Vorst Keith L, Todd Ewen C D, Rysert Elliot T
Department of Food Science and Human Nutrition, 2108 South Anthony Hall, Michigan State University, East Lansing, Michigan 48824, USA.
J Food Prot. 2004 Oct;67(10):2212-7. doi: 10.4315/0362-028x-67.10.2212.
Four sampling devices, a sterile environmental sponge (ES), a sterile cotton-tipped swab (CS), a sterile calcium alginate fiber-tipped swab (CAS), and a one-ply composite tissue (CT), were evaluated for quantitative recovery of Listeria monocytogenes from a food-grade stainless steel surface. Sterile 304-grade stainless steel plates (6 by 6 cm) were inoculated with approximately 106 CFU/cm2 L. monocytogenes strain Scott A and dried for 1 h. The ES and CT sampling devices were rehydrated in phosphate buffer solution. After plate swabbing, ES and CT were placed in 40 ml of phosphate buffer solution, stomached for 1 min and hand massaged for 30 s. Each CS and CAS device was rehydrated in 0.1% peptone before swabbing. After swabbing, CS and CAS were vortexed in 0.1% peptone for 1 min. Samples were spiral plated on modified Oxford agar with modified Oxford agar Rodac Contact plates used to recover any remaining cells from the stainless steel surface. Potential inhibition from CT was examined in both phosphate buffer solution and in a modified disc-diffusion assay. Recovery was 2.70, 1.34, and 0.62 log greater using CT compared with ES, CS, and CAS, respectively, with these differences statistically significant (P < 0.001) for ES and CT and for CAS, CS, and CT (P < 0.05). Rodac plates were typically overgrown following ES, positive after CS and CAS, and negative after CT sampling. CT was noninhibitory in both phosphate buffer solution and the modified disc-diffusion assay. Using scanning electron microscopy, Listeria cells were observed on stainless steel plates sampled with each sampling device except CT. The CT device, which is inexpensive and easy to use, represents a major improvement over other methods in quantifying L. monocytogenes on stainless steel surfaces and is likely applicable to enrichment of environmental samples.
对四种采样装置进行了评估,包括无菌环境海绵(ES)、无菌棉拭子(CS)、无菌海藻酸钙纤维拭子(CAS)和单层复合组织(CT),用于从食品级不锈钢表面定量回收单核细胞增生李斯特菌。将无菌304级不锈钢板(6×6厘米)接种约106 CFU/cm2的单核细胞增生李斯特菌菌株Scott A,并干燥1小时。ES和CT采样装置在磷酸盐缓冲溶液中复水。平板擦拭后,将ES和CT置于40毫升磷酸盐缓冲溶液中,均质1分钟并手动按摩30秒。每个CS和CAS装置在擦拭前在0.1%蛋白胨中复水。擦拭后,将CS和CAS在0.1%蛋白胨中涡旋1分钟。将样品螺旋接种在改良牛津琼脂上,使用改良牛津琼脂Rodac接触平板从不锈钢表面回收任何残留细胞。在磷酸盐缓冲溶液和改良纸片扩散试验中都检测了CT的潜在抑制作用。与ES、CS和CAS相比,使用CT的回收率分别高2.70、1.34和0.62个对数,ES与CT以及CAS、CS与CT之间的这些差异具有统计学意义(P<0.001)(P<0.05)。ES采样后Rodac平板通常长满菌苔,CS和CAS采样后呈阳性,CT采样后呈阴性。CT在磷酸盐缓冲溶液和改良纸片扩散试验中均无抑制作用。使用扫描电子显微镜,在除CT外的每个采样装置采样的不锈钢板上均观察到李斯特菌细胞。CT装置价格低廉且易于使用,在定量不锈钢表面的单核细胞增生李斯特菌方面比其他方法有重大改进,并且可能适用于环境样品的富集。