Metal-resistant Bacteria Research Articles

Full Copper Resistance in Cupriavidus metallidurans Requires the Interplay of Many Resistance Systems.

The metal-resistant bacterium Cupriavidus metallidurans uses its copper resistance components to survive the synergistic toxicity of copper ions and gold complexes in auriferous soils. The cup, cop, cus, and gig determinants encode as central component the Cu(I)-exporting PIB1-type ATPase CupA, the periplasmic Cu(I)-oxidase CopA, the transenvelope efflux system CusCBA, and the Gig system with unknown function, respectively. The interplay of these systems with each other and with glutathione (GSH) was analyzed. Copper resistance in single and multiple mutants up to the quintuple mutant was characterized in dose-response curves, Live/Dead-staining, and atomic copper and glutathione content of the cells. The regulation of the cus and gig determinants was studied using reporter gene fusions and in case of gig also RT-PCR studies, which verified the operon structure of gigPABT. All five systems contributed to copper resistance in the order of importance: Cup, Cop, Cus, GSH, and Gig. Only Cup was able to increase copper resistance of the Δcop Δcup Δcus Δgig ΔgshA quintuple mutant but the other systems were required to increase copper resistance of the Δcop Δcus Δgig ΔgshA quadruple mutant to the parent level. Removal of the Cop system resulted in a clear decrease of copper resistance in most strain backgrounds. Cus cooperated with and partially substituted Cop. Gig and GSH cooperated with Cop, Cus, and Cup. Copper resistance is thus the result of an interplay of many systems. IMPORTANCE The ability of bacteria to maintain homeostasis of the essential-but-toxic "Janus"-faced element copper is important for their survival in many natural environments but also in case of pathogenic bacteria in their respective host. The most important contributors to copper homeostasis have been identified in the last decades and comprise PIB1-type ATPases, periplasmic copper- and oxygen-dependent copper oxidases, transenvelope efflux systems, and glutathione; however, it is not known how all these players interact. This publication investigates this interplay and describes copper homeostasis as a trait emerging from a network of interacting resistance systems.

Arsenic and other heavy metals resistant bacteria in rice ecosystem: Potential role in promoting plant growth and tolerance to heavy metal stress

The role of microorganisms in arsenic (As) uptake in association with terrestrial plants is identified, though their rhizosphere specific association and function in rice and barnyard grass in As contaminated soil are not elucidated. We investigated the abundance and diversity of arsenic and other heavy metal tolerant bacteria isolated from barnyard grass and rice rhizospheres, and the bulk soil from an As-contaminated paddy field. Among 83 isolated As resistant bacteria, 22 strains were screened and characterized based on their growth potential in 100 mM arsenate. One strain oxidized and reduced arsenic, whereas five isolates oxidized arsenite to arsenate and seven isolates reduced arsenate to arsenite. All the isolates were tolerant to the highest levels of Zn, Cd, Cr, Cu, Co, and Hg tested and except one, all also survived at the highest level of Pb. Molecular characterization by 16s rRNA gene analysis revealed that the selected bacteria belong to Sphingomonadaceae (13), Moraxellaceae (2), Enterobacteriaceae (2), Planococcaceae (2), Pseudomonadaceae (1), Xanthomonadaceae (1) and Cyanobacteria (1). Bacterial abundances were more in the rhizosphere than in bulk soil and diversity was more in barnyard grass rhizosphere than in rice. Among the isolates, 21 strains solubilized phosphate, produced indole acetic acid and grew in N-free medium. All the isolates that were tested, enhanced the germination and root fresh weight of rice seedlings. These isolates, based on their As and other metals resistance, As detoxification capacity, and plant growth promotion ability, especially the members of Sphingomonadaceae, could be conclusively used for plant growth promotion and microbe assisted remediation of the As and other heavy metal contaminated soil employing barnyard grass and rice.

