江苏省鸡肉产品中弯曲菌耐药特征及Ⅰ型整合子分析

为探讨鸡肉产品中弯曲菌的耐药特征,本研究以2016~2017年分离于江苏省市售鸡肉中弯曲菌分离株为研究对象,采用K-B纸片扩散法测定分离株对9大类21种抗生素的敏感性,同时应用PCR法检测Ⅰ型整合子的整合酶基因、3’-CS以及可变区耐药基因。研究显示:弯曲菌分离株对环丙沙星、头孢噻肟、氧氟沙星、卡那霉素、氨苄西林耐药率较高,分别为83.16%、80.0%、68.0%、65.0%和64.2%。47.3%(45/95)的菌株为Ⅰ型整合酶基因阳性,10株分离株具有3’-CS端,其中5株分离株为可变区阳性,其耐药基因盒为aad A1。结论:鸡肉产品中分离到的弯曲菌多重耐药现象严重,整合子介导的多重耐药在江苏地区并不是主导的耐药机制,但其携带的耐药基因可在不同细菌间传递从而通过食物链传递给人类,应严格监测鸡肉产品中弯曲菌的耐药特征及Ⅰ型整合子的流行情况。     

Current and future interventions for improving poultry health and poultry food safety and security: A comprehensive review

Poultry is thriving across the globe. Chicken meat is the most preferred poultry worldwide, and its popularity is increasing. However, poultry also threatens human hygiene, especially as a fomite of infectious diseases caused by the major foodborne pathogens (Campylobacter, Salmonella, and Listeria). Preventing pathogenic bacterial biofilm is crucial in the chicken industry due to increasing food safety hazards caused by recurring contamination and the rapid degradation of meat, as well as the increased resistance of bacteria to cleaning and disinfection procedures commonly used in chicken processing plants. To address this, various innovative and promising strategies to combat bacterial resistance and biofilm are emerging to improve food safety and quality and extend shelf-life. In particular, natural compounds are attractive because of their potential antimicrobial activities. Natural compounds can also boost the immune system and improve poultry health and performance. In addition to phytochemicals, bacteriophages, nanoparticles, coatings, enzymes, and probiotics represent unique and environmentally friendly strategies in the poultry processing industry to prevent foodborne pathogens from reaching the consumer. Lactoferrin, bacteriocin, antimicrobial peptides, cell-free supernatants, and biosurfactants are also of considerable interest for their prospective application as natural antimicrobials for improving the safety of raw poultry meat. This review aims to describe the feasibility of these proposed strategies and provide an overview of recent published evidences to control microorganisms in the poultry industry, considering the human health, food safety, and economic aspects of poultry production.     

空肠和结肠弯曲菌鞭毛蛋白基因fla A的检测

目的 运用一种快速、敏感、特异的检测空肠和结肠弯曲菌的方法.方法 以空肠和结肠弯曲菌所共有特异的鞭毛蛋白基因 fla A的一段高度保守序列为引物,用PCR法扩增fla A基因上的一段约1 700 bp的片断.用该引物对空肠和结肠弯曲菌的标准株、福建省的食品分离株进行PER扩增检测,并同时检测该PCR方法的敏感性.结果 扩增片断表现出极好的特异性,2株空肠和结肠弯曲菌标准菌株、8株分离自不同食品样品的空肠穹曲菌和结肠弯曲菌菌株均为阳性,且敏感性实验显示该PCR方法的反应体系最低检出菌量为6 CFU.结论 该方法快速、敏感、特异,可用于突发性食物中毒和暴发感染的调查.     

Adhesion of Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET) used for bottled waters

Adhesion of the bacteria Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET), a polymer widely used within the bottled water industry was measured in two different groundwater solutions. From this, it was found that whilst the percentage cell adhesion for a given strain did not change between groundwater types, substantial variation was obtained between the two bacterial species tested: M. avium (10-30% adhered cells) and C. jejuni (1-2%) and no major variations were measured as a function of groundwater composition for a given strain. To explain this, the interfacial electro-hydrodynamic properties of the bacteria were investigated by microelectrophoresis, with the resultant data analysed on the basis of electrokinetic theory for soft biocolloidal particles. The results obtained showed that M. avium carries a significant volume charge density and that its peripheral layer exhibits limited hydrodynamic flow permeation compared to that of C. jejuni. It was also demonstrated that steric hindrance to flow penetration and the degree of hydrophobicity within/of the outer bacterial interface are larger for M. avium cells. In line with this, the larger amount of M. avium cells deposited onto PET substrates as compared to that of C. jejuni can be explained by hydrophobic attraction and chemical binding between hydrophobic PET and outer soft surface layer of the bacteria. Hydrophobicity of PET was addressed by combining contact angle analyses and force spectroscopy using CH₃-terminated AFM tip.     

