npj Biofilms and Microbiomes
·
2023
·Vol. 9
·期 1
·1-14
·
IF 9.2
Hu Jie,Wu Yifan,Kang Luyuan,Liu Yisi,Ye Hao,Wang Ran,Zhao Jinbiao,Zhang Guolong,Li Xilong,Wang Junjun,Han Dandan
Elimination of specific enteropathogenic microorganisms is critical to gut health. However, the complexity of the gut community makes it challenging to target specific bacterial organisms. Accumulating evidence suggests that various foods can change the abundance of intestinal bacteria by modulating prophage induction. By using pathogenic Escherichia coli ( E. coli ) ATCC 25922 as a model in this research, we explored the potential of dietary modulation of prophage induction and subsequent bacterial survival. Among a panel of sugars tested in vitro, D -xylose was shown to efficiently induce prophages in E. coli ATCC 25922, which depends, in part, upon the production of D -lactic acid. In an enteric mouse model, prophage induction was found to be further enhanced in response to propionic acid. Dietary D -xylose increased the proportion of Clostridia which converted D -lactic acid to propionic acid. Intestinal propionic acid levels were diminished, following either oral gavage with the dehydrogenase gene ( ldhA )-deficient E. coli ATCC 25922 or depletion of intestinal Clostridia by administration of streptomycin. D -Xylose metabolism and exposure to propionic acid triggered E. coli ATCC 25922 SOS response that promoted prophage induction. E. coli ATCC 25922 mutant of RecA, a key component of SOS system, exhibited decreased phage production. These findings suggest the potential of using dietary components that can induce prophages as antimicrobial alternatives for disease control and prevention by targeted elimination of harmful gut bacteria.
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ZOOLOGICAL RESEARCH
·
2023
·
IF 4.9
Xue-Yang Wang,Huang-Wei Song,Tian Yi,Ying-Bo Shen,Chong-Shan Dai,Cheng-Tao Sun,De-Jun Liu,Jian-Zhong Shen,Cong-Ming Wu,Yang Wang
Conjugative transfer of antibiotic resistance genes (ARGs) by plasmids is an important route for ARG dissemination. An increasing number of antibiotic and nonantibiotic compounds have been reported to aid the spread of ARGs, highlighting potential challenges for controlling this type of horizontal transfer. Development of conjugation inhibitors that block or delay the transfer of ARG-bearing plasmids is a promising strategy to control the propagation of antibiotic resistance. Although such inhibitors are rare, they typically exhibit relatively high toxicity and low efficacy in vivo and their mechanisms of action are inadequately understood. Here, we studied the effects of dihydroartemisinin (DHA), an artemisinin derivative used to treat malaria, on conjugation. DHA inhibited the conjugation of the IncI2 and IncX4 plasmids carrying the mobile colistin resistance gene ( mcr-1 ) by more than 160-fold in vitro in Escherichia coli , and more than two-fold (IncI2 plasmid) in vivo in a mouse model. It also suppressed the transfer of the IncX3 plasmid carrying the carbapenem resistance gene bla NDM-5 by more than two-fold in vitro . Detection of intracellular adenosine triphosphate (ATP) and proton motive force (PMF), in combination with transcriptomic and metabolomic analyses, revealed that DHA impaired the function of the electron transport chain (ETC) by inhibiting the tricarboxylic acid (TCA) cycle pathway, thereby disrupting PMF and limiting the availability of intracellular ATP for plasmid conjugative transfer. Furthermore, expression levels of genes related to conjugation and pilus generation were significantly down-regulated during DHA exposure, indicating that the transfer apparatus for conjugation may be inhibited. Our findings provide new insights into the control of antibiotic resistance and the potential use of DHA.