Screening and evaluation of Cr(VI) biosorption potential of metal-resistant bacteria isolated from Sukinda chromite mine soil of Odisha

Screening and evaluation of Cr(VI) biosorption potential of metal-resistant bacteria isolated from Sukinda chromite mine soil of Odisha

Physicochemical Properties, Heavy Metal Composition, and Metal Resistant Microorganisms Associated with Gold Mine Tailings in Jebba, Nigeria

Mine tailings are a source of unique microorganisms with novel characteristics. This study aims to determine the chemical composition of the soil and water from different locations of an active artisanal gold mine and the microorganisms associated with the mine tailings. The physiochemical analysis of tailings soil and water, HM species, and their carcinogenic and non-carcinogenic effects were determined. Heavy metal resistant bacteria (HMRB) were then isolated using nutrient agar supplemented with metals. The results showed that pH and temperature of the soil and water samples ranged between 6.2- 6.6 and 29–30 °C respectively. The estimated incremental lifetime cancer risk (ILCR) from one location was 7.9E − 6 was the highest. The HMs detected includes Arsenic (As), Mercury (Hg), Cadmium (Cd), and Lead (Pb). Twenty-one HMRB were isolated and they are putatively identified as Staphylococcus spp., Bacillus sp., Salinicoccus halodurans, Staphylococcus chromogenes, Paenibacillus cellulositrophicus, Paenibacillus residui, Sporosarcina globispora, and Streptococcus sp. Analysis of the 16S RNA genes sequences revealed that isolate MS2 has 98.01% similarity to Aneurinibacillus aneurinilyticus. The presence and levels of HMs elucidates the high risk of cancerous and non-cancerous effects borne by artisan gold miners. The isolated HMRB can be further analyzed for possible bioremediation of the site.

Mechanisms of Bacterial Resistance to Heavy Metals: A Mini Review

Because of rising levels of heavy metal pollution in the environment, microbial resistance to heavy metals has become an increasing concern. Heavy metal resistance in bacteria is typically achieved through a combination of passive and active mechanisms, including heavy metal sequestration, efflux, or transformation within the microbial cell. During the efflux mechanism, a membrane protein's energy-dependent ion efflux from the cell is necessary for heavy metal removal. Understanding the physicochemical parameters of the environment, structure and diversity of microbial communities, nature and concentration of heavy metals is critical for developing effective strategies for the remediation of heavy metal-contaminated sites. Many microbes play a significant part on functioning ecosystem more especially in the biogeochemical cycling of heavy metals by removing the metals from the environment. As, Pb, Cd, and Hg are among heavy metals that are associated with the most common ecologically hazardous metals that can be toxic to microbes and still nature has evolved few groups of microbes that were found to resist the effect of heavy metals while thriving within their ecosystem such as Pseudomonas sp., Escherichia coli and Serratia marcescens that can resist Hg. Pseudomonas putida, Cupriavidus necator, Exiguobacterium sp., Bacillus aquimaris, Bacillus cereus and Alcaligenes sp. can also resist Cu, Cd, Pb, Cr and Ni. The exposure of local and regional soil with heavy metal pollution due to smelting causes which poses major environmental issues that is currently on rise in human ecosystem. Therefore, studying the mechanisms of bacterial resistance to heavy metal is critical for developing strategies to reduce the environmental impact of heavy metal pollution.

Potential roles of the rhizospheric bacterial community in assisting Miscanthus floridulus in remediating multi-metal(loid)s contaminated soils

Phytoremediation technology is an important approach applied to heavy metal remediation, and how to improve its remediation efficiency is the key. In this study, we compared the rhizospheric bacterial communities and metals contents in Miscanthus floridulus (M. floridulus) of four towns, including Huayuan Town (HY), Longtan Town (LT), Maoer Village (ME), and Minle Town (ML) around the lead-zinc mining area in Huayuan County, China. The roles of rhizospheric bacterial communities in assisting the phytoremediation of M. floridulus were explored. It was found that the compositions of the rhizospheric bacterial community of M. floridulus differed in four regions, but majority of them were heavy metal-resistant bacteria that could promote plant growth. Results of bioconcentration factors showed the enrichment of Cu, Zn, and Pb by M. floridulus in these four regions were significantly different. The Zn enrichment capacity of ML was the strongest for Cu and stronger than LT and ME for Pb. The enrichment capacity of LT and ML was stronger than HY and ME. These bacteria may influence the different heavy metals uptake of M. floridulus by altering the soil physiochemical properties (e.g., soil peroxidase, pH and moisture content). In addition, co-occurrence network analysis also showed that LT and ML had higher network stability and complexity than HY and ME. Functional prediction analysis of the rhizospheric bacterial community showed that genes related to protein synthesis (e.g., zinc-binding alcohol dehydrogenase/oxidoreductase, Dtx R family transcriptional regulators and ACC deaminase) also contributed to phytoremediation in various ways. This study provides theoretical guidance for selecting suitable microorganisms to assist in the phytoremediation of heavy metals.

