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873300.9951Enhanced anti-herbivore defense of tomato plants against Spodoptera litura by their rhizosphere bacteria. BACKGROUND: The use of beneficial microorganisms as an alternative for pest control has gained increasing attention. The objective of this study was to screen beneficial rhizosphere bacteria with the ability to enhance tomato anti-herbivore resistance. RESULTS: Rhizosphere bacteria in tomato field from Fuqing, one of the four locations where rhizosphere bacteria were collected in Fujian, China, enhanced tomato resistance against the tobacco cutworm Spodoptera litura, an important polyphagous pest. Inoculation with the isolate T6-4 obtained from the rhizosphere of tomato field in Fuqing reduced leaf damage and weight gain of S. litura larvae fed on the leaves of inoculated tomato plants by 27% in relative to control. Analysis of 16S rRNA gene sequence identities indicated that the isolate T6-4 was closely related to Stenotrophomonas rhizophila supported with 99.37% sequence similarity. In the presence of S. litura infestation, inoculation with the bacterium led to increases by a 66.9% increase in protease inhibitor activity, 53% in peroxidase activity and 80% in polyphenol oxidase activity in the leaves of inoculated plants as compared to the un-inoculated control. Moreover, the expression levels of defense-related genes encoding allene oxide cyclase (AOC), allene oxide synthase (AOS), lipoxygenase D (LOXD) and proteinase inhibitor (PI-II) in tomato leaves were induced 2.2-, 1.7-, 1.4- and 2.7-fold, respectively by T6-4 inoculation. CONCLUSION: These results showed that the tomato rhizosphere soils harbor beneficial bacteria that can systemically induce jasmonate-dependent anti-herbivore resistance in tomato plants.202235606741
783610.9948Efficient Degradation of Intracellular Antibiotic Resistance Genes by Photosensitized Erythrosine-Produced (1)O(2). Intracellular antibiotic resistance genes (iARGs) constitute the important part of wastewater ARGs and need to be efficiently removed. However, due to the dual protection of intracellular DNA by bacterial membranes and the cytoplasm, present disinfection technologies are largely inefficient in iARG degradation. Herein, we for the first time found that erythrosine (ERY, an edible dye) could efficiently degrade iARGs by producing abundant (1)O(2) under visible light. Seven log antibiotic-resistant bacteria were inactivated within only 1.5 min, and 6 log iARGs were completely degraded within 40 min by photosensitized ERY (5.0 mg/L). A linear relationship was established between ARG degradation rate constants and (1)O(2) concentrations in the ERY photosensitizing system. Surprisingly, a 3.2-fold faster degradation of iARGs than extracellular ARGs was observed, which was attributed to the unique indirect oxidation of iARGs induced by (1)O(2). Furthermore, ERY photosensitizing was effective for iARG degradation in real wastewater and other photosensitizers (including Rose Bengal and Phloxine B) of high (1)O(2) yields could also achieve efficient iARG degradation. The findings increase our knowledge of the iARG degradation preference by (1)O(2) and provide a new strategy of developing technologies with high (1)O(2) yield, like ERY photosensitizing, for efficient iARG removal.202337531556
878220.9947Antagonistic bacterium Bacillus amyloliquefaciens induces resistance and controls the bacterial wilt of tomato. BACKGROUND: Bacterial wilt caused by Ralstonia solanacearum (RS) is a serious threat for agricultural production. In this study, Bacillus amyloliquefaciens strains CM-2 and T-5 antagonistic to RS were used to create bioorganic fertilisers to control tomato wilt under greenhouse conditions. The possible mechanism of resistance inducement by the antagonistic bacteria was also evaluated. RESULTS: The application of bioorganic fertilisers significantly reduced incidences of tomato wilt (by 63-74%), promoted plant growth and significantly reduced the RS populations in rhizosphere compared with the control. Both strains CM-2 and T-5 applied with bioorganic fertilisers survived well in the tomato rhizosphere. Tomato seedlings treated with cell suspension of T-5 followed by challenge inoculation with RS increased the activities of polyphenol oxidase, phenylalanine ammonia lyase and peroxidase compared with the untreated control. Furthermore, the expressions of the marker genes responsible for synthesis of phytohormones salicylic acid, ethylene and jasmonic acid in seedlings treated with T-5 in response to inoculated pathogen were significantly higher. CONCLUSIONS: This study suggests that strains CM-2 and T-5 containing bioorganic fertilisers effectively control tomato wilt. Increased enzyme activities and expression of defence genes in plants indicated that the antagonistic bacteria induced plant resistance, which was the potential biocontrol mechanism of tomato wilt.201323519834
