Evaluation of compost quality and the environmental effects of semipermeable membrane composting with poultry manure using sawdust or mushroom residue as the bulking agent.

Herein, the effects of different bulking agents (sawdust and mushroom residue), on compost quality and the environmental benefits of semipermeable film composting with poultry manure were investigated. The results show that composting with sawdust as the bulking agent resulted in greater efficiency and more cost benefits than composting with mushroom residue, and the cost of sawdust for treating an equal volume of manure was only 1/6 of that of mushroom residue. Additionally, lignin degradation and potential carbon emission reduction in the sawdust group were better than those in the mushroom residue group, and the lignin degradation efficiency of the bottom sample in the sawdust group was 48.57 %. Coupling between lignin degradation and potential carbon emission reduction was also closer in sawdust piles than in mushroom residue piles, and sawdust is more environmentally friendly. The abundance of key functional genes was higher at the bottom of each pile relative to the top and middle. Limnochordaceae, Lactobacillus and Enterococcus were the core microorganisms involved in coupling between lignin degradation and potential carbon emission reduction, and the coupled relationship was influenced by electric conductivity, ammonia nitrogen and total nitrogen in the compost piles. This study provides important data for supporting bulking agent selection in semipermeable film composting and for improving the composting process. The results have high value for compost production and process application.

A sustainable and economic strategy to reduce risk antibiotic resistance genes during poultry manure bioconversion by black soldier fly Hermetia illucens larvae: Larval density adjustment.

Black soldier fly (Hermetia illucens) larvae (BSFL) are common insects that are known for bioconversion of organic waste into a sustainable utilization resource. However, a strategy to increase antibiotic resistance gene (ARG) elimination in sustainable and economic ways through BSFL is lacking. In the present study, different larval densities were employed to assess the mcr-1 and tetX elimination abilities, and potential mechanisms were investigated. The application and economic value of each larval density were also analyzed. The results showed that the 100 larvae cultured in 100 g of manure group had the best density because the comprehensive disadvantage evaluation ratio was the lowest (14.97%, good bioconversion manure quality, low ARG deposition risk and reasonable larvae input cost). Further investigation showed that mcr-1 could be significantly decreased by BSFL bioconversion (4.42 ×107 copies/g reduced to 4.79 ×106-2.14 ×105 copies/g)(P＜0.05); however, mcr-1 was increasingly deposited in the larval gut with increasing larval density. The tetX abundance was stabilized by BSFL bioconversion, except that the abundance at the lowest larval density increased (1.22 ×1010 copies/g increase, 34-fold). Escherichia was the host of mcr-1 and tetX in all samples, especially in fresh manure; Alcaligenes was the host of tetX in bioconversion manure; and the abundance of Alcaligenes was highly correlated with the pH of bioconversion manure. The pH of bioconversion manure was extremely correlated with the density of larvae. Klebsiella and Providencia were both hosts of tetX in the BSF larval gut, and Providencia was also the host of mcr-1 in the BSF larval gut. The density of larvae influenced the bioconversion manure quality and caused the ARG host abundance to change to control the abundance of ARGs, suggesting that larval density adjustment was a useful strategy to manage the ARG risk during BSFL manure bioconversion.

The relationship between culturable doxycycline-resistant bacterial communities and antibiotic resistance gene hosts in pig farm wastewater treatment plants.

Pig farm wastewater treatment plants (WWTPs) are an important repository for resistant bacterial communities (RBCs) and antibiotic resistance genes (ARGs). However, the relationship between RBCs and ARG hosts has not been well characterized. In this study, water samples from influent and effluent from five pig farm WWTPs were collected. Gradient concentrations of doxycycline (DOX) were used to screen the culturable RBCs. The abundance of 21 subtypes of ARGs and the bacterial community were investigated. This study detected a large number of culturable DOX-RBCs and ARGs in the influent and effluent of pig farm WWTPs. The abundances of ARGs and RBCs in all effluent samples was significantly lower than that in the influent samples (P < 0.05), which indicated that the WWTPs can effectively remove most ARGs and RBCs in pig farm wastewater. The main potential culturable RBCs in pig farm wastewater were the dominant bacteria Proteobacteria, Actinobacteria, Pseudomonas, and Rheinheimera. However, most of the ARGs were mainly present in Bacteroidetes, Actinobacteria, Corynebacteriaceae, Macellibacteroides, Acinetobacter, and Enterobacteriaceae, which are considered potential ARG hosts. The results presented here showed that there were obvious differences between the species of culturable DOX-RBCs and ARG hosts in the pig farm WWTPs, which may be due to various environmental factors. This highlights the urgent need for further research on the relationship between RBCs and ARG hosts.

