Arboviruses represent a serious problem to public health and agriculture worldwide. Fast, accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation. We developed a cost-effective, rapid, and highly sensitive one-step “triplex RT-PCR enzyme hybridization” assay for simultaneous detections of Japanese Encephallitis virus (JEV, Flaviviridae), Getah virus (GETV, Togaviridae), and Tahyna virus (TAHV, Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction. The analytical sensitivity of this assay was 1 PFU/mL for JEV, 10 PFU/mL for GETV, and 10 PFU/mL for TAHV. This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods. When “triplex RT-PCR enzyme hybridization” was applied to 29 cerebrospinal fluid (CSF) samples that were JEV-positive by normal RT-PCR assay, all samples were strongly positive for JEV, but negative for GETV and TAHV, demonstrating a good sensitivity, specificity, and performance at CSF specimen detection.
A chemolithoautotrophic arsenite-oxidizing bacterium, designated strain KGO-5, was isolated from arsenic-contaminated industrial soil. Strain KGO-5 was phylogenetically closely related with Sinorhizobium meliloti with 16S rRNA gene similarity of more than 99%, and oxidized 5 mM arsenite under autotrophic condition within 60 h with a doubling time of 3.0 h. Additions of 0.01–0.1% yeast extract enhanced the growth significantly, and the strain still oxidized arsenite efficiently with much lower doubling times of approximately 1.0 h. Arsenite-oxidizing capacities (11.2–54.1 μmol h−1 mg dry cells−1) as well as arsenite oxidase (Aio) activities (1.76–10.0 mU mg protein−1) were found in the cells grown with arsenite, but neither could be detected in the cells grown without arsenite. Strain KGO-5 possessed putative aioA gene, which is closely related with AioA of Ensifer adhaerens. These results suggest that strain KGO-5 is a facultative chemolithoautotrophic arsenite oxidizer, and its Aio is induced by arsenic.
T . This strain harboured meso-diaminopimelic acid, alanine and glycine as the major cell-wall amino acids, xylose and glucose as the characteristic whole-cell sugars, and iso-C 15 : 0 (20.53 %),iso-C 17 : 0 (12.74 %), iso-C 16 : 0 (12.15 %), anteiso-C 17 : 0 (7.97 %), C 17 : 1 v8c (7.49 %) and C 17 : 0 (6.63 %) as the dominant fatty acids. The major menaquinones were MK-10(H 4 ) and MK-10(H 6 ). The phospholipid profile comprised phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol and unknown phosphoglycolipids. The DNA G+C content was 71.4 mol%. A comprehensive analysis of several physiological and biochemical traits and DNA-DNA relatedness indicated that strain K55T was different from closely related species. These phenotypic, genotypic and chemotaxonomic data suggest that strain K55 T represents a novel species of the genus Micromonospora, for which the name Micromonospora nickelidurans sp. nov., is proposed. The type strain is K55 T (5JCM 30559 T 5ACCC19713 T ).
A Gram-positive, facultative anaerobic, rod-shaped, and endospore-forming strain, designated 53-2(T) was isolated from the root nodule of Oxytropis ochrocephala Bunge growing on Qilian mountain, China. The strain can grow at pH 7.0-8.0, 10-50 °C and tolerate up to 11% NaCl. Optimal growth occurred at pH 7.2 and 37 °C. The result of BLASTn search based on 16S rRNA gene sequence revealed that strain 53-2(T) , being closest related to Bacillus acidicola 105-2(T) , possessed remote similarity (less than 95.64%) to the species within genus Bacillus. The DNA G + C content was 37.8%. Chemotaxonomic data (major quinone is MK-7; major polar lipids are diphosphatidylglycerol, phosphatidylglycerol, unknown phospholipid, and aminoglycophospholipid; fatty acids are anteiso-C15: 0 , iso-C15:0 and anteiso-C17: 0 ) supported the affiliation of the isolate to the genus Bacillus. On the basis of physiological, phylogenetic, and biochemical properties, strain 53-2(T) represents a novel species within genus Bacillus, for which the name Bacillus radicibacter is proposed. The type strain is 53-2(T) (=DSM27302(T) =ACCC06115(T) =CCNWQLS5(T) ).
An actinomycete, strain D34T , was isolated from a soil sample collected from the rhizosphere T (98.6 %). The strain contained meso-diaminopimelic acid, alanine, glycine and glutamic acid as major cell-wall amino acids. Mannose, rhamnose and galactose were the characteristic whole-cell sugars. The fatty acid profile consisted predominantly of iso-C 15 : 0 , iso-C 16 : 0 , iso-C 16 : 1 , C 17 : 1 v6c, anteiso-C 17 : 0 and anteiso-C 15 : 0 . The phospholipid profile included phosphatidylethanolamine (typical of phospholipid pattern type II). Furthermore, a combination of some physiological and biochemical properties and low DNA-DNA relatedness values indicated that strain D34 T was differentiated from members of closely related species. On the basis of these phenotypic, genotypic and chemotaxonomic data, strain D34
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