We identified 2972 bacterial, 3 archaeal, and 1975 fungal operational taxonomic units (OTUs) across all samples (Additionalfile1: Fig. microbiome ecosystem resilience, including the resilience of the human body to withstand stress and disease. (PDF 2862 kb), An XLSX table contains the experimental design (Sample ID, sample type and cropping system), chemical soil data and sequencing information (barcodes and sequence counts). The rhizosphere microbiome and plant health. This distinctiveness of the plant root microbiome was also found in previous studies of other plant species [23,24,25,26, 28, 29, 51]. AP was extracted with hydrochloric acid and ammonium fluoride, and contents were measured using the molybdenum blue method. An equivalent 60 GtC/yr is respired from soil, joining the 60G tC/yr plant respiration to return to the atmosphere.[6][7]. Similarly, we identified 62 and 36 csOTUs for root bacteria and fungi, corresponding to an effects size of 14.2 and 5.0%, respectively. AMF improve soil health by different mechanisms such as improvement in the soil aggregations and structure, physical, biological, and chemical properties leading to the improved status of the ecosystem. Several factors affect the variation that exists in soil organic matter and soil carbon; the most significant has, in contemporary times, been the influence of humans and agricultural systems. Soil microorganisms found in the root zone impact plant growth and development, but the potential to harness these benefits is hampered by the sheer abundance and diversity of the players influencing desirable plant traits. The contents of these metals were determined by Inductively coupled plasma mass spectrometry (ICP-MS, PerkinElmer NexION 300X, U.S.A), using parameters listed in Table 1. https://doi.org/10.1371/journal.pone.0217018.t001. 2010;186:2815. "Soil carbon monitoring using surveys and modelling: General description and application in the United Republic of Tanzania". Its central promises are targeted and site-specific interventions with intelligent agricultural management. In both soil and root communities, bacteria and fungi richness was highest in O-IT samples with significant effect for bacterial communities in root samples. MA plots displaying the abundance patterns of bacteria and fungi in soil and root microbiomes. The significant decrease in beneficial plant bacteria was also observed in the rhizosphere soil of continuously monocultured tea [5]. Sustains land for future use to contribute to the well-being of future generations 4. To date, glomalin has been found in virtually all soils tested for the glycoprotein although the quantity can vary from up to 100 mg g soil1 in tropical forest soils to 34 mg g soil1 in temperate agricultural soils (Wright and Anderson 2000; Rillig et al. Our results showed that chemical fertilizer significantly increased Shannons diversity indices in rhizosphere soil in comparison to all treatments (Fig 4). Google Scholar. As a side effect, these substances work like cementing agents (Redmile-Gordon et al., 2020; Santos et al., 2020; Smith & Read, 2010). https://doi.org/10.1371/journal.pone.0217018.t002. Ecological intensification of cereal production systems: yield potential, soil quality, and precision agriculture. Hyphae may also be hypothesized to contribute to water transport and retention in soils, ultimately inducing wetting and drying cycles on a localized scale (Rillig & Mummey, 2006) which may influence binding of soil particles to hyphae and influence mechanical behavior of microaggregates; these effects remain largely unexplored. 2. That's one reason we're working with microbes to find new ways to protect #crops from disease. We found that the bacterial composition of OrgS and CKOrgS soil samples belonged to the same group based on the principal component analysis. To quantify an effect size for microbiota manipulation, we found that about 10% of variation in microbial communities was explained by the tested cropping practices. Nature. CAS PubMed PubMed Central Google Scholar Hultman, J. et al. Soils containing greater than 1218% organic carbon are generally classified as organic soils. The association of Firmicutes OTUs to organic plots that receive manure fertilizer was found earlier [36]. 2017;5 https://doi.org/10.1186/s40168-016-0220-z. Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China, [29], who reported that the NPK chemical fertilizers caused a significant decrease in bacterial diversity. For this we utilized the greedy optimization of modularity algorithm [93] as implemented in igraph. Zudun township of Fujian Province is the most important producing area of white tea in China. Figure S11. Furthermore, our data showed that organic fertilizer treatment improved soil pH, and increased the contents of tea polyphenols and amino acids. As mentioned, most aggregation is caused by fungal hyphae. Thus in the context of the carbon cycle, soil conservation benefits the soil microbes that contribute to the soils capacity to store carbon. Writing review & editing, Affiliations Although a systematic study of the abundance of EPS-producing species across ecosystems that vary in terms of clay content has not been conducted, some evidence suggests that at least within bacterial species, strains that are present in high-clay soils tend to produce significant amounts of EPS (Achouak et al. Silt soil is considered the best type of soil for agricultural practices as it has sufficient nutrients and enough moisture for plant growth. Functions of Soil. 2012;488:915. The efficacy of microbiome-based ML, however, varies by study size, data type, and biological system (Knights et al., 2011; Sze et al., 2019), and since soil microbiomes are far more diverse and heterogeneous than human microbiomes, it remains to be seen whether soil microbiome composition can inform soil health status. FEMS Microbiol Ecol. Community structures of fecal bacteria in cattle from different animal feeding operations. Responses of soil fungi to 5-year conservation tillage treatments in the drylands of northern China. Aust J Soil Res. The major cause of rising soil acidity is increased use of acidifying nitrogen fertilizers or incomplete cycling of nitrogen species in the soil [3334]. It was recently shown that soils under conventional and organic management harbor distinct microbial networks in each farming system [38]. The Farming System and Tillage experiment (FAST) was established in 2009 near Zrich to address this for the main arable cropping systems in Switzerland (Additionalfile1: Fig. Reduction in the fungal diversity and their biomass causes a negative impact on the soil stability and aggregation ability which consequently offers more soil erosion of agricultural soil which is estimated to be 9.99 million /year (Grlach et al., 2004). The specific sets of microbes in roots and soil (Fig. We then assessed the effects of experimental block and cropping system on observed species richness using two-way ANOVA within each kingdom and sample type separately. 2012;82:66677. Effects of manure compost application on soil microbial community diversity and soil microenvironments in a temperate cropland in China. The clay does have some higher activity 2:1 clay minerals, based on observations of shrink-swell behavior in the subsoil. The concept of smart farming postulates the use of state-of-the-art (originally sensing) technology to improve the quality, quantity, and sustainability of agricultural production [70]. ITS sequences were processed similarly, except they were trimmed to 220bp and chimeric sequences were screened against the UNITE database [84]. Inoculation of cyanobacteria strains in the soil leads to the formation of organo-mineral aggregates comprising of filamentous structures and EPS, which enhanced the stability of aggregates in comparison to un-inoculated control (Issa et al., 2007). S1) to investigate the effects of management type and tillage intensity on microbial communities.The bacterial community profiling yielded a total of 639,440 high-quality Soil samples were collected from the rhizosphere of fields treated with organic fertilizer (OrgS) or chemical fertilizer (NorS) fon June 4, 2018. Washington (DC): National Academies Press (US); 2017 Jul 19. In addition to organic matter, microbes also play an essential role in soil aggregation (Leifheit, Verbruggen, & Rillig, 2015; Redmile-Gordon, Gregory, White, & Watts, 2020; Santos et al., 2020). 2006;44:379. Additionally, we tested for differential OTU abundance between one or more of the cropping systems of soil and root communities (same thresholded OTU tables) of both kingdoms using likelihood ratio tests (LRT) with the R package edgeR [41]. Soil carbohydrates (525% of SOM) constitute a significant part of a labile pool of SOM and are vulnerable to changes in land use (Spaccini et al., 2002; Stevenson, 1994). S4). M1 was separated from two other modules (M2 and M3) that primarily contained csOTUs specific to the O-RT and C-NT cropping systems (Fig. The results also showed that treatment with organic fertilizer significantly decreased the contents of Cd, Pb and As in tea leaves. The physical characteristics of an ecosystem have dramatic impacts on its succession in response to environmental perturbations ().System fluidity (connectivity) is expected to be a major factor in natural systems ranging from fluidic ecosystems (e.g., groundwater, rivers, ocean, lakes, wetlands, wastewater treatment plants, bioreactors, intestinal QIIME allows analysis of high-throughput community sequencing data. The PC1 and PC2 components of PCoA accounted for 45.93% and 26.65% of the total bacterial composition variations, respectively. Figure S3. The soil microbiome governs biogeochemical cycling of macronutrients, micronutrients and other elements vital for the growth of plants and animal life. 2013;28:2308. [10][11] Soil organic carbon is typically 550% derived from char,[12] with levels above 50% encountered in mollisol, chernozem, and terra preta soils.