaulot, Daniel; Galois, Robert; Spano, Anna-maria; Robert, Rene.
The ingestion of 8 algal species by oyster larvae (Crassostrea gigas Thunberg, 1793) was measured by flow cytometry (FCM). In a preliminary experiment, cell size (estimated by light scatter) and chlorophyll fluorescence of 30 algal species were evaluated to select those species which could be mixed together and still be easily discriminated by FCM. Grazing experiments were carried out over 48 h with 6 and 15-day old larvae fed on 3 algal mixtures, each containing 3 different algal species. The concentration of each species was estimated at 0, 6, 12, 24 and 48 h by FCM. Grazing pressure on a given algal species was dependent upon the age of the larvae, the time of the day and the composition of the mixture. Grazing rates of older larvae were about twice...
Tipo: Text Palavras-chave: Oyster larvae; Crassostrea gigas; Grazing; Algae; Selective feeding; Flow cytometry.
Ano: 1991 URL: http://archimer.ifremer.fr/doc/00090/20164/17817.pdf
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Analysis of nuclear DNA content and chromosome number for screening genotypes and crosses in Annual Ryegrass (Lolium multiflorum Lam.). Repositório Alice
BUSTAMANTE, F. de O.; ROCHA, L. C.; SANTOS, N. de S.; SILVEIRA, R. A. D.; NUNES, R. de C.; MITTELMANN, A.; TECHIO, V. H..
2015
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: Annual ryegrass; Cytogenetics; Flow cytometry; Plant breeding; Polyploidy.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1034882
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Antioxidant Effects of Protocatechuic Acid, Ferulic Acid, and Caffeic Acid in Human Neutrophils Using a Fluorescent Substance International Journal of Morphology
Hyogo,Akiyo; Kobayashi,Toshihiro; Garcia del Saz,Eva; Seguchi,Harumichi.
Human neutrophils stimulated by phorbol myristate acetate (PMA), an activator of protein kinase C, produce active oxygen by NADPH oxidase in intracellular structures. We added succinimidyl ester of dichlorodihydrofluorescein diacetate (H2DCFDA), which first emits fluorescence when oxidized with active oxygen species, to neutrophils to produce active oxygen, in order to investigate the antioxidant effects of protocatechuic acid, ferulic acid, and caffeic acid which belong to polyphenols and are widely distributed among plants. Particularly, we focused on examining whether these substances capture and eliminate active oxygen inside or outside the neutrophil cytoplasm and whether these substances inhibit NADPH oxidase. Fluorescence microscopy demonstrated...
Tipo: Journal article Palavras-chave: Antioxidant effects; Polyphenols; Human neutrophils; Flow cytometry.
Ano: 2010 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022010000300040
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Asynchronous expression of myeloid antigens in leukemic cells in a PML/RARalpha transgenic mouse model BJMBR
Santana,B.A.A.; Pintão,M.C.; Lima,R.S. Abreu e; Scheucher,P.S.; Santos,G.A.S.; Garcia,A.B.; Falcão,R.P.; Rego,E.M..
Acute promyelocytic leukemia (APL) is characterized by the expansion of blasts that resemble morphologically promyelocytes and harbor a chromosomal translocation involving the retinoic acid receptor a (RARa) and the promyelocytic leukemia (PML) genes on chromosomes 17 and 15, respectively. The expression of the PML/RARa fusion gene is essential for APL genesis. In fact, transgenic mice (TM) expressing PML/RARa develop a form of leukemia that mimics the hematological findings of human APL. Leukemia is diagnosed after a long latency (approximately 12 months) during which no hematological abnormality is detected in peripheral blood (pre-leukemic phase). In humans, immunophenotypic analysis of APL blasts revealed distinct features; however, the precise...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Acute promyelocytic leukemia; Acute myelogenous leukemia; Transgenic mice; Flow cytometry; Leukemogenesis.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000500008
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Atorvastatin-Medicated Dentifrice Significantly Inhibits CD4+ T Cell Proliferation: In vitro Pilot Study International Journal of Morphology
Rosenberg,David R; Kernitsky,Jeremy R; Andrade,Catherine X; Ramirez,Valeria; Violant,Deborah; Nart,José.
Reports indicate that statins (cholesterol-lowering drugs), in addition to lowering cholesterol, have an immunomodulatory effect. This effect may be beneficial for the treatment of several diseases, including periodontal disease. The aim of the present study was to evaluate the immunomodulatory effect of an atorvastatin-medicated dentifrice on CD4+ T cell proliferation. CD4+ T cell proliferation assays and peripheral blood mononuclear cell (PBMC) viability assays were conducted on PBMCs from healthy donors cultured under the following conditions: control, atorvastatin solution, atorvastatin-medicated dentifrice, and dentifrice without atorvastatin at concentrations of 1, 5, 10, 50 and 100 µM. A Generalized Equation Estimation (GEE) model was used to...
Tipo: Journal article Palavras-chave: Atorvastatin calcium; Dentifrices; T cells; Flow cytometry.
Ano: 2017 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022017000200002
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Bacterial and Archaeal Communities Variability Associated with Upwelling and Anthropogenic Pressures in the Protection Area of Arraial do Cabo (Cabo Frio region - RJ) Anais da ABC (AABC)
COELHO-SOUZA,SERGIO A.; ARAÚJO,FÁBIO V.; CURY,JULIANO C.; JESUS,HUGO E.; PEREIRA,GILBERTO C.; GUIMARÃES,JEAN R.D.; PEIXOTO,RAQUEL S.; DÁVILA,ALBERTO M.R.; ROSADO,ALEXANDRE S..
