Effects of micro-nano bubble water addition on maturation degree and microbial community during aerobic composting.

In order to explore the effects of micro-nano bubble water (MNBW) on compost maturation and the microbial community in cow manure and straw during aerobic composting, we conducted composting tests using tap water with 12 mg/L (O12), 15 mg/L (O15), 18 mg/L (O18), and 21 mg/L (O21) dissolved oxygen in MNBW, as well as tap water with 9 mg/L dissolved oxygen as a control (CK). The results showed that O21 increased the maximum compost temperature to 64 °C, which was higher than the other treatments. All treatments met the harmless standards for compost. The seed germination index (GI) was largest under O21 and 15.1% higher than that under CK, and the non-toxic compost degree was higher. Redundancy analysis showed that the temperature, C/N, pH, and GI were important factors that affected the microbial community composition. The temperature, C/N, and pH were significantly positively correlated with Firmicutes and Actinobacteria (p < 0.05). Firmicutes was the dominant phylum in the mesophilic stage (2-6 days) and it accounted for a large proportion under O21, where the strong thermophilic metabolism increased the production of heat and prolonged the high temperature period. The bacterial genus Ammoniibacillus in Firmicutes accounted for a large proportion under O21 and it accelerated the decomposition of substrates. Therefore, the addition of MNBW changed the microbial community to affect the maturation of the compost, and the quality of the compost was higher under O21.

Combination of Biochar and Trichoderma harzianum Can Improve the Phytoremediation Efficiency of Brassica juncea and the Rhizosphere Micro-Ecology in Cadmium and Arsenic Contaminated Soil.

Phytoremediation is an environment-friendly method for toxic elements remediation. The aim of this study was to improve the phytoremediation efficiency of Brassica juncea and the rhizosphere soil micro-ecology in cadmium (Cd) and arsenic (As) contaminated soil. A field experiment was conducted with six treatments, including a control treatment (CK), two treatments with two contents of Trichoderma harzianum (T1: 4.5 g m-2; T2: 9 g m-2), one biochar treatment (B: 750 g m-2), and two combined treatments of T1B and T2B. The results showed Trichoderma harzianum promoted the total chlorophyll and translocation factor of Brassica juncea, while biochar promoted plant biomass compared to CK. T2B treatment showed the best results, which significantly increased Cd accumulation by 187.49-308.92%, and As accumulation by 125.74-221.43%. As a result, the soil's total Cd content was reduced by 19.04% to 49.64% and total As contents by 38.76% to 53.77%. The combined amendment increased the contents of soil available potassium, phosphorus, nitrogen, and organic matter. Meanwhile, both the activity of glutathione and peroxidase enzymes in plants, together with urease and sucrase enzymes in soil, were increased. Firmicutes (dominant bacterial phylum) and Ascomycota (dominant fungal phylum) showed positive and close correlation with soil nutrients and plant potentially toxic elements contents. This study demonstrated that phytoremediation assisted by biochar and Trichoderma harzianum is an effective method of soil remediation and provides a new strategy for enhancing plant remediation efficiency.

Mechanism for combined application of biochar and Bacillus cereus to reduce antibiotic resistance genes in copper contaminated soil and lettuce.

The remediation of agricultural soil contaminated by antibiotic resistance genes (ARGs) is of great significance for protecting food safety and human health. Reducing the availability of copper in soil may control coresistance to ARGs. However, the feasibility of applying nano-biochar and Bacillus cereus to mitigate the spread of ARGs in Cu contaminated soil remains unclear. Therefore, this study investigated the use of biochar with different particle sizes (2 % apple branch biochar and 0.5 % nano-biochar) and 3 g m-2B. cereus in a 60-day pot experiment with growing lettuce. The effects of single and combined application on the abundances of ARGs in Cu-contaminated soil (Cu = 200 mg kg-1) were compared, and the related mechanisms were explored. Studies have shown that the addition of biochar alone is detrimental to mitigating ARGs in soil-lettuce systems. The combined application of 3 g m-2B. cereus and 0.5 % nano-biochar effectively inhibited the proliferation of ARGs in Cu-contaminated soil, and 3 g m-2B. cereus effectively inhibited the proliferation of ARGs in lettuce. Partial least squares-path modeling and network analysis showed that bacterial communities and mobile genetic elements were the key factors that affected the abundances of ARGs in rhizosphere soil, and Cu resistance genes and bioavailable copper (acid extractable state Cu (F1) + reducing state Cu (F2)) had less direct impacts. The bacterial community was the key factor that affected the abundances of ARGs in lettuce. Rhodobacter (Proteobacteria), Corynebacterium (Actinobacteria), and Methylobacterium (Proteobacteria) may have been hosts of ARGs in lettuce plants. B. cereus and nano-biochar affected the abundances of ARGs by improving the soil properties and reducing the soil bioavailability of Cu, as well as directly or indirectly changing the bacterial community composition in soil and lettuce, thereby impeding the transport of ARGs to aboveground plant parts.

