Assessment of metal pollution and effects of physicochemical factors on soil microbial communities around a landfill.

The physicochemical properties, chemical fractions of six metals (Cu, Zn, Pb, Cd, Cr, and Mn), and microbial communities of soil around a typical sanitary landfill were analyzed. The results indicate that soils around the landfill were from neutral to weak alkalinity. The contents of organic matter (OM), total nitrogen (TN), total phosphorous (TP), and activities of catalase, cellulase, and urease were significantly higher in landfill soils than those in background soils. Negative correlations were found between pH and metals. Cr was the dominant metal. Cu, Pb, Cr, and Mn were accumulated in the nearby farmland soils. Cd had the highest percentage of exchangeable fraction (33.7%-51.8%) in landfill and farmland soils, suggesting a high bioavailability to the soil environment affected by the landfill. Pb, Cr, and Mn existed mostly in oxidable fraction, and Cu and Zn were dominant in residual fraction. There was a low risk of soil metals around the landfill based on the RI values, while according to RAC classification, Cd had high to very high environmental risk. The MisSeq sequencing results showed that Actinobacteria, Proteobacteria, Chloroflexi, and Acidobacteria were the dominant phyla of bacteria, and the most abundant phylum of fungi was Ascomycota. The NMDS analysis revealed that the landfill could influence soil fungal communities more intensely than bacterial communities. TN, cellulase, and bioavailable metals (Pb-Bio and Cr-Bio) were identified to have main influences on microbial communities. Pb-Bio was the most dominant driving factor for bacterial community structures. For fungi, Pb-Bio was significantly negatively related to Olpidiomycota and Cr-Bio had a significantly negative correlation with Ascomycota. It manifests that bioavailable metals play important roles in assessing environmental risks and microbial community structures of soil around landfill.

Health risk assessment and potential sources of metals in riparian soils of the Wujiang River, China.

In order to understand the pollution status of metals in the riparian soils along the Wujiang River, 26 sampling sites in the mainstream and tributary streams were selected for investigation. The geo-accumulation index (Igeo), Nemerow integrated pollution index, and potential ecological risk index were applied to evaluate the contamination status and ecological risks of metals. Results revealed that the average concentrations of As, Cd, Cr, Cu, Mn, Ni, Pb, and Zn were 12.20, 0.51, 84.01, 57.42, 922.57, 38.37, 38.06, and 127.82 mg/kg, respectively. The metal contamination degree and ecological risks in the upper reaches were significantly higher than those in the middle and lower reaches of the Wujiang River. Cd was the dominant contamination metal. Significant non-carcinogenic and carcinogenic risks of metals were found in children based on the hazard index and carcinogenic risk. As was the main non-carcinogenic and carcinogenic pollutant metal in both adults and children. According to principal component analysis, hierarchical clustering analysis, and absolute principal component scores-multiple linear regression, anthropogenic sources (mining and agricultural activities) contributed most to Zn, Pb, Cr, Cd, Cu, and Ni, with contribution rates of 89.14, 82.32, 74.46, 72.12, 68.52, and 61.02%, respectively. Natural sources contributed most to Mn, with a contribution rate of 83.07%. Unidentified sources contributed most to As, with a contribution rate of 47.27%.

Speciation and Bioavailability of Metals in Sediments from a Stream Impacted by Abandoned Mines in Maoshi Town, Southwest of China.

The speciation of metals in surface sediments, as well as metal concentrations in muscle and liver of sharpbelly Hemiculter leucisculus from a stream near the Mo-Ni polymetallic mines in Maoshi town were investigated. The results indicated that metal concentrations were generally highest in the most upstream sampling location that is closest to the former mine operation. The total concentrations of Mn in sediments were higher than other metals. The Cr was mainly associated with residual fraction, while the order of bioavailability of metals (sum of the concentrations of exchangeable, reducible, and oxidizable fractions) was as follows: Cd (89.95%) > Mn (82.32%) > Ni (45.58%) > Mo (29.39%) > Hg (29.23%) > As (22.60%) > Pb (17.38%) > Cr (6.21%). The Cd concentration in muscle of H. leucisculus exceeded the permissible limit which indicated that the fish from this area are not safe for human consumption. The potential ecological risks of Cd cannot be ignored in this study area.

Differential expression of vacuolar and defective cell wall invertase genes in roots and seeds of metalliferous and non-metalliferous populations of Rumex dentatus under copper stress.

