Detoxification of phoxim by a gut bacterium of Delia antiqua.

The presence of certain associated bacteria has been reported to increase pest resistance to pesticides, which poses a serious threat to food security and the environment. Researches on the above microbe-derived pesticide resistance would bring innovative approaches for pest management. Investigations into the phoxim resistance of Delia antiqua, one Liliaceae crop pests, revealed the contribution of a phoxim-degrading gut bacterium, D39, to this resistance. However, how the strain degraded phoxim was unknown. In this study, the role of D39 in phoxim degradation and resistance was first confirmed. DT, which had an identical taxonomy but lacked phoxim-degrading activity, was analyzed alongside D39 via comparative genomics to identify the potential phoxim degrading genes. In addition, degradation metabolites were identified, and a potential degradation pathway was proposed. Furthermore, the main gene responsible for degradation and the metabolites of phoxim were further validated via prokaryotic expression. The results showed that D39 contributed to resistance in D. antiqua larva by degrading phoxim. Phoxim was degraded by an enzyme encoded by the novel gene phoD in D39 to O,O-diethyl hydrogen phosphorothioate and 2-hydroxyimino-2-phenylacetonitrile. Finally, downstream products were metabolized in the tricarboxylic acid cycle. Further analysis via prokaryotic expression of phoD confirmed its degradation activity. The mechanisms through which gut microbes promote pesticide resistance are elucidated in this study. These results could aid in the development of innovative pest control methods. In addition, this information could also be used to identify microbial agents that could be applied for the remediation of pesticide contamination.

Effects of Salinity on the Biodegradation of Polycyclic Aromatic Hydrocarbons in Oilfield Soils Emphasizing Degradation Genes and Soil Enzymes.

The biodegradation of organic pollutants is the main pathway for the natural dissipation and anthropogenic remediation of polycyclic aromatic hydrocarbons (PAHs) in the environment. However, in the saline soils, the PAH biodegradation could be influenced by soil salts through altering the structures of microbial communities and physiological metabolism of degradation bacteria. In the worldwide, soils from oilfields are commonly threated by both soil salinity and PAH contamination, while the influence mechanism of soil salinity on PAH biodegradation were still unclear, especially the shifts of degradation genes and soil enzyme activities. In order to explain the responses of soils and bacterial communities, analysis was conducted including soil properties, structures of bacterial community, PAH degradation genes and soil enzyme activities during a biodegradation process of PAHs in oilfield soils. The results showed that, though low soil salinity (1% NaCl, w/w) could slightly increase PAH degradation rate, the biodegradation in high salt condition (3% NaCl, w/w) were restrained significantly. The higher the soil salinity, the lower the bacterial community diversity, copy number of degradation gene and soil enzyme activity, which could be the reason for reductions of degradation rates in saline soils. Analysis of bacterial community structure showed that, the additions of NaCl increase the abundance of salt-tolerant and halophilic genera, especially in high salt treatments where the halophilic genera dominant, such as Acinetobacter and Halomonas. Picrust2 and redundancy analysis (RDA) both revealed suppression of PAH degradation genes by soil salts, which meant the decrease of degradation microbes and should be the primary cause of reduction of PAH removal. The soil enzyme activities could be indicators for microorganisms when they are facing adverse environmental conditions.

Enzyme cocktail containing NADH regeneration system for efficient bioremediation of oil sludge contamination.

Oil sludge is one kind of toxic and persistent contamination to ecology system from petroleum industry. In order to recycle contaminated sands and reduce environmental impacts at a lower operating cost, enzyme cocktail 21/CbFDH including NADH regeneration system for oily sludge bioremediation was constructed for the first time. The intracellular enzymes of oil-degrading strain Acinetobacter calcoaceticus 21 were prepared and the formate dehydrogenase gene Cbfdh from Candida boidinii was cloned and functionally expressed in E.coli BL21 induced by lactose. The activity and stability of CbFDH was enhanced through self-induction medium optimization using Box-Behnken design. The CbFDH activity was 12.2 times increased and was only decreased 3.9% upon storage at 30 °C for 5 d. The CbFDH increased the degradation rate of oil in high concentration. For the sludge with 10% oil (w/w), the degradation rate achieved 35.6% after 12 h using enzyme 21/CbFDH with the protein ratio of 1:4. The results will provide novel perspectives for creation and operation of petroleum-degrading enzymes involving formate dehydrogenase with higher efficiency and lower cost comparing to current microbial strains or consortium.