Integrated microbiological and metabolomics analyses to understand the mechanism that allows modified biochar to affect the alkalinity of saline soil and winter wheat growth.

In order to understand the mechanism that allows modified biochar (BC) to enhance the salt tolerance and growth of crops in saline-alkali soil, we tested the effects of ordinary BC, nanoparticle-size BC, acidified BC (HBC), and acidified nanoparticle-size BC on winter wheat growth and the soil properties by combining microbiological and metabolomics analyses. The results showed that compared with the control with no BC, the plant height increased by 17.33 % under HBC and the proportion of large soil aggregates increased by 1.25-2.83 times. HBC increased the relative abundances of some dominant genera of bacteria (e.g., Streptococcus) and fungi (e.g., Mycothermus), as well as functions such as bacterial metabolic genetic information processing and cellular processes, and reduced the abundance of pathotrophic fungi. Metabolomics analysis showed that HBC upregulated various metabolites (including amino acids and their derivatives, lipids, flavonoids, and organic acids) and five main metabolic pathways. Among the KEGG pathways, the pyrimidine metabolism pathway was significantly upregulated, as well as crop leaf metabolism, ß-alanine metabolism, and valine, leucine, and isoleucine metabolism, and the antioxidant levels and resistance to salt-alkali stress were enhanced in winter wheat leaves. Partial least squares-path modeling suggested that HBC affected the growth of winter wheat by significantly changing the soil physicochemical properties and microbial structure (path coefficients of 0.566 and 0.512, respectively).

Mechanism that allows manno-oligosaccharide to promote cellulose degradation by the bacterial community and the composting of cow manure with straw.

The new sugar source manno-oligosaccharide can regulate the structure of the microbial community. This study investigated the effects of adding manno-oligosaccharide at four different levels (0, 0.1%, 0.5%, and 1% w/w compost) to composting cow manure and straw on lignocellulose degradation and the bacterial community. Adding 0.5% manno-oligosaccharide had the greatest effects on accelerating the composting process, reducing its toxicity, and improving the stability of the product. After composting for 25 days, adding 0.5% manno-oligosaccharide decreased the hemicellulose, cellulose, and lignin contents to 2.25%, 11.25%, and 7.07%, respectively, compared with those under CK. Manno-oligosaccharide promoted the degradation of lignocellulose by increasing the abundances of Thermobifida, Streptomyces, and Luteimonas. In addition, manno-oligosaccharide inhibited pathogenic bacteria and increased the abundances of functional genes related to metabolism. Finally, adding 0.5% manno-oligosaccharide mainly affected the degradation of lignocellulose by enhancing the C/N ratio and the abundances of Streptomyces and the secretion system during composting according to redundancy analysis.

Bamboo charcoal enhances cellulase and urease activities during chicken manure composting: Roles of the bacterial community and metabolic functions.

