目录 Chapter 1 Microfluidics for rapid antibiotic susceptibility testing 11.1 Microfluidic platform design 21.1.1 System design of microfluidic platform 2 1.1.2 Construction of microfluidic platform 31.1.3 Preparation of microfluidic agarose chip 41.1.4 Characterization of concentration gradient on the chip 81.2 Observation and data processing 91.2.1 Image processing and characterization for bacterial quantification 91.2.2 Microbial counting and growth model 101.2.3 Calculation of bacterial growth rate 111.2.4 Microbe inhibition dynamic model 111.3 Comparison of bacterial culture by chips and traditional methods 111.3.1 Growth of Escherichia coli in the chip and well plate 111.3.2 Growth of ammonia-oxidizing bacteria in the chips and shaking flasks 141.4 Amoxicillin susceptibility testing of different bacterial strains 161.4.1 Escherichia coli 161.4.2 Nitrosomonas europaea 221.4.3 Comamonas denitrifican 25References 29Chapter 2 Bacterial persistence to antibiotics revealed by single cell tracking 312.1Discovery of opportunistic antibiotic resistance bacteria 322.1.1Bacterial growth process based on single-cell tracking 322.1.2Opportunistic antibiotic resistance in Escherichia coli 342.1.3Opportunistic antibiotic resistance in Comamonas denitrifican 402.2Development of bacterial resistance under long-term antibiotic pressure 422.2.1In situ recovery growth of Nitrosomonas europaea after inhibition 432.2.2Metabolites during in situ recovery of Nitrosomonas europaea 442.3Effect of lag phase on bacterial resistance to antibiotics 452.3.1Pure strains 452.3.2Activated sludge bacterial community 512.3.3Predicting bacterial antibiotic resistance based on IC50 and lag time 562.3.4Extended lag time promotes bacteria regrowth after removal of antibiotics 58References 59Chapter 3 Transfer characteristics of antibiotic resistance genes in biofilmsbased on microfluidics 633.1Mating assays based on microfluidics method 633.1.1Bacterial strains and amplification 633.1.2Single-channel microfluidic experiments 643.1.3Plasmid transfer frequency anaysis by cell sorting 673.1.4Cell sequencing and conjugative potential analysis 713.1.5Advantages of microfluidics over filter mating 733.2Transfer characteristics of ARGs in pure strains 733.2.1Cascading plasmid transfer in Escherichia coli biofilm 733.2.2The impact of recipient bacterial species on the gene transfer 783.3Transfer characteristics of ARGs in bacterial community 823.3.1Gene transfer in environmental community biofilms 823.3.2The effect of donor bacterial species on ARG transfer in bacterial community 823.3.3Transfer frequency in recipients from different activated sludgecommunities 873.3.4Community structure analysis of recipients and transconjugants 903.3.5Plasmid-host susceptible genera in recipients 933.3.6Conjugative potential of plasmid-host susceptible genera 95References 98Chapter 4 Direct observation and dissection of horizontal and verticalgene transfer in bacterial community 1024.1Single-cell tracking microfluidic chip 1034.2Dissecting of horizontal and vertical transfer of ARGs in the bacterial community 1044.2.1Methods for tracking HGT and VGT 1044.2.2Characteristics of HGT and VGT 1074.3The effect of antibiotics on HGT and VGT in bacterial communit y1114.3.1The effect of antibiotics on ARG transmission processes 1124.3.2The dynamics of ARG spread under different antibiotics 116References 118Chapter 5 The influence of typical environmental factors on antibioticresistance gene transfer 1215.1Porous PDMS-agarose chip 1225.2The influence of heavy metals on the transfer process of ARGs inbacterial community 1235.2.1Effects of six heavy metals on the growth of donor and recipientbacteria 1235.2.2Screening of heavy metals promoting ARG transfer bacterialcommunities 1265.2.3ARG transfer dynamics under different heavy metal stress 1305.2.4ARG transfer frequencies under selected concentrations of Pb,As and Hg 1345.2.5Mechanisms of selected heavy metals promoting ARG transfer 1365.3The dissemination potential of ARGs by DRB after chlorinationdisinfection 1385.3.1The growth characteristics of DRB by re-culture 1385.3.2ARG transfer in DRB after different chlorination disinfections 1405.3.3Visual analysis of ARG transfer in DRB based on microfluidic chip 145References 146Chapter 6 Microplastics-associated biofilm formation and antibiotic resistance gene spread 1496.1Microplastic interceptor chip 1506.2Small-sized microplastics 1516.2.1Bacterial growth exposed to microplastics 1516.2.2Effects of microplastics on ARG transfer 1656.2.3Effects of microplastics on biochemical indices of bacteria 1716.3Large-sized microplastics 1816.3.1Bacterial growth and biofilm formation 1816.3.2ARG transfer in the biofilm on microplastics 182References 183
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