Biochemical, Catabolic, and PGP Activity of Microbial Communities and Bacterial Strains from the Root Zone of Baccharis linearis in a Mediterranean Mine Tailing.

The management of mine tailings (MT) is commonly workload heavy, intrusive, and expensive. Phytostabilization offers a promising approach for MT management; however, it poses challenges due to the unfavorable physicochemical properties of these wastes. Nevertheless, native microorganisms capable of supporting plant growth and development could enhance the efficacy of phytostabilization. This study assesses the biological activity of microbial communities from the root zone of Baccharis linearis, which is naturally present in MT, in order to evaluate their biotechnological potential for phytostabilization. The root zone and bulk samples were collected from B. linearis plants located within a MT in the Mediterranean zone of Chile. Enzyme activities related to the cycling of C, N, and P were assessed. The community-level physiological profile was evaluated using the MicroRespTM system. Bacterial plant growth-promoting (PGP) traits and colony forming units (CFU) were evaluated through qualitative and microbiological methods, respectively. CFU, enzyme activities, and CLPP were higher in the root zone compared with the bulk samples. Five bacterial strains from the root zone exhibited PGP traits such as P solubilization and N acquisition, among others. The presence of microbial communities in the root zone of B. linearis with PGP traits suggests their potential to enhance the ecological management of MT through phytostabilization programs.

Environmental conditions and plant physiology modulate Cu phytotoxicity in field-contaminated soils.

Foliar Cu concentration has been widely used as a biomarker of plant growth in phytotoxicity bioassays. This relation has helped find plant processes altered by Cu in dose-response experiments (a bivariate approach). However, when plants are grown in field conditions, their responses can vary in function of multiple variables, such as the environment, plant physiology, and other elements in plant (plant ionome). These sources of variability are commonly unreported, which could limit bioassays' utility. Thus, the present study aimed to assess and integrate the mentioned sources of variability on Cu phytotoxicity. Lettuce was used as plant model. Lettuces were grown in growth chambers with contrasting light and air humidity conditions and on two different field-contaminated soils (sandy and loamy soils). Results showed that environmental conditions significantly affected foliar Cu and plant growth, but this effect differed in the two studied soils. Foliar Cu was not a good biomarker of plant growth. In contrast, integrating the potential phytotoxicity effect with the plant's nutritional status allowed a better understanding of plant growth. We remarked on using a structural equation modeling approach (SEM) to integrate plant physiology and plant ionome as moderators of plant growth. Results showed that plant growth was primarily related to plant nutritional status rather than Cu phytotoxicity. Also, the foliar Cu concentration would affect plant nutritional status due to photosynthesis-related plant processes and cation balance. Finally, this research invites to state and include sources of variability when assessing phytotoxicity. This way, it is possible to advance toward understanding complex linked processes occurring in field conditions.

Enzyme activities and microbial functional diversity in metal(loid) contaminated soils near to a copper smelter.

The monitoring of soil metal(loid) contamination is of global significance due to deleterious effects that metal(loid)s have on living organisms. Soil biological properties such as enzyme activities (EAs) are good indicators of metal(loid) contamination due to their high sensitivity, fast response, and low-cost. Here, the effect of metal(loid) contamination on physicochemical properties and microbial functionality in soils sampled from within 10 km of a Cu smelter is investigated. Soil composite samples were randomly taken within 2, 4, 6, 8 and10 km zones from a mining industry Cu smelter. The EAs of dehydrogenase (DHA), arylsulfatase (ARY), ß-glucosidase, urease, and arginine ammonification (AA) were studied as indicators of metal(loid) contamination, which included the ecological dose (ED50) with respect to Cu and As contents. The community level physiological profile (CLPP), functional diversity, and catabolic evenness were evaluated based on the C-substrate utilisation. All EAs decreased in zones with high degrees of metal(loid) contamination, which also had low TOC and clay contents, reflecting long term processes of soil degradation. Positive and strong relationships between EAs and TOC were found. DHA and ARY activities decreased by approximately 85-90% in highly metal(loid) contaminated soils. DHA and AA showed significant ED50 values associated with available Cu (112.8 and 121.6 mg CuDTPA kg-1, respectively) and total As contents (30.8 and 31.8 mg As kg-1, respectively). The CLPP showed different metabolic profiles along the metal(loid) contamination gradients. Long-term stress conditions in soils close to industrial areas resulted in the decreasing of general biological activity, catabolic capacity, and functional diversity.