Potential regulation of small RNAs on bacterial function activities in pig farm wastewater treatment plants.

Small RNAs (sRNAs) are key players in the regulation of bacterial gene expression. However, the distribution and regulatory functions of sRNA in pig farm wastewater treatment plants (WWTPs) remains unknown. In this study, the wastewaters in anoxic and oxic tanks of the WWTPs were collected. The profiles of the community structure, mRNA expression, and sRNA expression of bacteria in pig farm wastewater were investigated using transcriptome sequencing and qPCR. This study demonstrated that there was a higher abundance of sRNA in the pig farm WWTPs and 52 sRNAs were detected. The sRNAs were mainly present in Proteobacteria and Firmicutes, including the potential human pathogenic bacteria (HPB) (Escherichia, Shigella, Bordetella and Morganella), crop pathogen (Pectobacterium) and denitrifying bacteria (Zobellella). And the sRNAs were involved in the bacterial functional activities such as translation, transcription, drug resistance, membrane transport and amino acid metabolism. In addition, most sRNAs had a higher abundance in anoxic tanks which contained a higher abundance of the genes associated with infectious diseases and drug resistance than that in oxic tanks. The results presented here show that in pig farm WWTPs, sRNA played an important role in bacterial function activities, especially the infectious diseases, drug resistance and denitrification, which can provide a new point of penetration for improving the pig farm WWTPs.

The microbiota structure in the cecum of laying hens contributes to dissimilar H2S production.

BACKGROUND: Host genotype plays a crucial role in microbial composition of laying hens, which may lead to dissimilar odor gas production. The objective of this study was to investigate the relationship among layer breed, microbial structure and odor production. RESULTS: Thirty Hy-Line Gray and thirty Lohmann Pink laying hens were used in this study to determine the impact of cecal microbial structure on odor production of laying hens. The hens were managed under the same husbandry and dietary regimes. Results of in vivo experiments showed a lower hydrogen sulfide (H2S) production from Hy-Line hens and a lower concentration of soluble sulfide (S2-) but a higher concentration of butyrate in the cecal content of the Hy-Line hens compared to Lohmann Pink hens (P < 0.05), which was consistent with the in vitro experiments (P < 0.05). However, ammonia (NH3) production was not different between genotypes (P > 0.05). Significant microbial structural differences existed between the two breed groups. The relative abundance of some butyrate producers (including Butyricicoccus, Butyricimonas and Roseburia) and sulfate-reducing bacteria (including Mailhella and Lawsonia) were found to be significantly correlated with odor production and were shown to be different in the 16S rRNA and PCR data between two breed groups. Furthermore, some bacterial metabolism pathways associated with energy extraction and carbohydrate utilization (oxidative phosphorylation, pyruvate metabolism, energy metabolism, two component system and secretion system) were overrepresented in the Hy-Line hens, while several amino acid metabolism-associated pathways (amino acid related enzymes, arginine and proline metabolism, and alanine-aspartate and glutamate metabolism) were more prevalent in the Lohmann hens. CONCLUSION: The results of this study suggest that genotype of laying hens influence cecal microbiota, which in turn modulates their odor production. Our study provides references for breeding and enteric manipulation for defined microbiota to reduce odor gas emission.

The correlation between antibiotic resistance gene abundance and microbial community resistance in pig farm wastewater and surrounding rivers.

Antimicrobial resistance gene (ARG) abundance and microbial resistance (MR) are often used as important indicators of pollution risk; however, the relationship between ARGs abundance and MR in pig farm wastewater remains unknown. In this study, the raw pig farm wastewater, effluent water, upstream river water, domestic wastewater and downstream river water samples were collected. The concentration of 20 subtypes of ARGs and 2 integrons, minimal inhibit concentration (MIC), and bacterial communities were investigated. In this study, 20 subtypes of ARGs and integrons were detected in all sampling sites. The highest abundance of 17 of the 20 subtypes of ARGs was detected in raw pig farm wastewater, and ermA had the maximum average abundance of 108 copies/mL, with up to 2.41 ±â€¯0.12 × 108 copies/mL. There was no significant correlation between MR to three antibiotics (ciprofloxacin, streptomycin and tetracycline hydrochloride) and the abundance of their corresponding ARGs (P > 0.05), and a large difference was detected between the types of ARGs co-occur bacteria and resistance co-occur bacteria in the 5 sampling sites. And the pig farm wastewater treatment (WWT) could effectively reduce the ARGs and MR to the 3 antibiotics. The results presented here show that there may be no obvious correlation between ARGs and MCR in pig farm wastewater and surrounding rivers, which may be due to various environmental factors, highlighting the urgent need for a comprehensive evaluation of relationship between ARGs abundance and MR.

