Effective immobilization of heavy metals via reactive barrier by rhizosphere bacteria and their biofilms.

As the portal of plants, rhizosphere microorganisms play an essential role in controlling the species, transformation, and bioavailability of heavy metals, yet the potential passivation mechanism is still unclear. In this study, two heavy metal resistant and growth-promoting rhizosphere bacteria were screened, and their mechanisms in dealing with external stress and immobilizing heavy metal were explored. The results showed that heavy metals inhibited the ability of Pseudomonas sp. H13 and Brevundomonas sp. H16 to promote plant growth, but stimulated the production of extracellular polysaccharides and inorganic labile sulfide, and enhanced biofilm formation, thereby significantly improved the removal efficiency of Cu2+, Zn2+, Cd2+, and Pb2+. Compared with H16, the biofilm of H13 disintegrated rapidly in the later stage, so more metal ions were adsorbed on the planktonic cells. The C-OH and PO groups related to polysaccharides play a crucial role in heavy metal adsorption, and the immobilization mechanism of the planktonic cell is mainly ion exchange and group complex, but for H16, intracellular enrichment cannot be ignored. Functional group complexes played a dominant role in biofilm, and the immobilized heavy metals were more difficult to release into the environment. This study highlighted the potential application prospects of biofilm bacteria in heavy metal remediation and explained the reactive barrier of rhizosphere bacteria to heavy metals.

Degradation of methyl orange by pyrite activated persulfate oxidation: mechanism, pathway and influences of water substrates.

Degradation mechanism of methyl orange (MO), a typical azo dye, with pyrite (FeS2) activated persulfate (PS) was explored. The results showed that when the initial concentration of MO was 0.1 mM, FeS2 was 1.6 g/L and PS was 1.0 mM, the removal rate of MO could reach 92.9% in 150 min, and the removal rate of total organic carbon could reach 14.1%. In addition, both pH ≤ 2 and pH ≥ 10 could have an inhibitory effect in the FeS2/PS system. Furthermore, Cl- and low concentrations of HCO-3 had little effect on the degradation of MO with FeS2/PS. However, H2PO-4 and high concentrations of HCO-3 could inhibit the degradation of MO in the system. Besides, MO in river water and tap water were not degraded in FeS2/PS system, but acidification (pH = 4) would greatly promote the degradation. In addition, the removal rate of MO with FeS2/PS could still reach about 90% after five cycles of FeS2. Furthermore, the intermediates and possible degradation pathways were speculated by LC-MS, and the degradation mechanism of MO by FeS2/PS was that the cycle of Fe(III)/Fe(II) could continuously activate persulfate to produce SO4•-. The results could provide technical support for azo dye degradation in the FeS2/PS system.

Calcium-crosslinked alginate-encapsulated bacteria for remediating of cadmium-polluted water and production of CdS nanoparticles.

Pollution with the heavy metal cadmium (Cd2+) is a global problem. Cadmium adversely affects living organisms, highlighting the need to develop new methods for removal of this pollutant from the environment. In this study, we used a novel biomaterial based on calcium-crosslinked alginate-encapsulated bacteria to precipitate Cd2+ in polluted water. Our results show that calcium-crosslinked alginate-encapsulated bacteria effectively removed Cd2+ ions from cadmium-polluted water. Approximately 100% of Cd2+ ions were removed by 10 g (wet weight) of this biomaterial when the loading concentration of Cd2+ reached 1 mM in a volume of 50 ml water. During this process, a CdS nanoparticle, showing good crystallinity in the quantum range, was simultaneously produced. To validate the activity and stability of this biomaterial, we measured cysteine desulfhydrase activity in the stored biomaterial and whether this biomaterial could be recycled. The encapsulated bacteria maintained catalytic activity for at least 2 weeks. The capsules were easily regenerated and possessed good recyclability. Our results indicated that calcium-crosslinked alginate-encapsulated bacteria are suitable for depletion of Cd2+ in polluted water and for production of CdS nanoparticles. These calcium-crosslinked alginate-encapsulated bacteria are safe for biological manipulation and can be widely used to produce CdS nanoparticles during bioremediation of Cd2+-polluted water. KEY POINTS: • Calcium-crosslinked alginate-encapsulated bacteria can effectively precipitate Cd2+ in water coupled with production of CdS quantum dots. • The encapsulated bacteria maintained catalytic activity for at least 2 weeks. • The capsules were easily regenerated and possessed good recyclability.