Microbial enzymes are crucial for material biotransformation during the composting process. In this study, we investigated the effects of adding bamboo charcoal (BC) (i.e., at 5%, 10%, and 20% corresponding to BC5, BC10, and BC20, respectively) on the enzyme activity levels during chicken manure composting. The results showed that BC10 could increase the cellulose and urease activities by 56% and 96%, respectively. The bacterial community structure in BC10 differed from those in the other treatments, and Luteivirga, Lactobacillus, Paenalcaligenes, Ulvibacter, Bacillus, Facklamia, Pelagibacterium, Sporosarcina, Cellvibrio, and Corynebacterium had the most important roles in composting. Compared with other treatments, BC10 significantly enhanced the average rates of degradation of carbohydrates (D-xylose (40%) and α-D-lactose (44%)) and amino acids (L-arginine (16%), L-asparagine (14%), and L-threonine (52%)). We also explored the associations among the bacterial community and their metabolic functions with the changes in the activities of enzymes. Network analysis demonstrated that BC10 altered the co-occurrence patterns of the bacterial communities, where Ulvibacter and class Bacilli were the keystone bacterial taxa with high capacities for degrading carbon source, and they were related to increases in the activities of cellulase and urease, respectively. The results obtained in this study may help to further enhance the efficiency of composting.

Research progress and prospects for using biochar to mitigate greenhouse gas emissions during composting: A review.

Biochar possesses a unique porous structure and abundant surface functional groups, which can potentially help mitigate greenhouse gas (GHG) emissions from compost. This review summarizes the properties and functions of biochar, and the effects of biochar on common GHGs (methane (CH4), carbon dioxide (CO2), and nitrous oxide (N2O)) and ammonia (NH3, an indirect GHG) during composting. Studies have shown that it is possible to improve the mitigation of GHG emissions during composting by adjusting the biochar amount, type of raw material, pyrolysis temperature, and particle size. Biochar produced from crop residues and woody biomass has a greater effect on mitigating CH4, N2O, and NH3 emissions during composting, and GHG emissions can be reduced significantly by adding about 10% (w/w) biochar. Biochar produced by high temperature pyrolysis (500-900 °C) has a greater effect on mitigating CH4 and N2O emissions, whereas biochar generated by low temperature pyrolysis (200-500 °C) is more effective at reducing NH3 emissions. Interestingly, adding granular biochar is more beneficial for mitigating CH4 emissions, whereas adding powdered biochar is better at reducing NH3 emissions. According to the current research status, developing new methods for producing and using biochar (e.g., modified or combined with other additives) should be the focus of future research into mitigating GHG emissions during composting. The findings summarized in this review may provide a reference to allow the establishment of standards for using biochar to mitigate GHG emissions from compost.

Effects of Bacillus subtilis on carbon components and microbial functional metabolism during cow manure-straw composting.

This study is the first to investigate the changes in the composting process and carbon conversion in a cow manure-straw compost matrix with Bacillus subtilis added at four different levels (0, 0.5%, 1%, and 2% w/w compost), and to explain the mechanism responsible for carbon conversion through microbial functional metabolism. Inoculation with Bacillus subtilis at 2% had the best effect on fermentation among all treatments, but it inhibited the synthesis of total organic carbon and humus. Bacillus subtilis at 0.5% reduced mineralization in the cooling and maturity stages of composting, and enhanced the humification of carbon. The total organic carbon and humic sequence contents were significantly higher with Bacillus subtilis at 0.5% (12.5% and 20.2%, respectively) than Bacillus subtilis at 2% (P < 0.05). Redundancy analysis demonstrated that the pH and microbial functional metabolism were closely related to carbon sequestration during composting.

Changes in antibiotic resistance genes and mobile genetic elements during cattle manure composting after inoculation with Bacillus subtilis.

This study explored the effects of Bacillus subtilis at four levels (0, 0.5%, 1%, and 2% w/w compost) on the variations in ARGs, mobile genetic elements (MGEs), and the bacterial community during composting. The composting process had a greater impact on ARGs than Bacillus subtilis. The main ARG detected was sul1. The addition of Bacillus subtilis at 0.5% reduced the relative abundances of ARGs, MGEs, and human pathogenic bacteria (by 2-3 logs) in the mature products. Network and redundancy analyses suggested that intI1, Firmicutes, and pH were mainly responsible for the changes in ARGs, thus controlling these factors might help to inhibit the spread of ARGs.

Factors that affect the occurrence and distribution of antibiotic resistance genes in soils from livestock and poultry farms.

