15 research outputs found
Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods
QUANTIDADE DE FÓSFORO EXTRAÍDO PELAS SOLUÇÕES DE MEHLICH-1 E MEHLICH-3 EM RAZÃO DE DIFERENTES VELOCIDADES DE AGITAÇÃO, TEMPOS DE CONTATO E TEMPERATURAS
Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods
The aim of this study was to compare two nucleic acid extraction
methods for the recovery of enteric viruses from activated sludge. Test
samples were inoculated with human adenovirus (AdV), hepatitis A virus
(HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an
adsorption-elution-precipitation method. Two extraction methods were
used: an organic solvent-based method and a silica method. The
organic-based method was able to recoup 20% of the AdV, 90% of the RV
and 100% of both the PV and HAV from seeded samples. The silica method
was able to recoup 1.8% of the AdV and 90% of the RV. These results
indicate that the organic-based method is more suitable for detecting
viruses in sewage sludge
Identification of differentially transcribed genes in shrimp Litopenaeus vannamei exposed to osmotic stress and challenged with WSSV virus
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The effects of hyposmotic stress and white spot syndrome virus (WSSV) challenge in expression was studied in the marine shrimp Litopenaeus vannamei. Messenger RNA from gills of shrimp submitted to osmotic stress was isolated to identify genes differentially expressed through the suppressive subtractive hybridization (SSH) method. Two subtractive libraries forward and two reverse were constructed to identify up and down-regulated genes under these conditions. About 192 clones were sequenced, of which 46 genes were identified. These genes encode proteins corresponding to a wide range of biological roles, including defense, cell signaling, electron transfer, cell proliferation and differentiation, apoptosis, intermediary metabolism, cytoskeleton and digestion. Among the identified genes, 19 were up-regulated and 27 were down-regulated in the animals kept at a lower ion concentration. We evaluated the expression of eight genes by RT-qPCR in shrimp submitted to hyposmotic conditions with and without WSSV challenge. The SSH enabled the identification of genes that are influenced by hyposmotic stress. A significant up-regulation was observed in lectin-C, QM, TGF beta inducible nuclear protein 1, ciclophilin, malate dehydrogenase, mitochondrial ATP synthase F chain and ATP synthase subunit 9 precursor transcripts. However, the expression of these genes in L. vannamei was not affected by WSSV infection both at isosmotic and hyposmotic conditions. (C) 2011 Elsevier Inc. All rights reserved.717381FINEP-Brazil [01070625-00/336-2007]Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FINEP-Brazil [01070625-00/336-2007