Heavy Metal Pollution Impacts Soil Bacterial Community Structure and Antimicrobial Resistance at the Birmingham 35th Avenue Superfund Site.

Heavy metals (HMs) are known to modify bacterial communities both in the laboratory and in situ. Consequently, soils in HM-contaminated sites such as the U.S. Environmental Protection Agency (EPA) Superfund sites are predicted to have altered ecosystem functioning, with potential ramifications for the health of organisms, including humans, that live nearby. Further, several studies have shown that heavy metal-resistant (HMR) bacteria often also display antimicrobial resistance (AMR), and therefore HM-contaminated soils could potentially act as reservoirs that could disseminate AMR genes into human-associated pathogenic bacteria. To explore this possibility, topsoil samples were collected from six public locations in the zip code 35207 (the home of the North Birmingham 35th Avenue Superfund Site) and in six public areas in the neighboring zip code, 35214. 35027 soils had significantly elevated levels of the HMs As, Mn, Pb, and Zn, and sequencing of the V4 region of the bacterial 16S rRNA gene revealed that elevated HM concentrations correlated with reduced microbial diversity and altered community structure. While there was no difference between zip codes in the proportion of total culturable HMR bacteria, bacterial isolates with HMR almost always also exhibited AMR. Metagenomes inferred using PICRUSt2 also predicted significantly higher mean relative frequencies in 35207 for several AMR genes related to both specific and broad-spectrum AMR phenotypes. Together, these results support the hypothesis that chronic HM pollution alters the soil bacterial community structure in ecologically meaningful ways and may also select for bacteria with increased potential to contribute to AMR in human disease. IMPORTANCE Heavy metals cross-select for antimicrobial resistance in laboratory experiments, but few studies have documented this effect in polluted soils. Moreover, despite decades of awareness of heavy metal contamination at the EPA Superfund site in North Birmingham, Alabama, this is the first analysis of the impact of this pollution on the soil microbiome. Specifically, this work advances the understanding of the relationship between heavy metals, microbial diversity, and patterns of antibiotic resistance in North Birmingham soils. Our results suggest that polluted soils carry a risk of increased exposure to antibiotic-resistant infections in addition to the direct health consequences of heavy metals. Our work provides important information relevant to both political and scientific efforts to advance environmental justice for the communities that call Superfund neighborhoods home.

BON domain-containing protein-mediated co-selection of antibiotic and heavy metal resistance in bacteria

The widespread antibiotic resistance of bacteria has become one of the most severe threats to public health. However, the mechanisms that allow microbial acquisition of resistance are still poorly understood. In the present study, a novel BON domain-containing protein was heterologously expressed in Escherichia coli. It functions as an efflux pump-like to confer resistance to various antibiotics, especially for ceftazidime, with a >32-fold increase in minimum inhibitory concentration (MIC). The fluorescence spectroscopy experiment indicated that BON protein could interact with several metal ions, such as copper and silver, which has been associated with the induced co-regulation of antibiotic and heavy metal resistance in bacteria. Furthermore, the BON protein was demonstrated to spontaneously self-assemble into a trimer and generate a central pore-like architecture for antibiotic transporting. A WXG motif as a molecular switch is essential for forming the transmembrane oligomeric pores and controls the interaction between BON protein and cell membrane. Based on these findings, a mechanism termed “one-in, one-out”, was proposed for the first time. The present study provides new insights into the structure and function of BON protein and a previously unidentified antibiotic resistance mechanism, filling the knowledge gap in understanding BON protein-mediated intrinsic antibiotic resistance.

Biofilm-Forming Heavy Metal Resistance Bacteria From Bungus Ocean Fisheries Port (PPS) West Sumatra as a Waters Bioremediation Agent.