Acid invertase activities in roots and young seeds of a metalliferous population (MP) of Rumex dentatus were previously observed to be significantly higher than those of a non-metalliferous population (NMP) under Cu stress. To date, no acid invertase gene has been cloned from R. dentatus. Here, we isolated four full-length cDNAs from the two populations of R. dentatus, presumably encoding cell wall (RdnCIN1 and RdmCIN1 from the NMP and MP, respectively) and vacuolar invertases (RdnVIN1 and RdmVIN1 from the NMP and MP, respectively). Unexpectedly, RdnCIN1 and RdmCIN1 most likely encode special defective invertases with highly attenuated sucrose-hydrolyzing capacity. The transcript levels of RdmCIN1 were significantly higher than those of RdnCIN1 in roots and young seeds under Cu stress, whereas under control conditions, the former was initially lower than the latter. Unexpected high correlations were observed between the transcript levels of RdnCIN1 and RdmCIN1 and the activity of cell wall invertase, even though RdnCIN1 and RdmCIN1 do not encode catalytically active invertases. Similarly, the transcript levels of RdmVIN1 in roots and young seeds were increased under Cu stress, whereas those of RdnVIN1 were decreased. The high correlations between the transcript levels of RdnVIN1 and RdmVIN1 and the activity of vacuolar invertase indicate that RdnVIN1 and RdmVIN1 might control distinct vacuolar invertase activities in the two populations. Moreover, a possible indirect role for acid invertases in Cu tolerance, mediated by generating a range of sugars used as nutrients and signaling molecules, is discussed.

Heterologous expression and comparative characterization of vacuolar invertases from Cu-tolerant and non-tolerant populations of Elsholtzia haichowensis.

KEY MESSAGE: Vacuolar invertases (VINs) from Cu-tolerant and non-tolerant populations of Elsholtzia haichowensis have similar enzyme properties, and the enzyme protein divergences contribute little to the varied VIN activities between the contrasting populations. In our previous studies of Elsholtzia haichowensis, vacuolar invertase (VIN) activity in roots of a Cu-tolerant population was found to be significantly higher than that of a non-tolerant population under Cu stress. Divergences of amino acid residues in a sucrose-binding box and other regions of the VINs were detected. To test whether the amino acid divergences influence the enzyme properties of VINs, and thus are relevant to the differences in enzyme activities between the contrasting populations of E. haichowensis, two VIN genes from the Cu-tolerant population (EhCvINV) and non-tolerant population (EhNvINV) were heterologously expressed in Pichia pastoris, and the enzyme properties of the recombinants were characterized and compared. Both of the recombinant enzymes showed temperature optima of 70 °C and pH optima of 4.5-5.5. Copper as well as other heavy metals caused almost the same inhibition to EhNvINV and EhCvINV. No statistically significant differences were observed between EhNvINV and EhCvINV in K m and k cat values for sucrose. The results provided evidence that the observed residue divergences had little influence on the enzyme properties of VIN in E. haichowensis, and the varied VIN activities between the contrasting populations under Cu stress were not relevant to the amino acid divergences in the proteins. Also, some other possible reasons accounting for this difference in invertase activities were discussed, such as up-regulation of expression of the EhCvINV gene under Cu stress, as Cu tolerance mechanisms in Cu-mine plants.

Cloning and characterization of acid invertase genes in the roots of the metallophyte Kummerowia stipulacea (Maxim.) Makino from two populations: Differential expression under copper stress.

The roots of metallophytes serve as the key interface between plants and heavy metal-contaminated underground environments. It is known that the roots of metallicolous plants show a higher activity of acid invertase enzymes than those of non-metallicolous plants when under copper stress. To test whether the higher activity of acid invertases is the result of increased expression of acid invertase genes or variations in the amino acid sequences between the two population types, we isolated full cDNAs for acid invertases from two populations of Kummerowia stipulacea (from metalliferous and non-metalliferous soils), determined their nucleotide sequences, expressed them in Pichia pastoris, and conducted real-time PCR to determine differences in transcript levels during Cu stress. Heterologous expression of acid invertase cDNAs in P. pastoris indicated that variations in the amino acid sequences of acid invertases between the two populations played no significant role in determining enzyme characteristics. Seedlings of K. stipulacea were exposed to 0.3µM Cu(2+) (control) and 10µM Cu(2+) for 7 days under hydroponics׳ conditions. The transcript levels of acid invertases in metallicolous plants were significantly higher than in non-metallicolous plants when under copper stress. The results suggest that the expression of acid invertase genes in metallicolous plants of K. stipulacea differed from those in non-metallicolous plants under such conditions. In addition, the sugars may play an important role in regulating the transcript level of acid invertase genes and acid invertase genes may also be involved in root/shoot biomass allocation.

Differential responses of root growth, acid invertase activity and transcript level to copper stress in two contrasting populations of Elsholtzia haichowensis.