Livestock manure is generally dumped directly onto open soil or used to enhance the soil fertility. However, there are growing concerns regarding the impact of these practices on the development and dissemination of antibiotic resistance genes (ARGs) in soil. In this study, we sampled soils treated with manure from 10 large-scale farms (pig, beef cattle, and chicken farms) and those from farmland without manure. The results showed that the abundance of ARGs was more than 2.62 times higher in the soil samples treated with livestock manure than the farmland soil without manure. The abundances of ARGs and intI1 in all samples were in the following order: pig farms > chicken farms > beef cattle farms. tetX, sul1, sul2, and tetG were the dominant ARGs in farm soil. The concentrations of tetracycline antibiotics and sulfonamide antibiotics were 0.15-4.76 mg/kg and 0-2.62 mg/kg, respectively, in the soils treated with manure, which were higher than those in farmland soils without manure. Redundancy analysis (P < 0.05) and network analysis (P < 0.01, R > 0.80) demonstrated that copper, zinc, actinomycetes, and tetracycline antibiotics were the main factors that affected the distribution of ARGs in soils treated with livestock manure.

Prevalence of quinolone resistance genes, copper resistance genes, and the bacterial communities in a soil-ryegrass system co-polluted with copper and ciprofloxacin.

The presence of high concentrations of residual antibiotics and antibiotic resistance genes (ARGs) in soil may pose potential health and environmental risks. This study investigated the prevalence of plasmid-mediated quinolone resistance (PMQR) genes, copper resistance genes (CRGs), and the bacterial communities in a soil-ryegrass pot system co-polluted with copper and ciprofloxacin (CIP; 0, 20, or 80 mg kg-1 dry soil). Compared with the samples on day 0, the total relative abundances of the PMQR genes and mobile genetic elements (MGEs) were reduced significantly by 80-89% in the ryegrass and soil by the cutting stage (after 75 days). The abundances of PMQR genes and MGEs were reduced by 63-81% in soil treated with 20 mg kg-1 CIP compared with the other treatments, but the abundances of CRGs increased by 18-42%. The presence of 80 mg kg-1 CIP affected the microbial community structure in the soil by increasing the abundances of Acidobacteria and Thaumarchaeota, but decreasing those of Firmicutes. Redundancy analysis indicated that the pH and microbial composition were the main factors that affected the variations in PMQR genes, MGEs, and CRGs, where they could explain 42.2% and 33.3% of the variation, respectively. Furthermore, intI2 may play an important role in the transfer of ARGs. We found that 80 mg kg-1 CIP could increase the abundances of ARGs and CRGs in a soil-ryegrass pot system.

Effects of genetically modified cotton stalks on antibiotic resistance genes, intI1, and intI2 during pig manure composting.

Genetically modified (GM) cotton production generates a large yield of stalks and their disposal is difficult. In order to study the feasibility of using GM cotton stalks for composting and the changes that occur in antibiotic resistance genes (ARGs) during composting, we supplemented pig manure with GM or non-GM cotton stalks during composting and we compared their effects on the absolute abundances (AA) of intI1, intI2, and ARGs under the two treatments. The compost was mature after processing based on the germination index and C/N ratio. After composting, the AAs of ARGs, intI1, and intI2 were reduced by 41.7% and 45.0% in the non-GM and GM treatments, respectively. The ARG profiles were affected significantly by temperature and ammonia nitrogen. In addition, excluding tetC, GM cotton stalks had no significant effects on ARGs, intI1, and intI2 compared with the non-GM treatment (p < 0.05). Thus, similar to non-GM cotton stalks, GM cotton stalks can be used for aerobic composting with livestock manure, and the AAs of ARGs can be reduced. Furthermore, the results of this study provide a theoretical basis for the harmless utilization of GM cotton stalks.

Effects of biochar on reducing the abundance of oxytetracycline, antibiotic resistance genes, and human pathogenic bacteria in soil and lettuce.

