Concentrations of arsenic and heavy metals selleck chemical are extremely high in water, soil and sediments of this area [36]. The SY soil was collected from a pig farm, Shayang County, Jingmen City, Hubei Province where there are certain levels of arsenic in the soil due to the long-term usage

of arsenic in feed material to resist disease and stimulate pig growth. The other two samples, LY and YC, were collected from the low arsenic-contaminated soils near the Yellow Sea of Lianyungang and Yancheng Cities Jiangsu Province, eastern China, respectively. Several soil samples from each site were collected from the surface horizon (0–15 cm), stored at 4°C and mixed together for bacterial isolation. The total arsenic concentrations of the four soils (determined by atomic absorption spectrometry) were 337.2 mg kg-1 (183.4–882.2 mg kg-1, SD = 184.58), 72.1 mg kg-1 (43.4–94.6 mg kg-1, SD = 18.31), 24.1 mg kg-1 (15.7–40.1, mg kg-1, SD = 8.24) and 34.6 mg kg-1 (22.0–48.8 mg kg-1, SD = 8.96) for TS, SY, LY and YC, respectively. Isolation and identification PF01367338 of arsenite-resistant and arsenite-oxidizing bacteria One hundred grams of each soil sample was amended with NaAsO2 to a final concentration of 500 mg kg-1 and incubated at

28°C for a week. Alvocidib During incubation sterilized H2O was added to the jars to reach the original moisture value. Isolation of arsenite-resistant bacteria was performed by adding 10 g (triplicates) of each soil to 90 mL 0.85% NaCl in a 250 mL Erlenmeyer flask and shaken at 160 rpm for 30 min. 1 mL of the above mixture was added to 9 mL 0.85% NaCl for serial dilution and plated on chemically defined medium (CDM) plates [9] with a final concentration of 800 μM NaAsO2 and incubated at 28°C for another week. Single colonies were picked and restreaked several times to obtain pure check details isolates. The obtained arsenite-resistant bacteria were tested for their abilities to oxidize As(III) (NaAsO2) using a qualitative KMnO4 screening method [10]. Each arsenite-resistant bacterium was inoculated in CDM broth with a final concentration of 800 μM NaAsO2

and then shaken at 160 rpm for 5 days at 28°C. For each isolate 1 mL culture was added to a 1.5 mL centrifuge tube containing 30 μL of 0.01 M KMnO4 and the color change of KMnO4 was monitored. A pink color of the mixture indicated a positive arsenite oxidation reaction [formation of As(V)]. The sterile CDM medium containing the same amount of NaAsO2 was used as an abiotic control. The arsenite oxidizing phenotype was also detected using the molybdene blue method with a spectrophotometer (DU800, BeckMan, CA, USA) [48]. Total DNA of each strain was extracted using standard molecular genetic methods. Nearly full-length 16S rDNA of the bacteria was amplified by PCR using universal primers Uni-27F and Uni-1492R (Table 1) [49].