778330.9945Heterologous expression of the tetracycline resistance gene tetX to enhance degradability and safety in doxycycline degradation. 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 tetX gene products in ETD-1 cell extracts also exhibited an efficient DOX degradation ability, with a degradation rate of 80.5 ± 1.2% at 168 h. Furthermore, there was no significant proliferation of the tetX resistance gene during DOX degradation (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.202031968275
774840.9945Bacillus subtilis reduces antibiotic resistance genes of animal sludge in vermicomposting by improving heat stress tolerance of Eisenia foetida and bacterial community adjustment. Antibiotic resistance genes (ARGs) in livestock industry have been recognized as a kind of pollutant. The effect of Bacillus subtilis (B. subtilis) as an additive for the reduction of ARGs in animal sludge from livestock and poultry wastewater treatment plant during vermicomposting was investigated. We also evaluated the oxidative stress level and growth of earthworms, Eisenia foetida, bacterial community succession, and the quality of the end products. Two treatments were conducted using B. subtilis, one at 18 °C and another at 28 °C. Controls were setup without the bacteria. The results showed that inoculation of B. subtilis promoted the degradation of organics at 28 °C and increased the germination index to 236%. The increased activities of the superoxide dismutase (1.69 U/mg pr) and catalase (8.05 U/mg pr) and the decreased activity of malondialdehyde (0.02 nmol/mg pr) by B. subtilis at 28 °C showed that the earthworms were relieved of heat stress. The addition of B. subtilis reduced the abundance of 32 target ARGs, including integron (intI-1), transposase (IS613) and resistant genes, such as sulfonamide (sul2), quinolone (oprJ), macrolide-lincosamide-streptogramin group B (ermF, ermB), tetracycline (tetL-02, tetX), β-lactama (blaOXA10-01) and aminoglycoside [strB, aac(6')-Ib(aka aacA4)-01, aac(6')-Ib(aka aacA4)-02]. Organic matter degrading Membranicola, Paludisphaera, Sphingorhabdus and uncultured bacterium belonging to the order Chitinophagales, nitrifying and nitrogen-fixing Singulisphaera and Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, soil remediating Achromobacter, and plant growth promoting Kaistia, Galbibacter and Ilumatobacter were increased significantly (P < 0.05). However, the growth of harmful bacteria such as Burkholderiaceae was inhibited in the vermicompost. In earthworm guts, the probiotic Mesorhizobium was promoted, while the pathogenic uncultured bacterium belonging to the family Enterobacteriaceae was reduced. Besides, B. subtilis enhanced the host relationships between bacteria and ARGs. These findings might be helpful in the removal of ARGs in animal wastes and in understanding the synergy between earthworms and microorganisms.202336529325
795150.9945Proliferation of antibiotic resistance genes in microbial consortia of sequencing batch reactors (SBRs) upon exposure to trace erythromycin or erythromycin-H2O. A variety of antibiotics and their metabolites at sub-inhibitory level concentrations are suspected to expand resistance genes in the environment. However, knowledge is limited on the causal correlation of trace antibiotics or their metabolites with resistance proliferation. In this study, erythromycin (ERY) resistance genes were screened on microbial consortia of sequencing batch reactors (SBRs) after one year acclimation to ERY (100 μg/L) or dehydrated erythromycin (ERY-H(2)O, 50 μg/L). The identified esterase gene ereA explains that ERY could be degraded to six products by microbes acclimated to ERY (100 μg/L). However, ERY could