Monitoring of Fasciola Species Contamination in Water Dropwort by cox1 Mitochondrial and ITS-2 rDNA Sequencing Analysis

Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.

Protective Effect of Yellow-Green Vegetable Juices on DNA Damage in Chinese Hamster Lung Cell Using Comet Assay

The present study was attempted to investigate the antioxidant capacity of popular yellow-green vegetable juices (kale, Angelica keishei, carrot, small water dropwort) and to investigate the effect of vegetable juices on protecting oxidative damage to DNA in cultured Chinese hamster lung (CHL) cells. Antioxidant capacity was analyzed by TRAP assay (Total radical-trapping antioxidant potential). Cellular DNA dmamage was measured by SCGE (single-cell gel electrophoresis, also known as comet assay. Cells incubated in medium with PBS (negative control) or with various concentration of the freeze dried green juices (25, 50, 100, 250 ug/mL) resuspended in PBS were treated with H2O2 (200 micrometer) as an oxidative stimulus for 5 min at 4 degrees C. The physiological function of each vegetable juice on oxidative DNA damage was analyzed and expressed as tail moment (tail length X percentage migrated DNA in tail). Kale juice had the highest TRAP value suggesting that kale has the highest antioxidant capacity followed by Angelica keishei, small water dropwort and carrot. Cells treated with H2O2 had extensive DNA damage compared with cells treated with PBS or pre-treated with vegetable juice extracts. All green juices inhibited H2O2-induced DNA damage with kale being the most effective juice among the tested juices. These results indicate that green juice supplementation to CHL cells followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.