RESUMO
DNA double strand breaks (DSBs) are induced by external genotoxic agents (ionizing radiation or genotoxins) or by internal processes (recombination intermediates in lymphocytes or by replication errors). The DNA ends induced by these genotoxic processes are often not ligatable, requiring potentially mutagenic end-processing to render ends compatible for ligation by non-homologous end-joining (NHEJ). Using single molecule approaches, Loparo et al. propose that NHEJ fidelity can be maintained by restricting end-processing to a ligation competent short-range NHEJ complex that 'maximizes the fidelity of DNA repair'. These in vitro studies show that although this short-range NHEJ complex requires DNA ligase IV (Lig4), its catalytic activity is dispensable. Here using cellular models, we show that inactive Lig4 robustly promotes DNA repair in living cells. Compared to repair products from wild-type cells, those isolated from cells with inactive Lig4 show a somewhat increased fraction that utilize micro-homology (MH) at the joining site consistent with alternative end-joining (a-EJ). But unlike a-EJ in the absence of NHEJ, a large percentage of joints isolated from cells with inactive Lig4 occur with no MH - thus, clearly distinct from a-EJ. Finally, biochemical assays demonstrate that the inactive Lig4 complex promotes the activity of DNA ligase III (Lig3).
Assuntos
Reparo do DNA por Junção de Extremidades , Reparo do DNA , DNA/genética , Quebras de DNA de Cadeia Dupla , DNA Ligase Dependente de ATP/genética , DNA Ligases/genética , DNA Ligases/metabolismo , BiocatáliseRESUMO
The COVID-19 pandemic has caused serious social and public health problems. In the field of personal protection, the facial masks can prevent infectious respiratory diseases, safeguard human health, and promote public safety. Herein, we focused on preparing a core filter layer for masks using electrospun polyvinyl butyral/apocynum venetum extract nanofibrous membranes (PVB/AVE NMs), with durable interception efficiency and antibacterial properties. In the spinning solution, AVE acted as a salt to improve electrical conductivity, and achieve long-lasting interception efficiency with adjustable pore size. It also played the role of an antibacterial agent in PVB/AVE NMs to achieve win-win effects. The hydrophobicity of PVB-AVE-6% was 120.9° whereas its filterability reached 98.3% when the pressure drop resistance was 142 Pa. PVB-AVE-6% exhibited intriguing properties with great antibacterial rates of 99.38% and 98.96% against S. aureus and E. coli, respectively. After a prolonged usability test of 8 h, the filtration efficiency of the PVB/AVE masks remained stable at over 97.7%. Furthermore, the antibacterial rates of the PVB/AVE masks on S. aureus and E. coli were 96.87% and 96.20% respectively, after using for 2 d. These results indicate that PVB/AVE NMs improve the protective performance of ordinary disposable masks, which has certain application in air filtration.
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Activation-induced cytidine deaminase (AID) initiates somatic hypermutation of immunoglobulin (Ig) gene variable regions and class switch recombination (CSR) of Ig heavy chain constant regions. Two decades of intensive research has greatly expanded our knowledge of how AID functions in peripheral B cells to optimize antibody responses against infections, while maintaining tight regulation of AID to restrain its activity to protect B cell genomic integrity. The many exciting recent advances in the field include: 1) the first description of AID's molecular structure, 2) remarkable advances in high throughput approaches that precisely track AID targeting genome-wide, and 3) the discovery that the cohesion-mediate loop extrusion mechanism [initially discovered in V(D)J recombination studies] also governs AID-medicated CSR. These advances have significantly advanced our understanding of AID's biochemical properties in vitro and AID's function and regulation in vivo. This mini review will discuss these recent discoveries and outline the challenges and questions that remain to be addressed.
