# | Rank | Similarity | Title + Abs. | Year | PMID |
|---|---|---|---|---|---|
| 0 | 1 | 2 | 3 | 4 | 5 |
| 7547 | 0 | 1.0000 | Mechanism concerning the occurrence and removal of antibiotic resistance genes in composting product with ozone post-treatment. The soil application of composting product will probably cause the spread of antibiotic resistance genes (ARGs) to environment, thereby it is crucial to remove ARGs in composting product. Ozone was adopted for the first time as a post-treatment method to remove the ARGs in composting product in this study. Ozone treatment significantly removed the total ARGs and mobile genetic elements (MGEs) once ozonation process finished. After 10-day storage stage, although the amount of total intracellular ARGs and MGEs increased, the total extracellular ARGs and MGEs decreased in the ozone-treated compost product. Correlation analysis revealed that the reduction in intracellular 16S rRNA contributed to intracellular tetQ and tetW removal, while the variations of other ARGs after ozonation related to MGEs abundance. Network analysis suggested that the reduction of potential host bacteria, as well as the decline in NH(4)(+)-N and TOC after the ozonation, contributed to the intracellular ARGs removal. | 2021 | 33257169 |
| 7548 | 1 | 0.9999 | Maturity phase is crucial for removing antibiotic resistance genes during composting: novel insights into dissolved organic matter-microbial symbiosis system. Composting is widely regarded as an effective method for reducing antibiotic resistance genes (ARGs) in livestock and poultry manure. However, the critical mechanisms of ARGs in different composting phase are still unclear. In this study, normal composting and two types of rapid composting (without mature phase) were used to analyze the removal of ARGs and the succession of dissolved organic matter (DOM). Compared to normal composting, rapid composting reactivated tetracyclines, sulfonamide, and quinolones resistance genes during the maturation phase and reduced the total ARGs removal rates by 45.58 %-57.87 %. Humus-like components could inhibit the proliferation of ARGs, and the enrichment of protein-like components increased abundances of Pusillimonas, Persicitalea, and Pseudomonas, indirectly reducing the removal. This study is the first to demonstrate the contribution of DOM and microbial community to ARGs removal, emphasizing the importance of the maturation phase for ARGs elimination. This research provides guidance for producing safe compost products. | 2025 | 40311709 |
| 7550 | 2 | 0.9999 | Evaluating the effects of activated carbon on methane generation and the fate of antibiotic resistant genes and class I integrons during anaerobic digestion of solid organic wastes. The effects of activated carbon (AC) on methane production and the fate of antibiotic resistance genes (ARGs) were evaluated through comparing the anaerobic digestion performance and transformation of ARGs among anaerobic mono-digestion of food waste, co-digestion of food waste and chicken manure, and co-digestion of food waste and waste activated sludge. Results showed that adding AC in anaerobic digesters improved methane yield by at least double through the enrichment of bacteria and archaea. Conventional digestion process showed ability in removing certain types of ARGs, such as tetA, tetX, sul1, sul2, cmlA, floR, and intl1. Supplementing AC in anaerobic digester enhanced the removal of most of the ARGs in mono-digestion of food waste. The effects tended to be minimal in co-digestion of co-substrates such as chicken manure and waste activated sludge, both of which contain a certain amount of antibiotics. | 2018 | 29096147 |
| 7551 | 3 | 0.9999 | Microbial community evolution and fate of antibiotic resistance genes during sludge treatment in two full-scale anaerobic digestion plants with thermal hydrolysis pretreatment. Anaerobic digestion (AD) with thermal hydrolysis pretreatment is widely used as an efficient sludge treatment nowadays. However, the evolution of microbial community (especially for the archaea community), the fate of antibiotic resistance genes (ARGs), and their associations during such process in full-scale sludge treatment plants are rarely reported. Therefore, these scientific questions were explored at two full-scale sludge treatment plants through high-throughput sequencing and quantitative PCR. Results showed that Methanobacterium and Methanosphaera were the dominant archaea in thermal hydrolyzed sludge. The predominant bacteria in the sludge first shifted from nutrients removal functional bacteria to spore-forming bacteria after thermal hydrolysis, and then shifted to fermentative bacteria after AD. The full-scale plants could select ermB, ermF, mefA/E, qnrS and tetM. Though the bacteria and archaea biomass and community largely influenced the fate of ARGs, multiple linear regression analysis showed that the total ARGs were mainly affected by mobile genetic elements (MGEs). | 2019 | 31158777 |
