Antibiotic residues that enter the soil through swine manure could disturb the number, community structure and functions of microbiota which could also degrade antibiotics in soil. Five different concentrations of doxycycline (DOX) incorporated into swine manure were added to soil to explore the effects of DOX on microbiota in soil and degradation itself. The results showed that the soil microbiome evolved an adaptation to the soil containing DOX by generating resistance genes. Moreover, some of the organisms within the soil microbiome played crucial roles in the degradation of DOX. The average degradation half-life of DOX in non-sterile groups was 13.85 ± 0.45 days, which was significantly shorter than the 29.26 ± 0.98 days in the group with sterilized soil (P < 0.01), indicating that the soil microbiome promoted DOX degradation. DOX addition affected the number of tetracycline resistance genes, depending on the type of gene and the DOX concentration. Among these genes, tetA, tetM, tetW, and tetX had significantly higher copy numbers when the concentration of DOX was higher. In contrast, a lower concentration of DOX had an inhibitory effect on tetG. At the same time, the microbial compositions were affected by the initial concentration of DOX and the different experimental periods. The soil chemical indicators also affected the microbial diversity changes, mainly because some microorganisms could survive in adversity and become dominant bacterial groups, such as the genera Vagococcus and Enterococcus (which were associated with electrical conductivity) and Caldicoprobacter spp. (which were positively correlated with pH). Our study mainly revealed soil microbiota and DOX degradation answered differently under variable concentrations of DOX mixed with swine manure in soil.
The Zn-Al mixed metal oxide (ZnAl-MMO) with a plate-like structure was derived from Zn-Al layered double hydroxide. The ZnAl-MMO with a Zn/Al molar ratio of 3:1 exhibits superior absorption ability for H2S in a simulated coal gas at 600 ℃ due to the special structure of the ZnAl-MMO. Besides ZnS, elemental sulfur is also produced during the desulfurization process. The deactivation model could well simulate the absorption behavior of H2S. The sulfidation reaction over the sorbent shows large initial reaction rate constants (1110-5390 m3 min-1 kg-1) and low activation energy (29.5 kJ mol-1). The regeneration rate of the used sorbent can reach 99.8% under the optimum conditions. The regenerated sorbents still show high sulfur capacity (ca. 30%), implying its great application potential for industrial-scale desulfurization of the hot coal gas.
Despite our continuous improvement in understanding the evolution of antibiotic resistance, the changes in the carbon metabolism during the evolution of antibiotic resistance remains unclear. To investigate the evolution of antibiotic resistance and the changes in carbon metabolism under antibiotic pressure, Escherichia coli K-12 was evolved for 38 passages under a concentration gradient of doxycycline (DOX). The 0th-passage sensitive strain W0, the 20th-passage moderately resistant strain M20, and the 38th-passage highly resistant strain E38 were selected for the determination of biofilm formation, colony area, and carbon metabolism levels, as well as genome and transcriptome sequencing. The MIC of DOX with E. coli significantly increased from 4 to 96 μg/ml, and the IC50 increased from 2.18 ± 0.08 to 64.79 ± 0.75 μg/ml after 38 passages of domestication. Compared with the sensitive strain W0, the biofilm formation amount of the resistant strains M20 and E38 was significantly increased (p < 0.05). Single-nucleotide polymorphisms (SNPs) were distributed in antibiotic resistance-related genes such as ribosome targets, cell membranes, and multiple efflux pumps. In addition, there were no mutated genes related to carbon metabolism. However, the genes involved in the biosynthesis of secondary metabolites and carbon metabolism pathway were downregulated, showing a significant decrease in the metabolic intensity of 23 carbon sources (p < 0.05). The results presented here show that there may be a correlation between the evolution of E. coli DOX resistance and the decrease of carbon metabolism, and the mechanism was worthy of further research, providing a theoretical basis for the prevention and control of microbial resistance.