Assuntos
Citidina Desaminase , Hipermutação Somática de Imunoglobulina , Linfócitos B , Citidina Desaminase/química , Citidina Desaminase/genética , Switching de Imunoglobulina/genética , Hipermutação Somática de Imunoglobulina/genéticaRESUMO
Immunoglobulin (Ig) class switch recombination (CSR) is the gene rearrangement process by which B lymphocytes change the Ig heavy chain constant region to permit a switch of Ig isotype from IgM to IgG, IgA, or IgE. At the DNA level, CSR occurs via generation and joining of DNA double strand breaks (DSBs) at intronic switch regions located just upstream of each of the heavy chain constant regions. Activation-induced deaminase (AID), a B cell specific enzyme, catalyzes cytosine deaminations (converting cytosines to uracils) as the initial DNA lesions that eventually lead to DSBs and CSR. Progress on AID structure integrates very well with knowledge about Ig class switch region nucleic acid structures that are supported by functional studies. It is an ideal time to review what is known about the mechanism of Ig CSR and its relation to somatic hypermutation. There have been many comprehensive reviews on various aspects of the CSR reaction and regulation of AID expression and activity. This review is focused on the relation between AID and switch region nucleic acid structures, with a particular emphasis on R-loops.
Assuntos
Citidina Desaminase/genética , Citidina Desaminase/metabolismo , Switching de Imunoglobulina/genética , Região de Troca de Imunoglobulinas/genética , Recombinação Genética , Hipermutação Somática de Imunoglobulina/genética , Animais , Linfócitos B/metabolismo , DNA/genética , DNA/metabolismo , Quebras de DNA de Cadeia Dupla , Humanos , Imunoglobulinas/genética , Estruturas R-Loop/genética , Translocação GenéticaRESUMO
Proteoglycans (PGs) play important roles in many biological processes including tumor progression, cell adhesion, and regulation of growth factor activities. With glycosaminoglycan chains attached to the core proteins in nature, PGs are highly challenging synthetic targets due to the difficulties in integrating the sulfated glycans with the peptide backbone. To expedite the synthesis, herein, the utility of human xylosyltransferase I (XT-I), the enzyme responsible for initiating PG synthesis, has been explored. XT-I was found to be capable of efficiently installing the xylose unit onto a variety of peptide structures on mg scales. Furthermore, an unnatural sugar, i.e., 6-azidoglucose can be transferred by XT-I introducing a reactive handle onto the glycopeptide for selective functionalization. XT-I can be coupled with ß-4-galactosyl transferase-7 for one pot synthesis of glycopeptides bearing galactose-xylose disaccharide, paving the way toward efficient chemoenzymatic synthesis of PG glycopeptides and glycoproteins.
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Pentosiltransferases/metabolismo , Proteoglicanas/biossíntese , Humanos , Conformação Proteica , Proteoglicanas/química , UDP Xilose-Proteína XilosiltransferaseRESUMO
The base excision repair (BER) pathway is an important DNA repair pathway and is essential for immune responses. In fact, it regulates both the antigen-stimulated somatic hypermutation (SHM) process and plays a central function in the process of class switch recombination (CSR). For both processes, a central role for apurinic/apyrimidinic endonuclease 1 (APE1) has been demonstrated. APE1 acts also as a master regulator of gene expression through its redox activity. APE1's redox activity stimulates the DNA-binding activity of several transcription factors, including NF-κB and a few others involved in inflammation and in immune responses. Therefore, it is possible that APE1 has a role in regulating the CSR through its function as a redox coactivator. The present study was undertaken to address this question. Using the CSR-competent mouse B-cell line CH12F3 and a combination of specific inhibitors of APE1's redox (APX3330) and repair (compound 3) activities, APE1-deficient or -reconstituted cell lines expressing redox-deficient or endonuclease-deficient proteins, and APX3330-treated mice, we determined the contributions of both endonuclease and redox functions of APE1 in CSR. We found that APE1's endonuclease activity is essential for IgA-class switch recombination. We provide evidence that the redox function of APE1 appears to play a role in regulating CSR through the interleukin-6 signaling pathway and in proper IgA expression. Our results shed light on APE1's redox function in the control of cancer growth through modulation of the IgA CSR process.