not be degraded by microbes acclimated to ERY-H(2)O (50 μg/L), which may be due to the less proliferated ereA gene. Biodegradation of ERY required the presence of exogenous carbon source (e.g., glucose) and nutrients (e.g., nitrogen, phosphorus) for assimilation, but overdosed ammonium-N (>40 mg/L) inhibited degradation of ERY. Zoogloea, a kind of biofilm formation bacteria, became predominant in the ERY degradation consortia, suggesting that the input of ERY could induce biofilm resistance to antibiotics. Our study highlights that lower μg/L level of ERY or ERY-H(2)O in the environment encourages expansion of resistance genes in microbes.201121482429
780860.9944Visible light-driven C/O-g-C(3)N(4) activating peroxydisulfate to effectively inactivate antibiotic resistant bacteria and inhibit the transformation of antibiotic resistance genes: Insights on the mechanism. Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) dissemination within water pose a serious threat to public health. Herein, C and O dual-doped g-C(3)N(4) (C/O-g-C(3)N(4)) photocatalyst, fabricated via calcination treatment, was utilized to activate peroxydisulfate (PDS) to investigate the disinfection effect on tetracycline-resistant Escherichia coli and the transformation frequency of ARGs. As a result, approximately 7.08 log E. coli were inactivated, and 72.36 % and 53.96 % of antibiotics resistance gene (tetB) and 16 S rRNA were degraded respectively within 80 min. Futhermore, the transformation frequency was reduced to 0.8. Characterization and theoretical results indicated that C and O doping in g-C(3)N(4) might lead to the electronic structure modulation and band gap energy reduction, resulting in the production of more free radicals. The mechanism analysis revealed that C/O-g-C(3)N(4) exhibited a lower adsorption energy and reaction energy barrier for PDS compared to g-C(3)N(4). This was beneficial for the homolysis of O-O bonds, forming SO(4)(•-) radicals. The attack of the generated active species led to oxidative stress in cells, resulting in damage to the electron transport chain and inhibition of ATP production. Our findings disclose a valuable insight for inactivating ARB, and provide a prospective strategy for ARGs dissemination in water contamination.202437976858
784370.9944Inactivation of chlorine-resistant bacteria (CRB) via various disinfection methods: Resistance mechanism and relation with carbon source metabolism. With the widespread use of chlorine disinfection, chlorine-resistant bacteria (CRB) in water treatment systems have gained public attention. Bacterial chlorine resistance has been found positively correlated with extracellular polymeric substance (EPS) secretion. In this study, we selected the most suitable CRB controlling method against eight bacterial strains with different chlorine resistance among chloramine, ozone, and ultraviolet (UV) disinfection, analyzed the resistance mechanisms, clarified the contribution of EPS to disinfection resistance, and explored the role of carbon source metabolism capacity. Among all the disinfectants, UV disinfection showed the highest disinfection capacity by achieving the highest average and median log inactivation rates for the tested strains. For Bacillus cereus CR19, the strain with the highest chlorine resistance, 40 mJ/cm(2) UV showed a 1.90 log inactivation, which was much higher than that of 2 mg-Cl(2)/L chlorine (0.67 log), 2 mg-Cl(2)/L chloramine (1.68 log), and 2 mg/L ozone (0.19 log). Meanwhile, the UV resistance of the bacteria did not correlate with EPS secretion. These characteristics render UV irradiation the best CRB controlling disinfection method. Chloramine was found to have a generally high inactivation efficiency for bacteria with high chlorine-resistance, but a low inactivation efficiency for low chlorine-resistant ones. Although EPS consumed up to 56.7% of chloramine which an intact bacterial cell consumed, EPS secretion could not explain chloramine resistance. Thus, chloramine is an acceptable CRB control method. Similar to chlorine, ozone generally selected high EPS-secreting bacteria, with EPS consuming up to 100% ozone. Therefore, ozone is not an appropriate method for controlling CRB with high EPS secretion. EPS played an important role in all types of disinfection resistance, and can be considered the