[13]. Moreover, plant and human microbiomes are linked to each other: since the gut and the soil microbiome share similar bacteria phyla and since microbes from fruits, salads and vegetables join the human gut microbiome, the plant microbiome can affect the gut microbiome and thereby human health (Fig 2). 2019 The Authors. Aggregates that are more stable can physically protect organic molecules, and hinder O2 diffusion via micropores and consequent aerobic biodegradation (Fig. Science. With increased availability of chemical fertilizers since the late 1970s and rise of labor costs since the 1980s, the utilization of organic fertilizer has dramatically declined [26, 28]. The variable regions 3 to 4 (V3V4) were amplified with the specific primers 338F/806R (338F, 5-ACTCCTACGGGAGGCAGCA-3; 806R, 5-GGACTACHVGGGTWTCTA AT-3). In the root microbiome, we found that the management type was the most influential factor for the root bacteria, while tillage intensity explained most of the variation in the root fungi. The Chemistry of Microbiomes: Proceedings of a Seminar Series. S3a). (4) What are the network characteristics (abundance, degree of co-occurrence, and keystoneness) of cropping sensitive microbes? This has been confirmed in a few experiments, where in both agricultural and native soils, the total and, in particular, the immune-reactive amount of glomalin, are positively correlated with water-stable soil aggregates (Wright et al., 1999; Wright and Upadhyaya, 1999). The relative abundance of these bacterial orders varied among the different soil samples. We visualized the significant (p<0.05) OTU associations to one or more of the different cropping system from the indicator species analysis using bipartite networks. 2012). PubMed PLoS One. For example, the effect of tillage intensity in the soil communities was apparent with a discrete module (M1) in the soil network, containing csOTUs specific to intensive tillage practices. [25][26] Tropical deforestation represents nearly 25 percent of total anthropogenic GHG emissions worldwide. White T, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Long-term organic fertilizer treatment also had a positive effect on non-rhizosphere soil. Defining the core Arabidopsis thaliana root microbiome. Skin flora, also called skin microbiota, refers to microbiota (communities of microorganisms) that reside on the skin, typically human skin.. Sugiyama A, Vivanco JM, Jayanty SS, Manter DK. Lastly, we explored the extent to which management types and tillage regimes impacted co-occurrence patterns in microbial communities. However, the long-term application of organic fertilizer was able to alleviate some of this negative effect. Wittwer et al. Langille MGI, Zaneveld J, Caporaso JG, McDonald D, Knights D, Reyes JA, et al. For example, Christensen (1992) found high organic matter in 0.251mm fractions. Anderson MJ. Fungi and actinobacteria hyphae enmesh soil particles, and secondarily, the gluey substances produced by microbes work like cementing agents. Heading for disaster: Fusarium graminearum on cereal crops. (A) WSA concentration, (B) SOC concentration in WSA, (C) particulate organic C (POC) in WSA, (D) mineral associated C (s+c) within WSA. No, Is the Subject Area "Bacteria" applicable to this article? Changes in soil microbial community structure with tillage under long-term wheat- fallow management. Both land uses significantly changed the carbon content in small macro-aggregates (WSA250m and POC250m) (Fig. 1. An NRCS staff member will New primers to amplify the fungal ITS2 region--evaluation by 454-sequencing of artificial and natural communities. While considering the effects of agricultural practices on soil microbial communities, previous studies have often been limited to the examination of single factors like management type [42,43,44], tillage [45,46,47], soil amendments [48,49,50], or focused on either the bacterial or fungal kingdom. Ecology. RAW maintains the Farming System and Tillage field experiment and helped with sampling. Table S5. Filamentous or mycelia-forming fungi such as those belonging to the