The present study aimed to test a hypothesis that acid invertases in root of metallophytes might play important roles in root growth under heavy metal stress. Plants of two contrasting populations, one from an ancient Cu mine (MP) and the other from a non-contaminated site (NMP), of metallophyte Elsholtzia haichowensis were treated with Cu in controlled experiments. The results showed that MP was Cu tolerant under 10 µM Cu2+ treatment. Cu treatment resulted in a higher root/shoot biomass ratio in MP compared to NMP. Scaling exponent in root/shoot allometric function in MP was lower than NMP. More complicated root architecture was observed in MP under Cu stress. Four full-length cDNAs (EhNcwINV, EhCcwINV, EhNvINV and EhCvINV) encoding cell wall and vacuolar invertases were cloned. Both of the transcript level and activity of the acid invertase in MP elevated under Cu treatment. There were positive correlations between root acid invertase transcript level, activity and root/shoot biomass ratio. The results indicated important roles of acid invertase in governing root growth under Cu stress. It also suggested that there was a possible interrelation between acid invertases and Cu tolerance mechanisms in MP of E. haichowensis.

Comparative study of root growth and sucrose-cleaving enzymes in metallicolous and non-metallicolous populations of Rumex dentatus under copper stress.

Sucrose metabolism in roots of metallophytes is very important for root growth and maintenance of heavy metal tolerance. However, rare researches have been carried out on this topic so far. We tested here a hypothesis that roots of copper-tolerant plants should manifest higher activities of sucrose-cleaving enzymes than non-tolerant plants for maintaining root growth under Cu stress. Plants of two contrasting populations of metallophyte Rumex dentatus, one from an ancient Cu mine (MP) and the other from a non-mine site (NMP), were treated with Cu in controlled experiments. Cu treatment resulted in a higher root biomass and root/shoot biomass ratio in MP compared to NMP. More complicated root system architecture was showed in MP under Cu stress. Activities and transcript levels of acid invertase as well as contents of sucrose and reducing sugar in MP were elevated under Cu treatment, while activities of neutral/alkaline invertase and sucrose synthase showed no significant differences between two populations. The results indicate important roles of acid invertase in governing root growth under Cu stress.

Bioaccumulation and Health Risk Assessment of Metals in Small-Sized Fish (Rhodeus sinensis, Ctenogobius giurinus) and Mussel (Cristaria plicata) from a River Reservoir, Southwest China.

In order to assess the bioaccumulation and health risk of metals in a river reservoir, concentrations of copper (Cu), zinc (Zn), lead (Pb), cadmium (Cd), arsenic (As), and mercury (Hg) in the water, sediments, two small-sized fish, and a freshwater mussel from the Zhoubai reservoir were examined. The results indicated that all of these metals conform with class one of environmental quality standards for surface water (State Environmental Protection Administration of China, GB 3838-2002). There were no significant differences for total metal concentrations in sediment between the three sampling sites (p > 0.05), but the bioavailable concentrations in S3 were the lowest. The Cd was dominated with exchangeable fraction and showed considerable risk. All metal concentrations except for Pb in Rhodeus sinensis were significantly higher than those in Ctenogobius giurinus (p < 0.05). The metal concentrations in Cristaria plicata showed a similar pattern of bioavailable metals in sediment, indicating that the metal concentrations in aquatic organisms were determined by the bioavailable forms of metals. Negative correlations were observed between the size of fish and concentrations of Cu, Zn, Pb, Cd, and As. However, significant positive correlations were found between the size of mussel and concentrations of Cd (p < 0.01), As (p < 0.05), and Hg (p < 0.01). Zn had the highest BCF values in fish and mussel. The aquatic organisms showed lower ability of metal bioaccumulation from the sediment. Low values of target hazard quotient (THQ), hazard index (HI), and carcinogenic risk (CR) indicated that these metals do not pose a health risk to public through fish and mussel consumption in this study area.

Comparative study on metal concentrations in water, sediments, and two fish species (Cyprinus carpio and Pelteobagrus fulvidraco) from the Wujiang River, China.

In order to assess the metal pollution in the Wujiang River, concentrations of Cu, Zn, Fe, Mn, Pb, Cd, As, and Hg in the water, sediments, and two fish species Cyprinus carpio and Pelteobagrus fulvidraco from the middle reaches (Tuomugang, TMG) and lower reaches (Wulong, WL) of the Wujiang River were examined. The results indicated that all metal concentrations were lower than the values for grade one water quality according to the Environment Quality Standard for Surface Water of China (GB 3838-2002). The bioavailable fraction concentrations of Zn, Fe, Mn, Pb, and Hg in WL were significantly higher than those in TMG (p < 0.05), indicating that these metals in sediments of WL have higher bioavailability and mobility. The Cu, Zn, Fe, Pb, As, and Hg were mainly related to the residual fraction, while the Mn and Cd were mainly associated with the non-residual fraction. The risk assessment code (RAC) and the secondary phase to the primary phase (RSP) values indicated that Mn and Cd have a high risk of secondary release. The mean metal concentrations in the liver of the two fish species were higher than those in muscle. The higher metal concentrations of fish in WL suggested that bioaccumulation of metals in fish could be influenced by metal bioavailability. No identical relationships between metal concentrations and fish length were manifested in the present study. The values of target hazard quotient (THQ) and hazard index (HI), and carcinogenic risk (CR) of metals for the consumption of C. carpio and P. fulvidraco indicated that the anglers would likely not experience significant non-carcinogenic risk, but the carcinogenic risk of As cannot be ignored. Thanks to prohibited commercial fishing in the Wujiang River, the metal pollution will probably not pose a health risk to the general public for wild fish consumption.

