奶牛乳房炎源性产ESBL大肠杆菌的流行特征
发布时间：2021-07-22 11:11
奶牛乳房炎影响奶牛的奶产量和品质,能引起巨大的经济损失,并严重威胁人类的健康。抗生素是治疗奶牛乳房炎的常用药物,而细菌具有耐药的发展潜力,比如细菌代谢产物超广谱β-内酰胺酶（ESBL）可以使β内酰胺酶类药物失活。ESBL多重耐药的大肠杆菌日益流行,已受到广泛的关注。在兽医领域内,ESBL大肠杆菌的报道多来自于禽、猪,在奶牛方面的报道并不多见。因此,本研究旨在:1)调查患有临床型乳房炎的奶牛中产ESBL大肠杆菌的流行情况,并确定编码ESBL基因的亚型;2)确定ESBL大肠杆菌MCR-1基因的检出率3)使用纸片法和肉汤稀释法进行药敏试验,确定ESBL和MCR-1阳性大肠杆菌的耐药性;4)使用限制性片段长度多态性RFLP、构建系统发育树等方法对基因盒和插入序列IS进行序列分析;5)对ESBL和mcr-1阳性菌株进行PCR检测,如质粒复制子（PBRT）、多位点序列分型（MLST）,eBURST和geoBURST分析。本研究于2015-2017年从16个省2038份奶牛乳房炎的乳样中,检出249株（12%）大肠杆菌,其中产ESBL大肠杆菌49株,高达20%。对这些大肠杆菌进行双纸片协同试验和PC...
【文章来源】：中国农业大学北京市 211工程院校 985工程院校 教育部直属院校
【文章页数】：135 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER-Ⅰ: GENERAL INTRODUCTION AND REVIEW OF LITERATURE
1. INTRODUCTION
2. BOVINE MASTITIS
2.1. CLASSIFICATION OF BOVINE MASTITIS
2.2 ETIOLOGY OF BOVINE MASTITIS
2.3 ESCHERICHIA COLT AS A MASTITIS CAUSING PATHOGEN
2.4 EPIDEMIOLOGY OF ESCHERICHIA COLI MASTITIS
2.6 PATHOGENICITY AND PATHOGENESIS OF MASTITIS-ASSOCIATED ESCHERICHIA COLT
2.7 EFFECT OF ESCHERICHIA COLI INFECTION ON THE MAMMARY GLAND
2.8. TREATMENT OF ESCHERICHIA COLI MASTITIS
2.9. ECONOMIC LOSSES OF BOVINE MASTITIS
3. ANTIMICROBIALS
3.1 MECHANISMS OF ACTION OF ANTIMICROBIALS
3.2 USES OF ANTIMICROBIALS IN FOOD-PRODUCING ANIMALS
3.3 BETA-LACTAM ANTIMICROBIALS
3.4 ANTIMICROBIAL RESISTANCE
3.5 MECHANISMS OF ANTIMICROBIAL RESISTANCE
3.6 THE BETA-LACTAMASES
3.7 EXTENDED-SPECTRUM BETA-LACTAMASES(ESBLS)AND ESBL ENCODING GENES
3.8 EPIDEMIOLOGY OF ESBL-PRODUCING ESCHERICHIA COLT IN FOOD ANIMALS
3.9 COLISTIN RESISTANCE,MCR-1 GENE IN ESCHERICHIA COLT AND THE EPIDEMIOLOGY
4. PURPOSE AND SIGNIFICANCE OF THE STUDY
5. KEY QUESTIONS
6. AIMS OF THE STUDY
CHAPTER-Ⅱ: PREVALENCE AND CHARACTERISTICS OF ESBL-PRODUCINGESCHERICHIA COLI
1.INTRODUCTION
1.1 AIMS OF THE STUDY
2. MATERIALS AND METHODS
2.1 SAMPLE COLLECTION AND LOCATION
2.2 ISOLATION AND IDENTIFICATION OF ESCHERICHIA COLT
2.3 PHENOTYPIC SCREENING OF ESBL-PRODUCERS
2.4 GENOTYPIC SCREENING OF ESBL-PRODUCING ESCHERICHIA COLT ISOLATES
2.5 THE POLYMERASE CHAIN REACTION
2.6 ANTIBIOTIC SUSCEPTIBILITY TESTING
2.7 PHYLOGENETIC GROUPING
2.8 DETECTION OF INTEGRONS, GENE CASSETTES AND IS CRI
2.9 PCR-RFLP ASSAY FOR DETECTION OF GENE CASSETTES
2.10 NUCLEOTIDE SEQUENCING AND DATA ANALYSIS
3. RESULTS
3.1 PREVALENCE AND CHARACTERIZATION OF ESBL-PRODUCING ESCHERICHIA COLT
3.2 ANTIBIOTIC SUSCEPTIBILITY PROFILES
3.3 DETECTION OF INTEGRONS,GENE CASSETTES AND IS CRI
3.4 NUCLEOTIDE SEQUENCES ACCESSION NUMBER
4. DISCUSSION
5. CONCLUSION
CHAPTER-Ⅲ: MOLECULAR EPIDEMIOLOGY AND GENETIC DIVERSITY OFESBL-PRODUCING ESCHERICHIA COLL
1. INTRODUCTION
1.1 AIMS OF THE STUDY
2. MATERIALS AND METHOD
2.1 MILK SAMPLE COLLECTION