Antibiotics and antibiotic resistance genes (ARGs) in soil can affect human health via the food chain. Biochar is a soil amendment but its impacts on ARGs and the microbial communities associated with soil and vegetables are unclear. Therefore, we established three lettuce pot culture experiments, i.e., O300: 300 mg/kg oxytetracycline (OTC), BO300: 300 mg/kg OTC + 2% biochar, and a control without OTC or biochar. We found that under BO300, the relative abundances of ARGs were reduced by 51.8%, 43.4%, and 44.1% in lettuce leaves, roots, and soil, respectively, compared with O300. intI1 was highly abundant in soil and lettuce, and it co-occurred with some ARGs (tetW, ermF, and sul1). Redundancy analysis and network analysis indicated that the bacterial community succession was the main mechanism that affected the variations in ARGs and intI1. The reduction of Firmicutes due to the biochar treatment of soil and lettuce was the main factor responsible for the removal of tetracycline resistance genes in leaves. Biochar application led to the disappearance of human pathogenic bacteria (HPB), which was significantly correlated with the abundances of ermF and ermX. In summary, biochar is an effective farmland amendment for reducing the abundances of antibiotics, ARGs, and HPB in order to ensure the safety of vegetables and protect human health.

Effects of inoculation with organic-phosphorus-mineralizing bacteria on soybean (Glycine max) growth and indigenous bacterial community diversity.

Three different organic-phosphorus-mineralizing bacteria (OPMB) strains were inoculated to soil planted with soybean (Glycine max), and their effects on soybean growth and indigenous bacterial community diversity were investigated. Inoculation with Pseudomonas fluorescens Z4-1 and Brevibacillus agri L7-1 increased organic phosphorus degradation by 22% and 30%, respectively, compared with the control at the mature stage. Strains P. fluorescens Z4-1 and B. agri L7-1 significantly improved the soil alkaline phosphatase activity, average well color development, and the soybean root activity. Terminal restriction fragment length polymorphism analysis demonstrated that P. fluorescens Z4-1 and B. agri L7-1 could persist in the soil at relative abundances of 2.0%-6.4% throughout soybean growth. Thus, P. fluorescens Z4-1 and B. agri L7-1 could potentially be used in organic-phosphorus-mineralizing biofertilizers. OPMB inoculation altered the genetic structure of the soil bacterial communities but had no apparent influence on the carbon source utilization profiles of the soil bacterial communities. Principal components analysis showed that the changes in the carbon source utilization profiles of bacterial community depended mainly on the plant growth stages rather than inoculation with OPMB. The results help to understand the evolution of the soil bacterial community after OPMB inoculation.

Effects of bamboo charcoal on antibiotic resistance genes during chicken manure composting.

Composting is widely used for animal waste disposal, and bamboo charcoal (BC) can be used for nitrogen conservation during composting. However, the effects of BC on antibiotic resistance genes (ARGs) during chicken manure composting are still unclear. This study investigated the effects on ARGs of adding different proportions of BC (0%, 5%, 10%, and 20% w/w) to chicken manure compost. After 26 days, the relative abundances (RAs) of most ARGs (tetC, tetG, tetW, tetX, sul2, drfA1, drfA7, ermB, ermF, ermQ, and ermX) and intI1 declined by 21.6-99.5%, whereas sul1 increased by 7.5-17.7 times. The average RAs reductions with 0%, 5%, 10%, and 20% BC were 0.85, 1.05, 1.08, and 1.15 logs, respectively. The most important environmental factor for the ARG profiles was temperature according to redundancy analysis. Furthermore, BC significantly decreased the bio-Cu and bio-Zn levels, thereby reducing the co-selection pressure from heavy metals. Different proportions of BC had no significant effects on the removal of tetG, tetW, tetX, sul2, drfA1, and ermB. Supplementation with 10% BC was more effective at removing tetC and drfA7 compared with the other treatments. The results suggested that 10% BC supplementation is appropriate for reducing ARGs in chicken manure compost.

Relationships between sulfachloropyridazine sodium, zinc, and sulfonamide resistance genes during the anaerobic digestion of swine manure.