<b>Background and Objective:</b> Heavy metals are one of the most worrisome pollutants due to their toxicity. Prolonged exposure to heavy metals and their accumulation and biomagnification properties adversely affect aquatic biota and human health. The ability of microorganisms to bioremediate heavy metals into non-toxic forms is one solution. The research aims of the study were to find biofilm-forming heavy metal-resistant bacteria isolated from the waters of the Bungus Samudra Fishery Port (PPS), Padang City. <b>Materials and Methods:</b> This study used a marine agar medium modified with the addition of K<sub>2</sub>Cr<sub>2</sub>O<sub>7</sub>, Pb(NO<sub>3</sub>)<sub>2</sub> and CdSO<sub>4</sub>•H<sub>2</sub>O, Marine Broth medium and Congo Red Agar medium. The research methods include, the isolation of bacteria, isolate resistance test to heavy metals, testing the ability of isolates to form biofilms and determine the ability of isolates to reduce heavy metals. Furthermore, molecular identification of bacterial isolates was carried out to determine the type of species. <b>Results:</b> Five heavy metal-resistant bacterial isolates were found that were able to form biofilms, namely isolates B3Cd, B5Cr, B7Pb, B6Pb and B3Pb. The five isolates were able to reduce heavy metal content by 38.67-61.191%. Identification of the best bacterial isolates on each heavy metal tested, namely B3Cd, B5Cr and B7Pb, respectively, showed the type of <i>Acinetobacter schindleri</i>, <i>Acinetobacter</i> sp. and <i>Bacillus</i> sp. <b>Conclusion:</b> These three selected potential isolates can be used as bioremediation agents in metal-polluted waters in the future.

Interplay between Two-Component Regulatory Systems Is Involved in Control of Cupriavidus metallidurans Metal Resistance Genes.

Metal resistance of Cupriavidus metallidurans is based on determinants that were acquired in the past by horizontal gene transfer during evolution. Some of these determinants encode transmembrane metal efflux systems. Expression of most of the respective genes is controlled by two-component regulatory systems composed of a membrane-bound sensor/sensory histidine kinase (HK) and a cytoplasmic, DNA-binding response regulator (RR). Here, we investigated the interplay between the three closely related two-component regulatory systems CzcRS, CzcR2S2, and AgrRS. All three systems regulate the response regulator CzcR, while the RRs AgrR and CzcR2 were not involved in czc regulation. Target promoters were czcNp and czcPp for genes upstream and downstream of the central czc gene region. The two systems together repressed CzcRS-dependent upregulation of czcP-lacZ at low zinc concentrations in the presence of CzcS but activated this signal transmission at higher zinc concentrations. AgrRS and CzcR2S2 interacted to quench CzcRS-mediated expression of czcNp-lacZ and czcPp-lacZ. Together, cross talk between the three two-component regulatory systems enhanced the capabilities of the Czc systems by controlling expression of the additional genes czcN and czcP. IMPORTANCE Bacteria are able to acquire genes encoding resistance to metals and antibiotics by horizontal gene transfer. To bestow an evolutionary advantage on their host cell, new genes must be expressed, and their expression should be regulated so that resistance-mediating proteins are produced only when needed. Newly acquired regulators may interfere with those already present in a host cell. Such an event was studied here in the metal-resistant bacterium Cupriavidus metallidurans. The results demonstrate how regulation by the acquired genes interacts with the host's extant regulatory network. This leads to emergence of a new system level of complexity that optimizes the response of the cell to periplasmic signals.

Study of Cadmium Metal Resistance in Stenotrophomonas maltophilia.

Metal-resistant bacteria are recommended for metal removal applications due to their rapid multiplication and growth rates. To ensure safety replenishment in contaminated areas frequently hampered by heavy metal toxicity, it is crucial to comprehend their coping mechanisms under heavy metal stress. This study primarily examines the role of EPS (exopolysaccharide) in Stenotrophomonas maltophilia, a Gram-negative, aerobic, and rod-shaped bacteria, in response to Cd, as well as the binding behavior and biosorption mechanism between EPS and Cd, using SEM and FTIR. The studies showed that Stenotrophomonas maltophilia, can resist up to 150μM of Cd due to the binding of Cd to EPS. SEM analysis showed significant morphological changes and FTIR was to identify main structural groups like carboxyl and hydroxyl which confirms the presence of EPS. The study will also describe the mechanism of cross-reactivity between exopolysaccharide and siderophore production in metal-tolerant Stenotrophomonas maltophilia. This study proved that siderophore-mediated metal detoxification and effective absorption have been linked to metal chelation.

