扬州地区屠宰
发布时间：2019-03-20 16:38
【摘要】：沙门菌是重要的人兽共患病原菌,长期以来对人类和动物都造成了严重的危害。蛋类、禽肉和猪肉一直被认为是引起沙门菌病的三大来源。而随着蛋鸡中沙门菌防控项目的大规模开展,蛋类和禽肉中沙门菌的分离率均得到了显著的降低。因此,猪肉及猪肉制品中沙门菌的防控已成为当今发达国家关注的一大重点,而作为猪肉生产链中两个最重要的环节,屠宰场的屠宰流程和农贸市场猪肉零售点的卫生状况对于猪肉中沙门菌的防控有着深远的影响。本研究旨在了解扬州市屠宰场、农贸市场猪肉以及人群中沙门菌的分布状况、血清型特点以及耐药情况,并利用MLST、PFGE、CRISPR等分子亚分型方法对它们进行基因分型研究,以期探明屠宰场、农贸市场猪肉以及人来源沙门菌之间的内在联系并找出潜在的沙门菌传播途径,从而为食源性沙门菌的防控提供数据支持。一、扬州地区屠宰场、农贸市场以及人源沙门菌分离株的分离鉴定及药物敏感性分析2013年10月至2014年12月,从扬州市某屠宰场及其下游农贸市场采集环境样品和屠宰猪样品392份、猪肉样品178份,分别分离出沙门菌275株和130株,分离率为70.2%(275/392)和73.0%(130/178)。从扬州市和泰州市疾病预防与控制中心收集人源沙门菌49株。三种来源的沙门菌共计454株。统计学分析显示,屠宰场5次采样沙门菌分离率之间并无显著性差异(P0.05),且沙门菌的分离情况受季节因素影响不大。血清型鉴定结果显示,屠宰场和农贸市场样品中的最优势血清型均为德尔卑沙门菌,而在人源沙门菌中,德尔卑沙门菌位居第二,其他血清型如鼠伤寒沙门菌、鸭沙门菌、伦敦沙门菌在三种来源中也较为常见。对454株三种来源的沙门菌进行16种抗生素的药敏实验,结果显示屠宰场来源的沙门菌对四环素耐药率最高,为30.9%;而农贸市场猪肉来源的沙门菌对萘啶酸表现出最高的耐药性,达到了33.1%；人源沙门菌对氨苄西林抗性最大,为55.1%。同时,在p-内酰胺类抗生素中,三种来源的沙门菌对青霉素类的耐药性显著高于头孢菌素类(P0.05)；而在喹诺酮类抗生素中,对萘啶酸的耐药性也显著高于恩诺沙星、氧氟沙星等其他类抗生素(P0.05)。此外,三种来源的沙门菌对链霉素、复方新诺明等也表现出较高的耐药率。鼠伤寒沙门菌、伦敦沙门菌和德尔卑沙门菌均表现出较高的耐药性,其中鼠伤寒沙门菌多重耐药最为突出。值得注意的是,1株人源印第安纳沙门菌对本研究所用的16种抗生素均呈现耐药。三种来源沙门菌最常见的耐药型均为NAL,一定程度上说明了耐药沙门菌在屠宰场、农贸市场猪肉以及人三者之间的传播。二、扬州地区屠宰场、农贸市场以及人源沙门菌分离株的分子分型及相关性分析利用多位点序列分型方法(MLST)将屠宰场(275株)、农贸市场(130株)以及人群(49株)来源的沙门菌分离株分成了17、13和12种ST型,其中ST40均为主要ST型。屠宰场不同环节大量相同ST型的重复出现,说明了屠宰场中沙门菌沿屠宰线的传播。脉冲场凝胶电泳(PFGE)将50株德尔卑沙门菌和27株鼠伤寒沙门菌分成了46和17种PFGE型。屠宰场第一次采样过程中,胴体修饰环节、开膛环节以及保定环节分离的鼠伤寒沙门菌P2-D13、P2-E13和P2-S1,它们之间的相似性均为100%,说明这3株沙门菌可能为同一个克隆,显示沙门菌能够在屠宰场中沿屠宰线发生水平传播。分别来自于屠宰场、农贸市场以及人的鼠伤寒沙门菌P4-L1、N2-J3和YZJK2-17 , PFGE相似性超过90%,说明沙门菌可能通过猪肉这一媒介完成了在猪肉生产消费链中的传播。利用规律成簇间隔的短回文重复序列(CRIS(?) (?)R)分型方法可将27株来自屠宰场、农贸市场以及人的鼠伤寒沙门菌分成了7个CRISPR型。来自于屠宰场不同环节的沙门菌P2-D13、P2-E13和P2-S1的CRISPR间隔序列和数量完全一致,而来自于屠宰场、农贸市场猪肉以及人源的沙门菌P4-L1、N2-J3和YZJK2-17的间隔序列和数量也完全一致,证实了沙门菌不仅沿屠宰线发生了水平传播,同时也可能在整个猪肉生产消费链中传播。
[Abstract]:Salmonella is an important human animal and has a long history of serious harm to both humans and animals. Egg, poultry and pork have been considered to be the three major sources of Salmonella. And the separation rate of the salmonella in the egg and the poultry is obviously reduced with the large-scale development of the salmonella prevention and control project in the laying hen. Therefore, the prevention and control of Salmonella in pork and pork products has become a major focus of the attention of the developed countries, and as two of the most important links in the pork production chain, The slaughter process of the slaughterhouse and the health status of the retail outlets of the pork market pork have a far-reaching influence on the prevention and control of Salmonella in the pork. The purpose of this study was to understand the distribution status, serotype characteristics and drug resistance of Salmonella in the slaughterhouse, agricultural and trade market of Yangzhou, and the subtyping methods such as MLST, PFGE and CRISPR, in order to find out the slaughterhouse. The internal relation between the pork and the human source Salmonella in the market and the potential Salmonella transmission route are found, which can provide data support for the prevention and control of the food-borne Salmonella. 1. The isolation and identification of the slaughterhouse, the agricultural trade market and the human Salmonella isolates in Yangzhou area, and the drug sensitivity analysis from October 2013 to December 2014. The environmental samples and the slaughter pig samples are collected and the pork samples are 178 parts from a certain slaughterhouse in Yangzhou and its downstream agricultural trade market. The isolation rate was 70.2% (275/392) and 73.0% (130/178) respectively. 49 strains of human Salmonella were collected from the Centers for Disease Control and Control in Yangzhou and Taizhou. The total number of Salmonella in three sources was 454. The statistical analysis showed that there was no significant difference between the five samples of Salmonella in the slaughterhouse (P0.05), and the isolation of Salmonella was not affected by the seasonal factors. Serotype identification results showed that most of the most dominant serotypes in the slaughterhouse and the market for agricultural trade were the Salmonella, and that in the human Salmonella, the Salmonella, the second, and the other serotypes, such as Salmonella typhimurium, and the duck Salmonella, Salmonella in London are also common in three sources. The drug-sensitive experiments of 16 antibiotics from 454 strains of Salmonella were carried out. The results showed that the drug-resistant rate of Salmonella in the slaughterhouse was the highest, which was 30.9%. The maximum resistance