Cooley Michael B, Miller William G, Mandrell Robert E
Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, California 94710, USA.
Appl Environ Microbiol. 2003 Aug;69(8):4915-26. doi: 10.1128/AEM.69.8.4915-4926.2003.
Enteric pathogens, such as Salmonella enterica and Escherichia coli O157:H7, have been shown to contaminate fresh produce. Under appropriate conditions, these bacteria will grow on and invade the plant tissue. We have developed Arabidopsis thaliana (thale cress) as a model system with the intention of studying plant responses to human pathogens. Under sterile conditions and at 100% humidity, S. enterica serovar Newport and E. coli O157:H7 grew to 10(9) CFU g(-1) on A. thaliana roots and to 2 x 10(7) CFU g(-1) on shoots. Furthermore, root inoculation led to contamination of the entire plant, indicating that the pathogens are capable of moving on or within the plant in the absence of competition. Inoculation with green fluorescent protein-labeled S. enterica and E. coli O157:H7 showed invasion of the roots at lateral root junctions. Movement was eliminated and invasion decreased when nonmotile mutants of S. enterica were used. Survival of S. enterica serovar Newport and E. coli O157:H7 on soil-grown plants declined as the plants matured, but both pathogens were detectable for at least 21 days. Survival of the pathogen was reduced in unautoclaved soil and amended soil, suggesting competition from indigenous epiphytes from the soil. Enterobacter asburiae was isolated from soil-grown A. thaliana and shown to be effective at suppressing epiphytic growth of both pathogens under gnotobiotic conditions. Seed and chaff harvested from contaminated plants were occasionally contaminated. The rate of recovery of S. enterica and E. coli O157:H7 from seed varied from undetectable to 19% of the seed pools tested, depending on the method of inoculation. Seed contamination by these pathogens was undetectable in the presence of the competitor, Enterobacter asburiae. Sampling of 74 pools of chaff indicated a strong correlation between contamination of the chaff and seed (P = 0.025). This suggested that contamination of the seed occurred directly from contaminated chaff or by invasion of the flower or silique. However, contaminated seeds were not sanitized by extensive washing and chlorine treatment, indicating that some of the bacteria reside in a protected niche on the seed surface or under the seed coat.
肠道病原体,如肠炎沙门氏菌和大肠杆菌O157:H7,已被证明会污染新鲜农产品。在适当条件下,这些细菌会在植物组织上生长并侵入其中。我们开发了拟南芥作为一个模型系统,旨在研究植物对人类病原体的反应。在无菌条件和100%湿度下,肠炎沙门氏菌血清型纽波特和大肠杆菌O157:H7在拟南芥根部生长至10⁹CFU g⁻¹,在地上部分生长至2×10⁷CFU g⁻¹。此外,根部接种导致整株植物被污染,这表明在没有竞争的情况下,病原体能够在植物上或植物体内移动。用绿色荧光蛋白标记的肠炎沙门氏菌和大肠杆菌O157:H7接种显示,在侧根连接处根部受到侵袭。当使用肠炎沙门氏菌的非运动突变体时,运动被消除且侵袭减少。随着植物成熟,土壤种植植物上肠炎沙门氏菌血清型纽波特和大肠杆菌O157:H7的存活率下降,但两种病原体至少21天都可检测到。在未灭菌土壤和改良土壤中,病原体的存活率降低,这表明土壤中本土附生菌存在竞争。从土壤种植的拟南芥中分离出阿氏肠杆菌,结果表明在无菌条件下它能有效抑制两种病原体的附生生长。从受污染植物收获的种子和谷壳偶尔会被污染。根据接种方法的不同,从种子中回收肠炎沙门氏菌和大肠杆菌O157:H7的比率从检测不到到所测试种子库的19%不等。在存在竞争者阿氏肠杆菌的情况下,这些病原体对种子的污染检测不到。对74个谷壳样本的抽样表明,谷壳污染与种子污染之间存在很强的相关性(P = 0.025)。这表明种子污染直接来自受污染的谷壳,或者是通过花朵或角果的侵袭。然而,受污染的种子经过大量洗涤和氯处理后并未得到净化,这表明一些细菌存在于种子表面或种皮下方的受保护微环境中。