Soil pollution from heavy metals, especially mercury, is an environmental problem for human health. Biological approaches offer interesting tools, which necessarily involve the selection of organisms capable of transforming the environment via bioremediation. To evaluate the potential use of microorganisms in phytorhizoremediation, bacterial strains were isolated from rhizospheric and bulk soil under conditions of chronic natural mercury, which were identified and characterized by studying the following: (i) their plant growth promoting rhizobacteria (PGPR) activities; and (ii) their maximum bactericide concentration of mercury. Information regarding auxin production, phosphate solubilization, siderophore synthesis and 1-aminocyclopropane-1-carboxylic acid deaminase (ACCd) capacity of the isolates was compiled in order to select the strains that fit potential biotechnological use. To achieve this objective, the present work proposes the Bio-Mercury Remediation Suitability Index (BMR-SI), which reflects the integral behavior of the strains for heavy metal polluted soil bioremediation. Only those strains that rigorously fulfilled all of the established criteria were selected for further assays.

Soil contamination by mercury, which is one of the most toxic heavy metals due to its bioaccumulative capacity, poses a risk to the environment as well as health. The Almadén mining district in Ciudad Real, Spain is one of the most heavily-polluted sites in the world, making the soils unusable. Bioremediation, and more specifically phyto-rhizoremediation, based on the synergistic interaction established between plant and Plant Growth Promoting Rhizobacteria (PGPR), improves the plant's ability to grow, mobilize, accumulate, and extract contaminants from the soil. The objective of this study is to evaluate the plant growth-promoting ability of four PGPR strains (and mixtures), isolated from the bulk soil and rhizosphere of naturally grown plants in the Almadén mining district, when they are inoculated in emerged seeds of Lupinus albus, var. Dorado in the presence of high concentrations of mercury. After 20 days of incubation and subsequent harvesting of the seedlings, biometric measurements were carried out at the root and aerial levels. The results obtained show that the seeds treatment with PGPR strains improves plants biometry in the presence of mercury. Specifically, strain B2 (Pseudomonas baetica) and B1 (Pseudomonas moraviensis) were those that contributed the most to plant growth, both individually and as part of mixtures (CS5 and CS3). Thus, these are postulated to be good candidates for further in situ phyto-rhizoremediation tests of mercury-contaminated soils.

Heavy metal pollution of soil, particularly by mercury (Hg), is a problem that can seriously affect the environment and human health. For this reason, it is necessary to take steps to remediate these environments, prevent potential adverse effects, and restore these areas for subsequent use in agriculture, industry, ranching, and forestry. The present study has selected 40 bacterial strains from rhizosphere and bulk soil that grow naturally in high Hg-contaminated soils from the Almadén mining district in Ciudad Real, Spain. With the objective of evaluating the potential use of these strains in phyto-rhizoremediation, an evaluation and statistical analysis of their PGPR (Plant-Growth-Promoting Rhizobacteria) activity at different levels of Hg was carried out as the first condition of selection for their potential use in bioremediation. In addition, a Hg MBC (Maximum Bactericidal Concentration) was performed with the aim of selecting the strains with high Hg tolerance. Finally, strains with potential biotechnological use have been proposed according to the Bio-Mercury Remediation Suitability Index (BMRSI) criteria, which consider indole-3-acetic acid (IAA) production, acid 1- aminocyclopropane-1-carboxylic deaminase (ACCd) activity, phosphates solubilization, and siderophore production measured in the presence of Hg, as well as its MBC to Hg. The strains selected for further in vivo and in situ processes must reach at least an MBC (Hg) > 100 μg/mL and BMRSI ≥ 6.5.

Antibiotic resistance (AR) in the environment is of great global concern and a threat to public health. Soil bacteria, including Bacillus spp., could act as recipients and reservoirs of AR genes of clinical, livestock, or agricultural origin. These genes can be shared between bacteria, some of which could be potentially human pathogens. This process can be favored in conditions of abiotic stress, such as heavy metal contamination. The Almadén mining district (Ciudad Real, Spain) is one of the environments with the highest mercury (Hg) contamination worldwide. The link between heavy metal contamination and increased AR in environmental bacteria seems clear, due to co-resistance and co-selection phenomena. In the present study, 53 strains were isolated from rhizospheric and bulk soil samples in Almadén. AR was tested using Vitek®®2 and minimum inhibitory concentration (MIC) values were obtained and interpreted based on the criteria of the Clinical and Laboratory Standards Institute (CLSI) guidelines. Based on the resistance profiles, five different antibiotypes were established. The Hg minimum bactericidal concentration (MBC) of each strain was obtained using the plating method with increasing concentrations of HgCl2. A total of 72% of Bacillus spp. showed resistance to two or more commonly used antibiotics. A total of 38 isolates expressed AR to cephalosporins. Finally, the environmental co-selection of AR to cephalosporins and tetracyclines by selective pressure of Hg has been statistically demonstrated.

Soil contamination by heavy metals, particularly mercury (Hg), is a problem that can seriously affect the environment, animals, and human health. Hg has the capacity to biomagnify in the food chain. That fact can lead to pathologies, of those which affect the central nervous system being the most severe. It is convenient to know the biological environmental indicators that alert of the effects of Hg contamination as well as the biological mechanisms that can help in its remediation. To contribute to this knowledge, this study conducted comparative analysis by the use of Shotgun metagenomics of the microbial communities in rhizospheric soils and bulk soil of the mining region of Almadén (Ciudad Real, Spain), one of the most affected areas by Hg in the world The sequences obtained was analyzed with MetaPhlAn2 tool and SUPER-FOCUS. The most abundant taxa in the taxonomic analysis in bulk soil were those of Actinobateria and Alphaproteobacteria. On the contrary, in the rhizospheric soil microorganisms belonging to the phylum Proteobacteria were abundant, evidencing that roots have a selective effect on the rhizospheric communities. In order to analyze possible indicators of biological contamination, a functional potential analysis was performed. The results point to a co-selection of the mechanisms of resistance to Hg and the mechanisms of resistance to antibiotics or other toxic compounds in environments contaminated by Hg. Likewise, the finding of antibiotic resistance mechanisms typical of the human clinic, such as resistance to beta-lactams and glycopeptics (vancomycin), suggests that these environments can behave as reservoirs. The sequences involved in Hg resistance (operon mer and efflux pumps) have a similar abundance in both soil types. However, the response to abiotic stress (salinity, desiccation, and contaminants) is more prevalent in rhizospheric soil. Finally, sequences involved in nitrogen fixation and metabolism and plant growth promotion (PGP genes) were identified, with higher relative abundances in rhizospheric soils. These findings can be the starting point for the targeted search for microorganisms suitable for further use in bioremediation processes in Hg-contaminated environments.