ABSTRACTUpwelling systems contain a high diversity of pelagic microorganisms and their composition and activity are defined by factors like temperature and nutrient concentration. Denaturing gradient gel electrophoresis (DGGE) technique was used to verify the spatial and temporal genetic variability of Bacteria and Archaea in two stations of the Arraial do Cabo coastal region, one under upwelling pressure and another under anthropogenic pressure. In addition, biotic and abiotic variables were measured in surface and deep waters from three other stations between these stations. Six samplings were done during a year and adequately represented the degrees of upwelling and anthropogenic pressures to the system. Principal Component Analysis (PCA) showed...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Microbial diversity; Leucine incorporation; Flow cytometry; Temperature; Nutrients availability; Brazilian Upwelling System.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000401737
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Can widely used cell type markers predict the suitability of immortalized or primary mammary epithelial cell models? Biol. Res.
Ontsouka,Edgar Corneille; Bertschi,Janique Sabina; Huang,Xiao; Lüthi,Michael; Müller,Stefan; Albrecht,Christiane.
BACKGROUND: Mammary cell cultures are convenient tools for in vitro studies of mammary gland biology. However, the heterogeneity of mammary cell types, e.g., glandular milk secretory epithelial or myoepithelial cells, often complicates the interpretation of cell-based data. The present study was undertaken to determine the relevance of bovine primary mammary epithelial cells isolated from American Holstein (bMEC US) or Swiss Holstein-Friesian (bMEC CH) cows, and of primary bovine mammary alveolar epithelial cells stably transfected with simian virus-40 (SV-40) large T-antigen (MAC-T) for in vitro analyses. This was evaluated by testing their expression pattern of cytokeratin (CK) 7, 18, 19, vimentin, and α-smooth muscle actin (α-SMA....
Tipo: Journal article Palavras-chave: Cell characterization; Flow cytometry; Gene expression; Lactation marker; Mammalian; Mammary epithelial cell.
Ano: 2016 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602016000100001
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Caractérisation morphologique et fonctionnelle des hémocytes chez l'huître plate, Ostrea edulis ArchiMer
Xue, Quiggang.
The mechanism that bivalve molluscs react to infectious agents are important research subjects in the fIeld of disease control studies for these animals. While the selection of disease resistant populations have made decisive progression in some species, the mechanism behinds these resistant phenomena is not yet clear so far. However, hemocytes must represent one of the key factors in the reaction process. Based on this hypothesis, the structure and function of European flat oyster, Ostrea edulis, hemocytes have been investigated in aid of different techniques in the present study. ln addition, Pacific oyster, Crassostrea gigas, hemocytes have been analysed in certain aspects so as to provide some results for comparison. European flat oyster and Pacific...
Tipo: Text Palavras-chave: Bivalvia; Ostrea edulis; Crassostrea gigas; Mécanismes de défense; Hémocytes; Granulocytes; Hyalinocytes; Séparation; Anticorps monoclonaux; Enzymes; Cytométrie de flux; Bivalvia; Ostrea edulis; Crassostrea gigas; Defence mechanisms; Hemocytes; Granulocytes; Hyalinocytes; Separation; Monoclonal antibodies; Enzymes; Flow cytometry.
Ano: 1998 URL: http://archimer.ifremer.fr/doc/00107/21820/19406.pdf
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CD19+ B cell subsets in the peripheral blood and skin lesions of psoriasis patients and their correlations with disease severity BJMBR
Lu,J.; Ding,Y.; Yi,X.; Zheng,J..
T lymphocytes are important in the pathogenesis of psoriasis, and increasing evidence indicates that B cells also play an important role. The mechanisms of action, however, remain unclear. We evaluated the ratios of CD19+ B cells in peripheral blood mononuclear cells (PBMCs) from 157 patients with psoriasis (65 patients with psoriasis vulgaris, 32 patients with erythrodermic psoriasis, 30 patients with arthropathic psoriasis, and 30 patients with pustular psoriasis) and 35 healthy controls (HCs). Ratios of CD19+ B cells in skin lesions were compared with non-lesions in 7 erythrodermic psoriasis patients. The Psoriasis Area Severity Index (PASI) was used to measure disease severity. CD19+ B cell ratios in PBMCs from psoriasis vulgaris (at both the active...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Psoriasis; B-lymphocytes; B-lymphocyte subsets; Psoriasis Area Severity Index (PASI); Flow cytometry.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2016000900706
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Cellular and molecular responses of haemocytes from Ostrea edulis during in vitro infection by the parasite Bonamia ostreae ArchiMer
Morga, Benjamin; Renault, Tristan; Faury, Nicole; Chollet, Bruno; Arzul, Isabelle.
Bonamia ostreae is a protozoan, affiliated to the order Haplosporidia and to the phylum Cercozoa. This parasite is intracellular and infects haemocytes, cells notably involved in oyster defence mechanisms. Bonamiosis due to the parasite B. ostreae is a disease affecting the flat oyster, Ostrea edulis. The strategies used by protozoan parasites to circumvent host defence mechanisms remain largely unknown in marine bivalve molluscs. In the present work, in vitro experiments were carried out in order to study the interactions between haemocytes from O. edulis and purified parasite. B. ostreae. We monitored cellular and molecular responses of oyster haemocytes by light microscopy, flow cytometry and real-time PCR 1, 2, 4 and 8 h p.i. Light microscopy was used...
Tipo: Text Palavras-chave: Bonamia ostreae; Protozoan; Ostrea edulis; Haemocytes; Real-time PCR; Flow cytometry; Super oxide dismutase.
Ano: 2011 URL: http://archimer.ifremer.fr/doc/00037/14874/18001.pdf
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