| 7556 | 4 | 0.9999 | The fate and behavior mechanism of antibiotic resistance genes and microbial communities in anaerobic reactors treating oxytetracycline manufacturing wastewater. In this study, two parallel-operated expanded granular sludge bed (EGSB) reactors, one used to treat oxytetracycline (OTC) manufacturing wastewater with gradual increase of OTC concentration as experimental reactor and the other fed with the same wastewater without OTC as control reactor, were operated to investigate the behavior of antibiotics resistance genes (ARGs) and mobile genetic elements (MGEs) and their possible relationships with bacterial community among influent, sludge and effluent environments. Though the average absolute abundance of ARGs slightly decreased (0.26 - log), the ARGs' relative abundance normalized to 16S-rRNA gene copy numbers showed a significant upward trend in effluent (2 multiples - increase) and the absolute and relative abundances both extremely increased in anaerobic sludge, indicating that anaerobic treatment process cannot reduce ARGs efficiently, inversely can increase the risk of ARGs through the proliferation of antibiotics resistance bacteria (ARB) under the suppression of OTC. MGEs, bacterial communities and OTC concentration mainly impacted the ARGs profiles, which contributed 88.4% to the variation of ARGs. The differences and correlations of hosts in influent, effluent and sludge were further confirmed by network analysis. Overall, this study enhanced the understanding of the prevalence and transfer of ARGs in OTC production effluents during anaerobic treatment. | 2022 | 34740157 |
| 7546 | 5 | 0.9999 | Reduction of antibiotic resistance genes under different conditions during composting process of aerobic combined with anaerobic. Single aerobic or anaerobic composting is reportedly as inefficient for removing antibiotic resistance genes (ARGs) from animal manure. This study investigates the effects of ARG removal during aerobic combined with anaerobic composting (ACA) under different conditions. The results showed that a turning frequency of once a day, the proportion of swine manure and corn straw (3:1) and an anaerobic time of 6 days were the best operating conditions for ARG reduction (>95%, especially ermF: 99.78%) during ACA. Moreover, redundancy analysis demonstrated that the bacterial community (especially Truepera, Petrimonas and Ureibacillus) had a stronger effect on ARG removal than environmental factors did (especially temperature, total phosphorus, ammonia nitrogen and pH). Network analysis indicated the important effect of these potential host bacteria on the spread of ARGs through significant co-occurrence between individual ARGs and specific bacteria. These findings offer an effective technology to reduce and block ARGs spread from animal manure. | 2021 | 33493747 |
| 6919 | 6 | 0.9999 | Enhanced removal of antibiotic resistance genes during chicken manure composting after combined inoculation of Bacillus subtilis with biochar. This study explored the combined effects of Bacillus subtilis inoculation with biochar on the evolution of bacterial communities, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) during the composting of chicken manure. The results showed that B. subtilis inoculation combined with biochar increased bacterial abundance and diversity as well as prolonged the compost thermophilic period. Promoted organic matter biodegradation and facilitated the organic waste compost humification process, reduced the proliferation of ARGs by altering the bacterial composition. Firmicutes and Actinobacteriota were the main resistant bacteria related to ARGs and MGEs. The decrease in ARGs and MGEs was associated with the reduction in the abundance of related host bacteria. Compost inoculation with B. subtilis and the addition of biochar could promote nutrient transformation, reduce the increase in ARGs and MGEs, and increase the abundance of beneficial soil taxa. | 2024 | 37778803 |