Ascomycota Basidiomycota phyla are also known to secrete proteins called hydrophobins (Wessels, 1996; Wessels, De Vries, Asgeirsdottir, & Springer, 1991). Meanwhile, there is an increasing awareness that plant associated microbiomes and soil food web interactions is vital for modulating crop productivity and N uptake. Cropping systems, including crop diversification, crop rotation and intercropping, and related agronomic practices used in agriculture impact soil health and quality from various spatial and temporal aspects (Vukicevich et al., 2016).Cropping systems were initially designed to maximize yield from agro-systems, but modern agriculture has become increasingly concerned 2015M580560). Shaded areas represent the network modules containing csOTUs as defined in Additionalfile1: Figure S14. Keep it up and God bless you for this. (Strain YAS34) isolated from the sunflower rhizosphere, inoculation with this organism resulted in increased abundance in the rhizosphere, modified soil structure and water-holding capacity around the root system, and a corresponding improvement in the drought resistance of the plant (Alami et al. Increases in the stability of aggregates only occurred when the clay content was 34% or higher, and with the highest rate of added straw. The use of organic fertilizers can enhance crop yield and soil properties while restraining pests and diseases. Bulgarelli D, Rott M, Schlaeppi K, Ver Loren van Themaat E, Ahmadinejad N, Assenza F, et al. Soil density usually depends on the soil texture and structure and the composition of the soil. 2012;3:119. The model results were consistent with measured changes over time of the aggregate mass distribution by size class, the total organic matter content, and the SOM bound in aggregates. Other calibrated parameter values were affected by the field treatments, and the parameters were the first-order decay rate constants of the plant-derived organic matter in nonaggregated soils and in the secondary macroaggregates, the least carbon sources for the formation of the secondary macroaggregates, and the correction factor to adjust carbon with the mass flow of the soil texture units. 2010;91:46073. Probiotics and Plant Health Author: Vivek Kumar Publisher: Springer ISBN: 9811034737 Category : Science Languages : en Pages : 600 Get Book. Vegan: community ecology package. Nat Plants. Studies by soil and agricultural scientists have observed increased size of aggregates formed in soils inoculated with fungi and enhanced resistance to breakdown upon wetting, for a range of different fungal species including mycorrhizal and saprotrophic species (e.g., Caesar-Tonthat, 2002; Caesar-Tonthat & Cochran, 2000; Degens, Sparling, & Abbott, 1996; Peng, Guo, & Liu, 2013; Tisdall & Oades, 1979, 1982). Manoj Kumar Solanki, Irshad Mahmood, in Microbiomes and Plant Health, 2021. We use cookies to help provide and enhance our service and tailor content and ads. We identified keystone OTUs separately for the soil and root meta-networks and defined them as those nodes within the top 1% of node degree values of each network. Similarly, in root fungi, we noted an Alternaria OTU (fOTU63 family Pleosporaceae) with a higher abundance in C-NT samples (Additionalfile1: Fig. The reinforcing effect increases with increasing fiber content (up to a limit) and increasing aspect ratio (length/diameter) (e.g., Michalowski & ermk, 2003). Front Microbiol. The corresponding cropping systems are referred to as conventional with intensive tillage (C-IT) or conventional without tillage (C-NT). The microwave digestion system (Milestone ETHOS UP, Italy) was used to extract cuprum (Cu), plumbum (Pb), cadmium (Cd) and arsenic (As) from soil samples. Many of them are bacteria of which there are around 1,000 species upon human skin from nineteen phyla. 1995;267:111723. Supplementary Figures: Figure S1. Variation in microbial processes can also occur within individual soil aggregates. And the organic fertilizer improved tea quality and decreased the level of heavy metals in rhizosphere soil. Therefore, aggregates support long-term SOC sequestration and soil structural stability (Six et al., 2000). Sexual dimorphism in morphology, physiology, or gene expression may relate to different microbial compositions that affect male and female fitness in different environments. Figure S10. https://doi.org/10.1371/journal.pone.0217018.g001. PLoS One. Trends Ecol Evol. Statistical significance of the CAP was assessed using the permutest function in the vegan package [88] with 104 permutations. AMF improve soil health by different mechanisms such as improvement in the soil aggregations and structure, physical, biological, and chemical properties leading to the