In vitro assessment on effect of duodenal contents on the lead (Pb2+) binding capacity of two probiotic bacterial strains.

In vitro Lead (Pb2+) binding capacity of two probiotic bacteria strains, namely Bifidobacterium longumBB79 and Lactobacillus pentosusITA23, was assessed following incubation with the intestinal contents (IC) of laying hens. Results of this study demonstrated that IC treatment significantly enhanced (P<0.01) Pb2+ binding capacity of both bacterial strains. Fourier transform infrared analysis indicated that several functional groups (O-H or N-H, C-H, CË­O, C-O, and C-O-C) on the bacteria cell wall involved in metal ion binding were altered after IC incubation, and new groups appeared between the 3700cm-1 and 4000cm-1bands. Transmission electron microscopy demonstrated that after incubation with IC, unidentified IC components created new binding sites on the bacterial cell surface. These particles also changed the mechanism of Pb2+ binding of the two strains from intracellular accumulation to extracellular adsorption.

Multiomics analysis of the effects of manure-borne doxycycline combined with oversized fiber microplastics on pak choi growth and the risk of antibiotic resistance gene transmission.

In this study, oversized microplastics (OMPs) were intentionally introduced into soil containing manure-borne doxycycline (DOX). This strategic approach was used to systematically examine the effects of combined OMP and DOX pollution on the growth of pak choi, analyze alterations in soil environmental metabolites, and explore the potential migration of antibiotic resistance genes (ARGs). The results revealed a more pronounced impact of DOX than of OMPs. Slender-fiber OMPs (SF OMPs) had a more substantial influence on the growth of pak choi than did coarse-fiber OMPs (CF OMPs). Conversely, CF OMPs had a more significant effect on the migration of ARGs within the system. When DOX was combined with OMPs, the negative effects of DOX on pak choi growth were mitigated through the synthesis of indole through the adjustment of carbon metabolism and amino acid metabolism in pak choi roots. In this process, Pseudohongiellaceae and Xanthomonadaceae were key bacteria. During the migration of ARGs, the potential host bacterium Limnobacter should be considered. Additionally, the majority of potential host bacteria in the pak choi endophytic environment were associated with tetG. This study provides insights into the intricate interplay among DOX, OMPs, ARGs, plant growth, soil metabolism, and the microbiome.

Biosafety assessment of laying hens fed different treatments of black soldier flies (Hermetia illucens) under doxycycline stress.

The black soldier fly (BSF, Hermetia illucens) is a resource insect that can utilize livestock and poultry feces. However, BSFs may also increase the risk of transmission of antibiotic resistance genes (AGRs) that are widespread in livestock and poultry farm environments. Therefore, we aimed to evaluate the biosecurity risks of different BSF treatments in the laying chicken food chain using the "chicken manure-BSF-laying hens" model. Our results indicated that different BSF treatments significantly affected antibiotic residue, ARGs, MGEs, bacterial antibiotic resistance, and bacterial microbial community composition in the food chain of laying hens fed BSFs. These risks can be effectively reduced through starvation treatment and high-temperature grinding treatment. Comprehensive risk assessment analysis revealed that starvation combined with high-temperature milling (Group H) had the greatest effect.

Highly Stretchable, Tissue-like Ag Nanowire-Enhanced Ionogel Nanocomposites as an Ionogel-Based Wearable Sensor for Body Motion Monitoring.

The development of wearable electronic devices for human health monitoring requires materials with high mechanical performance and sensitivity. In this study, we present a novel transparent tissue-like ionogel-based wearable sensor based on silver nanowire-reinforced ionogel nanocomposites, P(AAm-co-AA) ionogel-Ag NWs composite. The composite exhibits a high stretchability of 605% strain and a moderate fracture stress of about 377 kPa. The sensor also demonstrates a sensitive response to temperature changes and electrostatic adsorption. By encapsulating the nanocomposite in a polyurethane transparent film dressing, we address issues such as skin irritation and enable multidirectional stretching. Measuring resistive changes of the ionogel nanocomposite in response to corresponding strain changes enables its utility as a highly stretchable wearable sensor with excellent performance in sensitivity, stability, and repeatability. The fabricated pressure sensor array exhibits great proficiency in stress distribution, capacitance sensing, and discernment of fluctuations in both external electric fields and stress. Our findings suggest that this material holds promise for applications in wearable and flexible strain sensors, temperature sensors, pressure sensors, and actuators.