Assuntos
DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , Imunoglobulina A/genética , Switching de Imunoglobulina , Animais , Linfócitos B/metabolismo , Linhagem Celular , Reparo do DNA , Humanos , Interleucina-6/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Oxirredução , Transdução de SinaisRESUMO
Nonhomologous end-joining (NHEJ) is the major DNA double-strand break (DSB) repair pathway in mammals and resolves the DSBs generated during both V(D)J recombination in developing lymphocytes and class switch recombination (CSR) in antigen-stimulated B cells. In contrast to the absolute requirement for NHEJ to resolve DSBs associated with V(D)J recombination, DSBs associated with CSR can be resolved in NHEJ-deficient cells (albeit at a reduced level) by a poorly defined alternative end-joining (A-EJ) pathway. Deletion of DNA ligase IV (Lig4), a core component of the NHEJ pathway, reduces CSR efficiency in a mouse B-cell line capable of robust cytokine-stimulated CSR in cell culture. Here, we report that CSR levels are not further reduced by deletion of either of the two remaining DNA ligases (Lig1 and nuclear Lig3) in Lig4(-/-) cells. We conclude that in the absence of Lig4, Lig1, and Lig3 function in a redundant manner in resolving switch region DSBs during CSR.
Assuntos
Reparo do DNA por Junção de Extremidades , DNA Ligases/metabolismo , Switching de Imunoglobulina/genética , Recombinação Genética , Animais , Linhagem Celular , Núcleo Celular/enzimologia , Dano ao DNA , DNA Ligase Dependente de ATP , DNA Ligases/genética , Camundongos , Proteínas de Ligação a Poli-ADP-Ribose , Proteínas de XenopusRESUMO
The recalcitrance of green waste, caused by its high lignocellulose content, is a technical challenge for accelerating green waste composting. However, because lignocellulose degradation in litter (similar to green waste) can be promoted during the freeze-thaw season, and the composting is difficult to implement in this period (due to the low temperature); seasonal freeze-thaw was intended to be used as a pretreatment strategy for the existing technical challenge in the winter of cold regions. In this process, green waste was pretreated with seasonal freeze-thaw to enhance its lignocellulose degradation for subsequent composting. To verify this assumption, two strategies for the pretreatment were used: the green waste was either drenched or immersed in water during the freeze-thaw season, and the effects on subsequent composting were evaluated. The results demonstrated that both strategies can significantly promote the mineralization of TOC (total organic carbon, by 2.73%-8.01% compared with the control, the following comparisons were all based on the control), TN (total nitrogen, by 0.21%-0.52%), and lignocellulose (lignin degradation was promoted by 3.52%-3.73%, cellulose degradation was promoted by 13.23%-14.26%) during composting and that the synthesis of humus was also enhanced (by 19.19%-21.43%). Furthermore, since the loss of NH4+N and NO3-N was significantly less in the drenched treatment than in the immersed treatment (by 9.15% for the loss of NH4+N and 7.66% for the loss of NO3-N), drenching the green waste during the freeze-thaw season might be a better strategy than immersing for nitrogen conservation. An additional advantage of drenching compared to immersing is water conservation.
Assuntos
Compostagem , Congelamento , Nitrogênio , Estações do Ano , Solo , TemperaturaRESUMO
The evidence that ATM affects resolution of RAG-induced DNA double-strand breaks is profuse and unequivocal; moreover, it is clear that the RAG complex itself cooperates (in an undetermined way) with ATM to facilitate repair of these double-strand breaks by the classical nonhomologous end-joining pathway. The mechanistic basis for the cooperation between ATM and the RAG complex has not been defined, although proposed models invoke ATM and RAG2's C terminus in maintaining the RAG postcleavage complex. In this study, we show that ATM reduces the rate of both coding and signal joining in a robust episomal assay; we suggest that this is the result of increased stability of the postcleavage complex. ATM's ability to inhibit VDJ joining requires its enzymatic activity. The noncore C termini of both RAG1 and RAG2 are also required for ATM's capacity to limit signal (but not coding) joining. Moreover, potential phosphorylation targets within the C terminus of RAG2 are also required for ATM's capacity to limit signal joining. These data suggest a model whereby the RAG signal end complex is stabilized by phosphorylation of RAG2 by ATM.
Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Proteínas de Ligação a DNA/metabolismo , Rearranjo Gênico do Linfócito B , Proteínas de Homeodomínio/metabolismo , Proteínas Nucleares/metabolismo , Éxons VDJ/genética , Proteínas Mutadas de Ataxia Telangiectasia/genética , Quebras de DNA de Cadeia Dupla , Proteínas de Ligação a DNA/genética , Células HEK293 , Proteínas de Homeodomínio/genética , Humanos , Complexos Multiproteicos/metabolismo , Proteínas Nucleares/genética , Fases de Leitura Aberta/genética , Fosforilação , Plasmídeos/genética , Sinais Direcionadores de Proteínas/genética , Estabilidade Proteica , Reparo de DNA por RecombinaçãoRESUMO
The â¼28-kb 3' regulatory region (3'RR), which is located at the most distal 3' region of the Ig H chain locus, has multiple regulatory functions that control IgH expression, class-switch recombination (CSR), and somatic hypermutation. In this article, we report that deletion of the entire 3'RR in a mouse B cell line that is capable of robust cytokine-dependent CSR to IgA results in reduced, but not abolished, CSR. These data suggest that 3'RR is not absolutely required for CSR and, thus, is not essential for targeting activation-induced cytidine deaminase to S regions, as was suggested. Moreover, replacing 3'RR with a DNA fragment including only its four DNase I hypersensitive sites (lacking the large spacer regions) restores CSR to a level equivalent to or even higher than in wild-type cells, suggesting that the four hypersensitive sites contain most of the CSR-promoting functions of 3'RR. Stimulated cells express abundant germline transcripts, with the presence or absence of 3'RR, providing evidence that 3'RR has a role in promoting CSR that is unique from enhancing S region transcription.
Assuntos
Imunoglobulina G/genética , Cadeias Pesadas de Imunoglobulinas/genética , Região de Troca de Imunoglobulinas/genética , Animais , Células Cultivadas , Imunoglobulina G/imunologia , Cadeias Pesadas de Imunoglobulinas/imunologia , Região de Troca de Imunoglobulinas/imunologia , CamundongosRESUMO
Free radical attack on the C1' position of DNA deoxyribose generates the oxidized abasic (AP) site 2-deoxyribonolactone (dL). Upon encountering dL, AP lyase enzymes such as DNA polymerase ß (Polß) form dead-end, covalent intermediates in vitro during attempted DNA repair. However, the conditions that lead to the in vivo formation of such DNA-protein cross-links (DPC), and their impact on cellular functions, have remained unknown. We adapted an immuno-slot blot approach to detect oxidative Polß-DPC in vivo. Treatment of mammalian cells with genotoxic oxidants that generate dL in DNA led to the formation of Polß-DPC in vivo. In a dose-dependent fashion, Polß-DPC were detected in MDA-MB-231 human cells treated with the antitumor drug tirapazamine (TPZ; much more Polß-DPC under 1% O2 than under 21% O2) and even more robustly with the "chemical nuclease" 1,10-copper-ortho-phenanthroline, Cu(OP)2. Mouse embryonic fibroblasts challenged with TPZ or Cu(OP)2 also incurred Polß-DPC. Nonoxidative agents did not generate Polß-DPC. The cross-linking in vivo was clearly a result of the base excision DNA repair pathway: oxidative Polß-DPC depended on the Ape1 AP endonuclease, which generates the Polß lyase substrate, and they required the essential lysine-72 in the Polß lyase active site. Oxidative Polß-DPC had an unexpectedly short half-life (â¼ 30 min) in both human and mouse cells, and their removal was dependent on the proteasome. Proteasome inhibition under Cu(OP)2 treatment was significantly more cytotoxic to cells expressing wild-type Polß than to cells with the lyase-defective form. That observation underscores the genotoxic potential of oxidative Polß-DPC and the biological pressure to repair them.
