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| Registros recuperados: 28 | |
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| Goarant, Cyrille; Merien, F. |
| Shrimp farming is a small but growing industry in New Caledonia. Since 1993, "Syndrome 93" has been affecting New Caledonian shrimp farming industry every cold season, causing severe epizootic mortalities in grow-out ponds and significant losses. Highly pathogenic strains of Vibrio penaeicida are considered the etiological agent of the disease in Litopenaeus stylirostris. On one hand, studies demonstrated that healthy shrimp may carry V penaeicida for weeks with a high overall prevalence, regardless of any seasonal pattern or temperature conditions. On the other hand, larvae are free of V penaeicida and are also resistant to experimental infection. V penaeicida is frequently detected in incoming water pumped from the bays, which was shown, by a molecular... |
| Tipo: Text |
Palavras-chave: Vibriosis; Vibrio; Real time PCR; Quantification; Mariculture; Extraction techniques. |
| Ano: 2006 |
URL: http://archimer.ifremer.fr/doc/2006/publication-1903.pdf |
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| Arraes,L.C.; de Souza,P.R.; Bruneska,D.; Castelo Filho,A.; de Souza Cavada,B.; de Lima Filho,J.L.; Crovella,S.. |
| We report a fast (less than 3 h) and cost-effective melting temperature assay method for the detection of single-nucleotide polymorphisms in the MBL2 gene. The protocol, which is based on the Corbett Rotor Gene real time PCR platform and SYBR Green I chemistry, yielded, in the cohorts studied, sensitive (100%) and specific (100%) PCR amplification without the use of costly fluorophore-labeled probes or post-PCR manipulation. At the end of the PCR, the dissociation protocol included a slow heating from 60º to 95ºC in 0.2ºC steps, with an 8-s interval between steps. Melting curve profiles were obtained using the dissociation software of the Rotor Gene-3000 apparatus. Samples were analyzed in duplicate and in different PCR runs to test the reproducibility of... |
| Tipo: Info:eu-repo/semantics/other |
Palavras-chave: Melting temperature assay; Real time PCR; MBL2; Polymorphisms; HIV-1-positive children. |
| Ano: 2006 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000600003 |
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| Ky, Chin Long; De Lorgeril, Julien; Hirtz, C; Sommerer, N; Rossignol, M; Bonhomme, F. |
| The European sea bass, Dicentrarchus labrax L., tolerates a range of salinities from freshwater to hyper-saline. To study differences in protein expression, fish were reared in both freshwater and seawater. After 3-month acclimation, gill and intestine epithelia were collected and the soluble protein extracted. In all, 362 spots were differentially expressed in the gills and intestines of fishes reared in seawater compared to those from freshwater. Fifty differential protein spots were excised from a colloidal Coomassie-stained gel. Nine separate protein spots were identified unambiguously by mass spectrometry and database searching. Among the six proteins over-expressed in gill cells in seawater, five were cytoskeleton proteins and one was the aromatase... |
| Tipo: Text |
Palavras-chave: Two dimensional gel electrophoresis; Sea bass; Salinity; Real time PCR; MALDI TOF mass spectrometry. |
| Ano: 2007 |
URL: http://archimer.ifremer.fr/doc/2007/publication-3952.pdf |
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| Goarant, Cyrille; Reynaud, Yann; Ansquer, Dominique; De Decker, Sophie; Merien, Fabrice. |
| In a previous study, we demonstrated the existence of an emerging cluster of Vibrio nigripulchritudo that proved to be associated with shrimp mortality events in New Caledonia. Using sequence polymorphisms evidenced in this previous MultiLocus Sequence Typing study, we developed two new quantitative PCR assays permitting the detection and quantification of V. nigripulchritudo at the genospecies level using SYBR Green I chemistry and at the emerging cluster level using Fluorescence Resonance Energy Transfer technology with hybridization probes. The use of this molecular diagnostic tool evidenced the colonization of the shrimp pond ecosystem by the pathogenic cluster at least at the onset of the disease. This new tool will allow better investigation of the... |