Digital automation of transdermal drug delivery with high spatiotemporal resolution.

Transdermal drug delivery is of vital importance for medical treatments. However, user adherence to long-term repetitive drug delivery poses a grand challenge. Furthermore, the dynamic and unpredictable disease progression demands a pharmaceutical treatment that can be actively controlled in real-time to ensure medical precision and personalization. Here, we report a spatiotemporal on-demand patch (SOP) that integrates drug-loaded microneedles with biocompatible metallic membranes to enable electrically triggered active control of drug release. Precise control of drug release to targeted locations (<1 mm2), rapid drug release response to electrical triggers (<30 s), and multi-modal operation involving both drug release and electrical stimulation highlight the novelty. Solution-based fabrication ensures high customizability and scalability to tailor the SOP for various pharmaceutical needs. The wireless-powered and digital-controlled SOP demonstrates great promise in achieving full automation of drug delivery, improving user adherence while ensuring medical precision. Based on these characteristics, we utilized SOPs in sleep studies. We revealed that programmed release of exogenous melatonin from SOPs improve sleep of mice, indicating potential values for basic research and clinical treatments.

Skin-inspired, sensory robots for electronic implants.

Drawing inspiration from cohesive integration of skeletal muscles and sensory skins in vertebrate animals, we present a design strategy of soft robots, primarily consisting of an electronic skin (e-skin) and an artificial muscle. These robots integrate multifunctional sensing and on-demand actuation into a biocompatible platform using an in-situ solution-based method. They feature biomimetic designs that enable adaptive motions and stress-free contact with tissues, supported by a battery-free wireless module for untethered operation. Demonstrations range from a robotic cuff for detecting blood pressure, to a robotic gripper for tracking bladder volume, an ingestible robot for pH sensing and on-site drug delivery, and a robotic patch for quantifying cardiac function and delivering electrotherapy, highlighting the application versatilities and potentials of the bio-inspired soft robots. Our designs establish a universal strategy with a broad range of sensing and responsive materials, to form integrated soft robots for medical technology and beyond.

Sodium butyrate reduces ammonia production in the cecum of laying hens by regulating ammonia-producing bacteria.

Sodium butyrate is a commonly used feed additive and can reduce ammonia (NH3) emissions from laying hens, but the mechanism of this effect is unknown. In this study, the sodium butyrate and cecal content of Lohmann pink laying hens were measured, and in vitro fermentation experiments and NH3-producing bacteria coculture experiments were carried out to explore the relationship between NH3 emissions and its associated microbiota metabolism. Sodium butyrate was found to significantly reduce NH3 emission from the cecal microbial fermentation of Lohmann pink laying hens (P < 0.05). The concentration of NO3--N in the fermentation broth of the sodium butyrate-supplemented group increased significantly, and the concentration of NH4+-N decreased significantly (P < 0.05). Moreover, sodium butyrate significantly reduced the abundance of harmful bacteria and increased the abundance of beneficial bacteria in the cecum. The culturable NH3-producing bacteria consisted mainly of Escherichia and Shigella, such as Escherichia fergusonii, Escherichia marmotae and Shigella flexnerii. Among them, E. fergusonii had the highest potential for NH3 production. The coculture experiment showed that sodium butyrate can significantly downregulate the expression of the lpdA, sdaA, gcvP, gcvH and gcvT genes of E. fergusonii (P < 0.05), thus reducing the NH3 emission produced by the bacteria during metabolism. In general, sodium butyrate regulated NH3-producing bacteria to reduce NH3 production in the cecum of laying hens. These results are of great significance for NH3 emission reduction in the layer breeding industry and for future research.

Effects of the oversized microplastic pollution layer on soil aggregates and organic carbon at different soil depths.