| 8121 | 7 | 0.9998 | Bioleaching rather than chemical conditioning using Fe[III]/CaO or polyacrylamide mitigates antibiotic resistance in sludge composting via pre-removing antibiotic resistance genes and limiting horizontal gene transfer. Conditioning can drastically improve the dewaterability of sewage sludge and is widely practiced in most wastewater treatment plants (WWTPs). Sludge conditioning was also reported as a crucial step in sludge treatment to attenuate antibiotic resistance, but it remains unclear whether the attenuated antibiotic resistance by conditioning treatments would guarantee low abundance of antibiotic resistance genes (ARGs) in the compost products of municipal sewage sludge. Herein, the impacts of three conditioning treatments, including bioleaching and chemical conditioning using Fe[III]/CaO or polyacrylamide (PAM), on the abundances of 20 ARGs and 4 mobile genetic elements (MGEs) during conventional aerobic composting of dewatered sludge were investigated. It was found that the absolute and relative abundances of total ARGs in compost product of bioleached sludge accounted for only 13.8%-28.8% of that in compost products of un-conditioned, Fe[III]/CaO-conditioned, or PAM-conditioned sludges. Besides, bioleaching conditioning resulted in the lowest abundances of ARG subtypes and ARG-associated bacteria in the sludge compost product. The shift of ARG profiles in the bioleached sludge composting can be mainly ascribed to the ARG-associated bacteria, while the MGEs drove the ARG profiles during conventional composting of un-conditioned sludge and the two chemically conditioned sludge. Thus, bioleaching conditioning is superior to the chemical conditioning using Fe[III]/CaO or PAM in mitigating antibiotic resistance in sludge compost products, which was contributed by the pre-removal of ARGs prior to composting treatment and the potential limitation of ARGs transfer during conventional composting. | 2022 | 34749181 |
| 7544 | 8 | 0.9998 | Antibiotic resistance reduction mechanisms during thermophilic anaerobic digestion of microalgae-bacteria aggregates. Microalgae-bacteria-based systems are an emerging and promising approach for wastewater treatment plants (WWTP), having nutrient and antibiotic resistance removal comparable to conventional technologies. Still, antibiotic-resistance genes and bacteria (ARG and ARB) can proliferate in microalga-bacteria aggregates (MABA), a concern to control. Different temperature regimes of MABA continuous anaerobic digestion (AD), thermophilic (55 °C), and mesophilic (35 °C) were evaluated in this study as a strategy to eliminate ARB and ARGs. Plate counting techniques and metagenomic-based analysis revealed that thermophilic temperature had a better performance, achieving ARB log reductions of 1.1 to 1.7 for various antibiotics and significantly reduced ARG abundance up to 19.5 ± 0.8 ppm. The microbiome selection, the mobilome restriction, and directed functionality to thermal stress resistance were the main mechanisms responsible for resistome reduction at thermophilic conditions. Thermophilic AD effectively manages antibiotic resistance in microalgae-bacteria aggregates, which has important implications for wastewater treatment and reduces environmental risks. | 2025 | 39756663 |
| 7549 | 9 | 0.9998 | Fate of antibiotic resistance genes during sludge anaerobic fermentation: Roles of different sludge pretreatment. Excess sludge, the primary by-product of wastewater treatment plants, is the source and sink of antibiotic resistance genes (ARGs). Sludge pretreatments are an indispensable pathway to improve the resource recovery and harmfulness for anaerobic digestion sludge. However, fewer studies have compared the effects of different pretreatment technologies on the distribution of ARGs during anaerobic sludge digestion. Here, this study established seven anaerobic digesters, and four typical ARGs and one integrase gene of class 1 integron (intI1) regarded as the representative mobile genetic elements (MGEs) were examined during the whole anaerobic digestion process. It was found anaerobic digestion could effectively remove ARGs with about 70.86% removal rate of total ARGs. Among these pretreatments, the reduce efficiency of ARGs was the highest in 50 °C pretreatment, followed by oxidant, and the last was acid-alkaline. The microbial community analysis demonstrated the microbial community structure, including ARGs hosts and antibiotic resistant bacteria, was significantly changed and influenced by high temperature pretreatment. In addition, high temperature and K(2)S(2)O(8) observably decrease the level of ROS production. Macro transcriptome analysis indicated that sludge pretreatment, except for 50 °C pretreatment, up-regulated the genes relevant to lyases and transferase, but down-regulated the genes responsible for peroxidase, antioxidant enzymes and T4SS gene. This study emphasized and compared the different sludge pretreatments on the fate of ARGs in anaerobic sludge, and highlighted concerns regarding the environmental and health risks to our society. | 2024 | 39393457 |