main mechanism for bacterial chlorine and ozone disinfection resistance. However, as EPS was not the main resistance mechanism in UV and chloramine disinfection, CRB with high EPS secretion were inactivated more effectively. Furthermore, carbon source metabolism was found related to the multiple resistance of bacteria. Those with low carbon source metabolism capacity tended to have higher multiple resistance, especially to chlorine, ozone, and UV light. Distinctively, among the tested gram-negative bacteria, in contrast to other disinfectants, chloramine resistance was negatively correlated with EPS secretion and positively correlated with carbon source metabolism capacity, suggesting a special disinfection mechanism.202337659185
879180.9944Synergistic biocontrol of Bacillus subtilis and Pseudomonas fluorescens against early blight disease in tomato. Early blight of tomato caused by Alternaria solani results in significant crop losses. In this study, Bacillus subtilis J3 and Pseudomonas fluorescens J8 were co-cultured as a synthetic microbial community (BCA) for synergistic biocontrol of A. solani, and the inhibition mechanism was investigated. BCA presented an inhibition ration against A. solani at 94.91%, which lowered the disease incidence by 38.26-42.87%; reduced peroxidase, catalase, superoxide dismutase activity of tomatoes by 73.11-90.22%; and promoted the biomass by 66.91-489.21%. With BCA protection, the relative expression of tomato resistance genes (including gPAL2, SWRKY, PR-10, and CHI) in roots and leaves was 12.83-90.70% lower than without protection. BCA also significantly altered the rhizosphere and phyllosphere microbial community. The abundance of potentially beneficial bacteria, including Bacillus, Pseudomonas, Arthrobacter, Lysobacter, and Rhizobium, elevated by 6.58-192.77%. They were negatively correlated with resistance gene expression, indicating their vital involvement in disease control. These results provided essential information on the synergistic biocontrol mechanism of bacteria against pathogens, which could contribute to developing novel biocontrol strategies. KEY POINTS: • Bacillus and Pseudomonas present a synergistic biocontrol effect against A. solani. • Biocontrol prevents pathogen damage and improves tomato growth and systemic resistance. • Beneficial bacteria thrive in the rhizosphere is the key to microbial regulation.202337540249
796990.9944Metagenomic insights into the influence of pH on antibiotic removal and antibiotic resistance during nitritation: Regulations on functional genus and genes. The changes in pH and the resulting presence of free nitrous acid (FNA) or free ammonia (FA) often inhibit antibiotic biodegradation during nitritation. However, the specific mechanisms through which pH, FNA and FA influence antibiotic removal and the fate of antibiotic resistance genes (ARGs) are not yet fully understood. In this study, the effects of pH, FNA, and FA on the removal of cefalexin and amoxicillin during nitritation were investigated. The results revealed that the decreased antibiotic removal under both acidic condition (pH 4.5) and alkaline condition (pH 9.5) was due to the inhibition of the expression of amoA in ammonia-oxidizing bacteria and functional genes (hydrolase-encoding genes, transferase-encoding genes, lyase-encoding genes, and oxidoreductase-encoding genes) in heterotrophs. Furthermore, acidity was the primary inhibitor of antibiotic removal at pH 4.5, followed by FNA. Antibiotic removal was primarily inhibited by alkalinity at pH 9.5, followed by FA. The proliferation of ARGs mediated by mobile genetic element was promoted under both acidic and alkaline conditions, attributed to the promotion of FNA and FA, respectively. Overall, this study highlights the inhibitory effects of acidity and alkalinity on antibiotic removal during nitritation.202439068965