Livestock farms are commonly regarded as the main sources of antibiotic resistance genes (ARGs), emerging pollutants with potential implications for human health, in the environment. This study investigated the occurrence and contamination profiles of nine ARGs of three types from swine manure to receiving environments (soil and water) in Guangdong Province, southern China. All ARGs occurred in 100% of swine manure samples. Moreover, the absolute concentration of total ARGs varied from 3.01 × 108 to 7.18 × 1014 copies/g, which was significantly higher than that in wastewater and manured soil (p 0.05). However, the number of ARGs (ermB, qnrS, acc(6')-Ib, tetM, tetO and tetQ) decreased but were not eliminated by wastewater treatment components (p
Ammonia emission is an important problem that needs to be solved in laying hen industries. Sodium butyrate (SB) is considered to have potential for reducing ammonia production because of its ability to improve nitrogen metabolism. In this in vitro fermentation study, we presented a correlation analysis of the metatranscriptome and metaproteome of lay hen cecal microorganisms, in order to identify important proteins and pathways involved in ammonia production reduction due to sodium butyrate supplementation. The results showed that sodium butyrate supplement decreased the production of ammonia by 26.22% as compared with the non-sodium butyrate supplementation (CK) group. The SB group exhibited a lower concentration of ammonium nitrogen (NH4+-N) and a decreased pH. Sodium butyrate promoted the uric acid concentration and lowered the uricase activity in the fermentation broth of laying hens cecal content. Notably, the 'alanine, aspartate and glutamate metabolism' category was more abundant in the SB group. The addition of sodium butyrate increased the expression of glutamate dehydrogenase (GDH) gene in cecal microbiota (e.g., Ruminococcus sp. and Bacteroides sp.) in vitro. The metaproteome analysis results showed that the expression of GDH with NADPH as coenzyme (NADPH-GDH) was up-regulated in cecal microbiota by sodium butyrate supplement. Our results indicate that sodium butyrate can affect glutamate metabolism through regulating the expression of glutamate dehydrogenase in cecal microorganisms, thereby reducing ammonia production. This study reveals that glutamate dehydrogenase-mediated glutamate metabolism play a key role in ammonia emission reduction in laying hen and provide theoretical basis for further developing ammonia production reduction approach.
Antibiotic residues in swine manure when entered the soil would most likely affect the complex composition and functions of the soil microbiome, which is also responsible for degrading these antibiotics. Three different methods of adding ciprofloxacin (CIP), a common antibiotic used in the swine industry, to the soil were used to investigate the effects of CIP on the soil microbiome and the degradation of CIP. Results of the study showed that the microbiome could promote the degradation of CIP in the soil when CIP was incorporated into the soil together with manure. However, the CIP degradation time was prolonged when adding the manure of swine fed with diet containing CIP in the soil. All treatments did not affect the copy number of the resistance genes, except for aac(6')-Ib-cr, as compared with the initial numbers of each treatment. MiSeq Illumina sequencing and Biolog-ECO microplates results showed that CIP had a significant effect on the abundance, structure, and function of the soil microbiome, but different addition methods resulted in distinct effects. Results of the present study demonstrated that the microbiome and fate of CIP responded differently to the different methods of adding CIP to the soil.
Ammonia emissions have become an important environmental challenge for the livestock industry. Probiotics are often used as additives to reduce ammonia, and the ammonia reduction efficiency of common probiotics is approximately 20-40%. In this study, we constructed a gdhA recombinant Pichia pastoris strain, glnA recombinant Pichia pastoris strain and gdhA-glnA Pichia pastoris recombinant strain using the gdhA and glnA genes, which have the potential function of reducing ammonia emissions. The results of in vitro fermentation showed that compared with the control, wild-type Pichia pastoris and pPICZA strains, the gdhA, glnA and gdhA-glnA recombinant strains significantly reduced ammonia emissions in laying hens (P
High concentrations of antibiotics may induce bacterial resistance mutations and further lead to fitness costs by reducing growth of resistant bacteria. However, antibiotic concentrations faced by bacteria are usually low in common environments, which leads to questions about how resistant bacteria with fitness costs regulate metabolism to coexist or compete with susceptible bacteria during sublethal challenge. Our study revealed that a low proportion (< 15%) of resistant bacteria coexisted with susceptible bacteria due to the fitness cost without doxycycline. However, the cost for the resistant strain decreased at a doxycycline concentration of 1 mg/L and even disappeared when the doxycycline concentration was 2 mg/L. Metabonomics analysis revealed that bypass carbon metabolism and biosynthesis of secondary metabolites were the primary metabolic pathways enriching various upregulated metabolites in resistant bacteria without doxycycline. Moreover, the alleviation of fitness cost for resistant bacteria competed with susceptible bacteria at 1 mg/L doxycycline was correlated with the downregulation of the biomarkers pyruvate and pilocarpine. Our study offered new insight into the metabolic mechanisms by which the fitness cost of resistant mutants was reduced at doxycycline concentrations as low as 1 mg/L and identified various potential metabolites to limit the spread of antimicrobial resistance in the environment.
In vitro Lead (Pb(2+)) 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) Pb(2+) binding capacity of both bacterial strains. Fourier transform infrared analysis indicated that several functional groups (O-H or N-H, C-H, C˭O, 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(-1)bands. 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 Pb(2+) binding of the two strains from intracellular accumulation to extracellular adsorption.