| 8062 | 10 | 0.9998 | Nanoscale zero-valent iron inhibits the horizontal gene transfer of antibiotic resistance genes in chicken manure compost. Livestock manure has been identified as a significant hotspot for antibiotic resistance genes (ARGs). However, the impact of nanoscale zero-valent iron (nZVI) on the fate of ARGs during livestock manure composting remains poorly understood. Here, we investigated the evolution of ARGs in chicken manure compost exposed to 100 and 600 mg kg(-1) nZVI. The results showed that nZVI addition reduced the concentration of some antibiotics such as doxycycline and sulfamethoxazole. Furthermore, nZVI addition decreased the abundances of most ARGs at the end of composting, but nZVI dosage did not have any significant effect. The abundances of the dominant ARGs (sul1 and sul2) were significantly correlated to the class 1 integron-integrase gene (intI1). A network analysis revealed that the genera Bacteroides, Bacillus, Corynebacterium, Thiopseudomonas and Pseudomonas were the main potential hosts for multiple ARGs, and the decreased abundance of these bacteria contributed to the removal of ARGs. Structural equation models demonstrated that the reduction in intI1 played a predominant role in ARG removal. The nZVI also had direct effects on the intI1 abundance. These findings suggest that the addition of nZVI is a promising strategy to minimize ARG release in chicken manure compost. | 2022 | 34416685 |
| 8061 | 11 | 0.9998 | The addition of nano zero-valent iron during compost maturation effectively removes intracellular and extracellular antibiotic resistance genes by reducing the abundance of potential host bacteria. Applying compost to soil may lead to the spread of antibiotic resistance genes (ARGs) in the environment. Therefore, removing ARGs from compost is critical. In this study, for the first time, nano zero-valent iron (nZVI) was added to compost during the maturation stage to remove ARGs. After adding 1 g/kg of nZVI, the abundance of total intracellular and total extracellular ARGs was decreased by 97.62% and 99.60%, and that of total intracellular and total extracellular mobile genetic elements (MGEs) was decreased by 92.39% and 99.31%, respectively. A Mantel test and network analysis indicated that the reduction in potential host bacteria and intI1 after nZVI treatment promoted the removal of intracellular and extracellular ARGs. The addition of nZVI during composting reduced the horizontal transfer of ARGs and improve the total nitrogen and germination index of compost, allowing it to meet the requirements for organic fertilizers. | 2023 | 37352990 |
| 8060 | 12 | 0.9998 | Responses of bacterial communities and antibiotic resistance genes to nano-cellulose addition during pig manure composting. Treatment with exogenous additives during composting can help to alleviate the accumulation of antibiotic resistance genes (ARGs) caused by the direct application of pig manure to farmland. In addition, nano-cellulose has an excellent capacity for adsorbing pollutants. Thus, the effects of adding 300, 600, and 900 mg/kg nano-cellulose to compost on the bacterial communities, mobile genetic elements (MGEs), and ARGs were determined in this study. After composting, treatment with nano-cellulose significantly reduced the relative abundance of ARGs, which was lowest in the compost product with 600 mg/kg added nano-cellulose. Nano-cellulose inhibited the rebound in ARGs from the cooling period to the maturity period, and weakened the selective pressure of heavy metals on microorganisms by passivating bio-Cu. The results also showed that MGEs explained most of the changes in the abundances of ARGs, and MGEs had direct effects on ARGs. The addition of 600 mg/kg nano-cellulose reduced the abundances of bacterial genera associated with ermQ, tetG, and other genes, and the number of links (16) between ARGs and MGEs was lowest in the treatment with 600 mg/kg added nano-cellulose. Therefore, adding 600 mg/kg nano-cellulose reduced the abundances of ARGs by affecting host bacteria and MGEs. The results obtained in this study demonstrate the positive effect of nano-cellulose on ARG pollution in poultry manure, where adding 600 mg/kg nano-cellulose was most effective at reducing the abundances of ARGs. | 2021 | 34649327 |
| 8076 | 13 | 0.9998 | Effects of chlortetracycline and copper on tetracyclines and copper resistance genes and microbial community during swine manure anaerobic digestion. As antibiotic and heavy metals are over used in the livestock industry, animal manure is a reservoir of antibiotic resistance genes (ARGs). Anaerobic digestion has been reported to have the potential to reduce ARGs. However, few studies investigated whether reduction of ARGs would be affected by different external pressures including antibiotics and heavy metals during anaerobic digestion. The purpose of this study was thus to investigate effects of both chlortetracycline (CTC) and Cu on reduction of ARGs, heavy metal resistance genes (HMRGs) and mobile genetic elements (MGEs) during the swine manure anaerobic digestion. The results showed that the predominant ARGs (tetO, tetW, tetX, tetL) could be effectively reduced (approximately 1.00 log copies/g TS) through mesophilic anaerobic digestion. Microbial community evolution was the main driver. It was interesting that Treponema might indicate the termination of anaerobic digestion and compete with ARGs host bacteria. Addition of CTC, Cu and CTC+Cu affected microbial community change and hindered removal of ARGs, especially, CTC+Cu seriously affected Treponema and ARGs during anaerobic digestion. | 2017 | 28432950 |