7807100.9943Copper oxide/peroxydisulfate system for urban wastewater disinfection: Performances, reactive species, and antibiotic resistance genes removal. In this study, copper oxide (CuO) catalyzed peroxydisulfate (PDS) system was investigated for the inactivation of a broad range of pathogenic microorganisms from urban wastewater. Complete inactivation of Escherichia coli, Enterococcus, F-specific RNA bacteriophages from secondary treated wastewater was achieved after a short time (15-30 min) treatment with CuO (10 g/L)/PDS (1 mM) system, but spores of sulfite-reducing bacteria took 120 min. No bacterial regrowth occurred during storage after treatment. Significant reduction of the pathogens was explained by the generation of the highly selective Cu(III) oxidant, as the predominant reactive species, which could quickly oxidize guanine through a one-electron oxidation pathway. Additionally, the potential of the CuO (10 g/L)/PDS (1 mM) system to inactivate antibiotic-resistant bacteria and antibiotic resistance genes (ARB&Gs) was explored. Sulfamethoxazole-resistant E. coli was used as the model ARB and a 3.2 log of reduction was observed after 10 min of treatment. A considerable reduction (0.7-2.3 log) of selected ARGs including blaTEM, qnrS, emrB, sul1, and genes related to the dissemination of antibiotic resistance, including the Class 1 integron-integrase (intI1), and the insertion sequence (IS613) was achieved after 60 min treatment. All these findings indicated the promising applicability of the CuO/PDS system as a disinfection technology for wastewater reuse in agriculture.202234648831
3534110.9943The Effects of Flavomycin and Colistin Sulfate Pre-Treatment on Ileal Bacterial Community Composition, the Response to Salmonella typhimurium and Host Gene Expression in Broiler Chickens. The composition of the bacterial community affects the intestinal health and growth performance of broiler chickens. The main purpose of this study was to explore the effects of flavomycin and colistin sulfate on the resistance to Salmonella typhimurium infection, ileal bacteria and intestinal health. In total, 396 1-day-old broiler chickens were randomly divided into six groups. Two groups were fed each one of the diets-the control diet (CON), the flavomycin at 10 mg/kg diet (AntiG+), and the colistin sulfate at 40 mg/kg diet (AntiG-), for 5 days. Then, one of each of the two groups was challenged with S. typhimurium on the 8th day; these were named CONS, AntiG+S and AntiG-S, respectively. The results showed that S. typhimurium significantly reduced the feed intake and body weight gain, and increased the feed conversion ratio (p < 0.05). It also increased the inflammatory expressions of NF-κB and MyD88 genes (p < 0.05); and reduced the expressions of claudin-1, occludin and mucin-2 (p < 0.05) tight junction genes in the intestines. S. typhimurium significantly reduced ileal bacterial diversity indexes of observed-species, chao1 and Shannon (p < 0.05). Compared with AntiG+S group, AntiG-S group increased the body weight gain of broiler chickens (p < 0.05), reduced the expression of inflammatory genes (p < 0.05) and intestinal permeability to fluorescein isothiocyanate (p < 0.05). AntiG-S group also improved the ileal bacterial diversity indexes of observed-species and Shannon (p < 0.05). There were many significant correlations between intestinal bacteria, intestinal gene expressions and intestinal morphology (p < 0.05). This study indicated that pre-constructed AntiG- bacteria could against a S. typhimurium infection by inhibiting the expressions of intestinal inflammation genes and increasing the diversity of intestinal bacteria.201931752202
7847120.9943Inactivation and change of tetracycline-resistant Escherichia coli in secondary effluent by visible light-driven photocatalytic process using Ag/AgBr/g-C(3)N(4). Control of antibiotic-resistant bacteria (ARB) and their related genes in secondary effluents has become a serious issue because of increased awareness of their health risks. A considerable number of techniques have been developed in recent years, particularly in relation to advanced oxidation. However, limited information is known about cellular behavior and resistance characteristic change during photocatalytic treatment. In this study, the inactivation of tetracycline (TC)-resistant Escherichia coli (TC-E. coli), removal of TC-resistant genes (TC-RGs), and antibiotic susceptibility were evaluated by employing photocatalytic treatment using Ag/AgBr/g-C(3)N(4) with visible light irradiation. The effects of light intensity, photocatalyst dosage, and reaction ambient temperature on photocatalysis were modelled and investigated. The rate of TC-E. coli removal was also optimized. Results demonstrated that the optimal conditions for TC-E. coli