Characterization and Initial Application of Endophytic Bacillus safensis Strain ZY16 for Improving Phytoremediation of Oil-Contaminated Saline Soils.

Hydrocarbon-degrading and plant-growth-promoting bacterial endophytes have proven useful for facilitating the phytoremediation of petroleum-contaminated soils with high salinity. In this study, we identified Bacillus safensis strain ZY16 as an endophytic bacterium that can degrade hydrocarbons, produce biosurfactants, tolerate salt, and promote plant growth. The strain was isolated from the root of Chloris virgata Sw., a halotolerant plant collected from the Yellow River Delta. ZY16 survived in Luria-Bertani (LB) broth with 0-16% (w/v) sodium chloride (NaCl) and grew well in LB broth supplemented with 0-8% NaCl, indicating its high salt tolerance. The endophytic strain ZY16 effectively degraded C12-C32 n-alkanes of diesel oil effectively, as well as common polycyclic aromatic hydrocarbons under hypersaline conditions. For example, in mineral salts (MS) liquid medium supplemented with 6% NaCl, ZY16 degraded n-undecane, n-hexadecane, n-octacosane, naphthalene, phenanthrene, and pyrene, with degradation percentages of 94.5, 98.2, 64.8, 72.1, 59.4, and 27.6%, respectively. In addition, ZY16 produced biosurfactant, as confirmed by the oil spreading technique, surface tension detection, and emulsification of para-xylene and paraffin. The biosurfactant production ability of ZY16 under hypersaline conditions was also determined. Moreover, ZY16 showed plant-growth-promoting attributes, such as siderophore and indole-3-acetic acid production, as well as phosphate solubilization. To assess the enhanced phytoremediation of saline soils polluted by hydrocarbons and the plant-growth-promotion ability of ZY16, a pot trial with and without inoculation of the endophyte was designed and performed. Inoculated and non-inoculated plantlets of C. virgata Sw. were grown in oil-polluted saline soil, with oil and salt contents of 10462 mg/kg and 0.51%, respectively. After 120 days of growth, significant enhancement of both the aerial and underground biomass of ZY16-inoculated plants was observed. The soil total petroleum hydrocarbon degradation percentage (a metric of phytoremediation) after incubation with ZY16 was 63.2%, representing an elevation of 25.7% over phytoremediation without ZY16 inoculation. Our study should promote the application of endophytic B. safensis ZY16 in phytoremediation by extending our understanding of the mutualistic interactions between endophytes and their host plants.

[Petroleum pollution and the microbial community structure in the soil of Shengli Oilfield].

Soils around a new oil well (2011- ) and an old oil well (1966-2003) were sampled to investigate the characteristics of petroleum pollution in the oilfield. The structure of soil microbial community was analyzed by PCR-DGGE and clone sequencing techniques. Results showed that the soils around the two oil wells were generally contaminated with petroleum, and the concentrations of total petroleum hydrocarbons mostly exceeded the threshold of the environmental quality standards of soil (500 mg x kg(-1)). The total petroleum hydrocarbons concentration of the polluted soil was significantly positively correlated with the contents of organic carbon, total nitrogen and available potassium, respectively. The microbial diversity index in the soil around the old oil well decreased with the increasing total petroleum hydrocarbons concentration, however, it was reversed for the soil around the new oil well. Sequence analysis of the prominent bands in DGGE profiles showed that some dominant species existed in the petroleum-contaminated soils around the oil wells and mostly were oil-associated and hydrocarbon degrading bacteria, including Microbacterium, Streptomyces, Dietzia, Flavobacterium, alpha-Proteobacteria, and gamma-Proteobacteria.

Effects of pollution sources and soil properties on distribution of polycyclic aromatic hydrocarbons and risk assessment.

