Document Type : Research articles

Authors

1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

3 Department of Biology, Roudehen Branch, Islamic Azad University, Roudehen, Iran

Abstract

Background: Over the past three decades, the use of probiotics has increased as growth promoters and effective supplements to reduce the pathogenicity of pathogens. In this regard, Lactobacillus bacteria are among the most common probiotics used, as they can help strengthening the digestive system and therefore reducing intestinal hystopathological damage when encountering pathogens.
Objectives: This study aimed to evaluate the histopathologic effects of Lactobacillus acidophilus as a dietary supplement in zebrafish ration, as an appropriate laboratory model, and in the exposure to Escherichia coli O157: H7.
Methods: In this project, 48 fish were grouped in 4 aquariums and monitored for 30 days; control group (C1A1) received basic ration; B1A1 group received control group ration and were exposed to E. coli O157: H7; Treatment 1 (T1A1) received basic ration containing L. acidophilus with no exposure to E. coliO157: H7, and Treatment 2 (T2A2) received basic ration containing L. acidophilus with exposure to E. coli O157: H7. During 30 days of the experiment, the samples were taken from the intestinal tissue in the days 15, 27, and 30 for the histopathological examinations.
Results: The results of the findings showed a significant increase in the length of the intestinal villi and the number of goblet cells in the studied tissue in the group treated with a ration containing probiotic supplements compared to the control group (P < 0.001, P < 0.01, P < 0.05). Also, in the group exposed to E. coli O157: H7, histopathological changes including mild edema, inflammatory cell accumulation in the intestinal mucosal tissue, severe necrosis and epithelium loss in the intestinal tissue were evident. These symptoms were much lower in the group fed with probiotic.
Conclusions: According to the obtained data, it can be concluded that feeding fish using L. acidophilus supplement can produce very beneficial effects in reducing tissue damage caused by E. coli O157: H7 infection in zebrafish intestines as a laboratory model.

