The role of gut microbiota in mediating increased toxicity of nano-sized polystyrene compared to micro-sized polystyrene in mice.

Microplastics (MPs) are widespread environmental contaminants that have been detected in animals and humans. However, their toxic effects on terrestrial mammals and the underlying mechanisms are still not well understood. Herein, we explored the role of gut microbiota in mediating the toxicity of micro- and nano-sized polystyrene plastics (PS-MPs/PS-NPs) using an antibiotic depleted mice model. The results showed that PS-MPs and PS-NPs exposure disrupted the composition and structure of the gut microbiota. Specifically, these particles led to an increase in pathogenic Esherichia-shigella, while depleting probiotics such as Akkermansia and Lactobacillus. Comparatively, PS-NPs particles had more pronounced effect, leading to obviously shifted the colon transcriptional profiles characterized by inducing the enrichment of colon metabolism and immune-related pathways (i.e., upregulated in genes like udgh, ugt1a1, ugt1a6a, ugt1a7c and ugt2b34). Additionally, both PS-MPs and PS-NPs induced oxidative stress, gut-liver damage and systemic inflammation in mice. Mechanistically, we confirmed that PS particles disturbed gut microbiota, activating TLR2-My88-NF-κB pathway to trigger the release of inflammatory cytokine IL-1ß and TNF-α. The damage and inflammation caused by both size of PS particles was alleviated when the gut microbiota was depleted. In conclusion, our findings deepen the understanding of the molecule mechanisms by which gut microbiota mediate the toxicity of PS particles, informing health implications of MPs pollution.

Landscape and risk assessment of microplastic contamination in farmed oysters and seawater along the coastline of China.

Microplastic (MP) pollution poses a significant threat to marine ecosystem and seafood safety. However, comprehensive and comparable assessments of MP profiles and their ecological and health in Chinese farming oysters are lacking. This study utilized laser infrared imaging spectrometer (LDIR) to quantify MPs in oysters and its farming seawater at 18 sites along Chinese coastlines. Results revealed a total of 3492 MPs in farmed oysters and seawater, representing 34 MP types, with 20-100 µm MP fragments being the dominant. Polyurethane (PU) emerged as the predominant MP type in oysters, while polysulfones were more commonly detected in seawater. Notably, oysters from the Bohai Sea exhibited a higher abundance of MPs (13.62 ± 2.02 items/g) and estimated daily microplastic intake (EDI, 2.14 ± 0.26 items/g/kg·bw/day), indicating a greater potential health risk in the area. Meanwhile, seawater from the Yellow Sea displayed a higher level (193.0 ± 110.7 items/L), indicating a greater ecological risk in this region. Given the pervasiveness and abundance of PU and its high correlation with other MP types, we proposed PU as a promising indicator for monitoring and assessing the risk MP pollution in mariculture in China. These findings provide valuable insights into the extent and characteristics of MP pollution in farmed oysters and seawater in China.

Dissemination of antibiotic resistance genes from aboveground sources to groundwater in livestock farms.

Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are prevalent in various environments on livestock farms, including livestock waste, soil, and groundwater. Contamination of groundwater by ARB and ARGs in livestock farms is a growing concern as it may have potentially huge risks to human health. However, the source of groundwater-borne ARB and ARGs in animal farms remains largely unknown. In this study, different types of samples including groundwater and its potential contamination sources from aboveground (pig feces, wastewater, and soil) from both working and abandoned swine feedlots in southern China were collected and subjected to metagenomic sequencing and ARB isolation. The source tracking based on metagenomic analysis revealed that 56-95 % of ARGs in groundwater was attributable to aboveground sources. Using metagenomic assembly, we found that 45 ARGs predominantly conferring resistance to aminoglycosides, sulfonamides, and tetracyclines could be transferred from the aboveground sources to groundwater, mostly through plasmid-mediated horizontal gene transfer. Furthermore, the full-length nucleotide sequences of sul1, tetA, and TEM-1 detected in ARB isolates exhibited the close evolutionary relationships between aboveground sources and groundwater. Some isolated strains of antibiotic-resistant Pseudomonas spp. from aboveground sources and groundwater had the high similarity (average nucleotide identity > 99 %). Notably, the groundwater-borne ARGs were identified as mainly carried by bacterial pathogens, potentially posing risks to human and animal health. Overall, this study underscores the dissemination of ARGs from aboveground sources to groundwater in animal farms and associated risks.