Soil aggregates (SAs) are the main site for soil organic carbon (SOC) fixation, and land plastic pollution is increasingly causing many soil problems. The effects of plastic on SAs and SOC seem to be significant, but there is still a lack of relevant research. This study investigated the effects of the "plastic contamination layer" (PCL) formed by the microplastic precursors (namely, oversized microplastics (OMPs)) on the content and properties of SAs of different particle sizes at different soil depths. The results showed that the PCL had an effect on SAs of different sizes at different depths: Compared with the control group, PCL mainly increased the content of SAs in 0-5 cm soil depth, about 28.08 mg macroaggregates, 13.79 mg microaggregates and 59.82 mg silt and clay aggregates per gram of soil. The presence of the PCL mainly down-regulates the organic carbon (OC) content in 0-5 cm macroaggregates, which is about 9.59 g/kg, the OC content in 10-20 cm microaggregates, which is about 16.41 g/kg, and the OC content in 0-5 cm silt and clay aggregates, which is about 4.16 g /kg, downregulated the expression of the key carbon metabolism genes (CMGs) coxL, and inhibited the contribution of the potential CMGs host bacteria Sphaerimonospora and Bacteroides to soil organic matter. This paper emphasizes that the presence of PCL reduced SOC sequestration.

The effect of manure-borne doxycycline combined with different types of oversized microplastic contamination layers on carbon and nitrogen metabolism in sandy loam.

The different forms and properties of microplastics (MPs) have different effects on the elemental cycles in soil ecosystems, and this is further complicated when the soil contains antibiotics; meanwhile, oversized microplastic (OMP) in soil is always ignored in studies of environmental behavior. In the context of antibiotic action, the effects of OMP on soil carbon (C) and nitrogen (N) cycling have rarely been explored. In this study, we created four types of oversized microplastic (thick fibers, thin fibers, large debris, and small debris) composite doxycycline (DOX) contamination layers (5-10 cm) in sandy loam, hoping to reveal the effects on soil C and N cycling and potential microbial mechanisms when exposed to the combination of manure-borne DOX and different types of OMP from the perspective of metagenomics in the longitudinal soil layer (0-30 cm). The results showed that all different forms of OMP, when combined with DOX, reduced the soil C content in each layer, but only reduced the soil N content in the upper layer of the OMP contamination layer. The microbial structure of the surface soil (0-10 cm) was more noteworthy than that of the deeper soil (10-30 cm). The genera Chryseolinea and Ohtaekwangia were key microbes involved in C and N cycling in the surface layer and regulated carbon fixation in photosynthetic organisms (K00134), carbon fixation pathways in prokaryotes (K00031), methane metabolism (K11212 and K14941), assimilatory nitrate reduction (K00367), and denitrification (K00376 and K04561). The present study is the first to reveal the potential microbial mechanism of C and N cycling under OMP combined with DOX in different layers, mainly the OMP contamination layer and its upper layer, and the OMP shape plays an important role in this process.

Effects of microplastics on functional genes related to CH4 and N2O metabolism in bacteriophages during manure composting and its planting applications.

Microplastics (MPs), as a new type of pollutant, widely exist in livestock and poultry breeding and agricultural soils. However, research on MPs pollution on greenhouse gas emissions in combined planting and breeding systems is lacking, especially from the perspective of phage horizontal gene transfer. Therefore, this paper explores the effects of MPs on functional genes related to CH4 and N2O metabolism in bacteriophages during manure composting and its planting applications. The results of the study indicated that the addition of MPs had an impact on both the physicochemical properties and microbial community structure of manure during the composting process and on the compost-applied rhizosphere soil of lactuca (Lactuca sativa). Specifically, on day 7 of composting, mcrA/pmoA and (nirS+nirK) levels in bacteria in the MP group significantly increased. Additionally, it was observed that the MP group had higher average temperatures during the high-temperature period of composting, which led to a rapid reduction in phages. However, the phage levels quickly recovered during the cooling period. Furthermore, the addition of MPs to the rhizosphere soil resulted in higher levels of nirK. These changes may affect greenhouse gas emissions.

Skin-inspired, sensory robots for electronic implants.