removal included light intensity of 96.0 mW/cm(2), photocatalyst dosage of 211.0 mg/L, and reaction ambient temperature of 23.7 °C. Under such conditions, the ARB removal rate was 6.1 log after 90 min and the related TC-RG removal rates were 49%, 86%, 69%, and 86% for tetA, tetM, tetQ, and intl1, respectively. The minimum inhibitory concentration test after photocatalysis shows that the antibiotic resistance of TC-E. coli was enhanced, which may be mainly due to the changes in the membrane potential and resulted in difficulty in destroying the bacteria through antibiotic contact. Hence, photocatalytic treatment could be an ideal method for ARB and antibiotic-resistant gene (ARG) control in wastewater, but the health risks of the remaining ARB and ARG should be investigated further.202031841919
6238130.9943A novel glutamate-dependent acid resistance among strains belonging to the Proteeae tribe of Enterobacteriaceae. Morganella, Providencia and Proteus strains were capable of surviving pH 2.0 for 1 h if glutamate was present. These strains did not have glutamic acid decarboxylase activity and the gadAB genes were not detected in any of these bacteria. When exposed to pH 2.0 acid shocks, the survival rate of these bacteria was significantly increased with glutamate concentrations as low as 0.3 mM in the acid media. Escherichia coli cells incubated at pH 3.4 consumed four times more glutamate and produced at least 7-fold more gamma-amino butyric acid than Morganella, Providencia and Proteus strains. These results indicate that strains belonging to the Proteeae tribe might have novel glutamate dependent acid-resistance mechanisms.200415321677
8045140.9943Correlation among extracellular polymeric substances, tetracycline resistant bacteria and tetracycline resistance genes under trace tetracycline. Antibiotic resistance occurrences and proliferation in activated sludge have attracted more and more attention nowadays. However, the role which extracellular polymeric substance (EPS) plays on the antibiotic resistance is not clear. The changes and correlation among EPS, tetracycline (TC) resistant bacteria (TRB) and TC resistance genes (TRGs) of sequencing batch reactors (SBRs) were investigated. Performance of SBR without TC was compared with two other SBRs to which different amounts of TC were added. Total average EPS contents were found to increase significantly from 66 mg g−1 VSS to 181 mg g−1 VSS as the TC concentrations increased from 0 to 100 μg L−1. As the EPS content increased, TRB in sludge of the three SBRs increased significantly from 105 to 106 colony forming unit mL−1 after being exposed to TC. In addition, the concentrations of three groups of TRGs (copies mL−1) were determined by real-time fluorescence quantitative polymerase chain reaction and followed the order: efflux pump genes > ribosome protected genes > degradation enzyme genes. The numbers of TRGs in the idle stage were larger than those in the aeration sludge. Correlation coefficients (R2) between EPS and TRB in sludge were 0.823 (p < 0.01) while the correlation between EPS and total TRGs was poor (R2 = 0.463, p > 0.05). But it showed the same tendency that EPS and TRGs in sludge increased with the increasing of TC.201425461932
8034150.9943Adding a complex microbial agent twice to the composting of laying-hen manure promoted doxycycline degradation with a low risk on spreading tetracycline resistance genes. Poultry manure is a reservoir for antibiotics and antibiotic resistance genes and composting is an effective biological treatment for manure. This study explored the effect of using two methods of adding a complex microbial agent to the composting of laying-hen manure on doxycycline degradation and tetracycline resistance genes elimination. The results showed that incorporating a complex microbial agent at 0.8% (w/w) on the 0(th) and 11th day (group MT2) effectively degraded doxycycline with a final degradation rate of 46.83 ± 0.55%. The half-life of doxycycline in this group was 21.90 ± 0.00 days and was significantly lower than that of group MT1 (1.6% (w/w) complex microbial agent added on the 0(th) day) and group DT (compost without complex microbial agent). But there was no significant difference in the final degradation rate of doxycycline between group DT and group MT1. The addictive with the