Metals in the tissues of two fish species from the rare and endemic fish nature reserve in the upper reaches of the Yangtze River, China.

Concentrations of copper, zinc, mercury, and arsenic were measured in the muscle, gill and liver tissues of Coreius heterodon and Pelteobagrus vachelli collected from the rare and endemic fish nature reserve in the upper reaches of the Yangtze River. The concentrations of copper and zinc in the tissues of these two fish species were higher than those of mercury and arsenic. Highest metal concentrations were generally found in fish samples from Yibin. The concentrations of copper, zinc, mercury, and arsenic in C. heterodon were higher than that in P. vachelli. The fish from this study area were not safe for human consumption.

Heavy metals speciation and distribution of microbial communities in sediments from the abandoned Mo-Ni polymetallic mines, southwest of China.

Chemical fractions of heavy metals (Mo, Ni, Cu, Zn, Fe, Mn, Pb, Cd, and Cr) and compositions of bacteria and fungi in surface sediments from the Mo-Ni polymetallic mine area were analyzed. The results indicated that the mean concentrations of Mo, Ni, Cu, Zn, and Cd were higher than their background values. The mean percentage of Cr in residual fraction was much higher than that of other heavy metals. Mo, Cu, Zn, Fe, and Pb were mainly associated with oxidizable fraction. The dominant proportions of Mn and Cd were found in exchangeable fraction with mean percentages of 93.46% and 54.50%, respectively. According to RAC classification and potential ecological risk index (PERI), the Cd with high bioavailability had a very high environmental risk. The MisSeq sequencing results of bacteria and fungi revealed that microbial communities discrepantly respond to different sampling sites. The most abundant phylum of bacteria and fungi were Proteobacteria and Ascomycota, respectively. The bioavailable heavy metals including Mo-B, Pb-B, and Cd-B were recognized to have important influences on both dominant bacterial and fungal communities. The present study manifested that the bioavailability of heavy metal is very important to assess the potential environmental risk and plays a key role in shaping microbial structure.

Impacts of environmental factors on the whole microbial communities in the rhizosphere of a metal-tolerant plant: Elsholtzia haichowensis Sun.

Rhizospheric microbes play important roles in plant growth and heavy metals (HMs) transformation, possessing great potential for the successful phytoremediation of environmental pollutants. In the present study, the rhizosphere of Elsholtzia haichowensis Sun was comprehensively studied to uncover the influence of environmental factors (EFs) on the whole microbial communities including bacteria, fungi and archaea, via quantitative polymerase chain reaction (qPCR) and high-throughput sequencing. By analyzing molecular ecological network and multivariate regression trees (MRT), we evaluated the distinct impacts of 37 EFs on soil microbial community. Of them, soil pH, HMs, soil texture and nitrogen were identified as the most influencing factors, and their roles varied across different domains. Soil pH was the main environmental variable on archaeal and bacterial community but not fungi, explaining 25.7%, 46.5% and 40.7% variation of bacterial taxonomic composition, archaeal taxonomic composition and a-diversity, respectively. HMs showed important roles in driving the whole microbial community and explained the major variation in different domains. Nitrogen (NH4-N, NO3-N, NO2-N and TN) explained 47.3% variation of microbial population composition and 15.9% of archaeal taxonomic composition, demonstrating its influence in structuring the rhizospheric microbiome, particularly archaeal and bacterial community. Soil texture accounted for 10.2% variation of population composition, 28.9% of fungal taxonomic composition, 19.2% of fungal a-diversity and 7.8% of archaeal a-diversity. Rhizosphere only showed strong impacts on fungi and bacteria, accounting for 14.7% and 4.9% variation of fungal taxonomic composition and bacterial a-diversity. Spatial distance had stronger influence on bacteria and archaea than fungi, but not as significant as other EFs. For the first time, our study provides a complete insight into key influential EFs on rhizospheric microbes and how their roles vary across microbial domains, giving a hand for understanding the construction of microbial communities in rhizosphere.