Role of novel bacterial Raoultella sp. strain X13 in plant growth promotion and cadmium bioremediation in soil.

Heavy metal pollution in agricultural soils has become a widespread serious problem with the rapid industrialization and urbanization in the past two decades. Cadmium (Cd2+) is of the most concern in soils due to its high toxicity. It is necessary to develop remediation strategies to remove or neutralize its toxic effects in Cd-contaminated soil. Microbial bioremediation is a promising technology to treat heavy metal-contaminated soils. In this study, Cd-resistant bacterium, isolated from heavy metal-polluted soil in Southern China, was characterized as Raoultella sp. strain X13 on the basis of its biochemical profile and 16S rRNA. We investigated the characterization of Cd2+ distribution in different cellular compartments after Cd2+ uptake. Cd2+ uptake by strain X13 was mainly by ion exchange and chelation binding tightly to the cell wall. In addition, X13 plant growth-promoting characteristics suggested that X13 could solubilize phosphate and produce indole acetic acid. Pot experiments for the remediation of Cd-contaminated soil in situ by X13 inoculation demonstrated that X13 application to Cd-contaminated soils significantly promoted pak choi growth and improved production. We also found that X13 substantially reduced the Cd2+ bioavailability for pak choi. Therefore, strain X13 is an effective treatment for potential application in Cd2+ remediation as well as for sustainable agronomic production programs in Cd-contaminated soils.

Effect of biochars with different particle sizes on fates of antibiotics and antibiotic resistance genes during composting of swine manure.

The impact of biochars with large particle sizes (LPB, 5-10 mm) and small particle sizes (SPB, <0.074 mm) on fates of antibiotics and antibiotic resistance genes (ARGs) during composting of swine manure with maize straw was explored. The results showed that antibiotics removal efficiencies were 40 %, 50 %, and 76 % for control treatment, control with LPB treatment, and control with SPB treatment after composting, respectively. The introduction of SPB reduced the total ARGs and mobile genetic elements (MGEs) levels by 28 % and 19 % after composting, respectively. The Mantel test results showed that organic matter, moisture content, and NH4+-N contributed the most to changes in antibiotics and ARGs. The significant effects of biochar specific surface area and antibiotics on MGEs further regulated the behavior of ARGs. Therefore, composting with SPB is more conducive to the removal of antibiotics and ARGs during composting of swine manure.

Effect of biochar on antibiotics and antibiotic resistance genes variations during co-composting of pig manure and corn straw.

Pig manure is a reservoir of antibiotics and antibiotic resistance genes (ARGs). The effect of biochar on the variations in physicochemical properties, bacterial communities, antibiotics, ARGs, and mobile genetic elements (MGEs) of compost product during co-composting of pig manure and corn straw have been investigated in this study. Compared with the control treatment (CK), biochar addition accelerated the increase in pile temperature and prolonged the high temperature period (>55°C) for 2 days. Under biochar influence, organic matter degradation, NH4 +-N conversion and NO3 --N production was accelerated, and dissolved total organic carbon (DOC) and dissolved total nitrogen (DTN) utilization by microorganisms were enhanced. Biochar addition altered the microbial community and promoted the vital activity of Actinobacteria in the later composting stage. The antibiotics removal efficiency (except danofloxacin and enrofloxacin) was accelerated in the early composting stage (1-14 days) by biochar addition, the pile temperature had a positive effect on antibiotics removal, and the total antibiotics removal efficiency in CK and CK+Biochar treatments was 69.58% and 78.67% at the end of the composting process, respectively. The absolute abundance of most of the ARGs in the CK+Biochar treatment was lower than that in the CK treatment during composting, and the ARGs removal mainly occurred in the early (1-14 days) and later (28-50 days) stages. Biochar addition reduced the absolute abundance of MGEs (intI1, intI2) in the compost product, and most of the ARGs had a significant positive correlation with MGEs. Network analysis and redundancy analysis showed that ARGs and MGEs occurred in various host bacteria (Firmicutes, Actinobacteria, Bacteroidetes, Proteobacteria, and Halanaerobiaeota), and that DTN and NH4 +-N are the main factors regulating the changes in bacterial communities, antibiotics, ARGs, and MGEs during composting. Moreover, MGEs contributed the most to the variation in ARGs. In summary, biochar addition during composting accelerated antibiotics removal and inhibited accumulation and transmission of ARGs. The results of this study could provide theoretical and technical support for biochar application for antibiotics and ARGs removal during livestock and poultry manure composting.