Assuntos
Dano ao DNA , DNA Polimerase beta/metabolismo , DNA/metabolismo , Animais , Linhagem Celular Tumoral , Humanos , Camundongos , OxirreduçãoRESUMO
The boundaries of R-loops are well-documented at immunoglobulin heavy chain loci in mammalian B cells. Within primary B cells or B cell lines, the upstream boundaries of R-loops typically begin early in the repetitive portion of the switch regions. Most R-loops terminate within the switch repetitive zone, but the remainder can extend a few hundred base pairs further, where G-density on the non-template DNA strand gradually drops to the genome average. Whether the G-density determines how far the R-loops extend is an important question. We previously studied the role of G-clusters in initiating R-loop formation, but we did not examine the role of G-density in permitting the elongation of the R-loop, after it had initiated. Here, we vary the G-density of different portions of the switch region in a murine B cell line. We find that both class switch recombination (CSR) and R-loop formation decrease significantly when the overall G-density is reduced from 46% to 29%. Short 50 bp insertions with low G-density within switch regions do not appear to affect either CSR or R-loop elongation, whereas a longer (150 bp) insertion impairs both. These results demonstrate that G-density is an important determinant of the length over which mammalian genomic R-loops extend.
Assuntos
Switching de Imunoglobulina , Região de Troca de Imunoglobulinas , Animais , Linhagem Celular , DNA/química , Camundongos , Recombinação Genética , Sequências Repetitivas de Ácido NucleicoRESUMO
Wolf-Hirschhorn syndrome (WHS) is a genetic disease with characteristic facial features and developmental disorders. Of interest, loss of the MMSET gene (also known as WHSC1) is considered to be responsible for the core phenotypes of this disease. Patients with WHS also display Ab deficiency, although the underlying cause of this deficiency is unclear. Recent studies suggest that the histone methyltransferase activity of MMSET plays an important role in the DNA damage response by facilitating the recruitment of 53BP1 to sites of DNA damage. We hypothesize that MMSET also regulates class switch recombination (CSR) through its effect on 53BP1. In this study, we show that MMSET indeed plays an important role in CSR through its histone methyltransferase activity. Knocking down MMSET expression impaired 53BP1 recruitment as well as the germline transcription of the Igh switch regions, resulting in defective CSR but no effect on cell growth and viability. These results suggest that defective CSR caused by MMSET deficiency could be a cause of Ab deficiency in WHS patients.
Assuntos
Histona-Lisina N-Metiltransferase/metabolismo , Switching de Imunoglobulina/genética , Proteínas Repressoras/metabolismo , Recombinação V(D)J/genética , Linhagem Celular , Regulação da Expressão Gênica , Loci Gênicos , Histonas/metabolismo , Humanos , Cadeias Pesadas de Imunoglobulinas/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Metilação , Ligação Proteica , Transcrição Gênica , Proteína 1 de Ligação à Proteína Supressora de Tumor p53 , Síndrome de Wolf-Hirschhorn/genéticaRESUMO
DNA double-strand breaks (DSBs) are essential intermediates in Ig gene rearrangements: V(D)J and class switch recombination (CSR). In contrast to V(D)J recombination, which is almost exclusively dependent on nonhomologous end joining (NHEJ), CSR can occur in NHEJ-deficient cells via a poorly understand backup pathway (or pathways) often termed alternative end joining (A-EJ). Recently, several components of the single-strand DNA break (SSB) repair machinery, including XRCC1, have been implicated in A-EJ. To determine its role in A-EJ and CSR, Xrcc1 was deleted by targeted mutation in the CSR proficient mouse B-cell line, CH12F3. Here we demonstrate that XRCC1 deficiency slightly increases the efficiency of CSR. More importantly, Lig4 and XRCC1 double-deficient cells switch as efficiently as Lig4-deficient cells, clearly indicating that XRCC1 is dispensable for A-EJ in CH12F3 cells during CSR.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Recombinação V(D)J , Alelos , Animais , Linfócitos B/citologia , Linhagem Celular , Núcleo Celular/metabolismo , Quebras de DNA de Cadeia Dupla , Marcação de Genes , Teste de Complementação Genética , Genômica , Ligantes , Camundongos , Modelos Genéticos , Fatores de Tempo , Proteína 1 Complementadora Cruzada de Reparo de Raio-XRESUMO
Ig class-switch recombination (CSR) is directed by the long and repetitive switch regions and requires activation-induced cytidine deaminase (AID). One of the conserved switch-region sequence motifs (AGCT) is a preferred site for AID-mediated DNA-cytosine deamination. By using somatic gene targeting and recombinase-mediated cassette exchange, we established a cell line-based CSR assay that allows manipulation of switch sequences at the endogenous locus. We show that AGCT is only one of a family of four WGCW motifs in the switch region that can facilitate CSR. We go on to show that it is the overlap of AID hotspots at WGCW sites on the top and bottom strands that is critical. This finding leads to a much clearer model for the difference between CSR and somatic hypermutation.