In this study, the concentrations of polycyclic aromatic hydrocarbons (PAHs) in soil profiles and the soil properties were analyzed in Hunpu, a typical wastewater irrigation area, northeast of China. The total concentrations of 16 priority control PAHs ranged from 7.88 to 2,231.42 µg/kg. Among 16 PAHs, the most abundant was Phenanthrene and the 3- or 4- ring PAHs were predominant. The PAH concentrations were higher in the upland fields near the oil wells, whereas leaching of PAH into the groundwater caused low concentrations in the paddy fields. The geochemical indices and the results from the principal component analysis of all 16 PAHs indicated that PAHs were mainly from atmospheric dusts in the top soil in I-1P/I-3P/I-7P and through soil profiles in I-4U/I-5P/I-8U, whereas those in the bottom layers were mainly from petroleum production and wastewater irrigation in I-1P/I-3P/I-7P and through soil profiles in I-2U and I-6U. In the redundancy analysis, PAHs exhibited negative correlation with pH, depth, silt, and clay, but had positive correlation with sand and organic matter. Finally, total toxic equivalent in the soil profiles and the calculated health risk of PAHs in the surface soil using contaminated land exposure assessment model elucidated the cancer risk that PAHs pose on human health in the Hunpu region.

Contamination, source, and input route of polycyclic aromatic hydrocarbons in historic wastewater-irrigated agricultural soils.

Contamination by polycyclic aromatic hydrocarbons (PAHs) of historic wastewater-irrigated agricultural topsoil (0-5 cm) and the contribution of groundwater irrigation and atmospheric deposition to soil PAHs were studied in a typical agricultural region, i.e. Hunpu region, Liaoning, China. Concentrations of total PAHs ranged from 0.43 to 2.64 mg kg⁻¹ in topsoil, being lower than those found in other wastewater-irrigated areas. The levels of PAHs in soil declined as the distance from a water source increased. Concentrations of individual PAHs were generally higher in upland than in paddy topsoils. The calculated nemerow composite index showed that agricultural soil in the region was "polluted" by PAHs. A human health risk assessment based on the total toxic equivalent concentration showed that the presence of elevated concentrations of PAHs in the soil might pose a great threat to the health of local residents. Ratios of pairs of PAHs and principal component analysis (PCA) showed that pyrogenesis, such as coal combustion, was the main source of PAHs, while petroleum, to some extent, also had a strong influence on PAHs contamination in upland soil. The distribution patterns of individual PAHs and composition of PAHs differed between irrigation groundwater and topsoil, but were similar between atmospheric deposition and topsoil. There were significant linear correlations (r = 0.90; p < 0.01) between atmospheric deposition rates and average concentrations of the 16 individual PAHs in soils, while no significant relationships were observed between irrigation groundwater and topsoil in levels of PAHs. These suggested that PAHs in agricultural soils were mainly introduced from atmospheric deposition, rather than from groundwater irrigation after the phasing out of wastewater irrigation in the region since 2002. This study provides a reference to ensure agricultural product safety, pollution control, and proper soil management.

[Soil environmental background concentrations in old course of the Yellow River in Shandong Province].

The environmental background concentrations of soil heavy metals, HCH, DDT and organic matter in old course of the Yellow River (Shandong area) were studied. Heavy metals vertical distribution, correlative analysis between organic matter and heavy metals, and the effect of different land use types were analyzed. The results showed that the background concentrations were 13.46 mg x kg(-1) for Cu, 16.23 mg x kg(-1) for Pb, 42.31 mg x kg(-1) for Zn, 30.97 mg x kg(-1) for Cr, 0.090 mg x kg(-1) for Hg, 3.90 mg x kg(-1) for As, 8.01 mg x kg(-1) for Co, 36.42 mg x kg(-1) for V, 426.83 mg x kg(-1) for Mn, 0.063 mg x kg(-1) for Cd, 18.71 mg x kg(-1) for Ni, and 0.74% for organic matter respectively. The detection rates of HCH and DDT were 100% and 60% respectively, and their concentrations were lower than Grade II of environmental quality standard for soils. All the heavy metals in soils were lower than Grade I of environmental quality standard for soils, and were lower than the background concentrations in the whole country, Shandong Province and the Loess Plateau area except for Hg. Mercury was significantly enriched in the surface soil, and the concentration of Cd was higher in the sublayer soil. The differences were significant between the layer 0-20 cm and 20-40 cm for Hg, Cd and organic matter, but not for the other heavy metals. The differences were not significant for environmental background concentrations among different land use types. Organic matter had highly positive correlations with Cu, As, Mn, Pb, Zn, Cr, Co and Ni.