| 7980 | 14 | 0.9998 | Effect of dissolved biochar on the transfer of antibiotic resistance genes between bacteria. The spread of antibiotic resistance genes (ARGs) is a global environmental issue. Dissolved biochar is more likely to contact bacteria in water, producing ecological risks. This study explored the effects of dissolved biochar on ARGs transfer in bacteria. Conjugative transfer efficiency was significantly different following treatment with different types of dissolved biochar. Typically, humic acid-like substance in dissolved biochar can significantly improve the transfer efficiency of ARGs between bacteria. When the concentration of dissolved biochar was ≤10 mg biochar/mL, humic acid-like substance substantially promoted ARGs transfer. An increase in dissolved biochar concentration weakened the ARGs transfer from humic acid-like substance. The inhibitory effects of small-molecule matters dominated, decreasing conjugative transfer frequency. At a concentration of 100 mg biochar/mL, the conjugative transfer efficiency of all treatments was lower than that of control. Compared with corn straw dissolved biochar, there were more transconjugants in pine sawdust dissolved biochar. Following treatment with 10 mg biochar/mL pine sawdust dissolved biochar, the number of transconjugants was at its maximum; approximately 7.3 folds higher than the control. We also explored mechanisms by which dissolved biochar impacts conjugative transfer. Due to the complex composition of dissolved biochar, its effects on the expression of conjugative transfer-related genes were also dynamic. This study investigates the ecological risk of biochar and guides its scientific application. | 2021 | 34274650 |
| 7553 | 15 | 0.9998 | Does the biological treatment or membrane separation reduce the antibiotic resistance genes from swine wastewater through a sequencing-batch membrane bioreactor treatment process. Swine wastes are the reservoir of antibiotic resistance genes (ARGs), which can potentially spread from swine farms to the environment. This study establishes a sequencing-batch membrane bioreactor (SMBR) for ARG removal from swine wastewater, and analyzes the effect of biological treatment and membrane separation on the ARG removal at different solid retention times (SRTs). The SMBR removed 2.91 logs (copy number) of ARGs at a short SRT (12 days). Raising the SRT reduced the removal rates of the detected genes by the biological treatment. Under the relative long SRT (30 days), ARGs and mobile genetic elements (MGEs) were maximized within the reactor and were well removed by membrane separation, with the average genes removal rate of 2.95 (copy number) and 1.18 logs (abundance). At the relatively low SRT, the biological treatment showed the dominant ARG removal effect, while the membrane separation took the advantages of ARG removal especially at the relatively long SRT. The ARG profile was related to the shift of the microbial community structure. The ARGs coexisted with the functional bacteria (ammonia oxidizing bacteria, nitrite oxidizing bacteria and denitrifiers), suggesting they are hosted by the functional bacteria. | 2018 | 29906721 |
| 8094 | 16 | 0.9998 | Additive quality influences the reservoir of antibiotic resistance genes during chicken manure composting. Aerobic composting is commonly used to dispose livestock manure and is an efficient way to reduce antibiotic resistance genes (ARGs). Here, the effects of different quality substrates on the fate of ARGs were assessed during manure composting. Results showed that the total relative abundances of ARGs and intI1 in additive treatments were lower than that in control, and high quality treatment with low C/N ratio and lignin significantly decreased the relative abundance of tetW, ermB, ermC, sul1 and sul2 at the end of composting. Additionally, higher quality treatment reduced the relative abundances of some pathogens such as Actinomadura and Pusillimonas, and some thermotolerant degrading-related bacteria comprising Pseudogracilibacillus and Sinibacillus on day 42, probably owing to the change of composting properties in piles. Structural equation models (SEMs) further verified that the physiochemical properties of composting were the dominant contributor to the variations in ARGs and they could also indirectly impact ARGs by influencing bacterial community and the abundance of intI1. Overall, these findings indicated that additives with high quality reduced the reservoir of antibiotic resistance genes of livestock manure compost. | 2021 | 34139628 |