Response characteristics of denitrifying bacteria and denitrifying functional genes to woody peat during pig manure composting.

This study aimed to explore the impacts of adding different proportions of woody peat (WP) (0%(CK), 5%(T1), and 15%(T2)) on denitrification during composting. The results demonstrated that compared with CK, T1 and T2 increased the total Kjeldahl nitrogen content (8% and 14%, respectively) and reduced the nitrate nitrogen (7% and 23%) content after composting. After composting, the abundances of nirK and nirS decreased by 4-9% and 33-35% under T1 and T2, respectively. Adding 15% WP reduced the abundances of key denitrifying bacteria such as Pseudomonas, Pusillimonas, Achromobacter, and Rhizobiales by 5-90%. The main factors that affected denitrification genes were the carbon content, nitrogen form (nitrite nitrogen and ammonium nitrogen), and denitrifying bacteria community. In summary, adding 15% WP has the best ability to reduce nitrogen loss by decreasing the abundances of denitrifying bacteria and denitrifying functional genes, thereby improving the agricultural value of composting products.

Exploring the mechanism associated with methane emissions during composting: Inoculation with lignocellulose-degrading microorganisms.

Inoculation with microorganisms is an effective strategy for improving traditional composting processes. This study explored the effects of inoculation with lignocellulose-degrading microorganisms (LDM) on the degradation of organic matter (OM), methane (CH4) emissions, and the microbial community (bacteria and methanogens) during composting. The results showed that LDM accelerated the degradation of OM (including the lignocellulose fraction) and increased the CH4 releases in the later thermophilic and cooling stages during composting. At the ending of composting, LDM increased the CH4 emissions by 38.6% compared with the control. Moreover, LDM significantly increased the abundances of members of the bacterial and methanogenic community during the later thermophilic period (P < 0.05). In addition, LDM promoted the growth and activity of major bacterial genera (e.g., Ureibacillus) with the ability to degrade macromolecular OM, as well as affecting key methanogens (e.g., Methanocorpusculum) in the composting system. Network analysis and variance partitioning analysis indicated that OM and temperature were the main factors that affected the bacterial and methanogen community structures. Structural equation modeling demonstrated that the higher CH4 emissions under LDM were related to the growth of methanogens, which was facilitated by the anaerobic environment produced by large amounts of CO2. Thus, aerobic conditions should be improved during the end of the thermophilic and cooling composting period when inoculating with lignocellulose-degrading microorganisms in order to reduce CH4 emissions.

Effects and microbial mechanisms of phosphogypsum and medical stone on organic matter degradation and methane emissions during swine manure composting.

The degradation of organic matter (OM) and CH4 emissions during composting greatly influence the composting efficiency and greenhouse effect. This study evaluated the effects of adding phosphogypsum (PPG) and medical stone (MS) on OM breakdown, CH4 emissions, and their underlying mechanisms. MS accelerated the breakdown of OM in the early composting stage, whereas PPG increased it in the cooling and maturation periods. At the ending of composting, humification was also significantly promoted by PPG and MS (P < 0.05). Moreover, MS and PPG reduced CH4 emissions by 27.64% and 23.12%, respectively, and significantly inhibited the activities of methanogens in terms of their abundance (mcrA) and composition (dominant genera such as Methanobrevibacter, Methanocorpusculum, and Methanothermus) (P < 0.05). Interestingly, MS enhanced the activity of enzymes and bacterial metabolism related to OM degradation in the early composting stage, whereas PPG promoted them during the cooling and maturity stages. MS and PPG inhibited the activities of enzymes related to CH4 release during the cooling and maturity stages. Therefore, PPG and MS may have influenced OM degradation and CH4 releases during composting via changes in bacterial metabolism and enzyme activity levels. PPG and MS could have altered the activities of methanogens to influence the transformation of carbon and CH4 emissions according to network analysis and partial least-squares path modeling analysis. These findings provide insights at the molecular level into the effects of adding PPG and MS on OM degradation and CH4 emissions during composting, thereby facilitating the application of PPG and MS in composting systems.