[Experiment study on the metamorphic amylum production wastewater treatment by anaerobic baffled reactor].

The present study reports the start-up of treating metamorphic amylum production wastewater by anaerobic baffled reactor (ABR) and the bio-chemical features of granular sludge. The optimal conditions in treatment of the metamorphic amylum production wastewater were proposed, and the running performance in treating salt-containing wastewater was studied. Our results show that the common anaerobic activated sludge can be acclimated by increasing the organic loading and salt-concentration at the same time, and the granular sludge tolerant to low salt can be achieved by the acclimation. When chloric ion was 8,500 mg/L and salinity was 1.6%, the anaerobic activated sludge could degrade organic materials normally in the wastewater after the acclimation, and the COD removal is over 85%. When the concentration of metamorphic amylum production wastewater in the experiment was 12,640 mg/L and the optimal hydraulic retention time was 48 h, the removal efficiency of COD was 85.9%. Effect of sharp decrease of chloric ion concentration on sludge microorganism is larger than that of the sharp increase in the system. The system can endure the change of chloric ion concentration by increasing from 8,500 mg/L to 12,500 mg/L or decreasing from 8,500 mg/L to 4,500 mg/L, and it is more tolerant to the sudden increase than that of the sudden decrease of chloric ion concentration. The ABR system can treat the wastewater with chloric ion below 15,000 mg/L and salinity of about 2.5%.

Effects of vegetation type on soil microbial community structure and catabolic diversity assessed by polyphasic methods in North China.

Soil microbes play a major role in ecological processes and are closely associated with the aboveground plant community. In order to understand the effects of vegetation type on the characteristics of soil microbial communities, the soil microbial communities were assessed by plate counts, phospholipid fatty acid (PLFA) and Biolog microplate techniques in five plant communities, i.e., soybean field (SF), artificial turf (AT), artificial shrub (AS), natural shrub (NS), and maize field (MF) in Jinan, Shandong Province, North China. The results showed that plant diversity had little discernible effect on microbial biomass but a positive impact on the evenness of utilized substrates in Biolog microplate. Legumes could significantly enhance the number of cultural microorganisms, microbial biomass, and community catabolic diversity. Except for SF dominated by legumes, the biomass of fungi and the catabolic diversity of microbial community were higher in less disturbed soil beneath NS than in frequently disturbed soils beneath the other vegetation types. These results confirmed that high number of plant species, legumes, and natural vegetation types tend to support soil microbial communities with higher function. The present study also found a significant correlation between the number of cultured bacteria and catabolic diversity of the bacterial community. Different research methods led to varied results in this study. The combination of several approaches is recommended for accurately describing the characteristics of microbial communities in many respects.

Plant community succession in modern Yellow River Delta, China.

Data were collected in different successional stages using a simultaneous sampling method and analyzed through quantitative classification method. Three large groups and 12 classes were made to represent the community patterns of three succession stages and 12 succession communities. The succession series of plant community in the study area was as follows: saline bare land-->community Suaeda salsa-->community Tamarix chinensis-->grassland. Succession degree and succession process of 12 succession communities were calculated. Most of these communities were in the lower succession stage, however, community Phragmites communis+Glycine soja and community Imperata cylindrica+G. soja were close to the succession stage of grassland climax. Five species diversity indices were used to study the changes in species richness, species evenness and diversity during succession of community. Heterogeneity index and richness index increased gradually during the community succession process, but species evenness tended to decrease with succession development. The relation between succession and environment was studied by ordination technique, and the results showed that the soil salt content was an important factor to halarch succession of the modern Yellow River Delta. It affected community structure, species composition and succession process.