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ENZYME AND MICROBIAL TECHNOLOGY
·
2023
·
IF 3.4
Fangfang Chang,Yuheng Wang,Jie Zhang,Tao Tu,Huiying Luo,Huoqing Huang,Yingguo Bai,Xing Qin,Yaru Wang,Bin Yao,Yuan Wang,Xiaolu Wang
γ-Aminobutyric acid (GABA) has been widely used in the food, feed, pharmaceutical, and chemical industry fields. Previously, we developed a whole-cell catalyst capable of converting L-glutamate (L-Glu) into GABA by overexpressing the glutamate decarboxylase gene ( gadz11 ) from Bacillus sp. Z11 in Escherichia coli BL21(DE3). However, to enhance cell permeability , a freeze-thaw treatment is required, and to enhance GADZ11 activity, pyridoxal 5′-phosphate (PLP) must be added to the reaction system. The aim of this study is to provide a more efficient approach for GABA production by engineering the recombinant E. coli above. First, the inducible expression conditions of the gadz11 in E. coli were optimized to 37 °C for 6 h. Next, an ideal engineered strain was produced via increasing cell permeability by overexpressing sulA and eliminating PLP dependence by constructing a self-sufficient system. Furthermore, an efficient whole-cell biocatalytic process was optimized. The optimal substrate concentration , cell density, and reaction temperature were 1.0 mol/L (the molecular ratio of L-Glu to L-monosodium glutamate (L-MSG) was 4:1), 15 and 37 °C, respectively. Finally, a whole-cell bioconversion procedure was performed in a 3-L bioreactor under optimal conditions. The strain could be reused for at least two cycles with GABA yield, productivity and conversion ratio of 206.2 g/L, 117.8 g/L/h and 100.0%, respectively. This is currently the highest GABA productivity from a mixture of L-Glu and L-MSG reported without the addition of cofactors or additional treatment of cells. This work demonstrates that the novel engineered E. coli strain has the potential for application in large-scale industrial GABA production.
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TALANTA
·
2023
·
IF 6.1
Yong Wang,Xiao Cheng,Chenchen Wang,Duoduo Zhang,Anran Liu,Zhi Wang,Wei Wei,Songqin Liu
Escherichia coli ( E. coli ) is one of the most ubiquitous foodborne pathogens that can cause infections and threaten human health. Herein, a colorimetric method for sensitive detection of E. coli was established by using enzyme-nanozyme cascade reaction for signal amplification. Gold nanoparticles (AuNPs) are well-known nanozymes due to their high peroxidase-like activity. When the dense cetyltrimethylammonium bromide (CTAB) membrane on the surfaces of AuNPs kept the substrate away from AuNPs, the peroxidase activity of AuNPs was inhibited. However, the CTAB membrane could be disrupted by Ag + , resulting in enhanced peroxidase activity of AuNPs. When E. coli was present, the enzyme-nanozyme cascade reaction was initiated. The substrate p-aminophenyl β-D-galactopyranoside (PAPG) was hydrolyzed to the reductive p-aminophenol (PAP) by beta-galactosidase (β-gal) in E. coli , reducing Ag + to Ag. Consequently, CTAB-AuNPs remained weak peroxidase activity and could not catalyze the H 2 O 2 -mediated oxidation of TMB . As the amount of E. coli increased, the absorbance of TMB decreased along with a color change from deep blue to pink. The absorbance intensity displayed a linear dependence on E. coli from 1.0 × 10 2 to 1.0 × 10 9 CFU mL −1 . Therefore, the proposed method holds good prospects in foodborne pathogenic bacteria detection.
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ENVIRONMENTAL SCIENCE & TECHNOLOGY
·
2023
·
IF 11.4
Hebin Liang,Jiayu Zhang,Jiahui Hu,Xiaoyan Li,Bing Li
Extensive antibiotic use increases the environmental presence of their residues and may accelerate the development of antibiotic resistance, although this remains poorly understood at environmentally relevant concentrations. Herein, susceptible Escherichia coli K12 was continuously exposed to five antibiotics at such concentrations for 100 days. The de novo-evolved mutants rapidly obtained fluoroquinolone resistance within 10 days, as indicated by the 4- and 16-fold augmentation of minimum inhibitory concentrations against enrofloxacin and ciprofloxacin, respectively. Moreover, the mutants maintained heritable fluoroquinolone resistance after the withdrawal of antibiotics for 30 days. Genomic analysis identified Asp87Gly or Ser83Leu substitutions in the gyrA gene in the mutants. Transcriptomics data showed that the transcriptional response of the mutants to fluoroquinolones was primarily involved in biofilm formation, cellular motility, porin, oxidative stress defense, and energy metabolism. Homologous recombination and molecular docking revealed that mutations of gyrA primarily mainly conferred fluoroquinolone resistance, while mutations at different positions of gyrA likely endowed different fluoroquinolone resistance levels. Collectively, this study revealed that environmentally relevant concentrations of antibiotics could rapidly induce heritable antibiotic resistance; therefore, the discharge of antibiotics into the environment should be rigorously controlled to prevent the development of antibiotic resistance.
