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Arsenic (As) poisoning in groundwater and rice paddy soil has increased globally, impacting human health and food security. There is an urgent need to deal with As-contaminated groundwater and soil. Biochar can be a useful remedy for toxic contaminants. This study explains the synthesis of pinecone-magnetic biochar (PC-MBC) by engineering the pinecone-pristine biochar with iron salts (FeCl3.6H2O and FeSO4.7H2O) to investigate its effects on As(V) adsorption and immobilization in water and soil, respectively. The results indicated that PC-MBC can remediate As(V)-contaminated water, with an adsorption capacity of 12.14 mg g-1 in water. Isotherm and kinetic modeling showed that the adsorption mechanism involved multilayer, monolayer, and diffusional processes, with chemisorption operating as the primary interface between As(V) and biochar. Post-adsorption analysis of PC-MBC, using FTIR and XRD, further revealed chemical fixing and outer-sphere complexation between As(V) and Fe, O, NH, and OH as the main reasons for As(V) adsorption onto PC-MBC. Recycling of PC-MBC also had excellent adsorption even after several regeneration cycles. Similarly, PC-MBC successfully immobilized As in paddy soil. Single and sequential extraction results showed the transformation of mobile forms of As to a more stable form, confirmed by non-destructive analysis using SEM, EDX, and elemental dot mapping. Thus, Fe-modified pine-cone biochar could be a suitable and cheap adsorbent for As-contaminated water and soil.

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This work revealed the profile of viral communities in paddy soils with different levels of arsenic (As) contamination during the flooded period. The structure of viral communities differed significantly in highly and moderately As-contaminated soils. The diversity of soil viral communities under high As contamination decreased. Siphoviridae, Podoviridae, Myoviridae, and Microviridae were the dominant viral families in all samples, and the relative abundances of five of the top 20 viral genera were significantly different between highly and moderately As-contaminated groups. Seventeen dissimilatory As(V)-reducing bacteria were predicted to host 161 viral operational taxonomic units (vOTUs), mainly affiliated with the genera of Sulfurospirillum, Deferribacter, Bacillus and Fusibacter. Among them, 28 vOTUs were also associated with Fe(III)-reducing bacteria, which belonged to different species of the genus Shewanella. Procrustes analysis showed that the community structure of soil viruses was strongly correlated with both prokaryotic community structure and geochemical properties. Random forest analyses revealed that the Total-Fe, DCB-Fe and oxalate-Fe were the most significant variables on viral community richness, while the total-As concentration was an important factor on the Shannon index. Furthermore, As resistance genes (ArsC, ArsR and ArsD), As methylation genes (arsM) and As transporter genes (Pst and Pit) were identified among the auxiliary metabolic genes (AMGs) of the virome. This work revealed that the viruses might influence microbial adaptation in response to As-induced stress, and provided a perspective on the potential virus-mediated biogeochemical cycling of As.

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Gene encoding the large subunit of As(III) oxidase (AioA), an important component of the microbial As(III) oxidation system, is a widely used biomarker to characterize As(III)-oxidizing communities in the environment. However, many studies were restricted to a few sequences generated by clone libraries and Sanger sequencing, which may have underestimated the diversity of As(III)-oxidizers in natural environments. In this study, we designed a primer pair, 1109F (5'-ATC TGG GGB AAY RAC AAY TA-3') and 1548R (5'-TTC ATB GAS GTS AGR TTC AT-3'), targeting gene sequence encoding for the conserved molybdopterin center of the AioA protein, yielding amplicons approximately 450 bp in size that are feasible for highly parallel amplicon sequencing. By utilizing in silico analyses and the experimental construction of clone libraries using Sanger sequencing, the specificity and resolution of 1109F/1548R are approximated with two other previously published and commonly used primers, i.e., M1-2F/M3-2R and deg1F/deg1R. With the use of the 1109F/1548R primer pair, the taxonomic composition of the aioA genes was similar both according to the Sanger and next-generation sequencing (NGS) platforms. Furthermore, high-throughput amplicon sequencing using the primer pair, 1109F/1548R, successfully identified the well-known As(III)-oxidizers in paddy soils and sediments, and they also revealed the differences in the community structure and composition of As(III)-oxidizers in above two biotopes. The random forest analysis showed that the dissolved As(III) had the highest relative influence on the Chao1 index of the aioA genes. These observations demonstrate that the newly designed PCR primers enhanced the ability to detect the diversity of aioA-encoding microorganisms in environments using highly parallel short amplicon sequencing.