Improvement of Karmali Agar by Addition of Polymyxin B for the Detection of Campylobacter jejuni and C. coli in Whole‐Chicken Carcass Rinse

The Karmali agar was modified by supplementation with a high concentration of polymyxin B. The goal of the study was to evaluate the effect of a high concentration of polymyxin B on the ability and selectivity of the modified Karmali agar to isolate Campylobacter jejuni and Campylobacter coli from whole chicken carcass rinse. A total of 80 whole chickens were rinsed with 400 mL of buffer peptone water. The rinsed samples were incubated with 2× blood‐free modified Bolton enrichment broth for 48 h, and then streaked onto unmodified Karmali agar and modified Karmali agar supplemented with 100000 IU/L polymixin B (P‐Karmali agar). The suspected colonies were finally confirmed by colony PCR. The P‐Karmali agar exhibited a significantly better (P < 0.05) isolation rate than the unmodified Karmali agar (P‐Karmali agar, 73.8%; unmodified Karmali agar, 33.8%). Moreover, the selectivity of the P‐Karmali agar was also better (P < 0.05) than that of the other selective agar when comparing the number of contaminated plates (P‐Karmali agar, 68.8%; unmodified Karmali agar, 87.5%) and growth index of competing flora (P‐Karmali agar, 1.4; unmodified Karmali agar, 2.7). The improved selective agar excluded competing flora resistant to antibiotic agents in unmodified Karmali agar, increasing isolation rate and selectivity for C. jejuni and C. coli.     

Improved Culturing Techniques for Campylobacter

ABSTRACT Improved techniques for culturing and enumerating Campylobacter jejuni were developed. Parameters included (1) growth vessel type, (2) growth atmosphere, (3) incubation time, and (4) frequency of subcul-turing. Improved growth and consistent morphology were obtained using a tissue culture flask compared with an Erlenmeyer flask or a test tube as well as exposure to modified atmosphere (10% CO2, 85% N2, and 5% O2). When cultures of C. jejuni were incubated more than 24 h, transformation to the coccoid increased, and fewer colony-forming units were detected. Excessive subculturing (>2 times) resulted in increased formation of coc-coid forms. Decreased concentrations of colony-forming units of C. jejuni commonly attributed to the production of a viable, nonculturable form also may be due to cellular clumping. These techniques should alleviate difficulty and reduce variation when working with Campylobacter .     

Dual enlargement of gold nanoparticles: from mechanism to scanometric detection of pathogenic bacteria

A mechanism of dual enlargement of gold nanoparticles (AuNPs) comprising two steps is described. In the first step, the AuNPs are enlarged by depositing Au atoms on their crystalline faces. In this process, the particles are not only enlarged but they are also observed to multiply: new Au nuclei are formed by the budding and division of the enlarged particles. In the second step, a silver enhancement is subsequently performed by the deposition of silver atoms on the enlarged and newly formed AuNPs to generate bimetallic Au@Ag core-shell structures. The dual nanocatalysis greatly enhances the electron density of the nanostructures, leading to a stronger intensity for colorimetric discrimination as well as better sensitivity for quantitative measurement. Based on this, a simple scanometric assay for the on-slide detection of the food-born pathogen Campylobacter jejuni is developed. After capturing the target bacteria, gold-tagged immunoprobes are added to create a signal on a solid substrate. The signal is then amplified by the dual enlargement process, resulting in a strong color intensity that can easily be recognized by the unaided eye, or measured by an inexpensive flatbed scanner. In this paper, dual nanocatalysis is reported for the first time. It provides a valuable mechanistic insight into the development of a simple and cost-effective detection format.     

一种检测活的非可培养状态空肠弯曲菌的新方法

为提高活的非可培养状态空肠弯曲菌的检出率,建立了一种免疫磁珠捕获-荧光聚合酶链式反应法。该法包括应用免疫磁珠技术制备弯曲菌免疫磁珠,以直接捕获受检样品中的空肠弯曲菌,并设计了检测空肠弯曲菌马尿酸酶(hipO)基因的引物与荧光标记探针。结果表明:磁捕获-荧光聚合酶链式反应法可直接检测空肠弯曲菌,无需增菌培养,具有灵敏度高、选择性好、简便、快速等特点,可在24h内完成样品中活的非可培养状态空肠弯曲菌的检测。     

Occurrence of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella and Campylobacter spp. on beef carcasses in Northern Ireland

A survey of beef carcasses was conducted in all 10 European community approved abattoirs in Northern Ireland to determine the incidence of Escherichia coli O157:H7. Analyses were based on excised samples of neck meat taken less than 48 h post-kill. Overall, 780 carcasses were sampled and all were negative for E. coli O157:H7. A sub-set of samples was analysed for the presence of Listeria monocytogenes (n=200), Salmonella (n=200) and Campylobacter spp.(n=100). L. monocytogenes was not detected but Listeria innocua was found on five carcasses and Listeria seeligeri on one. Three carcasses carried salmonellas; Salmonella Mbandaka was found on two and Salmonella Thompson on one. Campylobacter spp. were not detected on any carcasses. The results indicate that very few beef carcasses in Northern Ireland appear to carry any of the four pathogens sought, and this may help explain the low incidence of E. coli O157:H7 in the Northern Ireland human population, relative to the rest of the UK.     

