Responses of antibiotic resistance genes in the enhanced biological phosphorus removal system under various antibiotics: Mechanisms and implications.

The effects of antibiotics on the proliferation of antibiotic resistant genes (ARGs) in WWTPs have drawn great attention in recent years. The effects of antibiotics on ARGs in the enhanced biological phosphorus removal (EBPR) system and its mechanisms, however, are still not well understood. In this study, EBPR systems were constructed using activated sludge to investigate the effects of ten commonly detected antibiotics in the environment on the proliferation of ARGs and the mechanisms involved. The results showed that the total abundance of ARGs increased to varying degrees with the addition of different antibiotics (0.05 mmol/L), and the top 30 ARGs increased by 271.1 % to 370.0 %. Mobile genetic elements (MGEs), functional modules, and the bacteria community were consistently related to the changes in ARGs. Refractory antibiotics, in particular, have a stronger promoting effect on transduction in the EBPR system. The insertion sequence common region (ISCR) and transposon (Tnp) were identified as crucial factors in the proliferation of ARGs. Moreover, the risk of polyphosphate accumulating organisms (PAOs) carrying ARGs in the presence of antibiotics should not be ignored. Our findings emphasize the potential efficacy of employing strategies that target the reduction of MGEs, regulation of cellular communication, and management of microbial communities to effectively mitigate the risks associated with ARGs.

Ecological insight into antibiotic resistome of ion-adsorption rare earth mining soils from south China by metagenomic analysis.

Antibiotic resistome has led to growing global threat to public health. Rare earth elements play important roles in modern society and mining activity for them has caused serious impact on soil ecosystems. However, antibiotic resistome in, especially, ion-adsorption rare earth-related soils is still poorly understood. In this work, soils were collected from ion-adsorption rare earth mining areas and adjacent regions of south China and metagenomic analysis was employed for profile, driving factors and ecological assembly of antibiotic resistome in the soils. Results show prevalence of antibiotic resistance genes conferring resistance to tetracycline/fluoroquinolone (adeF), peptide (bcrA), aminoglycoside (rpsL), tetracycline (tet(A)) and mupirocin (mupB) in ion-adsorption rare earth mining soils. Profile of antibiotic resistome is accompanied by its driving factors, i.e., physicochemical properties (La, Ce, Pr, Nd and Y of rare earth elements in 12.50-487.90 mg kg-1), taxonomy (Proteobacteria, Actinobacteria) and mobile genetic elements (MGEs, plasmid pYP1, Transposase_20). Variation partitioning analysis and partial least-squares-path modeling demonstrate that taxonomy is the most important individual contributor and pose most direct/indirect effect to antibiotic resistome. Further, null model analysis reveals stochastic processes as dominant ecological assembly of antibiotic resistome. This work advances our knowledge on antibiotic resistome with emphasis on ecological assembly in ion-adsorption rare earth-related soils for ARGs mitigation, mining management and mine restoration.

Significant Differences in the Effects of Nitrogen Doping on Pristine Biochar and Graphene-like Biochar for the Adsorption of Tetracycline.

To improve the adsorption efficiency of pollutants by biochar, preparing graphene-like biochar (GBC) or nitrogen-doped biochar are two commonly used methods. However, the difference in the nitrogen doping (N-doping) effects upon the adsorption of pollutants by pristine biochar (PBC) and GBC, as well as the underlying mechanisms, are still unclear. Take the tetracycline (TC) as an example, the present study analyzed the characteristics of the adsorption of TCs on biochars (PBC, GBC, N-PBC, N-GBC), and significant differences in the effects of N-doping on the adsorption of TCs by PBC and GBC were consistently observed at different solution properties. Specifically, N-doping had varied effects on the adsorption performance of PBC, whereas it uniformly improved the adsorption performance of GBC. To interpret the phenomenon, the N-doping upon the adsorption was revealed by the QSAR model, which indicated that the pore filling (VM) and the interactions between TCs with biochars (Ead-v) were found to be the most important two factors. Furthermore, the density functional theory (DFT) results demonstrated that N-doping slightly affects biochar's chemical reactivity. The van der Waals (vdWs) and electrostatic interactions are the main forces for TCs-biochars interactions. Moreover, N-doping mostly strengthened the electrostatic interactions of TCs-biochars, but the vdWs interactions of most samples remained largely unaffected. Overall, the revealed mechanism of N-doping on TCs adsorption by biochars will enhance our knowledge of antibiotic pollution remediation.