Dynamics and key drivers of antibiotic resistance genes during aerobic composting amended with plant-derived and animal manure-derived biochars.

Plant-derived and animal manure-derived biochars have been used to improve the quality of compost but the differences in their effects on antibiotic resistance genes (ARGs) during composting are unclear. This study selected two types of biochar (RB and PB) produced from abundant agricultural waste to be added to the compost. Adding plant-derived RB performed better in ARGs, mobile genetic elements, and human pathogenic bacteria removal during aerobic composting, whereas adding manure-derived PB even increased ARGs abundance. Vertical gene transfer was possibly the key mechanism for persistent ARGs, and easily removed ARGs were regulated by horizontal and vertical gene transfer. Adding plant-derived RB reduced the abundances of persistent ARG hosts (e.g., Pseudomonas and Longispora) and ARG-related metabolic pathways and genes. The higher nitrogen content of manure-derived PB may have promoted the proliferation of ARG hosts. Overall, adding manure-derived biochar during composting may not be the optimal option for eliminating ARGs.

Risk of horizontal transfer of intracellular, extracellular, and bacteriophage antibiotic resistance genes during anaerobic digestion of cow manure.

The fate of intracellular antibiotic resistance genes (iARGs), extracellular ARGs (eARGs) and bacteriophage ARGs (bARGs) during anaerobic digestion (AD) of cow manure is unclear. Thus, the characteristics of iARGs, eARGs and bARGs during mesophilic AD (MAD) and thermophilic AD (TAD) of cow manure were investigated. The absolute abundances of iARGs decreased by 69.82% after TAD. After MAD and TAD, the total absolute abundances of eARGs increased by 63.5 times and 67.6 times, respectively, whereas those of the bARGs increased by 47.60% and 59.22%. eARGs were mainly derived from the non-specific lysis of Firmicutes, Bacteroidetes, while bacteriophages had a wide range of hosts. The variations in iARGs, eARGs and bARGs were affected by the microbial hosts but also directly driven by physicochemical factors (e.g., pH). Overall, the findings of this study revealed that there may be a risk of eARGs and bARGs disseminating during the AD of cow manure.

Elucidating the beneficial effects of diatomite for reducing abundances of antibiotic resistance genes during swine manure composting.

Diatomite (DE) has been used for nitrogen conservation during the composting of feces but its effects on antibiotic resistance genes (ARGs) and the associated mechanisms are still unclear. In this study, DE was added at three different proportions (0%, 4%, and 8%) to swine manure during composting. The results showed that adding DE helped to reduce the abundances of ARGs and the maximum decrease (88.99%) occurred with the highest dose. DE amendment promoted the transformation of reducible copper into a more stable form, i.e., the residual fraction, which reduced the selective pressure imposed by copper and further decreased the abundances of ARGs. Tn916/1545 and intI1 were critical genetic components related to ARGs, and thus the reductions in the abundances of ARGs may be attributed to the suppression of horizontal transfer due to the decreased abundances of mobile genetic elements (MGEs). The microbial community structure (bacterial abundance and diversity) played key role in the evolution of ARGs. DE could enhance the competition between hosts and non-hosts of ARGs by increasing the bacterial community diversity. Compared with CK, DE amendment optimized the bacterial community by reducing the abundances of the potential hosts of ARGs and pathogens such as Corynebacterium, thereby improving the safety of the compost product. In addition, KEGG function predictions revealed that adding DE inhibited the metabolic pathway and genes related to ARGs. Thus, composting with 8% DE can reduce the risk of ARG transmission and improve the practical value for agronomic applications.