弯曲菌及弯曲菌病的流行现状

为阐明弯曲菌在肠内外感染中的重要性 ,综述了弯曲菌病的发病率、流行特点、临床表现及弯曲菌对抗生素的耐药性。弯曲菌感染率在世界范围内呈普遍上升趋势 ,禽肉、水、牛奶、动物是主要传染源。空肠弯曲菌和结肠弯曲菌是弯曲菌感染最常见的两个种。格林巴利综合征是弯曲菌感染后最严重的并发症。近年来 ,弯曲菌对抗生素的耐药株迅速增多 ,最值得注意的是耐氟喹诺酮类弯曲菌。为更好地控制弯曲菌的感染与流行 ,各国 (特别是发展中国家 )有必要建立对弯曲菌的监测系统。     

Probabilistic inversion for chicken processing lines

We discuss an application of probabilistic inversion techniques to a model of campylobacter transmission in chicken processing lines. Such techniques are indicated when we wish to quantify a model which is new and perhaps unfamiliar to the expert community. In this case there are no measurements for estimating model parameters, and experts are typically unable to give a considered judgment. In such cases, experts are asked to quantify their uncertainty regarding variables which can be predicted by the model. The experts’ distributions (after combination) are then pulled back onto the parameter space of the model, a process termed “probabilistic inversion”. This study illustrates two such techniques, iterative proportional fitting (IPF) and PARmeter fitting for uncertain models (PARFUM). In addition, we illustrate how expert judgement on predicted observable quantities in combination with probabilistic inversion may be used for model validation and/or model criticism.     

叠氮溴乙锭-环介导等温扩增法快速检测空肠弯曲菌活菌

目的建立叠氮溴乙锭(ethidium monoazide bromide, EMA)-环介导等温扩增(loop-mediated isothermal amplification,LAMP)快速检测空肠弯曲菌活菌的方法。方法以空肠弯曲菌mapA基因为检测靶基因,纯培养物提取模板进行LAMP反应,检测空肠弯曲菌LAMP灵敏度、EMA曝光照射时间试验和使用浓度试验。结果空肠弯曲菌LAMP检测灵敏度为800 CFU/mL;曝光照射时间为5 min; EMA终浓度50μg/mL以下时不会抑制空肠弯曲菌活菌的DNA扩增,终浓度为1μg/mLEMA能有效抑制4×10~4CFU/mL空肠弯曲菌死菌扩增; 1%活菌混合体系中EMA-LAMP检测结果为阳性。结论 EMA-LAMP方法能有效快速检测空肠弯曲菌活菌。     

徐州市禽源弯曲菌的分离鉴定及耐药性分析

目的 了解徐州市市售禽肉中弯曲菌的污染状况及对抗生素的耐药情况。方法 随机采集农贸市场及超市新鲜或冷冻禽肉72份,采用滤膜法分离培养鉴定弯曲菌;采用琼脂稀释法测定分离出的菌株对6类11种抗生素的耐药性。结果 72份样品中共检出29份弯曲菌,总检出率为40.28%（29/72）,空肠弯曲菌和结肠弯曲菌检出率分别为19.44%（14/72）和20.83%（15/72）;新鲜禽肉和冷冻禽肉检出率分别为68.72%（22/32）和17.5%（7/40）,差异有统计学意义（χ2=19.412,P<0.05）,空肠弯曲菌在新鲜禽肉和冷冻禽肉中检出率分别为34.38%（11/32）和7.5%（3/40）差异有统计学意义（χ2=8.198,P<0.05）,结肠弯曲菌在新鲜禽肉和冷冻禽肉中检出率分别为34.38%（11/32）和10%（4/40）差异有统计学意义（χ2=6.404,P<0.05）;空肠弯曲菌耐药较高的五种抗生素为：萘啶酸100.00%(14/14)、环丙沙星100.00%(14/14)、四环素100.00%(14/14)、 氟苯尼考64.29%(9/14) ,庆大霉素28.57%(4/12);结肠弯曲菌耐药较高的五种抗生素为：萘啶酸100.00%(15/15)、环丙沙星100.00%(15/15)、四环素100.00%(15/15)、链霉素86.67%(13/15),庆大霉素80%(12/15) ;两种弯曲菌的多重耐药性均为100.00%。结论 徐州市市售禽肉中弯曲菌污染率较高,其中新鲜禽肉较冷冻禽肉中检出率高,徐州地区弯曲菌耐药情况严重