PCR-DGGE analysis of earthworm gut bacteria diversity in stress of Escherichia coli O157:H7

Yi Zhang, Gaochan Wang, Yupeng Wu, Hui Zhao, Yufeng Zhang, Zhenjun Sun

College of Resources and Environmental Sciences, China Agricultural University, Beijing, China.
Ohio Agricultural Research and Development Center, Department of Veterinary Preventive Medicine, The Ohio State University, Wooster, USA.

DOI: 10.4236/abb.2013.43A058                

Abstract

In order to test if the intestinal bacteria play an important role in antibacterial ability of earthworm, we chose Escherichia coli O157:H7, an anthropozoonosis pathogen, as a biological stressor and studied the change of intestinal bacteria community of earthworm by PCR-DGGE analysis. Results showed that the pathogen merely existed 1 - 3 days, then almost disappeared after through the earthworm’s gut. In this period, the diversity and abundance index of intestinal bacteria increased first, then decreased, and finally kept stably after 7 days. The result demonstrated that the intestinal bacteria of earthworm had ability of adjust community structure to eliminate the pathogen E. coli O157:H7, and the amount of bacteria Bacillus increased significantly, which might be the positive antagonism to E. coli O157:H7.

Keywords

PCR-DGGE; Earthworm Intestinal Bacteria Diversity; Escherichia coli O157:H7 Stress

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Pedersen, J.C. and Hendriksen, N.B. (1993) Effect of passage through the intestinal tract of detritivore earthworms (Lumbricus spp.) on the number of selected gram-negative and total bacteria. Biology and Fertility of Soils, 16, 227-232. doi:10.1007/BF00361413
[2]	McNeilly, T.N., Mitchell, M.C., Rosser, T., McAteer, S., Low, J.C., Smith, D.G.E., Huntley, J.F., Mahajan, A. and Gally, D.L. (2009) Immunization of cattle with a combination of purified intimin-531, EspA and Tir significantly reduces shedding of Escherichia coli O157:H7 following oral challenge. Vaccine, 28, 1422-1428. doi:10.1016/j.vaccine.2009.10.076
[3]	Wang, C., Sun, Z.J. and Zheng, D.M. (2006) Research advance in antibacterial immunity ecology of earthworm. Chinese Journal of Applied Ecology, 17, 525-529
[4]	Kumar, A.G. and Sekaran, G. (2005) Enteric pathogen modification by anaecic earthworm Lampito mauritii. Journal of Applied Sciences and Environmental Management, 9, 15-17.
[5]	Liu, X.L., Zhang, N., Cui, D.L., Wang, C. and Sun, Z.J. (2011) Lactic acid bacteria isolated from vermicompost and its antimicrobial activity in E. coli O157:H7. GAB, 1, 1-6
[6]	Organisation for Economic Cooperation and Development (1984) Guideline for testing of chemicals No. 207: Earthworm acute toxicity tests. OECD Publishing.
[7]	Stein, E.A. and Cooper, E.L. (1981) The role of opsonins in phagocytosis by celomocytes of the earthworm, Lumbricus terrestris. Developmental & Comparative Immunology, 5, 415-425.
[8]	Hutchison, M.L., Walters, L.D., Avery, S.M., Synge, B.A. and Moore, A. (2004) Levels of zoonotic agents in British livestock manures. Letters in Applied Microbiology, 39, 207-214. doi:10.1111/j.1472-765X.2004.01564.x
[9]	Jensen, J., Krogh, P.H. and Sverdrup, L.E. (2003) Effects of the antibacterial agents tiamulin, olanquindox and metronidazole and the anthelmintic ivermectin on the soil invertebrate species Folsomia fimetaria (Collembola) and Enchytraeus crypticus (Enchytraeidae). Chemosphere, 50, 437-443. doi:10.1016/S0045-6535(02)00336-3
[10]	Li, H.B., Song, X.L. and Li, Y. (1995) Progress on probiotic bacteria in aquiculture. Progress in Veterinary Medicine, 29, 94-99.
[11]	Liu, C.H., Chiu, C.S. and Ho, P.L. (2009). Improvement in the growth performance of white shrimp. Litopenaeus vannamei, by a protease-producing probiotic, Bacillus subtilis E20, from natto. Journal of Applied Microbiology, 107, 1031-l041. doi:10.1111/j.1365-2672.2009.04284.x
[12]	Kajimura, Y., Sugiyama, M. and Kaneda, M. (1995) Bacillopeptins, new cyclic lipopeptide antibiotics from Bacillus subtilis FR-2. The Journal of Antibiotics, 48, 1095-1110.