complex microbial agent changed the microbial community structure. Bacteroidetes, Firmicutes and Proteobacteria were the dominant phyla during composting. Aerococcus, Desemzia, Facklamia, Lactobacillus, Streptococcus, and Trichococcus were the bacteria related to the degradation of doxycycline. Moreover, the incorporation of a complex microbial agent could decrease the risk on spreading tetracycline resistance genes. The single addition promoted the elimination of tetM, whose possible hosts were Enterococcus, Lactobacillus, Staphylococcus, and Trichococcus. Adding the complex microbial agent twice promoted the elimination of tetX, which was related to the low abundance of Chryseobacterium, Flavobacterium and Neptunomonas in group MT2. Redundancy analysis showed that the bacterial community, residual doxycycline and physiochemical properties have a potential effect on the variation in tetracycline resistance genes levels. Overall, adding the complex microbial agent twice is an effective measure to degrade doxycycline.202032806409
7802160.9943Inactivating facultative pathogen bacteria and antibiotic resistance genes in wastewater using blue light irradiation combined with a photosensitizer and hydrogen peroxide. The effectiveness of antimicrobial blue light (aBL) irradiation in eliminating ten clinically significant antibiotic resistance genes (ARGs) and four taxonomic marker genes of the WHO-priority ESKAPE bacteria group from wastewater treatment plant (WWTP) effluent was examined. Experiments were conducted using an LED-driven continuous-flow photoreactor operating at wavelengths of 405 nm, 420 nm, and 460 nm. Irradiation with aBL alone was insufficient for effectively inactivating or eliminating ESKAPE bacteria and clinically relevant ARGs. The addition of the porphyrin-based photosensitizer TMPyP (10(-6) M) or the oxidative agent H₂O₂ (1 mM) resulted in several log(10) unit reductions of facultative pathogenic bacteria (FPB), their taxonomic gene markers, and target ARGs. However, the additional effects of TMPyP and H(2)O(2) were only noticeable in conjunction with aBL irradiation, as they were ineffective without it. The reduction of the different FPB and ARGs in WWTP effluents was analyzed using culturing and qPCR together with living/dead discrimination. Different FPB and ARGs showed varying susceptibility to aBL-mediated irradiation. Among the FPB, enterococci were the most sensitive, while among the ARGs bacteria carrying ermB, tetM, sul1, and bla(VIM) genes exhibited the strongest removal. This sensitivity may be due to the gene-carrying microorganism's response to aBL irradiation combined with TMPyP or H(2)O(2). Additionally, molecular biology results revealed that aBL irradiation induced up to 13 lesions per 10 kb DNA, which is hypothesized to contribute to the acute inactivation effect and prevent regrowth by inhibiting DNA repair activities.202540138903
7792170.9943Comparative removal of two antibiotic resistant bacteria and genes by the simultaneous use of chlorine and UV irradiation (UV/chlorine): Influence of free radicals on gene degradation. The research aimed to remove antibiotic resistance by the simultaneous use of UV irradiation and chlorine (UV/chlorine). The inactivations of tetracycline resistant bacteria (TRB) during chlorination, UV irradiation, and UV/chlorine was investigated and compared with those of amoxicillin resistant bacteria (AmRB). Similar examination was also conducted for comparing the removals of their resistant genes (i.e., tetM and blaTem). The removals of antibiotic resistance highly depended on chlorine doses and UV intensities. The sufficient chlorine dose (20 mg.L(-1)) in the chlorination and the UV/chlorine completely inactivated TRB and AmRB (>7.3 log), while the UV irradiation could not achieve the complete disinfection. Microorganisms resistant to different antibiotics exhibit different susceptibility to the disinfection processes. The removals of antibiotic resistant genes (i.e., tetM and blaTem) were more difficult than those of TRB and AmRB. The UV/chlorine was the greatest process for tetM and blaTem removals, followed by chlorination and UV irradiation, respectively. Chlorination decreased the tetM and blaTem by 0.40-1.45 log and 1.04-2.45 log, respectively. The blaTem gene was highly reactive to chlorine, compared