Lactobacillus plantarum metabolites reduce deoxynivalenol toxicity on jejunal explants of piglets.

The deterioration of food and feed stuffs and toxic intestinal effects due to fungal colonization and concomitant production of mycotoxins is an increasing concern. The development of fungi resistance to many commonly used chemical preservatives adds further alarm. Therefore, effective detoxification methods would be useful in counteracting this problem. Biotransformation/adsorption of mycotoxins by lactic acid bacteria and their metabolites is a promising approach to minimize the deleterious effects of mycotoxins. The objective of the present study was to evaluate the beneficial effects of Lactobacillus plantarum metabolites in reducing deoxynivalenol intestinal toxicity. To achieve this aim, histological, morphometrical and oxidative stress analyses were performed in the intestinal mucosa of piglets exposed to deoxynivalenol alone or associated with two strains (SN1 and SN2) of L. plantarum subsp. plantarum metabolites. Metabolites were obtained after dichloromethane (D) or ethyl acetate (A) extraction. Jejunal explants were exposed to the following treatments for 2 and 4 h a) culture medium (control group); b) deoxynivalenol (DON, 10 µM); c) L. plantarum metabolites DSN1; d) L. plantarum metabolites DSN1+DON; e) L. plantarum metabolites DSN2; f) L. plantarum metabolites DSN2+DON; g) L. plantarum metabolites ASN1; h) L. plantarum metabolites ASN1+DON; i) L. plantarum metabolites ASN2; j) L. plantarum metabolites ASN2+DON. The metabolites were incubated 1 h previously to DON challenge (one and 3 h of exposure). Histological assessment showed DON-treated explants with villi fusion and atrophy, multifocal apical necrosis and cuboid or flattened enterocytes with 2 and 4 h of exposure, while LP metabolites groups individually or associated with DON remained like control. The density of goblet cells in villi and crypts was reduced in DON explants compared to control group with 2 and 4 h of exposure; on the other hand, a significant increase in this parameter was achieved in LP metabolites groups compared to DON. Morphometric evaluation showed no difference in villi height or crypts depth in any treated explants. Overall, oxidative stress response assessments showed that explants exposed to SN1 extracted with dichloromethane and ethyl acetate, and SN2 extracted with dichloromethane reduced superoxide anion production. In conclusion, L. plantarum metabolites induced beneficial effects in intestinal mucosa, reducing the toxic effects of DON on intestinal morphology and oxidative response.

Histopathological and ultrastructural findings induced by heat-inactivated Lactobacillus plantarum and the culture supernatant on the intestinal mucosa of piglets: an ex vivo approach / Achados histopatológicos e ultraestruturais induzidos por Lactobacillus plantarum inativado pelo calor na mucosa intestinal de leitões: estudo ex-vivo

In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets jejunal explants exposed to two concentration of heat-inactivated Lactobacillus plantarum and their respective culture supernatants. Jejunal explants were incubated for 4 hours in DMEM culture medium with a) only culture medium (control group), b) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1 x 108CFU/ml), c) heat-inactivated Lactobacillus plantarum strain2 - LP2 (2.0 x 109CFU/ml), d) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1), and e) heat-inactivated Lactobacillus plantarum strain2 culture supernatant (CS2). Explants exposed to heat-inactivated L. plantarum strain 1 and 2 showed multifocal to difuse villi atrophy, villi apical necrosis and enterocyte flattening. Morphological assessment revealed similar results with bacterial adhesion to mucus and intestinal epithelial cells and, morphometric analysis showed a decreased villi height compared to the control group. Alterations in explants treated with the culture supernatant of both strains include mild villi atrophy and mild enterocyte apical necrosis. Morphological assesment reveled numerous well delineated villi and, morphometric analysis showed a significant increase in villi height compared to the control group. In general, exposure to the culture supernatants improved the intestinal morphology.(AU)

No presente estudo, foram realizadas análises histológica, morfométrica e ultraestrutural para investigar as alterações da mucosa intestinal em explantes jejunais de leitões expostos a duas cepas e concentrações de Lactobacillus plantarum inativado pelo calor e seus sobrenadantes de cultura. Os explantes jejunais foram incubados durante quatro horas, em meio de cultura DMEM com: a) meio de cultura (grupo controle); b) Lactobacillus plantarum, cepa 1 - LP1 (1,1 x 108CFU/mL); c) Lactobacillus plantarum, LP2 (2,0 x 109CFU/mL); d) sobrenadante da cultura do Lactobacillus plantarum, cepa 1 (SC1); e e) sobrenadante da cultura do Lactobacillus plantarum, cepa 2 (SC2). Os explantes expostos às cepas 1 e 2 do L. plantarum inativado pelo calor mostraram atrofia difusa de vilosidades, necrose apical das vilosidades e achatamento de enterócitos. A avaliação morfológica revelou resultados semelhantes, com adesão bacteriana ao muco e às células epiteliais intestinais, e a análise morfométrica mostrou uma diminuição da altura das vilosidades em relação ao grupo controle. Alterações nos explantes tratados com o sobrenadante da cultura de ambas as cepas caracterizaram-se por atrofia leve das vilosidades e necrose apical leve dos enterócitos. A avaliação morfológica revelou vilosidades bem delineadas, e a análise morfométrica mostrou um aumento significativo na altura das vilosidades em comparação ao grupo controle. Em geral, a exposição aos sobrenadantes da cultura melhora a morfologia intestinal.(AU)

Lactobacillus plantarum culture supernatants improve intestinal tissue exposed to deoxynivalenol.

In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets exposed to deoxynivalenol alone or associated with two strains of Lactobacillus plantarum and the respective culture supernatants. Jejunal explants were incubated for 4h in culture medium with a) only culture medium (DMEM, control group), b) deoxynivalenol (DON, 10µM), c) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1×108 CFU/ml) plus DON, d) heat-inactivated Lactobacillus plantarum strain2-LP2 (2.0×109 CFU/ml) plus DON, e) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON, and f) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON. Explants exposed to DON and DON plus LP1 and LP2 showed a significant increase in histological changes (mainly villi atrophy and apical necrosis) and a significant decrease in villi height when compared to unexposed explants. However, explants treated with CS1+DON and CS2+DON remained similar to the control group both in histological and morphometrical aspects. DON also induced a significant decrease in goblet cell density compared to control whereas CS1+DON treatment induced an increase in the number of goblet cells in comparison to DON explants. In addition, ultrastructural assessment showed control, CS1+DON and CS2+DON explants with well delineated finger shape villi, meanwhile DON-treated, LP1+DON and LP2+DON explants showed a severe villi atrophy with leukocytes exudation on the intestinal surface. Taken together, our results indicate that the culture supernatant treatment reduced the toxic effects induced by DON on intestinal tissue and may contribute as an alternative strategy to reduce mycotoxin toxicity.