Keywords

  1. Culligan EP, Hill C, Sleator RD. Probiotics and gastrointestinal disease: Successes, problems and future prospects. Gut Pathog. 2009;1(1):19. doi: 10.1186/1757-4749-1-19. [PubMed: 19930635]. [PubMed Central: PMC2789095].
  2. Medellin-Pena MJ, Wang H, Johnson R, Anand S, Griffiths MW. Probiotics affect virulence-related gene expression in Escherichia coli O157:H7. Appl Environ Microbiol. 2007;73(13):4259-67. doi: 10.1128/AEM.00159-07. [PubMed: 17496132]. [PubMed Central: PMC1932779].
  3. Karimi S, Azizi F, Nayeb-Aghaee M, Mahmoodnia L. The antimicrobial activity of probiotic bacteria Escherichia coli isolated from different natural sources against hemorrhagic E. coli O157:H7. Electron Physician. 2018;10(3):6548-53. doi: 10.19082/6548. [PubMed: 29765581]. [PubMed Central: PMC5942577].
  4. Ferens WA, Hovde CJ. Escherichia coli O157:H7: Animal reservoir and sources of human infection. Foodborne Pathog Dis. 2011;8(4):465-87. doi: 10.1089/fpd.2010.0673. [PubMed: 21117940]. [PubMed Central: PMC3123879].
  5. Maltby R, Leatham-Jensen MP, Gibson T, Cohen PS, Conway T. Nutritional basis for colonization resistance by human commensal Escherichia coli strains HS and Nissle 1917 against E. coli O157:H7 in the mouse intestine. PLoS One. 2013;8(1). e53957. doi: 10.1371/journal.pone.0053957. [PubMed: 23349773]. [PubMed Central: PMC3547972].
  6. Al-wgaa AA, Alwan MJ. Study the prevalence of Escherichia coli O157: H7 isolated from humans and sheep with histopathological study. J Entomol Zool Stud. 2017;5(6):2074-80.
  7. Stones DH, Fehr AGJ, Thompson L, Rocha J, Perez-Soto N, Madhavan VTP, et al. Zebrafish (danio rerio) as a vertebrate model host to study colonization, pathogenesis, and transmission of foodborne Escherichia coli O157. mSphere. 2017;2(5). doi: 10.1128/mSphereDirect.00365-17. [PubMed: 28959735]. [PubMed Central: PMC5607324].
  8. Rawls JF, Samuel BS, Gordon JI. Gnotobiotic zebrafish reveal evolutionarily conserved responses to the gut microbiota. Proc Natl Acad Sci USA. 2004;101(13):4596-601. doi: 10.1073/pnas.0400706101. [PubMed: 15070763]. [PubMed Central: PMC384792].
  9. Goldsmith JR, Jobin C. Think small: zebrafish as a model system of human pathology. J Biomed Biotechnol. 2012;2012:817341. doi: 10.1155/2012/817341. [PubMed: 22701308]. [PubMed Central: PMC3371824].
  10. Ayaz Ahmed KB, Raman T, Anbazhagan V. Platinum nanoparticles inhibit bacteria proliferation and rescue zebrafish from bacterial infection. RSC Advances. 2016;6(50):44415-24. doi: 10.1039/c6ra03732a.
  11. Nguyen-Chi M, Phan QT, Gonzalez C, Dubremetz JF, Levraud JP, Lutfalla G. Transient infection of the zebrafish notochord with E. coli induces chronic inflammation. Dis Model Mech. 2014;7(7):871-82. doi: 10.1242/dmm.014498. [PubMed: 24973754]. [PubMed Central: PMC4073276].
  12. Zhou Z, Wang W, Liu W, Gatlin DM, Zhang Y, Yao B, et al. Identification of highly-adhesive gut Lactobacillus strains in zebrafish (Danio rerio) by partial rpoB gene sequence analysis. Aquaculture. 2012;370:150-7. doi: 10.1016/j.aquaculture.2012.10.018.
  13. Aladame N. Bergey's manual of systematic bacteriology. Ann Institute Pasteur Microbiol. 1987;138(1):146. doi: 10.1016/0769-2609(87)90099-8.
  14. Wood BJ, Holzapfel W. The genera of lactic acid bacteria. Springer Science & Business Media; 1992.
  15. Safari R, Yaghoubzadeh Z. [Effects of Lactobacillus on growth parameters, survival rate and resistance of rainbow trout (Onchorhynchus mykiss) to Streptococcus iniae]. Iran Sci Fish J. 2018;26(5):131-8. Persian.
  16. Rahimpour Hesari M, Kazemi Darsanaki R, Salehzadeh A. Antagonistic activity of probiotic bacteria isolated from traditional dairy products against E. coli O157: H7. J Med Bacteriol. 2017;6(3-4):23-30.
  17. Pirarat N, Pinpimai K, Endo M, Katagiri T, Ponpornpisit A, Chansue N, et al. Modulation of intestinal morphology and immunity in nile tilapia (Oreochromis niloticus) by Lactobacillus rhamnosus GG. Res Vet Sci. 2011;91(3):e92-7. doi: 10.1016/j.rvsc.2011.02.014. [PubMed: 21536310].