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.

Insights into the beneficial effects of woody peat for reducing abundances of antibiotic resistance genes during composting.

Antibiotic resistance genes (ARGs) in manure endangered human health, while heavy metals in manure will pose selective pressure on ARGs. This study explored the effects on ARGs of adding woody peat during composting at different ratios (0 (CK), 5% (T1), and 15% (T2)). After composting, the relative abundances of 8/11 ARGs were 6.97-38.09% and 10.73-54.31% lower in T1 and T2, respectively, than CK. The bioavailable Cu content was 1.40% and 18.40% lower in T1 and T2, respectively, than CK. Network analysis showed that ARGs, mobile genetic elements (MGEs), and metal resistance genes possessed common potential host bacteria, such as Streptococcus, Dietzia, and Corynebacterium_1. Environmental factors, especially bioavailable Cu, and MGEs accounted for 80.75% of the changes in the abundances of ARGs. In conclusion, 15% Woody peat is beneficial to decrease the bioavailable Cu content and weaken horizontal gene transfer for controlling the spread of ARGs during composting.

Enhanced removal of antibiotic resistance genes and mobile genetic elements during swine manure composting inoculated with mature compost.

Livestock manure is a major source of antibiotic resistance genes (ARGs) that enter the environment. This study assessed the effects of inoculation with mature compost (MC) on the fates of ARGs and the bacterial community during swine manure composting. The results showed that MC prolonged the thermophilic period and promoted the decomposition of organic matter, which was due to the rapid growth and reproduction of thermophilic bacteria (Bacillus, Thermobifida, and Thermobacillus). MC significantly reduced the relative abundances of ARGs (1.02 logs) and mobile genetic elements (MGEs) (1.70 logs) after composting, especially sulfanilamide resistance genes. The total ARGs removal rate was 1.11 times higher in MC than the control. Redundancy analysis and structural equation modeling showed that horizontal gene transfer mediated by MGEs (ISCR1 and intI1) was the main direct factor related to the changes in ARGs during composting, whereas the C/N ratio and pH were the two most important indirect factors. Network analysis showed that members of Firmicutes comprising Romboutsia, Clostridisensu_stricto_1, and Terrisporobacter were the main bacterial hosts of ARGs and MGEs. MC reduced the risk of ARGs transmission by decreasing the abundances of bacterial hosts. Thus, MC is a promising strategy for reducing the proliferation risk of ARGs.

Effects of phosphogypsum and medical stone on nitrogen transformation, nitrogen functional genes, and bacterial community during aerobic composting.

This study explored the effects of adding phosphogypsum (PPG), medical stone (MS), and both (PPM) during composting on nitrogen transformation, nitrogen functional genes, the bacterial community, and their relationships with NH3 and N2O emissions. Adding MS and PPM reduced NH3 emissions by 25.78-68.37% and N2O emissions by 19.00-42.86%. PPG reduced NH3 emissions by 59.74% but slightly increased N2O emissions by 8.15%. MS was strongly correlated with the amoA-dominated nitrification process. PPG and PPM had strong correlations with nirS- and nirK-dominated, and nosZ-dominated denitrification processes, respectively. PPM promoted nitrification and denitrification processes more than PPG and MS. Different functional bacteria had key roles in nitrification and denitrification during different composting stages. Firmicutes probably contributed to the conversion and release of nitrogen in the thermophilic period, whereas Proteobacteria, Chloroflexi, Bacteroidetes, and other phyla might have played important roles in the cooling and maturation periods. PPM obtained the greatest reductions in NH3 and N2O release via the regulation of environmental variables, nitrogen functional genes, and the bacterial community. Overall, these results provide insights at a molecular level into the effects of PPG and MS on nitrogen transformation and NH3 and N2O emissions during composting.

Effects of shrimp shell powder on antibiotic resistance genes and the bacterial community during swine manure composting.