Spatio-temporal variation of the microbiome and resistome repertoire along an anthropogenically dynamic segment of the Ganges River, India

Aquatic ecosystems are regarded as a hub of antibiotic and metal resistance genes. River Ganges is a unique riverine system in India with socio-cultural and economic significance. However, it remains underexplored for its microbiome and associated resistomes along its anthropogenically impacted course. The present study utilized a nanopore sequencing approach to depict the microbial community structure in the sediments of the river Ganges harboring antibiotic and metal resistance genes (A/MRGs) in lower stretches known for anthropogenic impact. Comprehensive microbiome analyses revealed resistance genes against 23 different types of metals and 28 classes of antibiotics. The most dominant ARG category was multidrug resistance, while the most prevalent MRGs conferred resistance against copper and zinc. Seasonal differences dismally affected the microbiota of the Ganges. However, resistance genes for fosmidomycin and tetracycline varied with season ANOVA, p < 0.05. Interestingly, 333 and 334 ARG subtypes were observed at all the locations in pre-monsoon and post-monsoon, respectively. The taxa associated with the dominant ARGs and MRGs were Pseudomonas and Burkholderia, which are important nosocomial pathogens. A substantial phage diversity for pathogenic and putrefying bacteria at all locations attracts attention for its use to tackle the dissemination of antibiotic and metal-resistant bacteria. This study suggests the accumulation of antibiotics and metals as the driving force for the emergence of resistance genes and the affiliated bacteria trafficking them. The present metagenomic assessment highlights the need for comprehensive, long-term biological and physicochemical monitoring and mitigation strategies toward the contaminants associated with ARGs and MRGs in this nationally important river.

Copper and iron metal resistant rhizospheric bacteria boost the plant growth and bacoside A content in Bacopa monnieri under stress conditions.

Bacteria that enhance plant growth and development and are found in the vicinity of roots are referred to as plant growth-promoting rhizobacteria. Some beneficial bacteria help plant tolerance to many hazardous chemical elements. In this context, Cupriavidus basilensis, Novosphingobium humi, Bacillus zanthoxyli, Bacillus sp., Paenibacillus alvei, Ancylobacter aquaticus and Ralstonia syzygii metal-tolerant rhizospheric bacteria were isolated from rhizospheric soil associated with Bacopa monnieri. The beneficial effects of rhizospheric bacteria on B. monnieri plant physiology and biochemical responses were investigated under pot conditions at two levels (100μM and 500μM) of CuSO4 or FeCl3. N. humi, A. aquaticus and R. syzygii bacterial strains were associated with significantly increased height and biomass under normal and stress conditions. An assay for indole acetic acid in isolated rhizospheric bacteria found differential secretion except Bacillus zanthoxyli. Bacoside A is a major phytocompound in B. monnieri with medicinal value; maximum induction was observed in the R. syzygii treatment. High concentration of copper and iron salts negatively influenced height, biomass and photosynthetic pigments; however N. humi, A. aquaticus, Bacilllus sp. and R. syzygii beneficial bacterial helped plants under stress conditions. Moreover, a significant enhancement in chlorophyll a and b was noticed in C. basilensis, B. zanthoxyli, Bacilllus sp., P. alvei and R. syzygii treatments, without much influence on carotenoid levels. Therefore, the present study emphasises the importance of isolating plant growth-promoting rhizobacteria for use in bacopa plants exposed to metals such as copper and iron in soil.

Heavy metal and antibiotic resistance in four Indian and UK rivers with different levels and types of water pollution.

