Activation of aryl hydrocarbon receptor reduces carbendazim-induced cell death.

Carbendazim inhibits microtubule assembly, thus blocking mitosis and inhibiting cancer cell proliferation. Accordingly, carbendazim is being explored as an anticancer drug. Data show that carbendazim increased mRNA and protein expressions and promoter activity of CYP1A1. In addition, carbendazim activated transcriptional activity of the aryl hydrocarbon response element, and induced nuclear translocation of the aryl hydrocarbon receptor (AhR), a sign the AhR is activated. Carbendazim-induced CYP1A1 expression was blocked by AhR antagonists, and was abolished in AhR signal-deficient cells. Results demonstrated that carbendazim activated the AhR, thereby stimulating CYP1A1 expression. In order to understand whether AhR-induced metabolic enzymes turn carbendazim into less-toxic metabolites, Hoechst 33342 staining to reveal carbendazim-induced nuclear changes and flow cytometry to reveal the subG0/G1 population were applied to monitor carbendazim-induced cell apoptosis. Carbendazim induced less apoptosis in Hepa-1c1c7 cells than in AhR signal-deficient Hepa-1c1c7 mutant cells. Pretreatment with ß-NF, an AhR agonist that highly induces CYP1A1 expression, decreased carbendazim-induced cell death. In addition, the lower the level of AhR was, the lower the vitality present in carbendazim-treated cells, including hepatoma cells and their derivatives with AhR RNA interference, also embryonic kidney cells, bladder carcinoma cells, and AhR signal-deficient Hepa-1c1c7 cells. In summary, carbendazim is an AhR agonist. The toxicity of carbendazim was lower in cells with the AhR signal. This report provides clues indicating that carbendazim is more potent at inducing cell death in tissues without than in those with the AhR signal, an important reference for applying carbendazim in cancer chemotherapy.

Occurrence of Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus in the Aquacultural Environments of Taiwan.

The occurrence of Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae in a total of 72 samples from six aquaculture ponds for groupers, milk fish, and tilapia in southern Taiwan was examined by the membrane filtration and colony hybridization method. The halophilic V. parahaemolyticus was only recovered in seawater ponds, with a high isolation frequency of 86.1% and a mean density of 2.6 log CFU/g. V. cholerae was found in both the seawater and freshwater ponds but preferentially in freshwater ponds, with a frequency of 72.2% and a mean density of 1.65 log CFU/g. V. vulnificus was identified mainly in seawater ponds, with an isolation frequency of 27.8%. The density of V. parahaemolyticus in seawater ponds was positively related to water temperature (Pearson correlation coefficient, r = 0.555) and negatively related to salinity (r = 2 0.333). The density of V. cholerae in all six ponds was positively related to water temperature (r = 0.342) and negatively related to salinity (r = 2 0.432). Two putatively pathogenic tdh(+) V. parahaemolyticus isolates (1.4% of the samples) and no ctx(+) V. cholerae isolates were identified. The experimental results may facilitate assessments of the risk posed by these pathogenic Vibrio species in Taiwan, where aquaculture provides a large part of the seafood supply.

Molecular characterization of clinical and environmental Vibrio parahaemolyticus isolates in Taiwan.

Vibrio parahaemolyticus is the most prevalent foodborne pathogen in Taiwan and it is frequently recovered from seafood. In this study, V. parahaemolyticus that was isolated in recent years from aquacultural environments and clinical specimens were comparatively analyzed by NotI-restricted pulsed-field gel electrophoresis (PFGE) and polymerase chain reaction, targeting common toxin genes (tdh, trh, ureC), MTase gene, toxR regulator, markers for pandemic strains (ORF8, group-specific toxRS) and representative genes of type three secretion systems T3SS1 (vcrD1, VP1680, vopD) and T3SS2α (vcrD2, vopD2, vopB2, vopP, vopC, vopT). Among the 48 clinical isolates and 93 environmental isolates that were analyzed by PFGE, a total of 26 and 76 pulsetypes were identified and grouped into six and nine clusters, respectively, at 80% similarity. The pandemic O3:K6 clones and other clinical and environmental isolates were further characterized according to the distribution of these examined target genes. The MTase gene and the vopB2, vopP, vopC and vopT genes of T3SS2α were present at a significantly higher frequency (>90%) in the pandemic clones than in other clinical isolates. The MTase gene and some other virulence-associated genes were also present in a few of the environmental isolates, and these results suggest the horizontal transfer of these genes in the clinical and environmental isolates of this species.

Prevalence of Vibrio parahaemolyticus in oyster and clam culturing environments in Taiwan.

Vibrio parahaemolyticus is the most prevalent gastroenteritis pathogen in Taiwan and some other Asian countries, and it frequently occurs in oysters and other seafood. This study monitors changes in the density of V. parahaemolyticus and environmental parameters in oyster and hard clam aquacultural environments in Taiwan. Water, sediment and shellfish samples were collected from five sampling sites in 2008-2010, and analyzed for environmental physiochemical parameters, numbers of indicator bacteria (total aerobic counts, total coliforms and fecal coliforms), Vibrio and V. parahaemolyticus present. The results for open oyster farms and hard clam ponds did not differ significantly. V. parahaemolyticus was detected in 77.5, 77.5, 70.8 and 68.8% of the water, sediment, oyster and clam samples, respectively. The densities of V. parahaemolyticus were significantly higher in shellfish than in sediment or water samples, with mean values of 1.33, 1.04 and -0.02 Log CFU/g, respectively. Among these five sampling sites, Shengang and Fangyuan yielded significantly different data from those obtained at the other three sites. As determined by linear multiple regression, V. parahaemolyticus density in water samples depended significantly on the precipitation and Vibrio count, while the V. parahaemolyticus density in the sediment or shellfish samples depended significantly on the salinity of the seawater. Among 1076 isolates examined, a total of three putative pathogenic isolates were identified from 2.5% of the examined samples, and these isolates exhibited hemolytic or urease activities and the presence of gene markers for tdh, trh, type III secretion system (T3SS) 1 (vcrD1) or T3SS2α (vcrD2). The results herein may facilitate the assessment of risk associated with this pathogen in Taiwan and other geographically similar regions.