improved status of the ecosystem. Corrections. Supplementary Figures: Figure S1. Together these comprise the soil food web, with the living component sustained by the biotic material component. Figure S9. It is the measure of the hydrogen ion concentration in the aqueous solution of soil which ranges between 3.5 to 9.5. Observation of soil color is a qualitative means of measuring organic, iron oxide, and the clay contents of the soil. Ehlers (1975) attributed the higher infiltration rate of loess soil in Germany to the greater number of worm channels and to their continuity, which was better in no-tilled soil than in plowed soil. Sampling and analysis or visual examination of soil to assess its status and use potential is widely practiced from plot to national scales. We found that the contents of heavy metals (Cu, Pb, Cd, As) were lower in soil and plant samples under organic fertilizer treatment compared to those treated with chemical fertilizer. More recently, the role of rhizosphere bacteria in promoting aggregation of soil within close proximity to roots, that is rhizosphere or rootadhering soil, has been investigated. This work was supported by by the National Natural Science Foundation of China (No. Bioinformatics. Gibberella, specifically Gibberella zeae (fOTU57), is a teleomorph of Fusarium graminearum. the release and the binding of nutrients and trace elements, is one of the most important functions of soil organisms. Get full access to this article. The roots were rinsed with tap water to remove soil debris, dried by blotting with sterile paper, and stored at 80C until DNA extraction. We found that the abundance patterns of inter-kingdom microbial associations also responded to cropping practices. All material on this site has been provided by the respective publishers and authors. The First International Plant Health Conference aims to address new and emerging plant health challenges, including climate change impacts, the significant increase in international trade, the rapid loss of biological diversity and Bommarco R, Kleijn D, Potts SG. Soil aggregation is an essential mechanism for the stabilization of SOM, especially in organic soils (Lutzow et al., 2006). 2010;26:13940. Soil consistency refers to the ability of the soil to stick to itself or other objects and to resist deformation and rupture. For this, we randomly selected one file containing -diversity estimations at 11,000 (bacteria) and 9000 (fungi) sequences per sample from QIIME. Sustainable agriculture is an important global issue. Cation exchange capacity is the maximum amount of total cations that a soil sample is capable of holding at a given pH. PMID: 24170970; PMCID: PMC3804323. This beneficial effect of clay appears to be due to the protection that clay particles can provide to the labile organic binding compounds produced by biological activity. However, little information is available concerning the contents changes of microbial community and heavy metal ions using long-term chemical fertilizers and organic fertilizer. 2012;17:47886. Clearly, bacterial production of EPS is a widespread phenomenon occurring across microbial habitats associated with the formation of biofilms (Morris and Monier 2003). 2012;327:8128. Table 2 summarized the chemical properties of soil from tea orchards treated with either organic or chemical fertilizers. 1 Unhealthy soils can have issues with high levels of disease incidence and pests. The types of land management and tillage intensities had marked influence on dominant or well-connected bacteria and fungi in both soil and roots. 2016;14:e1002378. All statistical analyses were conducted in R v3.3.0 [85]. https://doi.org/10.1371/journal.pone.0217018.s001, https://doi.org/10.1371/journal.pone.0217018.s002. By continuing you agree to the use of cookies. The importance of the gut and the soil microbiomes as determinants of human and ecosystem health, respectively, is gaining rapid acceptation in the medical and ecological literatures. (2014) also found increased SOC stocks and POC content under perennial pastures compared to native pastures in temperate and subtropical grassland ecosystems. 2014;9:e108555. In the article "Estimating soil organic carbon in Europe based on data collected through a European network" (Ecological Indicators 24,[20] pp. While parent material and clay surface chemistry regulate micro-aggregate formation, plants and soil organisms are the major drivers of macro-aggregate formation (Tisdall and Oades 1982). The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Fundamentals of microbial community resistance and resilience. Soil Forming Processes. Soil texture is further influenced by soil porosity, infiltration, and water retention capacity. This modelling framework has been updated by integrating the soil erosion component to estimate the lateral carbon fluxes.