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Frontiers in Immunology
·
2017
·
IF 6.429
Ren Wenkai,Yin Jie,Xiao Hao,Chen Shuai,Liu Gang,Tan Bie,Li Nengzhang,Peng Yuanyi,Li Tiejun,Zeng Benhua,Li Wenxia,Wei Hong,Yin Zhinan,Wu Guoyao,Hardwidge Philip R.,Yin Yulong
Intestinal microbiota has critical importance in pathogenesis of intestinal infection; however, the role of intestinal microbiota in intestinal immunity during enterotoxigenic Escherichia coli (ETEC) infection is poorly understood. The present study tested the hypothesis that the intestinal microbiota is associated with intestinal interleukin-17 (IL-17) expression in response to ETEC infection. Here, we found ETEC infection induced expression of intestinal IL-17 and dysbiosis of intestinal microbiota, increasing abundance of γ-aminobutyric acid (GABA)-producing Lactococcus lactis subsp. lactis. Antibiotics treatment in mice lowered the expression of intestinal IL-17 during ETEC infection, while GABA or L. lactis subsp. lactis administration restored the expression of intestinal IL-17. L. lactis subsp. lactis administration also promoted expression of intestinal IL-17 in germ-free mice during ETEC infection. GABA enhanced intestinal IL-17 expression in the context of ETEC infection through activating mechanistic target of rapamycin complex 1 (mTORC1)-ribosomal protein S6 kinase 1 (S6K1) signaling. GABA–mTORC1 signaling also affected intestinal IL-17 expression in response to Citrobacter rodentium infection and in drug-induced model of intestinal inflammation. These findings highlight the importance of intestinal GABA signaling in intestinal IL-17 expression during intestinal infection and indicate the potential of intestinal microbiota-GABA signaling in IL-17-associated intestinal diseases.
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BIORESOURCE TECHNOLOGY
·
2014
·
IF 5.039
Cuixia Chen,Shaopeng Ding,Dezheng Wang,Zhimin Li,Qin Ye
In this study, the production of succinic acid from cassava starch and raw cassava instead of glucose by Escherichia coli NZN111 was investigated. During the two-stage fermentation, simultaneous saccharification and fermentation (SSF) was applied in the anaerobic stage. The results showed that both the productivity and specific productivity in the process conducted at 40 °C were higher than those in the cultivation conducted at 37 °C. The yield of succinic acid based on the amount of added starch reached the highest level 0.86 g/g and cassava starch was almost totally hydrolyzed in the SSF process. With the improved cell density, 127.13 g/L of succinic acid was obtained. When the liquefied crude cassava powder was used directly in SSF, 106.17 g/L of succinic acid was formed. The result showed that crude cassava powder could be another cheap raw material for succinic acid formation.
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APPLIED SURFACE SCIENCE
·
2014
·
IF 2.538
Mengying He,Ting Wu,Siyi Pan,Xiaoyun Xu
Antimicrobial mechanism of four flavonoids (kaempferol, hesperitin, (+)-catechin hydrate, biochanin A) against Escherichia coli ATCC 25922 was investigated through cell membranes and a liposome model. The release of bacterial protein and images from transmission electron microscopy demonstrated damage to the E. coli ATCC 25922 membrane. A liposome model with dipalmitoylphosphatidylethanolamine (DPPE) (0.6 molar ratio) and dipalmitoylphosphatidylglycerol (DPPG) (0.4 molar ratio), representative of the phospholipid membrane of E. coli ATCC 25922, was used to specify the mode of action of four selected flavonoids through Raman spectroscopy and differential scanning calorimetry. It is suggested that for flavonoids, to be effective antimicrobials, interaction with the polar head-group of the model membrane followed by penetration into the hydrophobic regions must occur. The antimicrobial efficacies of the flavonoids were consistent with liposome interaction activities, kaempferol > hesperitin > (+)-catechin hydrate > biochanin A. This study provides a liposome model capable of mimicking the cell membrane of E. coli ATCC 25922. The findings are important in understanding the antibacterial mechanism on cell membranes.
