Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Survival and Growth Characteristics of Foodborne Pathogen in Romaine Lettuce

로메인 상추에서 병원성미생물의 생존 및 증식 특성

  • Kim, Na-Ye Seul (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Kim, Chae Rin (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Kim, Da-Woon (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Jeong, Myung-In (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Oh, Kwang Kyo (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Kim, Bo-Eun (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Ryu, Jae Gee (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Jung, Jieun (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Jeon, Ik Sung (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA)) ;
  • Ryu, Kyoung-Yul (Microbial Safety Division, Agro-food Safety & Crop Protection Department, National Institute of Agricultural Science (NIAS), Rural Development Administration (RDA))
  • 김나예슬 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 김채린 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 김다운 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 정명인 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 오광교 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 김보은 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 류재기 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 정지은 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 전익성 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과) ;
  • 류경열 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물과)
  • Received : 2021.09.06
  • Accepted : 2021.10.29
  • Published : 2021.12.30
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Abstract

The purpose of this study was to provide safety management information by analyzing the survival and growth-related properties of foodborne pathogens from Romaine lettuce. After cultivating E. coli O157:H7 for 72 h on Romain lettuce via spray inoculation, the bacteria population increased by 2.0 log CFU/g from the initial population, confirming the possibility of survival and multiplication of the pathogen thereon. The study also revealed that there is no significant difference in the cultivation of E.coli O157:H7 after 72 h from inoculation on damaged and undamaged lettuce leaves. As a result of investigating distribution of E.coli O157:H7 on damaged lettuce leaves, it was found that the bacteria is unlikely to adhere on the smooth surface of undamaged leaves and, thus, results in a low population density, whereas the bacteria cluster on the rough surface of damaged leaves and easily enter through the damaged tissues. Furthermore, after 24 h of cultivation of the pathogenic microbe in the extract with concentrations of 10-100%, utilization of the lettuce extract by the pathogen was found to be 8.9 log CFU/mL E. coli O157:H7, 8.6 log CFU/mL L. monocytogenes, and 8.8 log CFU/mL P. carotovorum. The increase in the population of both the pathogenic microbe and foodborne pathogen reached over 4 log CFU/mL, implying the microbe can utilize the lettuce extract as a source of nutrition. Compared to the initial inoculation concentration in 0.1% lettuce extract, the final concentration has increased up to 2.7 log CFU/mL E. coli O157:H7, 1.3 log CFU/mL L. monocytogenes, and 2.9 log CFU/mL P. carotovorum. Accordingly, the study confirms that the minimal growth concentration of the pathogenic microbe is lower than 0.1% and that the pathogen possibly survive and multiply inside the lettuce leaves given the lettuce extract with concentration of 0.1% is consistently supplied through the damaged tissues.

본 연구는 로메인 상추에서 병원성미생물이 생존과 생육의 특성을 분석하여 안전관리 정보를 확보하고자 실시하였다. 로메인 상추에서 분무 접종한 E. coli O157:H7은 72시간 배양 후 초기균수 보다 2.0 log CFU/g 수준으로 증가하여 생존 및 증식이 가능한 것으로 판단되었다. 상추 잎의 상처 유무에 따른 E. coli O157:H7은 배양 72시간 후 유의적 차이가 없었다. 상추 잎에 인위적인 상처에 내어 E. coli O157:H7을 접종하고 병원균의 분포를 조사한 결과 상처가 없는 상추는 표면이 매끄러워 균이 부착하지 못하거나 균수가 매우 낮았고, 상처가 있는 상추 잎은 거친 표면에 균이 밀집되어 상처를 통해 상추 내부로 침입하는 것으로 판단되었다. 병원성미생물의 상추 추출물 이용 여부는 10-100% 농도에서 배양 24시간 이후에 E. coli O157:H7 8.9 log CFU/mL, L. monocytogenes 8.6 log CFU/mL, P. carotovorum 8.8 log CFU/mL로 나타났다. 이는 병원성미생물과 식물병원균이 유사한 4 log CFU/g 이상의 증가율을 나타내어 미생물이 상추 추출물을 영양원으로 사용할 수 있는 것으로 판단되었다. 상추 추출물 0.1%에서 초기 접종 농도와 비교하여 E. coli O157:H7 2.7, L. monocytogenes 1.3, P. carotovorum 2.9 log CFU/mL 수준으로 증가하였다. 이에 따라 병원성 미생물의 최소생육농도는 0.1%보다 낮은 것으로 판단되었고, 상처를 통해 지속적으로 0.1% 수준의 상추 추출물이 병원성미생물에 제공되면 상추 내부에서도 생존 및 증식이 가능할 것으로 확인하였다.

Keywords

Acknowledgement

본 연구는 2021년도 농촌진흥청 국가연구개발사업(PJ01516801)로 수행되었으며 이에 감사드립니다.