2.2 ISOLATION AND IDENTIFICATION OF ISOLATES AND ESBL PHENOTYPES
2.3 ANTIBIOTIC SUSCEPTIBILITY TESTING
2.4 MOLECULAR CHARACTERIZATION OF ESBL-PRODUCING ISOLATES
2.5 PLASMID REPLICON TYPING
2.6 RESISTANCE TRANSFER EXPERIMENT
2.7 MULTILOCUS SEQUENCE TYPING (MLST)OF ES BL-PRODUCING ESCHERICHIA COLI
2.8 POPULATION STRUCTURE ANALYSIS
2.9 SEQUENCE COMPOSITIONAL ANALYSIS
2.10 SPLIT NETWORK AND RECOMBINATION ANALYSIS
3. RESULTS
3.1 PREVALENCE AND GENOTYPES OF ESBL-PRODUCING ESCHERICHIA COLI
3.2 PHYLOGENETIC GROUPING
3.3 ANTIMICROBIAL RESISTANCE IN ESBL-PRODUCERS
3.4 PBRT OF ESBL-PRODUCING ESCHERICHIA COLT
3.5 RESISTANCE TRANSFER EXPERIMENT
3.6 MLST AND POPULATION STRUCTURE
3.7 SEQUENCE COMPOSITIONAL ANALYSIS
3.8 MULTI-LOCUS SEQUENCE ANALYSIS (MLSA)
3.9 RECOMBINATION ANALYSIS
4. DISCUSSION
5. CONCLUSION
CHAPTER-Ⅳ: CO-OCCURRENCE OF MCR-1 AND ESBL GENES INESCHERICHIA COLI
1. INTRODUCTION
1.1 AIMS OF THE STUDY
2. MATERIALS AND METHODS
2.1 THE STUDY AND ISOLATION OF ESCNERICHIA COLI
2.2 ANTIMICROBIAL RESISTANT PHENOTYPES
2.3 DETECTION OF MCR-1 GENE AND OTHER RESISTANCE GENES
2.4 INTEGRON AND PLASMID TYPING OF MCR-1 POSITIVE ISOLATES
2.5 CONJUGATION EXPERIMENT
2.6 MULTILOCUS SEQUENCE TYPING AND ANALYSIS
3. RESULTS
3.1 CO-OCCURRENCE OF MCR-1 AND ESBL ENCODING GENES
3.2 ANTIMICROBIAL SENSITIVITY PROFILE
3.3 CHARACTERIZATION OF INTEGRONS AND VARIABLE REGIONS
3.4 PLASMID REPLICON TYPING AND RESISTANCE TRANSFER EXPERIMENTS
3.5 MULTILOCUS SEQUENCE TYPING AND ANALYSIS
4. DISCUSSION
CHAPTER-Ⅴ: CONCLUSIONS AND RECOMMENDATIONS
NOVELTY OF THE STUDY
REFERENCES
ACKNOWLEDGEMENT
AUTHOR'S SHORT CV
Annexures
本文编号：3297041
【文章来源】：中国农业大学北京市 211工程院校 985工程院校 教育部直属院校
【文章页数】：135 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER-Ⅰ: GENERAL INTRODUCTION AND REVIEW OF LITERATURE
1. INTRODUCTION
2. BOVINE MASTITIS
2.1. CLASSIFICATION OF BOVINE MASTITIS
2.2 ETIOLOGY OF BOVINE MASTITIS
2.3 ESCHERICHIA COLT AS A MASTITIS CAUSING PATHOGEN
2.4 EPIDEMIOLOGY OF ESCHERICHIA COLI MASTITIS
2.6 PATHOGENICITY AND PATHOGENESIS OF MASTITIS-ASSOCIATED ESCHERICHIA COLT
2.7 EFFECT OF ESCHERICHIA COLI INFECTION ON THE MAMMARY GLAND
2.8. TREATMENT OF ESCHERICHIA COLI MASTITIS
2.9. ECONOMIC LOSSES OF BOVINE MASTITIS
3. ANTIMICROBIALS
3.1 MECHANISMS OF ACTION OF ANTIMICROBIALS
3.2 USES OF ANTIMICROBIALS IN FOOD-PRODUCING ANIMALS
3.3 BETA-LACTAM ANTIMICROBIALS
3.4 ANTIMICROBIAL RESISTANCE
3.5 MECHANISMS OF ANTIMICROBIAL RESISTANCE
3.6 THE BETA-LACTAMASES
3.7 EXTENDED-SPECTRUM BETA-LACTAMASES(ESBLS)AND ESBL ENCODING GENES
3.8 EPIDEMIOLOGY OF ESBL-PRODUCING ESCHERICHIA COLT IN FOOD ANIMALS
3.9 COLISTIN RESISTANCE,MCR-1 GENE IN ESCHERICHIA COLT AND THE EPIDEMIOLOGY
4. PURPOSE AND SIGNIFICANCE OF THE STUDY
5. KEY QUESTIONS
