Induction of Necrotizing Enterocolitis in Non-Premature Sprague-Dawley Rats and the Effect of Administering Breast Milk-Isolated Lactobacillus salivarius LPLM-O1


Due to an increasing incidence of necrotizing enterocolitis (NEC), as well as its associated mortality and long-term complications seen in surviving patients, the main focus of research in NEC has shifted to the prevention and treatment of the disease. The hypothesis of this work is that the strain Lactobacillus salivarius LPLM-O1 can decrease the intestinal injuries in a model of induced NEC. 26 newborn Sprague-Dawley pups were used in this study and randomized in three groups: control group (n = 6), which were fed with infant formula (Similac NeosureTM, Abbott); probiotic group (n = 10), which were fed with the same infant formula but fortified with 109 colony-forming units (CFU) of Lactobacillus salivarius LPLM-O1, and the NEC-induced group (n = 10). Each group was fed with 100 μl of food formula every three hours, using a modified syringe. The probiotic and NEC groups were exposed to asphyxia- and cold-induced stress to develop experimental NEC. At the end of the experiment (96 hrs), animals were sacrificed, and their small intestines were carefully removed and evaluated for typical signs of NEC, microbiological count and histological analyses. The histological analysis of the NEC-induced group showed transmural necrosis (grade 4); in the probiotic group, the grade was comparatively lower (grade 2). Survival ratewas higher in the probiotic group (83%) than in the NEC-induced group (46%); however, the difference in not statistically significant (p = 0.14). Lactic acid bacteria counts were higher in the probiotic group than in the NEC-induced group (8.4 × 108 and 6.1 × 107 CFU/intestine tissue gram, respectively). According to these results, the model of artificial induction of NEC was effectively establishedin all pups, and the probiotic strain slightly decreases the injuries’ grade in newborn pups.

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Castro, E. , Cofré, J. , Mellado, J. , Pardo, K. , Aguayo, M. , Monsalvez, E. , Montecinos, H. and González, M. (2014) Induction of Necrotizing Enterocolitis in Non-Premature Sprague-Dawley Rats and the Effect of Administering Breast Milk-Isolated Lactobacillus salivarius LPLM-O1. Food and Nutrition Sciences, 5, 1255-1260. doi: 10.4236/fns.2014.513136.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Fitzgibbons, S.C., Ching, Y., Yu, D., Carpenter, J., Kenny, M., Weldon, C., Lillehei, C., Valim, C., Horbar, J.D. and Jaksic, T. (2009) Mortality of Necrotizing Enterocolitis Expressed by Birth Weight Categories. Journal of Pediatric Surgery, 44, 1072-1075.
[2] Wu, S.F., Chiu, H.Y., Chen, A.C., Lin, H.Y., Lin, H.C. and Caplan, M. (2013) Efficacy of Different Probiotic Combinations on Death and Necrotizing Genterocolitis in a Premature Rat Model. Journal of Pediatric Gastroenterology and Nutrition, 57, 23-28.
[3] Panigrahi, P. (2014) Probiotics and Prebiotics in Neonatal Necrotizing Enterocolitis: New Opportu-nities for Translational Research. Pathophysiology, 21, 35-46.
[4] González-Crussi, F. and Hsueh, W. (1983) Experimental Model of Ischemic Bowel Necrosis. The Role of Platelet Activating Factor and Endotoxin. American Journal of Pathology, 112, 127-135.
[5] Sun, X., Caplan, M.S., Liu, Y. and Hsueh, W. (1995) Endotoxinresistant Mice Are Protected from PAF-Induced Bowel Injury and Death. Role of TNF, Complement Activation, and Endogenous PAF Production. Digestive Diseases and Sciences, 40, 495-502.
[6] Jilling, T., Simon, D., Lu, J., Meng, F.J., Li, D., Schy, R., Thomson, R.B., Soliman, A., Arditi, M. and Caplan, M.S. (2006) The Roles of Bacteria and TLR4 in Rat and Murine Models of Necrotizing Enterocolitis. The Journal of Immunology, 177, 3273-3282.
[7] Tian, R., Liu, S.X., Williams, C., Soltau, T.D., Dimmitt, R., Zheng, X. and De Plaen, I.G. (2010) Characterization of a Necrotizing Enterocolitis Model in Newborn Mice. International Journal of Clinical and Experimental Medicine, 3, 293-302.
[8] Caplan, M.S., Miller-Catchpole, R., Kaup, S., Russell,T., Lickerman, M., Amer, M., Xiao, Y. and Thomson, Jr., R. (1999) Bifidobacterial Supplementation Reduces the Incidence of Necrotizing Enterocolitis in a Neonatal Rat Model. Gastroenterology, 117, 577-583.
[9] Souza, D.G., Pinho, V., Soares, A.C., Shimizu, T., Ishii, S. and Teixeira, M.M. (2003) Role of PAF Receptors during Intestinal Ischemia and Reperfusion Injury. A Comparative Study between PAF Receptor-Deficient Mice and PAF Receptor Antagonist Treatment. British Journal of Pharmacology, 139, 733-740.
[10] Craig, A., Ling, L.N., Beardsley, D.J., Wingate-Pearse, N., Walker, D.W., Hohimer, A.R. and Back. S.A. (2003) Quantitative Analysis of Perinatal Rodent Oligodendrocyte Lineage Progression and Its Correlation with Human. Experimental Neurology, 181, 231-240.
[11] Petrosyan, M., Guner, Y.S., Williams, M., Grishin, A. and Ford, H.R. (2009) Current Concepts Regarding the Pathogenesis of Necrotizing Enterocolitis. Pediatric Surgery International, 25, 309-318.
[12] Lin, H.C., Su, B.H., Chen, A.C., et al. (2005) Oral Probiotics Reduce the Incidence and Severity of Necrotizing Enterocolitis in Very Low Birth Weight Infants. Pedriatrics, 115, 1-4
[13] Zani, A, Cordishi, L., Cananzi, M., De Coppi, P., Smith, V.V., Eaton, S. and Pierro, A. (2008) Assessment of a Neonatal Rat Model of Necrotizing Enterocolitis. European Journal of Pediatric Surgery, 18, 423-426.
[14] Braegger, C.P. (2010) Probiotics and the Prevention of Necrotizing Enterocolitis. Annals of Nutrition and Metabolism, 57, 14-15.
[15] Good, M., Sodhi, C., Ozolec, J., Buck, R.H., Morowitz, M.J., Fireck, B., Lu, P. and Hackman, D.J., (2014) Lactobacillus rhamnosus HN001 Decreases the Severity of Necrotizing Enterocolitis in Neonatal Mice and Preterm Piglets: Evidence in Mice for a Role of TLR9. American Journal of Physiology-Gastrointestinal and Liver Physiology, 306, G1021-G1032.

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