Influence of high gas production during thermophilic anaerobic digestion in pilot-scale and lab-scale reactors on survival of the thermotolerant pathogens Clostridium perfringens and Campylobacter jejuni in piggery wastewater

Safe reuse of animal wastes to capture energy and nutrients, through anaerobic digestion processes, is becoming an increasingly desirable solution to environmental pollution. Pathogen decay is the most important safety consideration and is in general, improved at elevated temperatures and longer hydraulic residence times. During routine sampling to assess pathogen decay in thermophilic digestion, an inversely proportional relationship between levels of Clostridium perfringens and gas production was observed. Further samples were collected from pilot-scale, bench-scale thermophilic reactors and batch scale vials to assess whether gas production (predominantly methane) could be a useful indicator of decay of the thermotolerant pathogens C. perfringens and Campylobacter jejuni. Pathogen levels did appear to be lower where gas production and levels of methanogens were higher. This was evident at each operating temperature (50, 57, 65 °C) in the pilot-scale thermophilic digesters, although higher temperatures also reduced the numbers of pathogens detected. When methane production was higher, either when feed rate was increased, or pH was lowered from 8.2 (piggery wastewater) to 6.5, lower numbers of pathogens were detected. Although a number of related factors are known to influence the amount and rate of methane production, it may be a useful indicator of the removal of the pathogens C. perfringens and C. jejuni.     

Recovery of thermophilic campylobacters from pond water and sediment and the problem of interference by background bacteria in enrichment culture

The aim of this study was to address problems in the determination of thermophilic campylobacters in turbid pond water and sediment. Thirty sets of three samples of pond water (volumes 10, 100, 1000 ml) or sediment (0.1, 1.0, 5.0 ml) were examined for the presence of thermophilic campylobacters. The different volumes of pond water were processed by membrane filtration followed by selective enrichment. The samples of sediment were subjected directly to selective enrichment. Presumptive isolates were confirmed by Gram stain, cell morphology, presence of oxidase and catalase, growth under microaerobic but not aerobic conditions, and PCR. Confirmed Campylobacter species were recovered only from 10 and 100 ml samples of water and from 0.1 and 1.0 ml samples of sediments. The 1000 ml samples of water and 5.0 ml samples of sediment never gave positive isolates. PCR indicated that the confirmed isolates were all either Campylobacter jejuni or C. coli. Enrichment cultures from 1000 ml filtrations contained the highest number of background bacteria. It is suggested that the processing of large volumes of turbid environmental water samples or of sediment is counterproductive and may not yield positive Campylobacter cultures. This is probably due to antagonistic effects of large numbers of background bacteria out-competing campylobacters during the enrichment stage. Pilot studies to establish appropriate volumes of pond water or sediment samples should be undertaken before routine determination of campylobacters is begun.     

Antibiotic resistance in Campylobacter jejuni and Campylobacter coli isolated from food in Poland

This study presents the results of investigations on the susceptibility of Campylobacter spp. strains isolated from chicken meat and giblets to fluorochinolones (ciprofloxacin), macrolides (erythromycin), tetracyclines (tetracycline) and aminoglycosides (gentamicin) andV an analysis of the molecular mechanisms of resistance to the selected antibiotics.Between January 2008 and December 2009 a total of 218 samples of chicken meat and giblets from retail trade in Poland were examined. Campylobacter bacteria were found in 143 samples, that is in 65.6% of the total number embraced by the study. Campylobacter coli was the most ubiquitous - its presence was determined in 108 samples out of 143 (75.5%), whereas Campylobacter jejuni was found in 35 of the contaminated samples (24.5%).The results obtained point to the high percentage (97.9%) of Campylobacter isolates resistant to ciprofloxacin. 92 strains (64.3%) were resistant to tetracycline, 14 (9.1%) to erythromycin and only 9 (6.3%) to gentamicin. Moreover, ten out of the 143 resistant Campylobacter strains (7.0%) were found to be resistant to at least three unrelated antibiotics.     

一种新的空肠弯曲菌外膜囊泡提取方法的建立

空肠弯曲菌（Campylobacter jejuni）外膜囊泡（outer membrane vesicles,OMVs）是其在生长过程中分泌到细胞外的球状小泡,主要含有外膜蛋白及一些周质空间的物质,对细菌的生存、定植、细菌与宿主细胞间的交流及致病机制发挥重要的作用。因此,开发一种高效提取方法是研究空肠弯曲菌OMVs生物学功能的关键。该研究发现在微需氧条件下,使用MH培养基培养空肠弯曲菌15 h后提取OMVs最为合适。在最优条件下,采用超滤浓缩法从菌液中提取OMVs,并使用0.22 μm的微孔滤头进一步过滤提取物除去鞭毛等杂质。为了表征所提取的OMVs质量,首先使用透射电镜技术,发现所提取的OMVs具有典型形态,大小在50~300 nm之间,所含杂质较少。SDS-PAGE电泳结果表明OMVs内部含有大量蛋白,含量可达40.50 mg/mL。最后利用外膜蛋白抗体成功证明所提取的物质的主要成分为OMVs。该研究成功建立了一种从液体培养基中提取空肠弯曲菌OMVs的提取体系,多种分析方法证明其提取效率高、质量较好,适用于后续空肠弯曲菌OMVs生物功能相关研究。