RESUMEN
OBJECTIVE: To study the characteristics of the strains of Salmonella enterica (S. enterica) serovar Senftenberg lacking Salmonella pathogenicity island 1 (SPI-1). METHODS: A total of 10 strains of S. enterica serovar Senftenberg were isolated from 10 cases of diarrhea patients. Pulsed field gel electrophoresis (PFGE), PCR, sequencing techniques and cell invasion test were adapted to study the molecular types and invasiveness of the genes and cells; and made a comparison between the 10 strains and the strains (C02013) isolated in Shenzhen in 2002. RESULTS: The 10 Senftenberg isolated (S09007-S09012, S09014-S09017) in Shanghai showed three PFGE patterns, which were significantly different from the strains isolated in Shenzhen. PCR-amplified results indicated the invasion gene (invA), secreted effector protein gene (sipA) and gene fragments as fhlA-hilA, hilA-spaP and spaP-invH in the 10 strains of SPI-1 were all negative. The sequencing results revealed that the 10 strains isolated in Shanghai lacked most parts of SPI-1 genes, as fragments from orgA to invH and parts of orgA gene itself; however, compared with strains isolated in Shenzhen, the sprB-orgC gene existed. The missing parts of genes were replaced by a simple insertion sequence (IS) of 1000 bp in the strains isolated both in Shenzhen in 2002 and in Shanghai in 2006. The invasiveness rates of the 10 strains (S09007-S09012, S09014-S09017) towards Hela cells were (0.0053 ± 0.0024)%, (0.0046 ± 0.0006)%, (0.0047 ± 0.0003)%, (0.0064 ± 0.0012)%, (0.0065 ± 0.0011)%, (0.0070 ± 0.0020)%, (0.0115 ± 0.0030)%, (0.0099 ± 0.0039)%, (0.0180 ± 0.0135)% and (0.0031 ± 0.0012)%, respectively; which were all significantly lower than the rate of invA-positive control strain STM1344 ((5.0800 ± 0.6333)%); lower or close to the rate of invA-lacked artificial-mutated strain STMinvA-((0.0193 ± 0.0045)%). CONCLUSION: SPI-1 genes are not essential for the diarrhea caused by S. enterica serovar Senftenberg.
Asunto(s)
Diarrea/microbiología , Heces/microbiología , Islas Genómicas , Salmonella enterica/genética , Adulto , Anciano , Técnicas de Tipificación Bacteriana , Femenino , Genes Bacterianos , Células HeLa , Humanos , Masculino , Persona de Mediana Edad , Salmonella enterica/aislamiento & purificación , Salmonella enterica/patogenicidadRESUMEN
Ever-growing application of engineering nanoparticles in many sectors of the society requires efficient methods to extract them from soil and sediment, for the sake of environmental protection. In this study, we develop a new method which uses sodium pyrophosphate solution (TSPP, Na4P2O7) as extratant to extract gold nanoparticle (AuNPs) from soil and sediment under optimized parameters through vortexing, water bath oscillation, ultrasonic bath and precipitation. SP-ICP-MS was used for the detection of number concentration, mass concentration and size distribution of AuNPs in soil. UV irradiation was innovatively used to directly degrade soil organic matter to improve the recovery of AuNPs due to their low recovery rate in rich organic soils. It could be found that the mass fraction recovery increased from 36% (without UV digestion) to 83% (with 48h UV digestion). The extraction method is versatile for different coating layers and wide-ranging particle sizes in real soil and sediment. Therefore, the rapid and efficient characterization and quantification of AuNPs in soil and sediment are achieved, and the researches on the extraction method of AuNPs and their behavior and toxicity assessment in soil environment can be enriched.
RESUMEN
OBJECTIVE: To determine the occurrence and distribution of specific clones of pathogenic Vibrio parahaemolyticus(VP)isolated in Shenzhen and to assess the relationship between serotype O3:K6 and the globally distributed pandemic clone. METHODS: A total of 1005 VPs isolated from diarrhea patients in 2002-2008 were sero-typed. Real-time PCR was used to detect the virulence genes tlh, toxR, tdh, trh and orf8 in 281 isolates from 68 different serotypes. The main serotypes were typed by pulsed field gel electrophoresis(PFGE). Strains with dominant serotypes and PFGE patterns were assayed by GS-PCR and toxRS sequencing for the identification of pandemic clone. Multilocus sequence typing(MLST)analysis was reserved for exemplary 41 O3 : K6 and O1 : K25 isolates. RESULTS: Seventy-nine serotypes were observed among the 1005 isolates, including O3 : K6(57.9%), O4 : K8(8.16%), O1 : KUT(5.87%), O1 : K25(5.27%), O4 : K68(1.39%), O1 : K56(1.39%) and O9 : K44(0.99%). Most of the strains(99.36%)showed PCR positive to tlh, toxR, and tdh but eleven strains were tdh negative. MLST showed that all the 36 O3 : K6 isolates belonged to ST3 and all the 5 O4 : K8 strains were ST189. These results matched the description of the pandemic VP clone. CONCLUSION: A recognizable burden of diarrheal illness caused by VP had been seen in Shenzhen. Results from serotyping indicated that although there existing a large variety of diversities, the dominant serotype appeared to be O3 : K6. VP isolates identified in Shenzhen mainly showed as tdh positive but trh negative, in consistent with the current pandemic O3 : K6 clone. The pandemic O3 : K6 clone did appear to co-exist with other clones of O3 : K6, as well as O4 : K8,O1 : K25. Potential outbreak of VP could be monitored through the laboratory-based surveillance programs, suggesting that the strategies related to prevention and control of VP should be prioritized in Shenzhen.