Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 3 de 3
Mais filtros

Base de dados
Intervalo de ano de publicação
J Fish Dis ; 2020 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-33340375


Rapid and user-friendly diagnostic tests are necessary for early diagnosis and immediate detection of diseases, particularly for on-site screening of pathogenic microorganisms in aquaculture. In this study, we developed a dual-sample microfluidic chip integrated with a real-time fluorogenic loop-mediated isothermal amplification assay (dual-sample on-chip LAMP) to simultaneously detect 10 pathogenic microorganisms, that is Aeromonas hydrophila, Edwardsiella tarda, Vibrio harveyi, V. alginolyticus, V. anguillarum, V. parahaemolyticus, V. vulnificus, infectious hypodermal and haematopoietic necrosis virus, infectious spleen and kidney necrosis virus, and white spot syndrome virus. This on-chip LAMP provided a nearly automated protocol that can analyse two samples simultaneously, and the tests achieved limits of detection (LOD) ranging from 100 to 10-1  pg/µl for genomic DNA of tested bacteria and 10-4 to 10-5  pg/µl for recombinant plasmid DNA of tested viruses, with run times averaging less than 30 min. The coefficient of variation for the time-to-positive value was less than 10%, reflecting a robust reproducibility. The clinical sensitivity and specificity were 93.52% and 85.53%, respectively, compared to conventional microbiological or clinical methods. The on-chip LAMP assay provides an effective dual-sample and multiple pathogen analysis, and thus would be applicable to on-site detection and routine monitoring of multiple pathogens in aquaculture.

Artigo em Inglês | MEDLINE | ID: mdl-27591837


Although iono-regulatory processes are critical for survival of crustaceans during the molt cycle, the mechanisms involved are still not clear. The Na+/K+/2Cl- cotransporter (NKCC), a SLC12A family protein that transports Na+, K+ and 2Cl- into cells, is essential for cell ionic and osmotic regulation. To better understand the role of NKCC in the molt osmoregulation, we cloned and characterized a NKCC gene from the mud crab, Scylla paramamosain (designated as SpNKCC). The predicted SpNKCC protein is well conserved, and phylogenetic analysis revealed that this protein was clustered with crustacean NKCC. Expression of SpNKCC was detected in all the tissues examined but was highest in the posterior gills. Transmission electron microscopy revealed that posterior gills had a thick type of epithelium for ion regulation while the anterior gills possessed a thin phenotype related to gas exchange. During the molting cycle, hemolymph osmolality and ion concentrations (Na+ and Cl-) increased significantly over the postmolt period, remained stable in the intermolt and premolt stages and then decreased at ecdysis. Meanwhile, the expression of SpNKCC mRNA was significantly elevated (26.7 to 338.8-fold) at the ion re-establishing stages (postmolt) as compared with baseline molt level. This pattern was consistent with the coordinated regulation of Na+/K+-ATPase α-subunit (NKA α), carbonic anhydrase cytoplasmic (CAc) isoform and Na+/H+ exchanger (NHE) genes in the posterior gills. These data suggest that SpNKCC may be important in mediating branchial ion uptake during the molt cycle, especially at the postmolt stages.

Crustáceos/metabolismo , DNA Complementar/genética , Brânquias/metabolismo , Simportadores de Cloreto de Sódio-Potássio/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Crustáceos/fisiologia , Muda , Concentração Osmolar , Homologia de Sequência de Aminoácidos , Simportadores de Cloreto de Sódio-Potássio/química
Dongwuxue Yanjiu ; 33(5): 503-9, 2012 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-23019033


Nibea albiflora was immunized by intraperitoneal injection with either Lipopolysaccharide (LPS) or outer membrane protein (OMP) extracted from Vibrio vulnificus or formalin killed Vibrio vulnificus (FKC). The influence of the three antigens on the immunological function of Nibea albiflora was determined at different time points following the injection by testing the agglutinating antibody titers of the serum, lysozyme activity of the serum, phagocytic activity of the blood and the relative survival percentage. The results showed that the three antigens have higher immunogenicity and antigenicity than the control group(injection with sterile saline). The agglutinating antibody titers of the immune challenged groups increased quickly, and were highest on the day 28. The lysozyme activity and phagocytic activity were raised significantly (P<0.01), reaching their top value on day 21, and then the index gradually reduced. The immunological indexes of three immune groups were higher than the control group (P<0.05). The agglutinating antibody titers of the LPS group or the OMP group were lower than the control group, but the relative survival percentage was adverse when challenged with a Vibrio vulnificus infection. The order of relative survival percentage was group LPS>OMP>FKC>Control.

Antígenos de Bactérias/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/prevenção & controle , Perciformes/imunologia , Vibrioses/veterinária , Vibrio vulnificus/imunologia , Animais , Antígenos de Bactérias/isolamento & purificação , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/isolamento & purificação , Doenças dos Peixes/microbiologia , Doenças dos Peixes/mortalidade , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/isolamento & purificação , Perciformes/microbiologia , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrioses/prevenção & controle , Vibrio vulnificus/química , Vibrio vulnificus/genética