  18. Kechagia M, Basoulis D, Konstantopoulou S, Dimitriadi D, Gyftopoulou K, Skarmoutsou N, et al. Health benefits of probiotics: A review. ISRN Nutr. 2013;2013:481651. doi: 10.5402/2013/481651. [PubMed: 24959545]. [PubMed Central: PMC4045285].
  19. Khodaii Z, Ghaderian SMH, Natanzi MM. Probiotic bacteria and their supernatants protect enterocyte cell lines from enteroinvasive Escherichia coli (EIEC) invasion. Int J Mol Cell Med. 2017;6(3):183-9. doi: 10.22088/acadpub.BUMS.6.3.183. [PubMed: 29682490]. [PubMed Central: PMC5898642].
  20. Rigobelo EE, Karapetkov N, Maesta SA, Avila FA, McIntosh D. Use of probiotics to reduce faecal shedding of Shiga toxin-producing Escherichia coli in sheep. Benef Microbes. 2015;6(1):53-60. doi: 10.3920/BM2013.0094. [PubMed: 25380795].
  21. Ota A. Protection against an infectious disease by enterohaemorrhagic E. coli 0-157. Med Hypotheses. 1999;53(1):87-8. doi: 10.1054/mehy.1998.0723. [PubMed: 10499834].
  22. Pennington H. Escherichia coli O157. Lancet. 2010;376(9750):1428-35. doi: 10.1016/S0140-6736(10)60963-4. [PubMed: 20971366].
  23. Pebody RG, Furtado C, Rojas A, McCarthy N, Nylen G, Ruutu P, et al. An international outbreak of Vero cytotoxin-producing Escherichia coli O157 infection amongst tourists; a challenge for the European infectious disease surveillance network. Epidemiol Infect. 1999;123(2):217-23. doi: 10.1017/s0950268899002940. [PubMed: 10579440]. [PubMed Central: PMC2810752].
  24. Sherman PM, Johnson-Henry KC, Yeung HP, Ngo PS, Goulet J, Tompkins TA. Probiotics reduce enterohemorrhagic Escherichia coli O157:H7- and enteropathogenic E. coli O127:H6-induced changes in polarized T84 epithelial cell monolayers by reducing bacterial adhesion and cytoskeletal rearrangements. Infect Immun. 2005;73(8):5183-8. doi: 10.1128/IAI.73.8.5183-5188.2005. [PubMed: 16041036]. [PubMed Central: PMC1201237].
  25. Putaala H, Salusjarvi T, Nordstrom M, Saarinen M, Ouwehand AC, Bech Hansen E, et al. Effect of four probiotic strains and Escherichia coli O157:H7 on tight junction integrity and cyclo-oxygenase expression. Res Microbiol. 2008;159(9-10):692-8. doi: 10.1016/j.resmic.2008.08.002. [PubMed: 18783733].
  26. Silva TFA, Petrillo TR, Yunis-Aguinaga J, Marcusso PF, da Silva Claudiano G, de Moraes FR, et al. Effects of the probiotic Bacillus amyloliquefaciens on growth performance, hematology and intestinal morphometry in cage-reared Nile tilapia. Latin Am J Aqua Res. 2015;43(5):963-71.
  27. Kim Y, Kim SH, Whang KY, Kim YJ, Oh S. Inhibition of Escherichia coli O157:H7 attachment by interactions between lactic acid bacteria and intestinal epithelial cells. J Microbiol Biotechnol. 2008;18(7):1278-85. [PubMed: 18667857].
  28. Wang Y, Ren Z, Fu L, Su X. Two highly adhesive lactic acid bacteria strains are protective in zebrafish infected with Aeromonas hydrophila by evocation of gut mucosal immunity. J Appl Microbiol. 2016;120(2):441-51. doi: 10.1111/jam.13002. [PubMed: 26555459].
  29. Gagnon M, Kheadr EE, Dabour N, Richard D, Fliss I. Effect of Bifidobacterium thermacidophilum probiotic feeding on enterohemorrhagic Escherichia coli O157:H7 infection in BALB/c mice. Int J Food Microbiol. 2006;111(1):26-33. doi: 10.1016/j.ijfoodmicro.2006.04.041. [PubMed: 16822570].
  30. Pagliaro G, Battino M. The use of probiotics in gastrointestinal diseases. Mediterr J Nutr Metabol. 2010;3(2):105-13. doi: 10.1007/s12349-010-0008-9.
  31. Ringo E, Gatesoupe FJ. Lactic acid bacteria in fish: A review. Aquaculture. 1998;160(3-4):177-203. doi: 10.1016/s0044-8486(97)00299-8.
  32. Kumar A, Alrefai WA, Borthakur A, Dudeja PK. Lactobacillus acidophilus counteracts enteropathogenic E. coli-induced inhibition of butyrate uptake in intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol. 2015;309(7):G602-7. doi: 10.1152/ajpgi.00186.2015. [PubMed: 26272259]. [PubMed Central: PMC4593819].