with tetM. The UV irradiation caused the tetM and blaTem reductions by 0.32-0.91 log and 0.59-0.96 log, respectively. The UV/chlorine improved the tetM and blaTem removals by 0.98-3.20 log and 1.28-3.36 log, respectively. The •OH contributed to the fraction of tetM and blaTem removals by 48% and 19%, respectively. The effect of reactive chlorine species on the tetM and blaTem removals was minor. The pseudo 1st-order kinetic constants (k') for tetM and blaTem removals by the UV/chlorine were highest. The •OH enhanced the k' values by 120% and 20% for the tetM and blaTem removals, respectively. The study showed the potential use of UV/chlorine for controlling antibiotic resistance.202133059146
8068180.9943Safety of composts consisting of hydrothermally treated penicillin fermentation residue: Degradation products, antibiotic resistance genes and bacterial diversity. Combining hydrothermal treatment and composting is an effective method to dispose of penicillin fermentation residue (PFR), but the safety and related mechanism are still unclear. In this study, penicillin solution was hydrothermally treated to decipher its degradation mechanism, and then hydrothermally treated PFR (HT-PFR) was mixed with bulking agents at ratios of 2:0 (CK), 2:1.5 (T1), and 2:5 (T2) to determine the absolute abundance of antibiotic resistance genes (ARGs) and the succession of bacterial community. Results showed that penicillin was degraded to several new compounds without the initial lactam structure after hydrothermal treatment. During composting, temperature and pH of the composts increased with the raising of HT-PFR proportion, except the pH at days 2. After 52 days of composting, the absolute copies of ARGs (blaTEM, blaCMY2, and blaSFO) and the relative abundance of bacteria related to pathogens were reduced significantly (P < 0.05). Especially, the total amount of ARGs in the samples of CK and T1 were decreased to equal level (around 5 log(10) copies/g), which indicated that more ARGs were degraded in the latter by the composting process. In the CK samples, Bacteroidetes and Proteobacteria accounted for ~69.8% of the total bacteria, but they were gradually replaced by Firmicutes with increasing proportions of HT-PFR, which can be caused by the high protein content in PFR. Consisting with bacterial community, more gram-positive bacteria were observed in T1 and T2, and most of them are related to manganese oxidation and chitinolysis. As composting proceeded, bacteria having symbiotic or pathogenic relationships with animals and plants were reduced, but those related to ureolysis and cellulolysis were enriched. Above all, hydrothermal treatment is effective in destroying the lactam structure of penicillin, which makes that most ARGs and pathogenic bacteria are eliminated in the subsequent composting.202134492529
6351190.9943Heterogeneous expression of DnaK gene from Alicyclobacillus acidoterrestris improves the resistance of Escherichia coli against heat and acid stress. Alicyclobacillus acidoterrestris, an acidophilic and thermophilic bacteria, is an important microbial resource for stress resistance genes screening. In this study, DnaK gene from A. acidoterrestris was subcloned to construct the recombinant plasmid pET28a-DnaK. The successful construction of the plasmid was verified by double-enzyme digestion and sequencing analysis. The recombinant plasmid was transformed into Escherichia coli BL21 and isopropy-β-D-thiogalactoside (IPTG) was used to induce recombinant E. coli to express DnaK gene. A 70 kD fusion protein was identified by SDS-PAGE, which suggested that DnaK gene from A. acidoterrestris was successfully expressed. The recombinant and wild BL21 were treated with high temperatures of 54, 56 and 58 °C at pH values of 5.0-7.0 to compare the effects of heterogeneous expression of the DnaK gene from A. acidoterrestris on the stress resistance. The experimental results showed that survival rate of recombinant BL21-DnaK has been improved considerably under heat and acid stresses in contrast with the wild BL21, and D-values of recombinant BL21 were 14.7-72% higher than that of wild BL21, which demonstrated that heterogeneous expression of DnaK gene from A. acidoterrestris could significantly enhance the resistance of host bacteria E. coli against heat and acid stresses.201728194744