Living organisms with motor and sensor units integrated seamlessly demonstrate effective adaptation to dynamically changing environments. Drawing inspiration from cohesive integration of skeletal muscles and sensory skins in these organisms, we present a design strategy of soft robots, primarily consisting of an electronic skin (e-skin) and an artificial muscle, that naturally couples multifunctional sensing and on-demand actuation in a biocompatible platform. We introduce an in situ solution-based method to create an e-skin layer with diverse sensing materials (e.g., silver nanowires, reduced graphene oxide, MXene, and conductive polymers) incorporated within a polymer matrix (e.g., polyimide), imitating complex skin receptors to perceive various stimuli. Biomimicry designs (e.g., starfish and chiral seedpods) of the robots enable various motions (e.g., bending, expanding, and twisting) on demand and realize good fixation and stress-free contact with tissues. Furthermore, integration of a battery-free wireless module into these robots enables operation and communication without tethering, thus enhancing the safety and biocompatibility as minimally invasive implants. Demonstrations range from a robotic cuff encircling a blood vessel for detecting blood pressure, to a robotic gripper holding onto a bladder for tracking bladder volume, an ingestible robot residing inside stomach for pH sensing and on-site drug delivery, and a robotic patch wrapping onto a beating heart for quantifying cardiac contractility, temperature and applying cardiac pacing, highlighting the application versatilities and potentials of the nature-inspired soft robots. Our designs establish a universal strategy with a broad range of sensing and responsive materials, to form integrated soft robots for medical technology and beyond.

Transmission of tetracycline resistance genes and microbiomes from manure-borne black soldier fly larvae frass to rhizosphere soil and pakchoi endophytes.

Manure treatment with black soldier fly larvae (BSFL) and BSFL frass application in crop land is a sustainable strategy; however, whether residual antibiotic resistance genes (ARGs) and their transmission risk are related to the manure BSFL treatment process is still unknown. In this paper, the effect of BSFL addition density on residual tetracycline resistance genes (TRGs) and transmission from frass to pakchoi was determined. The results showed that BSFL frass can provide sufficient nutrients for growth, improve the economic value of pakchoi, and reduce the risk of transmission of TRGs in chicken manure regardless of BSFL density. The potential hosts of the TRGs we detected were found in BSFL frass (Oblitimonas and Tissierella), rhizosphere soil (Mortierella and Fermentimonas), and pakchoi endophytes (Roseomonas). The present study concluded that BSFL frass produced by adding 100 BSFL per 100 g of chicken manure has the advantages of high value and low risk. These findings will provide important strategic guidance for animal manure disposal and theoretical support for preventing the transmission of TRGs in BSFL applications.

Abundance, diversity and diffusion of antibiotic resistance genes in cat feces and dog feces.

The ARG profiles in pet feces, such as cat and dog feces, and their potential threat to environmental safety are still unclear. In this study, ARGs in 45 cat and 28 dog fecal samples were detected, and a diffusion experiment was performed to assess the risk of ARGs diffusion into the air. The results showed that the abundances of ARGs in cat feces and dog feces were high, and the abundance in dog feces (0.89 ± 0.17 copies/bacterial cell) was significantly higher than that in cat feces (0.46 ± 0.09 copies/bacterial cell) (P < 0.05). The bacterial community, especially Firmicutes and Desulfobacterota in cat feces, and Proteobacteria in dog feces, was the main factor affecting the variation in the ARG profiles, contributing to 31.6% and 32.4% of the variation in cat feces and dog feces, respectively. Physicochemical factors (especially NH4+-N) and age also indirectly affected the variation in the ARG profiles by affecting the bacterial community. In addition, the ARGs in cat feces and dog feces diffused into the air, but there was no evidence that this diffusion posed a threat to environmental safety and human health. These results can provide reference data for healthy animal breeding and the prevention and control of ARG pollution.

The fate of antibiotic resistance genes and their association with bacterial and archaeal communities during advanced treatment of pig farm wastewater.

Advanced wastewater treatment plants are widely used in most large-scale pig farms in southern China. However, the fate of antibiotic resistance genes (ARGs) and their association with bacterial and archaeal communities during advanced wastewater treatment remain unclear. In this study, the profiles of ARGs in typical advanced wastewater treatment plants were surveyed using metagenomic analysis. The results showed that 279- 326 different subtypes of ARGs were detected in raw wastewater, with a total abundance of 5.98 ± 0.48 copies per bacterial cell. During the advanced wastewater treatment, the abundance and number of ARGs were significantly reduced. Microbial communities (bacteria and archaea) contributed the most to the variation in ARG abundance and composition (PCA axis_1), accounting for 10.8 % and 15.7 %, respectively, followed by mobile genetic elements (MGEs) and physicochemical factors. Special attention should be given to potential pathogenic bacteria such as Escherichia, Streptococcus, Enterococcus and Staphylococcus and archaea such as Methanocorpusculum, Candidatus Methanoplasma and Candidatus Methanomethylophilus, which were important potential ARG hosts. Bacterial communities may indirectly affect ARG variation by affecting archaeal communities. These findings indicated that ARG levels in pig farm wastewater can be effectively reduced during advanced treatment and highlighted the important role played by archaea, which should not be ignored.