Purification of total DNA extracted from activated sludge

Purification of the total DNA extracted from activated sludge samples was studied. The effects of extraction buffers and lysis treatments （lysozyme, sodium dodecyl sulfate （SDS）, sonication, mechanical mill and thermal shock） on yield and purity of the total DNA extracted from activated sludge were investigated. It was found that SDS and mechanical mill were the most effective ways for cell lysis, and both gave the highest DNA yields, while by SDS and thermal shock, the purest DNA extract could be obtained. The combination of SDS with other lysis treatment, such as sonication and thermal shock, could apparently increase the DNA yields but also result in severe shearing. For the purification of the crude DNA extract, polyvinyl polypyrrolidone was used for the removal of humic contaminants. Cetyltrimethyl ammonium bromide, potassium acetate and phenol/chloroform were used to remove proteins and polysaccharides from crude DNA. Crude DNA was further purified by isopropanol precipitation. Thus, a suitable protocol was proposed for DNA extraction, yielding about 49.9 mg （total DNA）/g volatile suspended solids, and the DNA extracts were successfully used in PCR amplifications for 16S rDNA and 16S rDNA V3 region. The PCR products of 16S rDNA V3 region allowed the DGGE analysis （denatured gradient gel electrophoresis） to be possible.

A Study of the Characteristics of Microorganisms for Effective Degradation of Marine Oil Spills

Four microorganism strains were isolated from coastal petroleum-polluted soil and sand samples of Bohai Sea oilfield; they were found to degrade marine oil spills effectively. The experimental results show that the degradation efficiency of crude oil with these four strains （XT-4, SZ-1-25, B-4-9, BS-3-12） is 95.97%, 96.01%, 97.99% and 98.99%, respectively, in their optimum conditions. The characteristics of bacterial biodegradation are investigated. The simulation biotreatment of oil-contaminated beach sand, with an initial oil content of 5,664mg/kg-dry-sand, shows that the residual oil content is 2,700 mg/kg-dry-sand and 2,679 mg/kg-dry-sand after 170 days＇ treatment with two bacteria （B-4-9, BS-3- 12）, respectively.

Recovering rare earth elements via immobilized red algae from ammonium-rich wastewater

Biotreatment of acidic rare earth mining wastewater via acidophilic living organisms is a promising approach owing to their high tolerance to high concentrations of rare earth elements(REEs);however,simultaneous removal of both REEs and ammonium is generally hindered since most acidophilic organisms are positively charged.Accordingly,immobilization of acidophilic Galdieria sulphuraria(G.sulphuraria)by calcium alginate to improve its affinity to positively charged REEs has been used for simultaneous bioremoval of REEs and ammonium.The results indicate that 97.19%,96.19%,and 98.87%of La,Y,and Sm,respectively,are removed by G.sulphuraria beads(GS-BDs).The adsorption of REEs by calcium alginate beads(BDs)and GS-BDs is well fitted by both pseudo first-order(PFO)and pseudo second-order(PSO)kinetic models,implying that adsorption of REEs involves both physical adsorption caused by affinity of functional groups such as-COO-and -OH and chemical adsorption based on ion exchange of Ca^(2+) with REEs.Notably,GS-BDs exhibit high tolerance to La,Y,and Sm with maximum removal efficiencies of 97.9%,96.6%,and 99.1%,respectively.Furthermore,the ammonium removal efficiency of GS-BDs is higher than that of free G.sulphuraria cells at an initial ammonium concentration of 100 mg L^(-1),while the efficiency decreases when initial concentration of ammonium is higher than 150 mg L^(-1).Last,small size of GS-BDs favors ammonium removal because of their lower mass transfer resistance.This study achieves simultaneous removal of REEs and ammonium from acidic mining drainage,providing a potential strategy for biotreatment of REE tailing wastewater.

Strength-increase mechanism and microstructural characteristics of a biotreated geomaterial

Microbially induced calcite precipitation(MICP)is a recently proposed method that is environmentally friendly and has considerable potential applications in artificial biotreated geomaterials.New artificial biotreated geomaterials are produced based on the MICP technology for different parent soils.The purpose of this study is to explore the strength-increase mechanism and microstmctural characteristics of the biotreated geomaterial through a series of experiments.The results show that longer mineralization time results in higher-strength biotreated geomaterial.The strength growth rate rapidly increases in the beginning and remains stable afterwards.The calcium ion content significantly increases with the extended mineralization time.When standard sand was used as a parent soil,the calcium ion content increased to a factor of 39 after 7 days.The bacterial cells with attached calcium ions serve as the nucleus of crystallization and fill the pore space.When fine sand was used as a parent soil,the calcium ion content increased to only a factor of 7 after 7 days of mineralization.The nucleus of crystallization could not normally grow because of the limited pore space.The porosity and variation in porosity are clearly affected by the parent soil.Therefore,the strength of the biotreated geomaterial is affected by the parent soil properties,mineralization time,and granular material pore space.This paper provides a basis for theory and experiments for biotreated geomaterials in future engineering practice.