VolumeVol. 54 • Number No. 2 • April - June 2016 Back
Histomorphologic Effects of Bacillus clausii Spores in Enteropathogenic E. coli O127:H21-infected Mice: A Pilot Study
Authors: Marc Gregory Yu, Ryan Torres, Diana Francesca Tuano, Raissa Marie Tud, Arlene Umali, Christine Hope Umandap, Mikhail Lew Ver, Ralph Elvi Villalobos, Ana Patricia Villanueva, Katrina Lenora Villarante, Liz Villasenor, Ada Lyn Yao, Mari Anne Yorobe, Anna Carmina Zantua, Wilson Zuniga
DOI: 10.65564/pjim.d8bacfa124
Date of Publication: June 2016
Abstract
Objective: The study aimed to determine the basic histomorphologic effects of Bacillus clausii (B. clausii) spores in enteropathogenic Escherichia coli (E. coli) O127:H21-infected mice by evaluating the spleen, mesenteric lymph nodes, and intestinal mucosa.
Methods: The study involved 46 apparently healthy Balb/c mice (Mus musculus) which were acclimatized for 19 days prior to any intervention. Sixteen mice were used to determine the sublethal dose of E. coli, which was performed by administering serially-diluted solutions and subsequent generation of a standard curve. From the remaining 30 mice, ten served as normal controls while the remaining 20 were randomized to receive either B. clausii or placebo of sterile water for a week. All mice were then challenged with E. coli for another week and euthanized, and the spleen, mesenteric lymph nodes, and small intestine harvested and examined microscopically. All study personnel were blinded of the treatment assignments.
Results: Histologic evaluation of the small intestine in E. coli only-fed mice exhibited prominent attachment-effacement lesions, with severely denuded mucosa, lymphocytic infiltration, and debris in the intestinal lumen. However, mice given B. clausii prior to E. coli infection displayed only minimal mucosal damage with less sloughing of villus tips, plus increased mucus-secreting goblet cells. In the spleen, E. coli only-fed mice showed moderate to severe lymphoid hyperplasia with blurred boundaries between red and white pulp. In contrast, mice which received B. clausii prior to E. coli infection had only mild degrees of lymphoid hyperplasia. Similar findings were seen in the mesenteric lymph nodes where E. coli only-fed mice showed moderate to severe lymphoid hyperplasia while those given B. clausii prior to E. coli infection merely had mild lymphoid hyperplasia.
Conclusion: B. clausii exerts a potential protective and immunomodulatory action in E. coli O127:H21-infected mice based on histomorphologic effects. However, additional studies are needed to fully characterize these mechanisms.mice based on histomorphologic effects.
References
- Nomoto, K. Prevention of infections by probiotics: Review. J Biosc and Bioeng. 100 (6): 583-92, 2005
- Tomasik, PJ and Tomasik, P. Probiotics and prebiotics. Cereal chem; 80(2): 113-7, 2003
- Isolauri, E; Sutas, Y; Kankaanpaa, P; Arvilommi, H; Salminen, S. Probiotics: effect on immunity. Am J Clin Nutr. 2001, 73(2):444S-450S, 2001
- Mazza, P. The use of Bacillus subtilis as an antidiarrheal microorganism. Boll Chim Farmaceutico; 133: 3-18, 1994
- Rolfe, RD. The role of probiotic cultures in the control of gastrointestinal health. J Nutr. 130: 396S-402S, 2000
- Leading causes of morbidity and mortality. [Online] [Cited: September 26, 2015.] Can be accessed at: www.doh.gov.ph/ node/198.html.