Soil and indoor dust as environmental media of human exposure to As, Cd, Cu, and Pb near a copper smelter in central Chile.

In the present study, we assessed the non-carcinogenic and carcinogenic human health risk due to exposure to trace elements in soil and indoor dust in Puchuncaví valley. We also determined the associations between trace element concentration in hair/toenails and the estimated chronic daily intake of trace elements in soil and indoor dust. We found statistically significant association between the trace element concentration in hair/toenails and the estimated chronic daily intake of soil and indoor dust. Indoor dust was more important than soil in terms of human exposure to trace elements in Puchuncaví, due to the high concentration of trace elements on this environmental media and long periods of time that the population spends at their households. With regards to non-carcinogenic risk, we found that there was no health risk associated to soil and indoor dust exposure in the Puchuncaví valley, because none of the hazard quotient values surpassed 1.0. However, carcinogenic risk due to arsenic exposure was above the threshold value of 1.0E-04 in the population of young children (from 1 to 5 years old) in all studied areas, including the control, and in the population of children (from 6 to <18 years old) in the exposed area. Such risk values are classified as unacceptable (US EPA, 2001), requiring some target intervention from the Chilean government.

Comparison of exposure to trace elements through vegetable consumption between a mining area and an agricultural area in central Chile.

Human exposure to trace elements has been a large concern due to the potential health issues. Accordingly, this study aimed to compare the concentrations of arsenic, copper, and zinc in the edible parts of vegetables grown in a mining-agricultural area and in an exclusively agricultural area and to compare the potential human health risks of consuming vegetables from both areas. The consumption habits of the studied population were extracted from the 2010 National Alimentary Survey of Chile. In most cases, the concentrations of trace elements in the edible tissues of vegetables (lettuce, spinach, garlic, onion, carrot, potato, sweet corn, and tomato) were higher in the mining-agricultural area than those in the control area. This difference was most pronounced for leafy vegetables, with arsenic being the trace element of concern. Specifically, the arsenic concentrations in the edible tissues of lettuce and spinach were 8.2- and 5.4-fold higher, respectively, in the mining-agricultural area than in the control area. Lettuce was the vegetable of concern due to its relatively high consumption and relatively high concentration of trace elements. Nevertheless, there was no health risk associated with vegetable consumption in either the mining area or the control area because none of the HQ values surpassed 1.0.

Proposed modification to avoidance test with Eisenia fetida to assess metal toxicity in agricultural soils affected by mining activities.

Use of avoidance tests is a quick and cost-effective method of assessing contaminants in soils. One option for assessing earthworm avoidance behavior is a two-section test, which consists of earthworms being given the choice to move between a test soil and a control substrate. For ecological relevance, tested soils should be field-contaminated soils. For practical reasons, artificial soils are commonly used as the control substrate. Interpretation of the test results compromised when the test soil and the artificial substrate differ in their physico-chemical properties other than just contaminants. In this study we identified the physico-chemical properties that influence avoidance response and evaluated the usefulness of adjusting these in the control substrate in order to isolate metal-driven avoidance of field soils by earthworms. A standardized two-section avoidance test with Eisenia fetida was performed on 52 uncontaminated and contaminated (Cu >155mgkg-1, As >19mgkg-1) agricultural soils from the Aconcagua River basin and the Puchuncaví Valley in Chile. Regression analysis indicated that the avoidance response was determined by soil organic matter (OM), electrical conductivity (EC) and total soil Cu. Organic matter content of the artificial substrate was altered by peat additions and EC by NaCl so that these properties matched those of the field soils. The resultant EC80 for avoidance (indicative of soils of "limited habitat") was 433mg Cu kg-1 (339 - 528mgkg-1 95% confidence intervals). The earthworm avoidance test can be used to assess metal toxicity in field-contaminated soils by adjusting physico-chemical properties (OM and EC) of the artificial control substrate in order to mimic those of the field-collected soil.