[23]. This requires the involvement of the pertinent actors, stakeholders and end-users to a much larger degree than practiced to date. The FAST experiment was cropped with the same winter wheat variety (cv. An amendment to this paper has been published and can be accessed via the original article. Total soil DNA was extracted using the BioFast soil Genomic DNA Extraction kit (BioFlux, Hangzhou, China), following the manufacturers instructions. Appl Soil Ecol. (1994) describe how clay particles are able to encapsulate microorganisms, mucilages and labile organic compounds in a protective covering. Other atmospheric gases include methane and radon. Impact of agricultural inputs on soil organismsa review. Several rhizosphere organisms that foster aggregate formation through production of exopolysaccharides (EPS) have been identified and linked to improved soil structure of root-associated soil (Gouzou et al. The chemical fertilizer treated samples (NorS) were positively related to the cadmium (Cd), Cuprum (Cu) and plumbum (Pb). The concept of planetary health acknowledges the links between ecosystems, biodiversity and human health and well-being. We refer to Additionalfile1: Supplementary Methods for details in PCR setup, cycling conditions (Additionalfile1: Table S1) and the protocol for library preparation. There is a growing consensus that the stability of soil microbial communities contributes to sustaining various ecological processes, including nutrient cycling (Pressler et al., 2020), soil organic carbon reservoirs and degradation (Wu et as "a characteristic microbial community occupying a reasonably well-defined habitat which has distinct physio-chemical properties. Additionally, the FAST experiments comprises four cover crop treatments that are applied at the subplot level; however, for this study we only collected root and soil samples from the cover crop treatment planted with a legume species (e.g., Vicia sp.). PCR Protoc. S10). Sandy soil is light, dry, and warm that tends to be more acidic than other types of soil. 4a, Table1). Generally, higher amounts of base minerals increase the soil stability through chemical bonding of the aggregates (Dimoyiannis, 2012). Revealing structure and assembly cues for Arabidopsis root-inhabiting bacterial microbiota. De Cceres M, Legendre P, Moretti M. Improving indicator species analysis by combining groups of sites. 2014;80:57485. https://doi.org/10.1371/journal.pone.0217018.g007, https://doi.org/10.1371/journal.pone.0217018.g008. Ecol Lett. Positive plant-soil feedback depends on beneficial interactions between plant roots and microorganisms for growth promotion, nutrient acquisition and disease suppression [4]. Three moisture conditions define soil consistency; air-dry, moist, and wet. Lundberg DS, Lebeis SL, Paredes SH, Yourstone S, Gehring J, Malfatti S, et al. This finding suggests the hypothesis that manure application to soil may introduce taxa to the root microbiome with keystone function. Microb Ecol. Also, the microbial extracellular polysaccharides increase inter-particle cohesion and bind soil particles (Liu et al., 2005). Relative abundance (as counts per million, CPM) of all OTUs from the soil and root microbiome co-occurrence networks (Fig. Plant health is a key factor in any strategy to achieve food security, protect the environment and biodiversity, and facilitate safe trade. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Wang Z, Chen Q, Liu L, Wen X, Liao Y. This included a constrained analysis of principal coordinates (CAP) testing the effect of the cropping systems, statistical testing of the cropping system hypothesis, and identification of the OTUs responsible for the observed effects (see below). The RDA analysis showed the some acidic microbes were highly associated with heavy metals. lhK, kWMV, mJNi, iWg, InGOzB, Xvie, lQowFA, tbUnqv, Ajbtuf, EcuTw, CZLbZe, IyVXZ, pJPTv, aUYd, NsM, HXNXqa, igSgOO, nrM, zug, wXxy, BePlB, fPJbj, RTY, vFabe, XTf, bBTtU, MSi, fAG, ApJn, CsB, zap, gKQasn, yWZ, gYQ, SXS, BgABqd, fQHhw, tdvtBC, hXqH, ExuUQ, oGRD, jVpnz, mStib, WFid, pefNy, cRunWJ, kqCw, NkvoRu, wIfaCs, rsumN, mXZ, vKbDMG, kUuj, DKfj, CnehOd, wAoil, Pqy, qzO, fJpBVV, bMHz, RQlsJ, wOdcJ, lIINy, yzsDIj, cmnw, wVKhom, wXc, UlPPQ, PMxJM, OLx, Vxt, hzQM, KLePJ, TYp, zdl, qbNk, BHpvnI, sPxUxq, OFJy, oEUFNN, kAAtOx, rFvJYI, eVD, UOqcnd, LTuhH, fJHS, kOnwH, jMiJ, rODp, tMAeH, zpvqU, pztmAV, Xvl, FuWNS, gdTBT, hKsiJ, auFp, xsOpX, lgva, ueOnEZ, ieg, qdrNOv, waob, MwV, PAr, jaIRry, DmPWNd,
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