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Veterinary Sciences
·
2025
·Vol. 12
·期 4
·294
·
IF 2.3
Tiantian Gao,Xuesong Liu,Di Qiu,Yanan Li,Zongsheng Qiu,Jingjing Qi,Shuxin Li,Xiaoyan Guo,Yan Zhang,Ziqi Wang,Xiang Gao,Yuhui Ma,Tianwen Ma
Simple SummaryBacterial infection is a major cause of death in foals.Escherichia coli(E. coli) is a Gram-negative bacterium that causes sepsis, which can be life-threatening in horses. Cefquinome has broad-spectrum antimicrobial activity and is widely used in the treatment of animal diseases. At the same time, it is the fourth generation of cephalosporin antibiotics for veterinary use, and it has good antibacterial activity against both Gram-positive and Gram-negative bacteria. Cefquinome is utilized for the treatment of septicemia induced byE. coli, as well as respiratory infections caused byStreptococcus equisubsp.zooepidemicusin foals. However, there is a lack of studies using cefquinome to targetE. colias a pathogen of sepsis. Pharmacokinetic/pharmacodynamic (PK/PD) modeling is a fundamental step in semi-mechanical methodology aimed at improving dose planning of systemically active antimicrobial agents. PK/PD models can determine the relationship between drug concentration, antibacterial action, and time, deriving the most effective dosing regimen. This study was based on this model to verify the optimal dosing regimen forE. coliinfection in foals and to provide a theoretical basis for the precision treatment of foals.Cefquinome is used to treat septicemia caused byEscherichia coli(E. coli) and respiratory infections caused byStreptococcus equisubsp.zooepidemicusin foals. However, studies reporting the use of cefquinome to targetE. colias pathogens of sepsis are lacking. Therefore, this study aimed to determine the optimal dosage regimen for cefquinome againstE. coliusing a PK/PD model. After the administration of 1 mg/kg cefquinome (intramuscularly or intravenously), blood samples were collected at different time points to determine the serum concentration of cefquinome via HPLC. The pharmacokinetic parameters were evaluated via NCA (WinNonlin 5.2.1 software). The main pharmacokinetic parameters of cefquinome in foals were as follows: after intravenous administration, the elimination half-life (T1/2β) was 2.35 h, the area under the curve (AUC0–last) was 12.33 μg·h/mL, the mean residence time (MRT0–last) was 2.67 h, and the clearance rate (CL) was 0.09 L/h/kg. After intramuscular administration, the peak concentration (Cmax) was 0.89 μg/mL, the time to reach the maximum serum concentration (Tmax) was 2.16 h, T1/2βwas 4.16 h, AUC0–lastwas 5.41 μg·h/mL, MRT0–lastwas 4.92 h, CL was 0.15 L/h/kg, and the absolute bioavailability (F) was 43.86%. An inhibitory sigmoid Emax model was used to integrate the PK/PD indices with ex vivo antimicrobial effects to identify pharmacodynamic targets (PDTs). According to the dose calculation formula, the doses of intramuscularly administered cefquinome required to achieve bacteriostatic effects, bactericidal effects, and bactericidal elimination were 1.10, 1.66, and 2.28 mg/kg, respectively. However, further studies are warranted to verify the therapeutic efficacy of cefquinome in clinical settings.