| 7552 | 17 | 0.9998 | Effect of temperature on removal of antibiotic resistance genes by anaerobic digestion of activated sludge revealed by metagenomic approach. As antibiotic resistance continues to spread globally, there is growing interest in the potential to limit the spread of antibiotic resistance genes (ARGs) from wastewater sources. In particular, operational conditions during sludge digestion may serve to discourage selection of resistant bacteria, reduce horizontal transfer of ARGs, and aid in hydrolysis of DNA. This study applied metagenomic analysis to examine the removal efficiency of ARGs through thermophilic and mesophilic anaerobic digestion using bench-scale reactors. Although the relative abundance of various ARGs shifted from influent to effluent sludge, there was no measureable change in the abundance of total ARGs or their diversity in either the thermophilic or mesophilic treatment. Among the 35 major ARG subtypes detected in feed sludge, substantial reductions (removal efficiency >90%) of 8 and 13 ARGs were achieved by thermophilic and mesophilic digestion, respectively. However, resistance genes of aadA, macB, and sul1 were enriched during the thermophilic anaerobic digestion, while resistance genes of erythromycin esterase type I, sul1, and tetM were enriched during the mesophilic anaerobic digestion. Efflux pump remained to be the major antibiotic resistance mechanism in sludge samples, but the portion of ARGs encoding resistance via target modification increased in the anaerobically digested sludge relative to the feed. Metagenomic analysis provided insight into the potential for anaerobic digestion to mitigate a broad array of ARGs. | 2015 | 25994259 |
| 7555 | 18 | 0.9998 | Deciphering the factors influencing the discrepant fate of antibiotic resistance genes in sludge and water phases during municipal wastewater treatment. The discrepant fate of antibiotic resistance genes (ARGs) in sludge and water phases was investigated in a municipal wastewater treatment plant, and a lab-scale A(2)O-MBR was operated to provide background value of ARGs. The influencing factors of ARGs including microbial community, co-selection from heavy metals, biomass and horizontal gene transfer were concerned. Results showed that iA(2)O (inversed A(2)O) showed better ARGs reduction, and longer SRT (sludge retention time) increased ARGs relative abundance while reduced the gene copies of ARGs in the effluent, but significantly increased the ARGs in sludge phase. Compared to background value, the most enriched ARG was tetX in water phase, while it was intI1 in sludge phase. There existed higher abundance of multi-resistant bacteria in sludge phase, and microbial community determined the fate of ARGs in both water and sludge phase, while the direct effects from horizontal gene transfer should not be overlooked especially in water phase. | 2018 | 29909361 |
| 6915 | 19 | 0.9998 | Industrial-scale aerobic composting of livestock manures with the addition of biochar: Variation of bacterial community and antibiotic resistance genes caused by various composting stages. The presence of large amounts of antibiotic resistance genes (ARGs) in livestock manures poses an impending, tough safety risk to ecosystems. To investigate more comprehensively the mechanisms of ARGs removal from industrial-scale composting of livestock manure based on biochar addition, we tracked the dynamics of bacterial community and ARGs at various stages of aerobic composting of livestock manures with 10% biochar. There were no significant effects of biochar on the bacterial community and the profiles of ARGs. During aerobic composting, the relative abundance of ARGs and mobile genetic elements (MGEs) showed overall trends of decreasing and then increasing. The key factor driving the dynamics of ARGs was bacterial community composition, and the potential hosts of ARGs were Caldicoprobacter, Tepidimicrobium, Ignatzschineria, Pseudogracilibacillus, Actinomadura, Flavobacterium and Planifilum. The retention of the thermophilic bacteria and the repopulation of the initial bacteria were the dominant reasons for the increase in ARGs at maturation stage. Additionally, among the MGEs, the relative abundance of transposon gene was substantially removed, while the integron genes remained at high relative abundance. Our results highlighted that the suitability of biochar addition to industrial-scale aerobic composting needs to be further explored and that effective measures are needed to prevent the increase of ARGs content on maturation stage. | 2022 | 36162559 |