Preparation of five high-purity iridoid glycosides from Gardenia jasminoides Eills by molecularly imprinted solid-phase extraction integrated with preparative liquid chromatography.

Five iridoid glycosides were prepared using molecularly imprinted solid-phase extraction combined with preparative high-performance liquid chromatography. Hydrophilic molecularly imprinted polymers were synthesized using α-1-allyl-2-N-acetyl glucosamine, which introduced an abundance of hydrophilic groups into the polymers. Using molecularly imprinted solid-phase extraction as the sample pretreatment procedure, five iridoid glycosides, gardenoside, geniposide, shanzhiside, geniposidic acid, and genipin-1-O-gentiobioside, were selectively enriched from Gardenia fructus extracts. Preparative high-performance liquid chromatography then provided iridoid glycosides with a purity >98%. The structures were elucidated by using nuclear magnetic resonance spectroscopy, optical rotation and melting point measurements, and mass spectrometry. The results demonstrate that molecularly imprinted solid-phase extraction combined with preparative high-performance liquid chromatography was an efficient, rapid, and economical method for the preparation of bioactive compounds from natural products.

Leaching behavior of heavy metals from sewage sludge solidified by cement-based binders.

Sewage sludge was stabilized and solidified (S/S) with cement-based binders prior to landfill in order to reduce the potential hazard of heavy metals. The leaching behavior and chemical speciation of heavy metals in the solidified sludge were studied by semi-dynamic leaching test, toxicity characteristic leaching procedure (TCLP) and sequential extraction procedures. The effectiveness of S/S treatment was evaluated by determining diffusion coefficients (De) calculated from the data achieved in semi-dynamic leaching test. High De values under low pH (4.0 compared to 7.0 and 10.0) and high temperature (35°C compared to 10°C and 25°C) conditions were always recorded. Acid-assisted S/S product showed lower De values and leaching tendency from TCLP test results for all the selected heavy metals (Ni, Cu, Zn and Pb) compared to traditional S/S product using the same binders. More heavy metals were converted into stable fractions by acid-assisted S/S than usual S/S according to sequential extraction test.

Dewaterability characteristics of sludge conditioned with surfactants pretreatment by electrolysis.

The potential benefits of electrolysis-conditioned sludge dewaterability treatments with surfactants were investigated in this study. Capillary suction time (CST) and specific resistance of filtration (SRF) were used to evaluate the sludge dewaterability. Extracellular polymeric substance (EPS) content, viscosity and zeta potential were determined in an attempt to explain the observed changes in the conditioning process. The results indicated that SDS (Sodium Dodecyl Sulphate) and Triton X-100 have negative effect on the dewaterability of sludge pretreated both with and without electrolysis. However, with a combination of CTAB (Cetyl Trimethyl Ammonium Bromide) and electrolysis pretreatment presented clear advantages over surfactant conditioning alone for improving sludge dewaterability. The optimal dosage of CTAB to give maximal dewaterability was found to be 2000 mg/L, which generated sludge with optimal EPS concentration (150-300 mg/L), viscosity (55-62 mpa s) and zeta potential (-2.12 to -1.19 mV).

Chemical and microbial changes during autothermal thermophilic aerobic digestion (ATAD) of sewage sludge.

Autothermal thermophilic aerobic digestion (ATAD) is a promising process for sewage sludge stabilization. Batch experiments were conducted on sewage sludge collected from a municipal wastewater treatment plant in Shanghai, China, to evaluate the effectiveness of the ATAD system by determining changes in volatile suspended solids (VSSs) and to study its microbial diversity by denaturing gradient gel electrophoresis of 16S rRNA gene sequences amplified by PCR. The digestion system achieved rapid degradation of the organic substrate at 55 degrees C. The VSS was removed by up to 45.3% and 50.4% at 216 h and 264 h, respectively, while NH(4)(+)-N, chemical oxidation demand and total organic carbon of supernatant as well as total nitrogen did not exhibit obvious declines after 168 h. The microbial diversity changed during the thermophilic process as thermophiles belonging to the Hydrogenophilaceae, Thermotogaceae, Clostridiaceae and the genus Ureibacillus replaced less temperature-tolerant microorganisms such as Sphingobacteriaceae and the genus Trichococcus.