6. AIMS OF THE STUDY
CHAPTER-Ⅱ: PREVALENCE AND CHARACTERISTICS OF ESBL-PRODUCINGESCHERICHIA COLI
1.INTRODUCTION
1.1 AIMS OF THE STUDY
2. MATERIALS AND METHODS
2.1 SAMPLE COLLECTION AND LOCATION
2.2 ISOLATION AND IDENTIFICATION OF ESCHERICHIA COLT
2.3 PHENOTYPIC SCREENING OF ESBL-PRODUCERS
2.4 GENOTYPIC SCREENING OF ESBL-PRODUCING ESCHERICHIA COLT ISOLATES
2.5 THE POLYMERASE CHAIN REACTION
2.6 ANTIBIOTIC SUSCEPTIBILITY TESTING
2.7 PHYLOGENETIC GROUPING
2.8 DETECTION OF INTEGRONS, GENE CASSETTES AND IS CRI
2.9 PCR-RFLP ASSAY FOR DETECTION OF GENE CASSETTES
2.10 NUCLEOTIDE SEQUENCING AND DATA ANALYSIS
3. RESULTS
3.1 PREVALENCE AND CHARACTERIZATION OF ESBL-PRODUCING ESCHERICHIA COLT
3.2 ANTIBIOTIC SUSCEPTIBILITY PROFILES
3.3 DETECTION OF INTEGRONS,GENE CASSETTES AND IS CRI
3.4 NUCLEOTIDE SEQUENCES ACCESSION NUMBER
4. DISCUSSION
5. CONCLUSION
CHAPTER-Ⅲ: MOLECULAR EPIDEMIOLOGY AND GENETIC DIVERSITY OFESBL-PRODUCING ESCHERICHIA COLL
1. INTRODUCTION
1.1 AIMS OF THE STUDY
2. MATERIALS AND METHOD
2.1 MILK SAMPLE COLLECTION
2.2 ISOLATION AND IDENTIFICATION OF ISOLATES AND ESBL PHENOTYPES
2.3 ANTIBIOTIC SUSCEPTIBILITY TESTING
2.4 MOLECULAR CHARACTERIZATION OF ESBL-PRODUCING ISOLATES
2.5 PLASMID REPLICON TYPING
2.6 RESISTANCE TRANSFER EXPERIMENT
2.7 MULTILOCUS SEQUENCE TYPING (MLST)OF ES BL-PRODUCING ESCHERICHIA COLI
2.8 POPULATION STRUCTURE ANALYSIS
2.9 SEQUENCE COMPOSITIONAL ANALYSIS
2.10 SPLIT NETWORK AND RECOMBINATION ANALYSIS
3. RESULTS
3.1 PREVALENCE AND GENOTYPES OF ESBL-PRODUCING ESCHERICHIA COLI
3.2 PHYLOGENETIC GROUPING
3.3 ANTIMICROBIAL RESISTANCE IN ESBL-PRODUCERS
3.4 PBRT OF ESBL-PRODUCING ESCHERICHIA COLT
3.5 RESISTANCE TRANSFER EXPERIMENT
3.6 MLST AND POPULATION STRUCTURE
3.7 SEQUENCE COMPOSITIONAL ANALYSIS
3.8 MULTI-LOCUS SEQUENCE ANALYSIS (MLSA)
3.9 RECOMBINATION ANALYSIS
4. DISCUSSION
5. CONCLUSION
CHAPTER-Ⅳ: CO-OCCURRENCE OF MCR-1 AND ESBL GENES INESCHERICHIA COLI
1. INTRODUCTION
1.1 AIMS OF THE STUDY
2. MATERIALS AND METHODS
2.1 THE STUDY AND ISOLATION OF ESCNERICHIA COLI
2.2 ANTIMICROBIAL RESISTANT PHENOTYPES
2.3 DETECTION OF MCR-1 GENE AND OTHER RESISTANCE GENES
2.4 INTEGRON AND PLASMID TYPING OF MCR-1 POSITIVE ISOLATES
2.5 CONJUGATION EXPERIMENT
2.6 MULTILOCUS SEQUENCE TYPING AND ANALYSIS
3. RESULTS
3.1 CO-OCCURRENCE OF MCR-1 AND ESBL ENCODING GENES
3.2 ANTIMICROBIAL SENSITIVITY PROFILE
3.3 CHARACTERIZATION OF INTEGRONS AND VARIABLE REGIONS
3.4 PLASMID REPLICON TYPING AND RESISTANCE TRANSFER EXPERIMENTS
3.5 MULTILOCUS SEQUENCE TYPING AND ANALYSIS
4. DISCUSSION
CHAPTER-Ⅴ: CONCLUSIONS AND RECOMMENDATIONS
NOVELTY OF THE STUDY
REFERENCES
ACKNOWLEDGEMENT
AUTHOR'S SHORT CV
Annexures
本文编号：3297041
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