| Tipo: Text |
Palavras-chave: Shrimp; Cluster specific; Vibrio; Pathogen; Mariculture; Real time PCR. |
| Ano: 2007 |
URL: http://archimer.ifremer.fr/doc/2007/publication-2729.pdf |
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| Martins,Polyana K.; Jordão,Berenice Q.; Yamanaka,Naoki; Farias,José R.B.; Beneventi,Magda A.; Binneck,Eliseu; Fuganti,Renata; Stolf,Renata; Nepomuceno,Alexandre L.. |
| Drought cause serious yield losses in soybean (Glycine max), roots being the first plant organ to detect the water-stress signals triggering defense mechanisms. We used two drought induction systems to identify genes differentially expressed in the roots of the drought-tolerant soybean cultivar MG/BR46 (Conquista) and characterize their expression levels during water deficit. Soybean plants grown in nutrient solution hydroponically and in sand-pots were submitted to water stress and gene expression analysis was conducted using the differential display (DD) and real time polymerase chain reaction (PCR) techniques. Three differentially expressed mRNA transcripts showed homology to the Antirrhinum majus basic helix-loop-helix transcription factor bHLH, the... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Cell division; Differential display; Gene expression; Real time PCR; Soybean; Water stress. |
| Ano: 2008 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000300019 |
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| De Lorgeril, Julien; Gueguen, Yannick; Goarant, Cyrille; Goyard, Emmanuel; Mugnier, Chantal; Fievet, Julie; Piquemal, D; Bachere, Evelyne. |
| Understanding of antimicrobial defence mechanisms of penaeid shrimp should help in the design of efficient strategies for the management and disease control in aquaculture. In this study, we have specifically analysed the expression in circulating hemocytes of antimicrobial peptides (AMPs) encoding genes, such as PEN2 and PEN3, ALF, crustin, lysozyme and a putative cysteine-rich peptide. We evidenced a relationship between the level of expression of some AMPs and the successful response of the shrimp, Litopenaeus stylirostris, to circumvent a pathogenic Vibrio penaeicida infection. Additionally, significant differences in some AMP transcript amounts are evidenced between control, non-selected shrimp line and the third generation breeding of shrimp selected... |
| Tipo: Text |
Palavras-chave: Real time PCR; Cysteine rich peptide; Crustin; Anti LPS factor; Lysozyme; Penaeidins; Immune response; Vibrio penaeicida; Penaeid; Decapoda; Crustacean. |
| Ano: 2008 |
URL: http://archimer.ifremer.fr/doc/2008/publication-4524.pdf |
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| Pepin, Jean-francois; Riou, Antoine; Renault, Tristan. |
| Herpes and herpes-like virus infections have been reported in various marine mollusc species associated with high mortality rates. Following the characterisation and genome sequencing of ostreid herpesvirus 1 (OsHV-1), specific diagnostic tools have been developed based on conventional PCR techniques or in situ hybridisation. We have now developed a real-time PCR assay for rapid, sensitive and quantitative detection of OsHV-1, and compared it with a conventional PCR technique described previously. The new assay utilised SYBR® Green chemistry with specific primers C9/C10 targeting the C region. The melt curve analysis of OsHV-1 DNA or DNA extracted from infected material showed only one melting temperature peak (75.75 ± 0.1 °C). The assay had a detection... |
| Tipo: Text |
Palavras-chave: SYBR® Green; Real time PCR; Crassostrea gigas; Ostreid herpesvirus 1; Herpesvirus. |
| Ano: 2008 |
URL: http://archimer.ifremer.fr/doc/2008/publication-3951.pdf |
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| Saulnier, Denis; De Decker, Sophie; Haffner, Philippe. |
| Because Vibrio aestuarianus is known to cause serious infections in Pacific oyster Crassostrea gigas, a real-time PCR assay was developed targeting the dnaJ gene of this bacterium. Only V. aestuarianus strains isolated from C. gigas mortality events in different geographic areas and the reference strain tested positive, whereas no amplification products was obtained with type strains belonging to 23 other species of Vibrio. Sensitivity and reproducibility of the method were assessed using either seawater or oyster homogenate samples spiked with one V aestuarianus strain. All these samples were stored at -20 degrees C in order to mimic retrospective or grouped natural sample analysis without quantification bias due to prolonged freezing. Analysis of... |