The present study compared the effects of adding shrimp shell powder (SSP) at four levels comprising 0% (CK), 5% (L), 10% (M), and 15% (H) on the abundance of antibiotic resistance genes (ARGs) and the bacterial community succession during swine manure composting. The relative abundances of 5/11 ARGs were reduced in CK, and 7/11 in H. Moreover, the removal rate was enhanced by adding SSP. Thus, H decreased the total abundance of ARGs by 32.68%, whereas CK increased it by 6.31%. Redundancy analysis indicated that mobile genetic elements (MGEs) (46.6%) and the bacterial community (31.1%) mainly explained the changes in ARGs. H enhanced the removal of MGEs, prolonged the thermophilic phase, stabilized copper and zinc, and retained nitrogen. LEfSe analysis and non-metric multidimensional scaling indicated that the bacterial community changed in the composting process, and it was optimized by H. The abundances of the potential bacterial co-hosts (such as Lactobacillus, Corynebacterium_1, and Ornithinicoccus) of ARGs and MGEs were lower and the decomposition of organic matter was higher in H compared with CK. Thus, composting with 15% SSP can reduce the risk of ARGs and improve the practical value for agronomic application.

Microbial mechanisms related to the effects of bamboo charcoal and bamboo vinegar on the degradation of organic matter and methane emissions during composting.

In this study, functional microbial sequencing, quantitative PCR, and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) were employed to understand the microbial mechanisms related to the effects of bamboo charcoal (BC) and bamboo vinegar (BV) on the degradation of organic matter (OM) and methane (CH4) emissions during composting. BC + BV resulted in the highest degradation of OM. BV was most effective treatment in controlling CH4 emissions and it significantly reduced the abundance of the mcrA gene. Methanobrevibacter, Methanosarcina, and Methanocorpusculum were closely related to CH4 emissions during the thermophilic composting period. PICRUSt analysis showed that BC and/or BV enhanced the metabolism associated with OM degradation and reduced CH4 metabolism. Structural equation modeling indicated that BC + BV strongly promoted the metabolic activity of microorganisms, which had a positive effect on CH4 emissions. Together these results suggest that BC + BV may be a suitable composting strategy if the aerobic conditions can be effectively improved during the thermophilic composting period.

Microbial succession and molecular ecological networks response to the addition of superphosphate and phosphogypsum during swine manure composting.

This study assessed the effects of superphosphate (SPP) and phosphogypsum (PPG) on the bacterial and fungal community succession and molecular ecological networks during composting. Adding SPP and PPG had positive effects on the bacterial richness and diversity, negative effects on the fungal richness and diversity. The microbial diversity and richness were higher in PPG than SPP. Non-metric multidimensional scaling analysis clearly separated SPP and PPG from the control treatment with no additives. The dominant genera comprised Turicibacter, Bacillus, norank_o_SBR1031, Thermobifida, norank_f_Limnochordaceae, Truepera, Thermopolyspora, Mycothermus, Dipodascus, Thermomyces, and unclassified_p_Ascomycota. In all treatments, the major bacterial species differed clearly in the later thermophilic, cooling, and maturation composting stages, whereas the main fungal species varied significantly in the thermophilic stage. The changes in the dominant microorganisms in SPP and PPG may have inhibited or promoted the degradation of organic matter during various composting stages. Adding SPP and PPG led to more complex bacterial networks and less complex fungal networks, where SPP had more adverse effects on the fungal networks than PPG. SPP and PPG could potentially alter the co-occurrence patterns of the bacterial and fungal communities by changing the most influential species. SPP and PPG changed the composition and succession of the microbial community by influencing different physiochemical properties during various composting stages where the pH was the main explanatory factor. Overall, this study provides new insights into the effects of SPP and PPG on the microbial community and its interactions during composting.

Effects of inoculation with lignocellulose-degrading microorganisms on nitrogen conversion and denitrifying bacterial community during aerobic composting.