Biochar, because of its unique physiochemical properties and sorption capacity, may be an ideal amendment in reducing gaseous emissions during composting process but there has been little information on the potential effects of different types of biochar on undesired gaseous emissions. The objective of this study was to examine the ability and mechanism of different types of biochar, as co-substrate, in mitigating gaseous emission from composting of layer hen manure. The study was conducted in small-scale laboratory composters with the addition of 10% of one of the following biochars: cornstalk biochar, bamboo biochar, woody biochar, layer manure biochar and coir biochar. The results showed that the cumulative NH3 production was significantly reduced by 24.8±2.9, 9.2±1.3, 20.1±2.6, 14.2±1.6, 11.8±1.7% (corrected for initial total N) in the cornstalk biochar, bamboo biochar, woody biochar, layer manure biochar and coir biochar treatments, respectively, compared to the control. Total CH4 emissions was significantly reduced by 26.1±2.3, 15.5±2.1, 22.4±3.1, 17.1±2.1% (corrected for the initial total carbon) for cornstalk biochar, bamboo biochar, woody biochar and coir biochar treatments than the control. Moreover, addition of cornstalk biochar increased the temperature and NO3(-)-N concentration and decreased the pH, NH4(+)-N and organic matter content throughout the composting process. The results suggested that total volatilization of NH3 and CH4 in cornstalk biochar treatment was lower than the other treatments; which could be due to (i) decrease of pH and higher nitrification, (ii) high sorption capacity for gases and their precursors, such as ammonium nitrogen from composting mixtures, because of the higher surface area, pore volumes, total acidic functional groups and CEC of cornstalk biochar.
Microbial remediation has the potential to inexpensively yet effectively decontaminate and restore contaminated environments, but the virulence of pathogens and risk of resistance gene transmission by microorganisms during antibiotic removal often limit its implementation. Here, a cloned tetX gene with clear evolutionary history was expressed to explore doxycycline (DOX) degradation and resistance variation during the degradation process. Phylogenetic analysis of tetX genes showed high similarity with those of pathogenic bacteria, such as Riemerella sp. and Acinetobacter sp. Successful tetX expression was performed in Escherichia coli and confirmed by SDS-PAGE and Western blot. Our results showed that 95.0 ± 1.0% of the DOX (50 mg/L) was degraded by the recombinant strain (ETD-1 with tetX) within 48 h, which was significantly higher than that for the control (38.9 ± 8.7%) and the empty plasmid bacteria (8.8 ± 5.1%) (P 0.05). The efficient and safe DOX-degrading capacity of the recombinant strain ETD-1 makes it valuable and promising for antibiotic removal in the environment.
Fermented soybean meal (FSM), which has lower anti-nutritional factors and higher active enzyme, probiotic and oligosaccharide contents than its unfermented form, has been reported to improve the feeding value of soybean meal, and hence, the growth performance of piglets. However, whether FSM can affect the bacterial and metabolites in the large intestine of piglets remains unknown. This study supplemented wet-FSM (WFSM) or dry-FSM (DFSM) (5% dry matter basis) in the diet of piglets and investigated its effects on carbon and nitrogen metabolism in the piglets' large intestines. A total of 75 41-day-old Duroc×Landrace×Yorkshire piglets with an initial BW of 13.14±0.22 kg were used in a 4-week feeding trial. Our results showed that the average daily gain of piglets in the WFSM and DFSM groups increased by 27.08% and 14.58% and that the feed conversion ratio improved by 18.18% and 7.27%, respectively, compared with the control group. Data from the prediction gene function of Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) based on 16S ribosomal RNA (rRNA) sequencing showed that carbohydrate metabolism function families in the WFSM and DFSM groups increased by 3.46% and 2.68% and that the amino acid metabolism function families decreased by 1.74% and 0.82%, respectively, compared with the control group. These results were consistent with those of other metabolism studies, which showed that dietary supplementation with WFSM and DFSM increased the level of carbohydrate-related metabolites (e.g. 4-aminobutanoate, 5-aminopentanoate, lactic acid, mannitol, threitol and β-alanine) and decreased the levels of those related to protein catabolism (e.g. 1,3-diaminopropane, creatine, glycine and inosine). In conclusion, supplementation with the two forms of FSM improved growth performance, increased metabolites of carbohydrate and reduced metabolites of protein in the large intestine of piglets, and WFSM exhibited a stronger effect than DFSM.