References

  1. Kim, S.H., Lee, K.I., Heo, S.Y., Lee, W.J., 2019. Research on fresh-cut fruits and vegetables. Korea Rural Economic Institute. Naju,Korea, pp. 253.
  2. Xu, A., Buchanan, R.L., Evaluation of sampling methods for the detection of pathogenic bacteria on pre-harvest leafy greens. Food Microbiol., 77, 137-145 (2019). https://doi.org/10.1016/j.fm.2018.09.007
  3. Franz, E., Visser, A.A., van Diepeningen, A.D., Klerks, M.M., Termorshuizen, A.J., van Bruggen A.H.C., Quantification of contamination of lettuce by GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. Food Microbiol., 24, 106-112 (2007). https://doi.org/10.1016/j.fm.2006.03.002
  4. Hora, R., Warriner, K., Shelp, B.J., Griffiths, M.W., Internalization of Escherichia coli O157:H7 following biological and mechanical disruption of growing spinach plants. J. Food Prot., 68, 2506-2509 (2005). https://doi.org/10.4315/0362-028X-68.12.2506
  5. Klerks, M.M., Franz, E., van Gent-Pelzer, M., Zijlstra, C., van Bruggen A.H.C., Differential interaction of Salmonella enterica serovars with lettuce cultivars and plant-microbe factors influencing the colonization efficiency. ISME J., 1, 620-631 (2007). https://doi.org/10.1038/ismej.2007.82
  6. Sharma, M., Ingram, D.T., Patel., J.R., Millner, P.D., Wang, X., Hull, A.E., Donnenberg, M.S., A novel approach to investigate the uptake and internalization of Escherichia coli O157:H7 in spinach cultivated in soil and hydroponic medium. J. Food Prot., 72, 1513-1520 (2009). https://doi.org/10.4315/0362-028X-72.7.1513
  7. Wright, K.M., Crozier, L., Marshall, J., Merget, B., Holmes, A., Holden N.J., Difference in internalization and growth of Escherichia coli 0157:H7 within the apoplast of edible plants spinach and lettuce, compared with the model species Nicotiana benthamiana. Microb. Biotechnol., 10, 555-569 (2017). https://doi.org/10.1111/1751-7915.12596
  8. Grinberg, M., Orevi, T., Steinberg, S., Kashtan, N., Bacterial survival in microscopic surface wetness. eLife, 8, e48508 (2019). https://doi.org/10.7554/eLife.48508
  9. Oliveira, M., Vinas, I., Usall, J., Anguera, M., Abadias, M., Presence and survival of Escherichia coli 0157:H7 on lettuce leaves and in soil treated with contaminated compost and irrigation water. Int. J. Food Microbiol., 156, 133-140 (2012). https://doi.org/10.1016/j.ijfoodmicro.2012.03.014
  10. Mulaosmanvic, E., Windstam, S.T., Vagsholm, I., Alsanius, B.W., Size matters: Biological and food safety relevance of leaf damage for colonization of Escherichia coli 0157:H7 gfp+. Front. Microbiol., 11, 608086 (2021). https://doi.org/10.3389/fmicb.2020.608086
  11. Kim, W.I., Jung, H.M., Kim, S.R., Park, K.H., Kim, B.S., Yun, J.C., Ryu, K.Y., Investigation of microbial contamination levels of leafy greens and its distributing conditions at different time - Focused on perilla leaf and lettuce. J. Food Hyg. Saf., 27, 277-284 (2012). https://doi.org/10.13103/JFHS.2012.27.3.277
  12. Koseki, S., Isobe, S., Growth of Listeria monocytogenes on iceberg lettuce and solid media. Int. J. Food Microbiol., 101, 217-225 (2005). https://doi.org/10.1016/j.ijfoodmicro.2004.11.008
  13. Rodriguez, E.G., Gundersen, A., Sbidio, A., Koike, S., Evaluation of post contamination survival and persistence of applied attenuated Escherichia coli 0157:H7 and naturally-contaminating Escherichia coli 0157:H7 on spinach under field conditions and following postharvest handling. Food Microbiol., 77, 173-184 (2019). https://doi.org/10.1016/j.fm.2018.08.013
  14. Park, W.J., Ryu, H.Y., Lim, G.Y., Lee, Y.D., Park, J.H., Microbial prevalence and quality of organic farm produce from various production sites. Korean J. Food Sci. Technol., 46, 262-267 (2014). https://doi.org/10.9721/KJFST.2014.46.2.262
  15. Wilson, M., Hirano, S.S., Lindow, S.E., Location and survival of leaf-associated bacteria in relation of pathogenicity and potential for growth within the leaf. Appl. Environ. Microbial., 65, 1435-1443 (1999). https://doi.org/10.1128/AEM.65.4.1435-1443.1999
  16. Kim, Y.S., Kim, S.Y., An, J.H., Sang, M.K., Weon, H.Y., Song J.K., Changes in resident soil bacterial communities in response to inoculation of soil with beneficial Bacillus spp.. Microbiol. Biotechnol. Lett., 46, 253-260 (2018). https://doi.org/10.4014/mbl.1807.07027