- Shornikova, A; Casas, I; Isolauri, E; Mykkanen, H; Vesikari, T. Lactobaciilus reuteri as a therapeutic agent in acute diarrhea in young children. J Ped Gastroenterol Nutr. 24: 399-404, 2007
- Kelleher, S; Casas, I; Carbajal, N; Lonnerdal, B. Supplementation of infant formula with the probiotic Lactobacillus reuteri and zinc: Impact on enteric infection and nutrition in infant rhesus monkeys. J Ped Gastroenterol Nutr; 35: 162-8, 2002
- Szymanski, H; Pejcz, J; Jawien, M; Chmielarczyk, A; Strus, M; Heczko, P, et al. Treatment of acute infectious diarrhea in infants and children with a mixture of three Lactobacillus rhamnosus strains - a randomized, double-blind, placebo-controlled trial. Aliment Pharmacol Ther. 23: 247-53, 2006
- Duc, LH; Hong, HA; Fairweather, N; Ricca, E; Cutting, SM. Bacterial spores as vaccine vehicles. Infect Immun. 71: 2810-18, 2003
- Duc, H, Hong, HA and Cutting, SM. Germination of the spore in the gastrointestinal tract provides a novel route for heterologous antigen delivery. Vaccine. 21: 4215-24, 2003
- Fuller, R. Probiotics in human medicine. Gut. 32: 439-42. 1991
- La Ragione, RM; Casula, G; Cutting, SM; Woodward, SM. Bacillus subtilis spores competitively exclude Escherichia coli 070:K80 in poultry. Vet Microbiol. 79: 133-42, 2001
- Rengpipat, S; Phianphak, W; Piyatiratitivorakul, S; Menasveta, P. Effects of a probiotic bacterium on black tiger shrimp Penaeus monodon survival and growth. Aquaculture. 167: 301-13, 1998
- Pinchuk, IV; Bressollier, P; Verneuli, B; Fenet, B; Sorokulova, IB; Megraud, F, et al. In vitro anti-Helicobacter pylori activity of the probiotic strain Bacillus subtilis 3 is due to secretion of antibiotics. Antimicrob Agents Chemother. 45: 3156-61, 2001
- Ghilardi, ACR; Gomes, TAT; Elias, WP; Trabulsi, LR. Virulence factors of E. coli strains belonging to serogroups O127 and O142. Epidemiol Infect. 131: 815-21, 2003
- Cheng, JC, Bahaman, AR and Zakaria, Z. Efficacy of a commercial probiotic in protecting mice against Salmonella infection. Conference proceedings. 5th Seminar on Veterinary Sciences. 2010. Serdang, Selangor, Malaysia.
- Andino, A; Zhang, N; Diaz-Sanchez, S; Yard, C; Pendleton, S; Hanning, I, et al. Characterization and specificity of probiotics to prevent salmonella infection in mice. Functional Foods in Health and Disease. 4 (8): 370-80, 2014
- Ruehl-Fehlert, C; Kittel, B; Morawietz, G; Bube, A; Keane, K; Halm, S, et al. Revised guides for organ sampling and trimming in rats and mice-part 1. Exp Toxicol Pathol. 55 (2-3): 91-106, 2003
- Bregman, CL; Adler, RR; Morton, DG; Regan, KS; Yano, BL. Recommended tissue list for histopathologic examination in repeat-dose toxicity and carcinogenicity studies: a proposal of the Society of Toxicologic Pathology (STP). Toxicol Pathol 31 (9): 252-3, 2003
- Frame, SR and Mann, PC [ed.] Principles of pathology for toxicology studies. In: Principles and methods of toxicology, 5th edition. Wallace, AH. [ed.] New York : Informa Healthcare. 591- 610,2008
- Knutton, S; Baldwin, T; Williams, PH; McNeish, AS. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun. 57: 1290-98, 1989
- Donnenberg, MS and Kaper, JB. Enteropathogenic Escherichia coli. Infect Immun. 60: 3953-61, 1992
- Slomiany, BL, Sarosiek, Y and Slomiany, A. Gastric mucus and the mucosal barrier. Dig Dis Sci. 5: 125-45, 1987
- Fang, H, Morita, H and Ouwehand, AC. Bifidobacteria and Lactobacilli exhibited different mitogenic activity on murine splenocytes. Int J Probiot and Prebiot. 1(1): 77-82, 2006
- Robinett, JP and Rank, RG. Splenomegaly in murine trypanosomiasis: T cell-dependent phenomenon. Infect Immun. 23 (2): 270-5, 1979
- Urdaci, MC, Bresollier, P and Pinchuk, I. Bacillus clausii probiotic strains: antimicrobial and immunomodulatory activities. J Clin Gastroenterol. 38: S86-90, 2004
- Narioka, J; Nishi, M; Ogata, Y; Kuwahara, T; Nakayama, H; Tashiro, S, et al. Promotion of bacterial translocation by major liver resection in obstructive jaundice in rats colonised predominantly with indigenous Escherichia coli. J Med Microbiol. 51: 687-94, 2002
- Costa-Ribeiro, H; Ribeiro, T; Mattos, A; Valois, S; Neri, D; Almeida, P, et al. Limitations of probiotic therapy in acute, severe dehydrating diarrhea. J Ped Gastroentero Nutr. 36: 112-5, 2003