The present study compared the effects of inoculation (WSD treatment) and non-inoculation (CK treatment) with lignocellulose-degrading microorganisms on nitrogen conversion, nitrogen functional genes, and the denitrifying bacterial community during aerobic composting, and their potential relations to NH3 and N2O emissions were also explored. Results showed that, WSD reduced the NH3 and N2O emissions by 25.9% and 34.98%, respectively, compared with CK. WSD also reduced the abundances of nitrifying (bacteria amoA) and denitrifying (nirS, nirK, and nosZ) genes during composting, which were significantly positively correlated with N2O emissions (P < 0.01). The most important nosZ denitrifying microorganisms belonged to Proteobacteria. Redundancy analysis showed that environmental factors could affect the succession of the denitrifying bacterial community during composting. Based on these results, structural equation modeling demonstrated that the reduction in N2O emissions under WSD was related to the lower accumulation of NO3--N utilized by denitrifying microorganisms during the compost maturation period.

Elucidating the effect of microbial inoculum and ferric chloride as additives on the removal of antibiotic resistance genes from chicken manure during aerobic composting.

This experiment investigated the effect of adding a microbial inoculum (M) and ferric chloride (F) on the fate of antibiotic resistance genes (ARGs) during chicken manure composting. Adding M and F improved the microbial activity in the compost and facilitated the removal of ARGs, whereas the combined treatment achieved the best results, especially in reducing the enrichment of sul resistance genes. Tn916/1545 and intI1 were important genetic elements that affected the transfer of ARGs, and Tn916/1545 was closely related to the transfer of tetM, tetW, and ermQ in Firmicutes. Kyoto Encyclopedia of Genes and Genomes functional predictions indicated that M and F could reduce the abundance of membrane transport and signal transduction molecules in the compost products. Thus, these findings suggest that the combined application of M and F is a promising strategy that could potentially inhibit the transfer of ARGs during composting.

Beneficial effects of bacterial agent/bentonite on nitrogen transformation and microbial community dynamics during aerobic composting of pig manure.

This study investigated the effects of adding a bacterial agent (B) and bentonite (BT) on nitrogen transformation, nitrogen functional genes, and the microbial community dynamics during the aerobic composting of pig manure, as well as their contributions to NH3 and N2O emissions. Treatments B, BT, and BT + B reduced the NH3 emissions by 31.34%, 18.82%, and 23.67%, respectively, and the N2O emissions by 53.16%, 72.56%, and 63.41%. N2O and NH3 emissions were strongly related to the functional genes. Adding bacterial agent promoted the ammonia oxidation process to reduce NH3 emissions, whereas the influence of bentonite on nitrogen conversion was mostly related to nirS and nirK in denitrification processes. Nitrification and denitrification were dominated by different functional microorganisms in various stages of composting, where Proteobacteria comprised the most important denitrifying microorganisms. Thus, the additives reduced NH3 and N2O emissions by regulating nitrification and denitrification processes, and adding both was highly advantageous.

Effects of inoculating with lignocellulose-degrading consortium on cellulose-degrading genes and fungal community during co-composting of spent mushroom substrate with swine manure.

Composting is used widely for recycling spent mushroom substrate (SMS). This study investigated the effects of inoculating a lignocellulose-degrading consortium at two levels comprising 0% (control: CK) and 10% (T) on the fungal community and cellulose-degrading genes during SMS co-composting with swine manure. Lignocellulose degradation rate in T was 8.77-34.45% higher compared with CK. Inoculation affected the distribution of the fungal community, increased the community diversity, and inhibited pathogens. Network analysis showed that inoculation changed the co-occurrence patterns of the fungal communities and made the co-composting system more stable. The relative abundances of glycoside hydrolase genes GH3E (fungal GH3), GH6, and GH7 were 0.45, 0.09, and 0.39 logs higher in T, respectively, than CK. Partial least-squares path modeling suggested that the variations in cellulose-degrading genes were driven mainly by changes in the fungal community during co-composting. Therefore, the lignocellulose-degrading consortium accelerated the transformation of lignocellulose to facilitate safer composting.