| Tipo: Text |
Palavras-chave: V. aestuarianus; Vibrio; Taqman; Real time PCR; Pathogen; Oyster; Crassostrea gigas. |
| Ano: 2009 |
URL: http://archimer.ifremer.fr/doc/2009/publication-6447.pdf |
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| Sauvage, Christopher; Pepin, Jean-francois; Lapegue, Sylvie; Boudry, Pierre; Renault, Tristan. |
| Ostreid herpes virus 1 (OsHV-1) infections, notably reported in Europe and the USA, are closely associated with significant mortalities of the Pacific oyster, Crassostrea gigas, especially during its early stages of life. In summer 2006, we monitored mortality by strict daily verification of three full-sib families of oysters reared under common conditions. We quantified OsHV-1 using real-time PCR in dead and living individuals during and after a mortality event. Mortality events were severe and brief, but significantly different between tested families (cumulative mortality ranging from 1.2 to 49%). Real-time PCR assays revealed different viral DNA loads in dead individuals from different families (P < 0.001). Moreover, the mean level of infection... |
| Tipo: Text |
Palavras-chave: Viral DNA quantification; Real time PCR; OsHV 1; Mortality; Crassostrea gigas. |
| Ano: 2009 |
URL: http://archimer.ifremer.fr/doc/2009/publication-6265.pdf |
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| Robert, Maeva; Garcia, Celine; Chollet, Bruno; Lopez-flores, Inmaculada; Ferrand, Sylvie; Francois, Cyrille; Joly, Jean-pierre; Arzul, Isabelle. |
| Bonamia ostreae is an intracellular protozoan which is recognized as a cause of mortality in European populations of flat oysters (Ostrea edulis). Based on the recent characterization of actin genes of B. ostreae, specific primers were designed for real-time PCR using SYBR® Green chemistry. Specificity was demonstrated by the unique melting temperature peak observed in positive samples and by the lack of amplification in samples of oysters infected by closely related parasites, including Bonamia exitiosa. A calibration curve using a cloned template was defined to estimate copy number. The assay had a 6 log- dynamic range, mean inter- and intra-assay variation coefficients of <1% and a minimum detection limit of 50 gene copies per reaction. Using... |
| Tipo: Text |
Palavras-chave: Heart imprints; Ostrea edulis; Quantification; Detection; Real time PCR; Bonamia ostreae. |
| Ano: 2009 |
URL: http://archimer.ifremer.fr/doc/2009/publication-6934.pdf |
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| Oda,Sandra Helena Inoue; Nepomuceno,Alexandre Lima; Ledur,Mônica Corrêa; Oliveira,Maria Cristina Neves de; Marin,Silvana Regina Rockenbach; Ida,Elza Iouko; Shimokomaki,Massami. |
| Total RNA isolated from Pectoralis major muscle from PSE (L*24h>53.0, pH<5.8) and non-PSE (44<"L*24h>"53) meats of two phenotypically distinct chicken lines, broiler and layer, was used to investigate the α-ryr and β-ryr gene expression by real-time RT-PCR approach. Mean relative quantification (RQ) values were lower (p<0.05) for β-ryr in PSE chickens from both lines when compared to non-PSE chickens, while there was no difference (p>0.05) in α-ryr gene expression regardless of line studied. The β-ryr RQ results suggested that in PSE samples an alteration might occur in the regular ratio (1:1) of α-RyR/β-RyR normally found in avian muscles. These results provided the first evidence of PSE meat occurrence as a result of the differential... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Malignant hyperthermia; Poultry; Gene expression; Real time PCR. |
| Ano: 2009 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000600024 |
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| Lunardi,Lígia Uno; Guembarovski,Roberta Losi; Hanai,Luiz Ricardo; Cristiano,Valderi; Vieira,Maria Lucia Carneiro; Sartori,Daniele; Fungaro,Maria Helena Pelegrinelli. |