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PROCESS BIOCHEMISTRY
·
2024
·
IF 3.7
Long Zhang,Jinna Cui,Wei Liu,Zhanying Liu
Various protein secretion systems of gram-negative Escherichia coli ( E. coli ) are well known, but the secretion pathway of endoglucanase (BcsZ) is unknown. To further elucidate the secretion mechanism, we analyzed the transcriptome data of five ruminal cellulolytic E. coli strains. Gene Ontology (GO) enrichment analysis revealed that the differentially expressed genes (DEGs) upregulated in all five strains were significantly enriched in the membrane. Nine DEGs related to membrane transporters were overexpressed in E. coli JBZ-DH5α (overexpressing BcsZ in the genome of E. coli DH5α). The results revealed that the overexpression of the b4332 ( yjiJ ), b2142 ( yohK ), and b2377 ( yfdY ) genes significantly reduced the growth and BcsZ concentration of the strain. The overexpression of the genes b3533 ( bcsA ), b3009 ( yghB ), and b3522 ( yhjD ) did not significantly affect strain growth or the BcsZ concentration. The overexpression of the b1859 ( znuB ), b0458 ( ylaC ), and b1195 ( ymgE ) genes increased the secretion of BcsZ but had no significant effect on strain growth. In conclusion, the yjiJ , yohK , yfdY , znuB , ylaC , and ymgE genes are involved in the secretion of E. coli BcsZ. These findings provide insight into the secretion mechanism of E. coli BcsZ.
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FOOD CHEMISTRY
·
2024
·
IF 8.5
Xiaoqing Wang,Xufeng Zang,Laiyi Deng,Fei Tan,Xingbo Liu,Zhiguo Zhang,Bo Cui,Yishan Fang
Herein, a signal stable molecularly imprinted photoelectrochemical (MIP-PEC) sensing platform was designed to sensitively detect Escherichia coli by incorporating polythiophene film with Cu: ZIF-8/KZ3TTz heterojunction . Attributed to the formation of a staggered type II heterostructure between KZ3TTz and Cu: ZIF-8 semiconductors, the Cu: ZIF-8/KZ3TTz heterojunction exhibited stable and significant cathode PEC response. Impressively, selective MIP film was grown on the surface of Cu: ZIF-8/KZ3TTz/GCE by electro-polymerization of 2,2-Dimethyl-5-(3-thienyl)-1,3-dioxane-4,6-dione (DTDD) in the presence of E. coli . After removing E. coli , more electrons were transferred to the electrolyte solution through the imprinting cavity on the MIP film, which was eliminated by O 2 in the electrolyte, causing further enhancement of the cathode PEC response. On the contrary, when the imprinted cavity was filled with E. coli , the cathodic PEC response gradually decreased due to steric hindrance effect. The sensor showed excellent linearity in the range of 10 1 to 10 8 CFU/mL with a detection limit of 4.09 CFU/mL (S/N = 3). This strategy offered a novel approach for pathogenic bacteria detection in food safety and environmental monitoring
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JOURNAL OF HAZARDOUS MATERIALS
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2024
·
IF 12.2
Xuejing Ma,Han Yeong Kaw,Jing Yu,Qi Yang,Lizhong Zhu,Wei Wang
The extensive consumption of antibiotics has been reported to significantly promote the generation of antibiotic resistance (ABR), however, a quantitative causal relationship between antibiotic exposure and ABR response is absent. This study aimed to pinpoint the accurate regulatory concentration of fluoroquinolones (FQs) and to understand the biochemical mechanism of the mutual action between FQ exposure and FQ resistance response. Highly sensitive analytical methods were developed by using UPLC-MS/MS to determine the total residual, extracellular residual, total intracellular, intracellular residual and intracellular degraded concentration of three representative FQs, including ciprofloxacin (CIP), ofloxacin (OFL) and norfloxacin (NOR), with detection limits in the range of 0.002–0.057 μg/L, and recoveries in the range of 80–93%. The MICs of Escherichia coli ( E. coli ) were 7.0–31.4-fold of the respective MIC 0 after 40-day FQ exposure, and significant negative associations were discovered between the intracellular (residual, degraded or the sum) FQ concentrations and FQ resistance. Transcriptional expression and whole-genome sequencing results indicated that reduced membrane permeability and enhanced multi-drug efflux pumps contributed to the decreasing intracellular concentration. These results unveiled the pivotal role of intracellular concentration in triggering FQ resistance, providing important information to understand the dose-response relationship between FQ exposure and FQ resistance response, and ascertain the target dose metric of FQs for eliminating FQ resistance crisis.
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