Characterization of Cd2+ biosorption by Pseudomonas sp. strain 375, a novel biosorbent isolated from soil polluted with heavy metals in Southern China.

Water pollution with heavy metals is a global problem. Using microbial adsorbents to remediate water bodies contaminated with heavy metals has been garnering considerable attention. In this study, a cadmium (Cd2+)-resistant bacterium, isolated from soil polluted with heavy metals, was characterized as Pseudomonas sp. 375 based on its biochemical characteristics and 16S rRNA gene. The minimum inhibitory concentration (MIC) of Cd2+ for strain 375 was 6 mM. We evaluated the effects of different parameters, such as initial pH, contact time, and initial Cd2+ concentration, on Cd2+ uptake. The data acquired using nonliving biomass were fitted to a Langmuir isotherm model; however, the Freundlich isotherm model showed better fit for data acquired using living biomass. The maximum biosorption capacities were 92.59 mg g-1 and 63.29 mg g-1 for living and nonliving cells, respectively. The kinetics of biosorption was described using a pseudo-second order kinetic model. The tightly bound Cd on the cell wall played a major role in Cd2+ adsorption for both biosorbents. SEM-EDX analysis also showed that Cd2+ was bound to the cell wall. FTIR spectral analysis showed that -CH2, -OH, -SO3, CO, N-H, C-N, phosphate, or sulfate functional groups were the main functional sites for the binding of Cd2+ ions. Effectively Cd2+ removal from Cd2+ contaminated water suggested Pseudomonas sp. 375 was an (a) inexpensive, effective, and promising biosorbent that can be used for bioremediation Cd2+-contaminated wastewater.

Co-effect of minerals and Cd(II) promoted the formation of bacterial biofilm and consequently enhanced the sorption of Cd(II).

Heavy metal pollution is very common in soils. Soils are complex systems including minerals, bacteria, and various other substances. In Cd(II) contaminated soil, the combined effects of clay minerals and heavy metals on bacterial biofilm and Cd(II) adsorption are unappreciated. Our study showed that the combination of clay minerals (goethite, kaolinite, and montmorillonite) and heavy metals promoted Serratia marcescens S14 biofilm development significantly more than clay minerals or Cd(II) alone. The amount of biofilm after binary treatment with clay minerals and Cd(II) was 2.3-7.3 times than that in control. Mineral-induced cell death and the expression of the fimA, bsmA, and eps were key players in biofilm formation. Binary treatment with montmorillonite and Cd(II) significantly enhanced biofilm development and consequently increased the adsorption of Cd(II). Cd(II) removal is the result of co-adsorption of bacteria and minerals. Bacterial biofilm played an important role in Cd(II) adsorption. FTIR spectroscopy showed the components of biofilm were not affected by minerals and revealed the functional groups -OH, -NH, -CH2, -SH, -COO participated in Cd(II) immobilization. Our findings are of fundamental significance for understanding how minerals and Cd(II) affect biofilms and thereby enhance Cd(II) adsorption and predicting the mobility and fate of heavy metals in heavy metal-contaminated soil.

Complete genome sequence of Raoultella sp. strain X13, a promising cell factory for the synthesis of CdS quantum dots.

A novel cadmium-resistant bacterium, Raoultella sp. strain X13, recently isolated from heavy metal-contaminated soil, and this strain can synthesize CdS quantum dots using cadmium nitrate [Cd(NO4)2] and l-cysteine. Biomineralization of CdS by strain X13 can efficiently remove cadmium from aqueous solution. To illuminate the molecular mechanisms for the biosynthesis of CdS nanoparticle, the complete genome of Raoultella sp. strain X13 was sequenced. The whole genome sequence comprises a circular chromosome and a circular plasmid. Cysteine desulfhydrase smCSE has been previously found to be associated with the synthesis of CdS quantum dots. Bioinformatics analysis indicated that the genome of Raoultella sp. strain X13 encodes five putative cysteine desulfhydrases and all of them are located in the chromosome. The genome information may help us to determine the molecular mechanisms of the synthesis of CdS quantum dots and potentially enable us to engineer this microorganism for applications in biotechnology.