of human Salmonella to methicillin was 55.1%. In the same time, the resistance of the three sources of Salmonella to Penicillin was significantly higher than that of the cephalosporin (P 0.05), and the drug resistance of the three kinds of antibiotics was higher than that of the other antibiotics (P0.05). In addition, the three sources of Salmonella showed high resistance to streptomycin and compound neomycin and the like. Salmonella typhimurium, Salmonella, and S. del are resistant to high resistance, and the multiple drug resistance of Salmonella typhimurium is the most prominent. It is worth noting that 1 human-derived Salmonella indiana is resistant to the 16 antibiotics used in the study. The most common drug-resistant type of the three sources of Salmonella is NAL, and the spread of the drug-resistant Salmonella among the slaughterhouse, the pork and the market pork and the human is explained to some extent. 2. The molecular typing and correlation analysis of the slaughterhouse, farm and trade market and human Salmonella isolates in Yangzhou area, using the multi-site sequence typing method (MLST), will be used to classify the slaughterhouse (275 strains). There were 17,13 and 12 ST-types of Salmonella isolates from the farmers' market (130 strains) and the population (49), of which ST40 was the main ST-type. The repeated appearance of the same ST-type in different parts of the slaughterhouse shows the spread of the Salmonella in the slaughter line along the slaughter line. Salmonella typhi and Salmonella typhimurium were divided into 46 and 17 PFGE types by pulse-field gel electrophoresis (PFGE). In the first sampling of the slaughterhouse, Salmonella typhimurium P2-D13, P2-E13 and P2-S1 isolated from the stage of the carcass modification, the opening of the carcass and the fixed link, the similarity between them was 100%, indicating that the three Salmonella strains could be the same clone, It is shown that Salmonella can be spread horizontally along the slaughter line in the slaughterhouse. The similarity of PFGE of Salmonella typhimurium P4-L1, N2-J3 and YZJK2-17 from the slaughterhouse, farm and trade market and human is more than 90%, which indicates that the transmission of Salmonella in the pork production consumption chain can be completed by the medium of pork. Short message repeat sequences (CRIS (?) using regular cluster intervals? ) (? ) R) The method can divide 27 strains of Salmonella typhimurium into 7 CRISPR patterns from the slaughterhouse, the agricultural market and the human. the sequence and the number of the CRISPR intervals of the Salmonella P2-D13, P2-E13 and P2-S1 from different parts of the slaughterhouse are identical, and the interval sequences and the number of the salmonella P4-L1, N2-J3 and YZJK2-17 from the slaughterhouse, the agricultural and trade market pork and the human are also identical, It is confirmed that the Salmonella not only has a horizontal spread along the slaughter line, but also can be spread throughout the pork production consumption chain.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.5
本文编号：2444402
[Abstract]:Salmonella is an important human animal and has a long history of serious harm to both humans and animals. Egg, poultry and pork have been considered to be the three major sources of Salmonella. And the separation rate of the salmonella in the egg and the poultry is obviously reduced with the large-scale development of the salmonella prevention and control project in the laying hen. Therefore, the prevention and control of Salmonella in pork and pork products has become a major focus of the attention of the developed countries, and as two of the most important links in the pork production chain, The slaughter process of the slaughterhouse and the health status of the retail outlets of the pork market pork have a far-reaching influence on the prevention and control of Salmonella in the pork. The purpose of this study was to understand the distribution status, serotype characteristics and drug resistance of Salmonella in the slaughterhouse, agricultural and trade market of Yangzhou, and the subtyping methods such as MLST, PFGE and CRISPR, in order to find out the slaughterhouse. The internal relation between the pork and the human source Salmonella in the market and the potential Salmonella transmission route are found, which can provide data support for the prevention and control of the food-borne Salmonella. 