Advances on the determination of thresholds of Cu phytotoxicity in field-contaminated soils in central Chile.

To better determine phytotoxicity thresholds for metals in the soil, studies should use actual field-contaminated soil samples rather than metal-spiked soil preparations. However, there are surprisingly few such data available for Cu phytotoxicity in field-contaminated soils. Moreover, these studies differ from each other with regards to soil characteristics and experimental setups. This study aimed at more accurately estimating Cu phytotoxicity thresholds using field-collected agricultural soils (Entisols) from areas exposed to contamination from Cu mining. For this purpose, the exposure to Cu was assessed by measuring total soil Cu, soluble Cu, free Cu2+ activity, and Cu in the plant aerial tissues. On the other hand, two bioassay durations (short-term and long-term), three plant species (Avena sativa L., Brassica rapa CrGC syn. Rbr, and Lolium perenne L.), and five biometric endpoints (shoot length and weight, root length and weight, and number of seed pods) were considered. Overall plant growth was best predicted by total Cu content of the soil. Despite some confounding factors, it was possible to determine EC10, EC25 and EC50 of total Cu in the soil. Brassica rapa was more sensitive than Avena sativa for all endpoints, while Lolium perenne was of intermediate sensitivity. For the short-term bioassay (21 days for all three species), the averaged EC10, EC25 and EC50 values of total soil Cu (in mg kg-1) were 356, 621, and 904, respectively. For the long-term bioassay (62 days for oat and 42 days for turnip), the averaged EC10, EC25 and EC50 values of total soil Cu (in mg kg-1) were 355, 513, and 688, respectively. The obtained results indicate that chronic test is a suitable method for assessing Cu phytotoxicity in field-contaminated soils.

Thresholds of arsenic toxicity to Eisenia fetida in field-collected agricultural soils exposed to copper mining activities in Chile.

Several previous studies highlighted the importance of using field-collected soils-and not artificially-contaminated soils-for ecotoxicity tests. However, the use of field-collected soils presents several difficulties for interpretation of results, due to the presence of various contaminants and unavoidable differences in the physicochemical properties of the tested soils. The objective of this study was to estimate thresholds of metal toxicity in topsoils of 24 agricultural areas historically contaminated by mining activities in Chile. We performed standardized earthworm reproduction tests (OECD 222 and ISO 11268-2) with Eisenia fetida. Total soil concentrations of Cu, As, Zn, and Pb were in the ranges of 82-1295 mg kg(-1), 7-41 mg kg(-1), 86-345 mg kg(-1), and 25-97 mg kg(-1), respectively. In order to differentiate between the effects of different metals, we used regression analysis between soil metal concentrations and earthworm responses, as well as between metal concentrations in earthworm tissues and earthworm responses. Based on regression analysis, we concluded that As was a metal of prime concern for Eisenia fetida in soils affected by Cu mining activities, while Cu exhibited a secondary effect. In contrast, the effects of Zn and Pb were not significant. Soil electrical conductivity was another significant contributor to reproduction toxicity in the studied soils, forcing its integration in the interpretation of the results. By using soils with electrical conductivity ≤ 0.29 dS m(-1) (which corresponds to EC50 of salt toxicity to Eisenia fetida), it was possible to isolate the effect of soil salinity on earthworm reproduction. Despite the confounding effects of Cu, it was possible to determine EC10, EC25 and EC50 values for total soil As at 8 mg kg(-1), 14 mg kg(-1) and 22 mg kg(-1), respectively, for the response of the cocoon production. However, it was not possible to determine these threshold values for juvenile production. Likewise, we were able to determine EC10, EC25 and EC50 of earthworm tissue As of 38 mg kg(-1), 47 mg kg(-1), and 57 mg kg(-1), respectively, for the response of the cocoon production. Finally, we determined the no-observed effect concentration of tissue As in E. fetida of 24 mg kg(-1). Thus, earthworm reproduction test is applicable for assessment of metal toxicity in field-collected soils with low electrical conductivity, while it might have a limited applicability in soils with high electrical conductivity because the salinity-induced toxicity will hinder the interpretation of the results.