In this study, swine manure containing sulfachloropyridazine sodium (SCPS) and zinc was subjected to mesophilic (37°C) anaerobic digestion (AD). The absolute abundances (AAs) of antibiotic resistance genes (ARGs) were evaluated, as well as intI1 and intI2, and the degradation of SCPS according to variation in the amount of bio-available zinc (bio-Zn). In digester that only contained SCPS, the concentrations of SCPS were lower than that digesters both contain SCPS and Zn. Compared with the control digester, the addition of SCPS increased the AAs of sul1, sul3, drfA1, and drfA7 by 1.3-13.1 times. However, compared with the digester with SCPS but no added Zn, the AAs of sul3, drfA1, and drfA7 were decreased by 21.4-70.3% in the presence of SCPS and Zn, whereas sul1 and sul2 increased 1.3-10.7 times. There were significant positive correlations (P<0.05) between the concentrations of SCPS with several ARGs and bio-Zn.

Reducing antibiotic resistance genes, integrons, and pathogens in dairy manure by continuous thermophilic composting.

This study explored the effects of composting using three temperature regimes, namely, insufficient thermophilic composting (ITC), normal thermophilic composting (NTC), and continuous thermophilic composting (CTC), on antibiotic resistance genes (ARGs), integrons, and human pathogenic bacteria (HPB), as well as the mechanisms involved. The NTC and CTC treatments led to greater decreases in 5/10 ARGs and two integrons than ITC, and the abundances of ARGs (tetC, tetG, and tetQ) and int1 only declined in the NTC and CTC treatments. The abundances of HPB decreased by 82.8%, 76.9%, and 96.9% under ITC, NTC, CTC, respectively. Redundancy analysis showed that both bacterial succession and horizontal gene transfer play important roles in the variation of ARGs, and the changes in different ARGs were due to diverse mechanisms. CTC performed significantly better at reducing ARGs, integrons, and HPB, thus it may be used to manage the public health risks of ARGs in animal manure.

Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure.

Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

Variable effects of oxytetracycline on antibiotic resistance gene abundance and the bacterial community during aerobic composting of cow manure.

Livestock manure is often subjected to aerobic composting but little is known about the variation in antibiotic resistance genes (ARGs) during the composting process under different concentrations of antibiotics. This study compared the effects of three concentrations of oxytetracycline (OTC; 10, 60, and 200mg/kg) on ARGs and the succession of the bacterial community during composting. Very similar trends were observed in the relative abundances (RAs) of each ARG among the OTC treatments and the control during composting. After composting, the RAs of tetC, tetX, sul1, sul2, and intI1 increased 2-43 times, whereas those of tetQ, tetM, and tetW declined by 44-99%. OTC addition significantly increased the absolute abundances and RAs of tetC and intI1, while 200mg/kg OTC also enhanced those of tetM, tetQ, and drfA7. The bacterial community could be grouped according to the composting time under different treatments. The highest concentration of OTC had a more persistent effect on the bacterial community. In the present study, the succession of the bacterial community appeared to have a greater influence on the variation of ARGs during composting than the presence of antibiotics. Aerobic composting was not effective in reducing most of the ARGs, and thus the compost product should be considered as an important reservoir for ARGs.

Effects of sulphamethazine and zinc on the functional diversity of microbial communities during composting.

The changes in the functional diversity of the microbial community in a compost matrix with a single or compound addition of zinc (Zn; 0, 600, and 1800 mg/kg) and sulphamethazine (SM2; 0, 1, and 25 mg/kg) were studied with the Biolog method during composting. The microbial community was extracted from the compost matrix comprising swine manure and wheat straw at day 6 (themophilic period) and day 25 (mature period) of composting. Results proved that the Shannon index, average well-colour development, and substrate utilization significantly decreased as the concentrations of SM2 and Zn increased on day 6. The negative effect of the combined addition of SM2 and Zn was lower than that of the individual addition of SM2 and Zn. On day 25, the inhibition effect disappeared, and microbial metabolic activities were higher than those on day 6. The effects of SM2 and Zn could be further differentiated via the principal component analysis (PCA) and cluster analysis. On day 6, the treatments were divided into three groups by PC1 and PC2. The separation of the different treatments in the PCA plots became increasingly apparent on day 25. In conclusion, the effects of SM2 and Zn on the microbial community during composting became evident in the themophilic period and that the microbial activity recovered in the mature period. The combination of SM2 and Zn decreased the inhibition with the addition of individual additive.