Heavy metal pollution can enhance the level of antibiotic resistance, posing concerns to ecosystem and public health. Here, we investigated heavy metal concentrations, heavy metal resistant bacteria and antibiotic resistant bacteria and their corresponding resistant genes, and integrons in four different river environments, i.e., low heavy metals and low wastewater, high heavy metals and low wastewater, low heavy metals and high wastewater, and high heavy metals and high wastewater levels. Heavy metals were found to show positive and significant correlations with heavy metal resistance and antibiotic resistance and integrons (r > 0.60, p < 0.05), indicating that heavy metal selective pressure can cause heavy metal and antibiotic resistance to be transmitted simultaneously via integrons, which can result in the development of multi-resistant bacteria in the heavy metal-polluted environments. Moreover, there were significant associations between heavy metal resistance and antibiotic resistance (r > 0.60, p < 0.05), demonstrating heavy metal and antibiotic resistance are connected via a same or related mechanism. Class 1 integrons were found to have strong correlations with heavy metals and heavy metal resistance and antibiotic resistance (r > 0.60, p < 0.05), indicating a higher occurrence of antibiotic resistance co-selection in the heavy metal-polluted environments.

In vitro and In silico analysis of heavy metal-resistant bacteria from cooum river for bioremediation approaches

In vitro and In silico analysis of heavy metal-resistant bacteria from cooum river for bioremediation approaches

Utilization of metal-resistant bacteria in the detoxification of contaminated dumpsites

The potentials of four metal-resistant bacterial strains to reduce the concentration of selected metals in dumpsite leachate were studied. The Environmental Microbiology and Biotechnology laboratory's culture collection of bacteria was used to extract the bacteria, which were then exposed to progressively higher metal concentrations in a medium enriched with metals. After the bacterial cells had been separated from the leachate by centrifugation at 10000 rpm for 15 min, the bacteria were then added to a batch culture biosorption set-up containing the culture medium and tyndallized leachate. The residual metal concentration was then determined after a 14-day incubation period using the atomic absorption spectrophotometer (AAS). The bacterial strains demonstrated high resistance to the four selected heavy metals [lead (Pb), chromium (Cr), cadmium (Cd) and nickel (Ni)] and their combination. The Minimum Inhibitory Concentration (MIC) value for the strains on the metal-incorporated medium for all the selected metals ranged from 700-1500 µg/ml. The resistance to the metals was in the order: Pb ˃ Ni ˃ Cr ˃ Cd. Pseudomonas aeruginosa had the highest MIC to the metal combination (1300 µg/ml) while the lowest was Proteus mirabilis (800 µg/ml); Paenalcaligenes faecalis and Bordetella petrii had MIC values of 1000 µg/ml and 1200 µg/ml respectively. Bordetella petrii removed the highest concentration of Cd and Ni from the leachate, with values of 32.81 percent and 34.91 percent, respectively, according to the biosorption setup, while Paenalcaligenes hominis had a higher percentage reduction for Pb in the leachate with a reduction of 35.77 percent. However, when the leachate was treated with a combination of the four bacterial strains, the largest percentage decrease for Cr (32.54 percent) was seen. According to this research, these metal-resistant bacteria might be highly helpful in the biological treatment of wastewater that contains metals.

Metal- and antibiotic-resistant heterotrophic plate count bacteria from a gold mine impacted river: the Mooi River system, South Africa

The Wonderfonteinspruit, South Africa, is highly impacted by a century of gold mining activities. The aim of this study was to investigate the physico-chemical properties of the Wonderfonteinspruit and the receiving Mooi River system, the levels of antimicrobial (metals and antibiotics) resistance characteristics and heterotrophic bacteria levels in these water systems. Various physico-chemical parameters were determined. R2A agar and R2A agar supplemented with antimicrobials were used to enumerate heterotrophic bacteria. Morphologically distinct antimicrobial-resistant isolates were purified and screened for antibiotic susceptibility by a disc diffusion method. Selected isolates were identified, and minimum inhibitory concentration ranges determined. Among the antimicrobial resistant isolates, 87% were resistant to at least one antibiotic. Of these, almost 50% were resistant to more than 3 antibiotic classes. A large proportion was resistant to all 7 antibiotics tested. Phyla detected were Proteobacteria, Firmicutes and Bacteriodetes. High MIC levels for metals and antibiotics were detected among all the genera. Results demonstrate potential impacts of physico-chemical properties on levels of antimicrobial-resistant bacteria. Metal-resistant bacteria were also resistant to multiple antibiotics, suggesting that metal pollution from mining may be responsible for co-selection and maintenance of antibiotic-resistant bacteria in this aquatic system.