Assuntos
Citidina Desaminase/imunologia , Citidina Desaminase/metabolismo , Switching de Imunoglobulina/fisiologia , Animais , Linfócitos B/enzimologia , Linfócitos B/imunologia , Sequência de Bases , Sítios de Ligação/genética , Linhagem Celular , DNA/genética , DNA/metabolismo , Camundongos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Mutação , Recombinação GenéticaRESUMO
In response to antigens, B cells undergo affinity maturation and class switching mediated by activation-induced cytidine deaminase (AID) in germinal centers (GCs) of secondary lymphoid organs, but uncontrolled AID activity can precipitate autoimmunity and cancer. The regulation of GC antibody diversification is of fundamental importance but not well understood. We found that autoimmune regulator (AIRE), the molecule essential for T cell tolerance, is expressed in GC B cells in a CD40-dependent manner, interacts with AID and negatively regulates antibody affinity maturation and class switching by inhibiting AID function. AIRE deficiency in B cells caused altered antibody repertoire, increased somatic hypermutations, elevated autoantibodies to T helper 17 effector cytokines and defective control of skin Candida albicans. These results define a GC B cell checkpoint of humoral immunity and illuminate new approaches of generating high-affinity neutralizing antibodies for immunotherapy.
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Food waste is a potential resource to prepare microbial fertilizer. However, functional microorganisms derived from the food waste compost (FWC) are relatively lacking. We have isolated, identified, characterized and optimized a high-yielding indole-3-acetic acid (IAA) strain from FWC and further evaluated its growth promoting effect on plants. A IAA high-yielding strain, Providencia sp.Y, with an initial IAA yield of 139.98â mg L-1, was obtained through high-throughput screening, and identified by 16S rRNA gene sequence. The novel strain Y may simultaneously involve the following three pathways from L-tryptophan to IAA, which were identified using liquid chromatography-tandem mass spectrometry: (1) L-tryptophan-indole-3-ethanol-indole-3-acetaldehyde-indole-3-acetic acid; (2) L-tryptophan-1-hydroxy-indole-3-ethanol-indole-3-acetic acid; (3) L-tryptophan-indole-3-acetamide-indole-3-acetic acid. The most suitable comprehensive conditions for IAA production, which were optimized by single factor experiment, were: culture time 12â h, inoculation amount 2% (v/v), NaCl concentration 4% (w/v), culture temperature 25â, initial pH = 5, and L-tryptophan concentration 3.0â g L-1. The yield of IAA after optimization was increased by 590.48%, from 139.98â mg L-1 (before optimization) to 966.54â mg L-1. Diluted 200-fold microbial suspension could significantly improve the growth of pakchoi seedlings. The seedling plant height, root length, leaf width, leaf length, and fresh weight with microbial suspension increased by 17.39%, 107.35%, 77.98%, 37.75%, and 215.38%, respectively, compared with those without microbial suspension. The increase was greater than that of commercial bacterial agents. In conclusion, this isolated strain can be used as an economical microbial inoculant and provides a new germplasm resource for developing microbial fertilizers.
