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鸭舍中可培养空气传播细菌的定量与鉴定

Quantification and identification of culturable airborne bacteria from duck houses.

作者信息

Martin Elena, Kämpfer Peter, Jäckel Udo

机构信息

Federal Institute for Occupational Safety and Health, Nöldnerstrasse 40-42, 10317 Berlin, Germany.

出版信息

Ann Occup Hyg. 2010 Mar;54(2):217-27. doi: 10.1093/annhyg/mep088. Epub 2009 Dec 30.

DOI:10.1093/annhyg/mep088
PMID:20042465
Abstract

Employees at agricultural working places are often exposed to complex bioaerosols. Investigations of bioaerosols in duck houses revealed concentrations of cultivable bacteria between 0.4 and 3 x 10(5) colony forming units (CFU) m(-3) on tryptone soy agar, 0.3 and 2 x 10(5) CFU m(-3) on actinomycetes isolation agar, and 0.8 and 5 x 10(3) CFU m(-3) on Middlebrook agar, respectively, when incubated at 25 degrees C. At an incubation temperature of 37 degrees C, 0.6-3 x 10(2) CFU m(-3) were counted on MacConkey agar and 0.3-2 x 10(3) CFU m(-3) on Middlebrook agar, and the concentrations of bacteria on glycerol-arginine agar and oatmeal agar incubated at 50 degrees C varied between 0.1 and 2 x 10(3) and 1 and 7 x 10(3) CFU m(-3), respectively. In addition, high concentrations of cells were observed by fluorescence microscope quantification of cell counts after 4',6-diamidino-2-phenylindol staining with 3-8 x 10(7) cells m(-3). A total of 213 colonies with different morphological appearance were selected and the isolated pure cultures were identified at the genus level using the 16S rRNA gene sequence analyses. In summary, 19 different genera of Actinobacteria, four genera of the Firmicutes, one genus of the Bacteroidetes, and five genera of the Proteobacteria were identified. Several isolates represent new phylogenetic lineages. Based on 16S rRNA gene analyses, some isolates were most closely related to Cellulosimicrobium funkei, Corynebacterium falsenii, Corynebacterium xerosis, Mycobacterium arupense, and Staphylococcus epidermidis, which have been grouped into Risk group 2 of biological agents and may cause negative pulmonary health effects. These bacterial species were present in high concentrations up to 10(4) CFU m(-3). For this reason, we recommend an adequate personal breathing protection at these working places.

摘要

农业工作场所的员工经常暴露于复杂的生物气溶胶中。对鸭舍生物气溶胶的调查显示,在25℃培养时,胰蛋白胨大豆琼脂上可培养细菌的浓度为0.4至3×10⁵菌落形成单位(CFU)/立方米,放线菌分离琼脂上为0.3至2×10⁵CFU/立方米,Middlebrook琼脂上为0.8至5×10³CFU/立方米。在37℃培养温度下,MacConkey琼脂上计数为0.6 - 3×10²CFU/立方米,Middlebrook琼脂上为0.3 - 2×10³CFU/立方米,在50℃培养的甘油 - 精氨酸琼脂和燕麦片琼脂上细菌浓度分别在0.1至2×10³CFU/立方米和1至7×10³CFU/立方米之间变化。此外,在用4',6 - 二脒基 - 2 - 苯基吲哚染色后通过荧光显微镜定量细胞计数观察到高浓度的细胞,为3 - 8×10⁷个细胞/立方米。总共挑选了213个具有不同形态外观的菌落,并使用16S rRNA基因序列分析在属水平鉴定分离出的纯培养物。总之,鉴定出了19个不同的放线菌属、4个厚壁菌门属、1个拟杆菌门属和5个变形菌门属。一些分离株代表新的系统发育谱系。基于16S rRNA基因分析,一些分离株与弗氏纤维微菌、假诺卡氏棒杆菌、干燥棒杆菌、阿鲁普分枝杆菌和表皮葡萄球菌关系最为密切,这些已被归类为生物制剂风险组2,可能对肺部健康产生负面影响。这些细菌物种的浓度高达10⁴CFU/立方米。因此,我们建议在这些工作场所提供足够的个人呼吸防护。

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