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用选定化学消毒剂处理鸡皮后空肠弯曲菌活力的直接显微镜观察

Direct microscopic observation of viability of Campylobacter jejuni on chicken skin treated with selected chemical sanitizing agents.

作者信息

Chantarapanont Walairut, Berrang Mark E, Frank Joseph F

机构信息

Center for Food Safety, Department of Food Science and Technology, University of Georgia, Athens, Georgia 30602-2106, USA.

出版信息

J Food Prot. 2004 Jun;67(6):1146-52. doi: 10.4315/0362-028x-67.6.1146.

Abstract

The objective of this research was to determine the effect of chlorine, acidified sodium chlorite, and peracetic acid treatments on viable Campylobacter jejuni located at various depths within follicles or folds of chicken skin. Chicken skin was inoculated with C. jejuni transformed with P(c)gfp plasmid (GFP-Campylobacter), which also codes for kanamycin resistance. Effectiveness of sanitizer treatments was determined by plate count. C. jejuni were also observed on chicken skin by confocal scanning laser microscopy, whereby viable and nonviable cells were differentiated by their ability to take up staining with 5-cyano-2,3-ditolyl tetrazolium chloride. Sodium hypochlorite, peracetic acid, and acidified sodium chlorite were each applied at 40 or 100 ppm for 2 or 15 min. Each sanitizer resulted in approximately a 1-log decrease (CFU) when used at 100 ppm for 15 min and no significant decrease when used at 40 ppm for 2 min. Numbers of viable cells observed on the skin by direct microscopic count were similar to numbers obtained by plate count. Although viable counts decreased with sanitizer treatments, the total number of Campylobacter cells (live plus dead) attached to the skin remained unchanged. After each chemical treatment, viable C. jejuni were observed at depths of 0 to 10, 11 to 20, and 21 to 30 microm in folds or follicles of chicken skin. Most of the C. jejuni that survived treatment were located at 0 to 10 microm depth, which is where most of the viable cells were located before treatment. The inability of chemical sanitizers to effectively eliminate C. jejuni on chicken skin does not appear to be a result of protection by location in feather follicles or other depressions in the skin.

摘要

本研究的目的是确定氯、酸化亚氯酸钠和过氧乙酸处理对位于鸡皮肤毛囊或褶皱不同深度处的空肠弯曲菌存活菌的影响。用携带P(c)gfp质粒(绿色荧光蛋白-空肠弯曲菌)转化的空肠弯曲菌接种鸡皮肤,该质粒还编码卡那霉素抗性。通过平板计数确定消毒剂处理的有效性。还通过共聚焦扫描激光显微镜在鸡皮肤上观察空肠弯曲菌,通过其摄取5-氰基-2,3-二苯基四氮唑氯化物染色的能力来区分存活细胞和非存活细胞。分别以40或100 ppm的浓度施加次氯酸钠、过氧乙酸和酸化亚氯酸钠2或15分钟。当以100 ppm使用15分钟时,每种消毒剂导致大约1个对数级的减少(菌落形成单位),而当以40 ppm使用2分钟时,没有显著减少。通过直接显微镜计数在皮肤上观察到的存活细胞数量与通过平板计数获得的数量相似。尽管消毒剂处理后存活菌数减少,但附着在皮肤上的空肠弯曲菌细胞总数(活的加死的)保持不变。每次化学处理后,在鸡皮肤褶皱或毛囊中0至10微米、11至20微米和21至30微米的深度处观察到存活的空肠弯曲菌。大多数存活处理的空肠弯曲菌位于0至10微米深度,这也是处理前大多数存活细胞所在的位置。化学消毒剂无法有效消除鸡皮肤上的空肠弯曲菌,这似乎不是由于位于毛囊或皮肤其他凹陷处的保护作用。

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