Effects of antibiotics on enhanced biological phosphorus removal and its mechanisms.

Many kinds of antibiotics are continuously discharged into wastewater and typically cause a great decrease in sewage treatment performance, whereas mechanisms of differences in the impacts of commonly used antibiotics on phosphate removal are still elusive. Thus, an enhanced biological phosphorus removal (EBPR) system, as an effective method of phosphate removal, was developed, and its performance in the treatment of artificial wastewater containing antibiotics at short- (8 h) and long-term (15 days) exposure was investigated. The results show that phosphorus removal was consistently inhibited by the addition of antibiotics with a significant difference (P < 0.05). To interpret the phenomena, mechanistic equations were developed, and the results indicate that for short-term tests, the difference was mainly caused by the suppression of polyhydroxyalkanoate (PHA) degradation and the activity of polyphosphate kinase (PPK), resulting in the different inhibition of the soluble orthophosphorus (SOP) uptake process. For long-term tests, the difference in SOP uptake was principally caused by the inhibition of PHA degradation and the activity of PPK, whereas the difference in SOP release resulted from the inhibition of activities of exopolyphosphatase (PPX) and adenylate kinase (ADK). Moreover, micro-mechanisms of such inhibition were identified from molecular docking and electrostatic potential.

Dissipation of Sulfonamides in Soil Emphasizing Taxonomy and Function of Microbiomes by Metagenomic Analysis.

Sulfonamides (SAs) are widespread in soils, and their dissipation behavior is important for their fate, risk assessment, and pollution control. In this work, we investigated the dissipation behavior of different SAs in a soil under aerobic condition, focusing on revealing the relationship between overall dissipation (without sterilization and in dark) and individual abiotic (sorption, hydrolysis)/biotic (with sterilization and in dark) factors and taxonomy/function of microbiomes. The results showed that dissipation of all SAs in the soil followed the pseudo-first-order kinetic model with dissipation time at 50% removal (DT50) of 2.16-15.27 days. Based on, experimentally, abiotic/biotic processes and, theoretically, partial least-squares modeling, a relationship between overall dissipation and individual abiotic/biotic factors was developed with microbial degradation as the dominant contributor. Metagenomic analysis showed that taxonomic genera like Bradyrhizobium/Sphingomonas/Methyloferula and functions like CAZy family GT51/GH23/GT2, eggNOG category S, KEGG pathway ko02024/ko02010, and KEGG ortholog K01999/K03088 are putatively involved in SA microbial degradation in soil. Spearman correlation suggests abundant genera being multifunctional. This study provides some new insights into SA dissipation and can be applied to other antibiotics/soils in the future.

New parameters for the quantitative assessment of the proliferation of antibiotic resistance genes dynamic in the environment and its application: A case of sulfonamides and sulfonamide resistance genes.

Antibiotic resistance genes (ARGs) have been widely detected around the world and are generally viewed as emerging pollutants with environmental persistence. The proliferation of ARGs can be easily promoted by antibiotics. However, the dynamics of ARGs in the environment are still unable to be quantified using a single parameter, which is vital to evaluating the ability of ARGs to spread by antibiotics and effectively controlling ARGs. A new parameter, termed the relative area ratio of sample to control (ΔAR), was developed based on the quantitative features determined by ARG-time curves in soils contaminated with sulfonamides (SAs) and verified by quantitative structure-activity relationships (QSARs) models. The results showed that ΔAR can not only be used to accurately quantify the characteristics of SAs resistance genes (Suls) over time but also be applied to reveal the relationships between the proliferation of Suls and important factors (i.e., concentrations and chemical structures). Moreover, the ΔAR-based QSARs model indicated that bioavailability and the frequency of conjugative transfer, rather than the ability of induced mutations in bacteria, tend to be key processes of the characteristics of the proliferation of Suls. Therefore, ΔAR is a useful parameter to perform environmental risk assessments of ARG proliferation in the environment.