Utilization of metal-resistant bacteria in the detoxification of contaminated dumpsites

Utilization of metal-resistant bacteria in the detoxification of contaminated dumpsites

Human Health and Soil Health Risks from Heavy Metals, Micro(nano)plastics, and Antibiotic Resistant Bacteria in Agricultural Soils

Humans are exposed to agricultural soils through inhalation, dermal contact, or the consumption of food. Human health may be at risk when soils are contaminated; while some soil contaminants such as heavy metals (HMs) have been extensively studied, others such as micro(nano)plastics (MNPs) or antibiotic-resistant bacteria (ARB) pose novel threats. This paper investigates the linkages between soil contamination and human health risk by reviewing the state of knowledge on HMs, MNPs, and ARB in agricultural soils. A keyword-based search in Web of Science, Scopus, and Google Scholar was conducted, complemented with a backward snowball search. We analysed main sources of contamination for agricultural soils, risks to human health differentiated by uptake pathway (ingestion, inhalation, dermal), and interactions of contaminants with microorganism, soil fauna, and plants. Results show that the emergence and spread of ARB and antibiotic resistant genes from agricultural soils and their contribution to antibiotic resistances of human pathogens is recognized as a significant threat. Likewise, a growing body of evidence indicates that MNPs are able to enter the food chain and to have potentially harmful effects on human health. For HM, knowledge of the effects on human health is well established. Multiple agricultural practices increase HM concentrations in soils, which may lead to adverse health effects from the ingestion of contaminated products or inhalation of contaminated soil particles. Severe knowledge gaps exist about the pathways of the contaminants, their behaviour in soil, and human uptake. Little is known about long-term exposure and impacts of MNPs, antibiotics and ARB on human health or about the possible combined effects of MNPs, ARB, and HMs. Missing monitoring systems inhibit a comprehensive assessment of human health risks. Our research demonstrates the need for human health risk assessment in the context of agricultural soils, in particular to be able to assess risks related to measures reinforcing the concept of the circular economy.

Integrative application of heavy metal-resistant bacteria, moringa extracts, and nano-silicon improves spinach yield and declines its contaminant contents on a heavy metal-contaminated soil.

Microorganism-related technologies are alternative and traditional methods of metal recovery or removal. We identified and described heavy metal-resistant bacteria isolated from polluted industrial soils collected from various sites at a depth of 0-200 mm. A total of 135 isolates were screened from polluted industrial soil. The three most abundant isolate strains resistant to heavy metals were selected: Paenibacillus jamilae DSM 13815T DSM (LA22), Bacillus subtilis ssp. spizizenii DSM 15029T DSM (MA3), and Pseudomonas aeruginosa A07_08_Pudu FLR (SN36). A test was conducted to evaluate the effect of (1) isolated heavy metal-resistant bacteria (soil application), (2) a foliar spray with silicon dioxide nanoparticles (Si-NPs), and (3) moringa leaf extract (MLE) on the production, antioxidant defense, and physio-biochemical characteristics of spinach grown on heavy metal-contaminated soil. Bacteria and MLE or Si-NPs have been applied in single or combined treatments. It was revealed that single or combined additions significantly increased plant height, shoot dry and fresh weight, leaf area, number of leaves in the plant, photosynthetic pigments content, total soluble sugars, free proline, membrane stability index, ascorbic acid, relative water content, α-tocopherol, glycine betaine, glutathione, and antioxidant enzyme activities (i.e., peroxidase, glutathione reductase, catalase, superoxide dismutase, and ascorbate peroxidase) compared with the control treatment. However, applying bacteria or foliar spray with MLE or Si-NPs significantly decreased the content of contaminants in plant leaves (e.g., Fe, Mn, Zn, Pb, Cd, Ni, and Cu), malondialdehyde, electrolyte leakage, superoxide radical , and hydrogen peroxide (H2O2). Integrative additions had a more significant effect than single applications. It was suggested in our study that the integrative addition of B. subtilis and MLE as a soil application and as a foliar spray, respectively, is a critical approach to increasing spinach plant performance and reducing its contaminant content under contaminated soil conditions.