Assuntos
Compostagem , Eliminação de Resíduos , Alimentos , Triptofano/metabolismo , Fermentação , RNA Ribossômico 16S/genética , Redes e Vias Metabólicas , Plantas/genética , Plantas/metabolismoRESUMO
ABSTRACTAerobic composting of food waste (FW) from rural China using a composting device results in a substantial financial burden on the government. This study aimed to assess the feasibility of mitigating this cost using vermicomposting of composted FW. The specific aims were to elucidate the effects of composted FW on earthworm growth and reproduction, reveal the changes in the physical and chemical properties of earthworm casts during vermicomposting, identify the microbial community structure associated with vermicomposting, and perform a financial analysis based on the yield of earthworms and earthworm casts. Mixing composted FW and mature cow dung in an equal ratio achieved the highest earthworm reproduction rate, where 100 adult earthworms produced 567 juvenile earthworms and 252 cocoons in 40 d. Earthworms reduce salt content of vermicomposting substrates by assimilating Na+ and promoting humification by transforming humin into humic and fulvic acid, thus producing earthworm casts with a high generation index > 80%. When composted FW was added to a vermicomposting substrate, a distinctive microbial community structure with alkaliphilic, halophilic, and lignocellulolytic microorganisms dominated the microflora. The dominant bacterial species was Saccharopolyspora rectivirgula, and the dominant fungal species changed from Kernia nitida to Coprinopsis scobicola. Furthermore, microbial genes for refractory organic matter and fat degradation were observed in Vibrio cholerae, Kernia nitida, and Coprinopsis scobicola. Financial analysis showed that vermicomposting has the potential to reduce the cost associated with FW disposal from $ 57 to $ 18/t.
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Microplastics (MPs) in soil have attracted extensive attention as an emerging pollutant, and the transport of MPs is affected by their own physical and chemical properties, the chemical composition of soil solutions, and soil minerals. However, in the presence of oxides, the underlying mechanism for the transport of MPs in different ionic types and ionic strengths is still not fully understood. In this study, the effects of ionic type, ionic strength, and iron oxide on the transport of polystyrene microplastics (PSMPs) with different functional groups were investigated through stability experiments and transport experiments. The colloid transport model, CD-MUSIC model, and DLVO theory were used to explore the transport mechanism. The results showed that normalized concentrations (c/c0) of PSMPs were 0.99 in the NaH2PO4 background and 0.94 in the CaCl2 background, respectively, which indicated that the strongest stability of PSMPs was observed in the former and the weakest in the latter. Different ionic types had different effects on the transport of PSMPs. For the cations Na+ and Ca2+, Ca2+ strongly inhibited PSMPs transport in pure quartz sand because of the bridging effect and strong charge neutralization effect; the recovery rate of the PSMPs in the effluent was (43.83±1.71)%, and a first-order retention coefficient on the second kinetic Site-2 (k2a) was 1.54 min-1. The presence of iron oxide enhanced the inhibition, the recovery rate of the PSMPs in the effluent decreased to (6.04±0.40)%, and k2a increased to 5.33 min-1. For the anions Cl- and PO43-, the transport of PSMPs in pure quartz sand was dominated by surface electronegativity of PSMPs, and PSMPs exhibited lower electronegativity under Cl- background and thus showed higher recovery[(92.95±0.63)%] and lower k2a (0.19 min-1). However, in the presence of iron oxides, the Zeta potential of the quartz sand surface was the controlling factor for PSMPs transport. According to results of the CD-MUSIC model, PO43- could be easily adsorbed on the iron oxide surface to form innersphere complexes, which reduced the surface electronegativity of the iron-loaded quartz sand and enhanced the transport of PSMPs, higher recovery[(76.22±1.39)%], and lower k2a (0.66 min-1). Moreover, the species of the formed innersphere complex was controlled by the PO43- concentration, and different species of innersphere complexes had distinct negative surface charges. Higher surface electronegativity of the iron-loaded quartz sand was observed under higher PO43- concentration, which was not conducive to the transport of PSMPs. Further, the transport ability of PSMPs decreased with the increase in ionic strength. Finally, the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory was used to calculate the variation in the primary barrier between PSMPs and the collector under the conducted experimental conditions, which helped better elucidate the transport behavior of PSMPs. The variation in the primary barrier was consistent with the transport ability of PSMPs, and a higher primary barrier indicated a larger repulsion between PSMPs and the collector, which was in favor of PSMPs transport.