1. The isolation and identification of the slaughterhouse, the agricultural trade market and the human Salmonella isolates in Yangzhou area, and the drug sensitivity analysis from October 2013 to December 2014. The environmental samples and the slaughter pig samples are collected and the pork samples are 178 parts from a certain slaughterhouse in Yangzhou and its downstream agricultural trade market. The isolation rate was 70.2% (275/392) and 73.0% (130/178) respectively. 49 strains of human Salmonella were collected from the Centers for Disease Control and Control in Yangzhou and Taizhou. The total number of Salmonella in three sources was 454. The statistical analysis showed that there was no significant difference between the five samples of Salmonella in the slaughterhouse (P0.05), and the isolation of Salmonella was not affected by the seasonal factors. Serotype identification results showed that most of the most dominant serotypes in the slaughterhouse and the market for agricultural trade were the Salmonella, and that in the human Salmonella, the Salmonella, the second, and the other serotypes, such as Salmonella typhimurium, and the duck Salmonella, Salmonella in London are also common in three sources. The drug-sensitive experiments of 16 antibiotics from 454 strains of Salmonella were carried out. The results showed that the drug-resistant rate of Salmonella in the slaughterhouse was the highest, which was 30.9%. The maximum resistance of human Salmonella to methicillin was 55.1%. In the same time, the resistance of the three sources of Salmonella to Penicillin was significantly higher than that of the cephalosporin (P 0.05), and the drug resistance of the three kinds of antibiotics was higher than that of the other antibiotics (P0.05). In addition, the three sources of Salmonella showed high resistance to streptomycin and compound neomycin and the like. Salmonella typhimurium, Salmonella, and S. del are resistant to high resistance, and the multiple drug resistance of Salmonella typhimurium is the most prominent. It is worth noting that 1 human-derived Salmonella indiana is resistant to the 16 antibiotics used in the study. The most common drug-resistant type of the three sources of Salmonella is NAL, and the spread of the drug-resistant Salmonella among the slaughterhouse, the pork and the market pork and the human is explained to some extent. 2. The molecular typing and correlation analysis of the slaughterhouse, farm and trade market and human Salmonella isolates in Yangzhou area, using the multi-site sequence typing method (MLST), will be used to classify the slaughterhouse (275 strains). There were 17,13 and 12 ST-types of Salmonella isolates from the farmers' market (130 strains) and the population (49), of which ST40 was the main ST-type. The repeated appearance of the same ST-type in different parts of the slaughterhouse shows the spread of the Salmonella in the slaughter line along the slaughter line. Salmonella typhi and Salmonella typhimurium were divided into 46 and 17 PFGE types by pulse-field gel electrophoresis (PFGE). In the first sampling of the slaughterhouse, Salmonella typhimurium P2-D13, P2-E13 and P2-S1 isolated from the stage of the carcass modification, the opening of the carcass and the fixed link, the similarity between them was 100%, indicating that the three Salmonella strains could be the same clone, It is shown that Salmonella can be spread horizontally along the slaughter line in the slaughterhouse. The similarity of PFGE of Salmonella typhimurium P4-L1, N2-J3 and YZJK2-17 from the slaughterhouse, farm and trade market and human is more than 90%, which indicates that the transmission of Salmonella in the pork production consumption chain can be completed by the medium of pork. Short message repeat sequences (CRIS (?) using regular cluster intervals? ) (? ) R) The method can divide 27 strains of Salmonella typhimurium into 7 CRISPR patterns from the slaughterhouse, the agricultural market and the human. the sequence and the number of the CRISPR intervals of the Salmonella P2-D13, P2-E13 and P2-S1 from different parts of the slaughterhouse are identical, and the interval sequences and the number of the salmonella P4-L1, N2-J3 and YZJK2-17 from the slaughterhouse, the agricultural and trade market pork and the human are also identical, It is confirmed that the Salmonella not only has a horizontal spread along the slaughter line, but also can be spread throughout the pork production consumption chain.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.5
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