| Ochratoxin A is a mycotoxin produced by some fungi species. Among them, Aspergillus carbonarius is considered a powerful producer. Genes involved in the ochratoxin A biosynthesis pathway have been identified in some producer species. However, there are few studies that purpose to identify these genes in A. carbonarius. The use of insertion mutants to identify genes associated with certain properties has been increased in the literature. In this work, the region of T-DNA integration was investigated in one A. carbonarius ochratoxin-defective mutant previously obtained by Agrobacterium tumefaciens-mediated transformation, in order to find an association between interrupted gene and the biosynthesis of ochratoxin A. The integration occurred in a gene that... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Ochratoxigenic fungi; Mycotoxin; Real time PCR; Splicing coactivator protein. |
| Ano: 2009 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700018 |
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| Morga, Benjamin; Arzul, Isabelle; Faury, Nicole; Renault, Tristan. |
| Bonamia ostreae is an intrahaemocytic protozoan affecting Ostrea edulis. The parasite multiplies within haemocytes without being degraded and involves changes in cellular activities. Studies aiming at better understanding host response to a pathogen at the transcriptome levels are frequently based on the use of real time PCR assays, which require some reference genes. However, very few sequence data is available for O. edulis in public databases. Subtracted cDNA libraries were constructed from the O. edulis haemocytes in order to identify genes involved in host reactions against the parasite and quantitative real time PCR assays were developed to study expression of these genes. In this context, identification of reference genes and study of their... |
| Tipo: Text |
Palavras-chave: Real time PCR; Housekeeping genes; Haemocytes; Ostrea edulis; Bonamia ostreae. |
| Ano: 2010 |
URL: http://archimer.ifremer.fr/doc/00018/12880/9829.pdf |
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| Borges,Andreia A.; Humann,Fernanda C.; Campos,Lucio A. Oliveira; Tavares,Mara G.; Hartfelder,Klaus. |
| In Hymenoptera, homozygosity at the sex locus results in the production of diploid males. In social species, these pose a double burden by having low fitness and drawing resources normally spent for increasing the work force of a colony. Yet, diploid males are of academic interest as they can elucidate effects of ploidy (normal males are haploid, whereas the female castes, the queens and workers, are diploid) on morphology and life history. Herein we investigated expression levels of ten caste-related genes in the stingless bee Melipona quadrifasciata, comparing newly emerged and 5-day-old diploid males with haploid males, queens and workers. In diploid males, transcript levels for dunce and paramyosin were increased during the first five days of adult... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Stingless bee; Real time PCR; Caste; Diploid male; Differential gene expression. |
| Ano: 2011 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000400025 |
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| Pribylova,Radka; Kralik,Petr; Donnelly,Neysan; Matiasovic,Jan; Pavlik,Ivo. |
| The aim of this work was to study the expression of selected Mycobacterium avium subsp. paratuberculosis (MAP) genes connected with MAP virulence, adhesion and stress response. The temperature of 6°C and 65°C were chosen with regard to the food industry, storage conditions (refrigerator) and low-temperature pasteurization. A pH of 2.0, using lactic acid, was selected to mimic the natural environment of the stomach. Expression of selected genes was studied using real time reverse transcription PCR on three different MAP isolates. MAP isolates were chosen according to the number of their preceding cultivations. While isolates 8672 and 8819 were previously cultivated only once, MAP isolate 12146 went through four passages. Different expression profiles were... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: MAP; Johne's disease; Stress; Treatment; Real time PCR. |
| Ano: 2011 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000200049 |
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| Registros recuperados: 28 | |
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