Identification and regulation of host genes related to Rice stripe virus symptom production.

Viral infections cause plant chlorosis, stunting, necrosis or other symptoms. The down-regulation of chloroplast-related genes (ChRGs) is assumed to be responsible for chlorosis. We identified the differentially expressed genes (DEGs) in Rice stripe virus (RSV)-infected Nicotiana benthamiana, and examined the contribution of 75 down-regulated DEGs to RSV symptoms by silencing them one by one using Tobacco rattle virus (TRV)-induced gene silencing. Silencing of 11 of the 75 down-regulated DEGs caused plant chlorosis, and nine of the 11 were ChRGs. Silencing of a down-regulated DEG encoding the eukaryotic translation initiation factor 4A (eIF4A) caused leaf-twisting and stunting that were visible on RSV-infected N. benthamiana. A region of RSV RNA4 was complementary to part of eIF4A mRNA and virus-derived small interfering (vsiRNAs) from that region were present in infected N. benthamiana. When expressed as artificial microRNAs, those vsiRNAs could target NbeIF4A mRNA for regulation. We provide experimental evidence supporting the association of ChRGs with chlorosis and show that eIF4A is involved in RSV symptom development. This is also the first report demonstrating that siRNA derived directly from a plant virus can target a host gene for regulation.

A transmembrane domain determines the localization of rice stripe virus pc4 to plasmodesmata and is essential for its function as a movement protein.

The pc4 protein encoded by rice stripe virus (RSV) is a viral movement protein (MP). A transmembrane (TM) domain spanning AAs 106-123 of pc4 was identified and shown to be essential for localization of pc4 to plasmodesmata (PD) (but not to chloroplasts) and for its ability to recover the movement of movement-deficient PVX. Analysis of alanine-scanning mutants showed that M116A and G120A had a similar localization to wild type pc4, being localized at PD and chloroplasts, but all other mutants were only localized at chloroplasts and not at PD. Mutants that could not be localized at PD had little (G123A) or no ability to recover PVX-GFPΔp25 movement, indicating that PD localization is crucial for the function of pc4 as a movement protein. Strangely, mutants M116A and G120A localized at PD and retained the ability to bind single-stranded RNA but did not support PVX-GFPΔp25 movement, indicating that properties other than PD localization and nucleotide binding ability may be needed for the function of pc4 as a movement protein.

Tobacco smoking, epilepsy, and seizures.

Tobacco smoking is considered the greatest risk factor for death caused by noncommunicable diseases. In contrast to extensive research on the association between tobacco smoking and diseases such as heart attack, stroke, and cancers, studies on the association between tobacco smoking and seizures or epilepsy are insufficient. The exact roles tobacco smoking and nicotine use play in seizures or epilepsy have not been well reviewed. We reviewed available literature and found that 1) there are vast differences between tobacco smoke and nicotine based on their components and their effects on seizures or epilepsy; 2) the seizure risk in acute active tobacco smokers, women who smoke during pregnancy, electronic cigarette smokers, and the role of smoking in sudden unexplained/unexpected death in epilepsy remain unclear; 3) seizure risks are higher in acute secondhand smokers, chronic active smokers, and babies whose mothers smoke; 4) tobacco smoke protects against seizures in animal models whereas nicotine exerts mixed effects in animals; and 5) tobacco smoking agents can be noneffective, proconvulsant, or anticonvulsant. Finally, the opportunities for future research on this topic is discussed.

Fluoradenes via palladium-catalyzed intramolecular arylation.

A novel approach towards 7b-aryl-indeno[1,2,3-jk]fluorene based on a nitrogen-containing core is reported. The acid-promoted Friedel-Crafts reaction of 9-(2-bromophenyl)-9-fluorenol with carbazole, triphenylamine or triindole afforded 9-(2-bromophenyl)fluorenyl-carbazole, -triphenylamine and -triindole derivatives, which were subsequently converted to 7b-aryl-fluoradenes via palladium-catalyzed intramolecular C-H direct arylation as a key step.