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Title: Leaky gut and mycotoxins: Aflatoxin B1 does not increase gut permeability in broiler chickens

Author
item GALARZA-SEEBER, ROSARIO - University Of Arkansas
item LATORRE, JUAN - University Of Arkansas
item BIELKE, LISA - University Of Arkansas
item KUTTAPPAN, VIVEK - University Of Arkansas
item WOLFENDEN, AMANDA - University Of Arkansas
item HERNANDEZ-VELASCO, XOCHITL - Universidad Nacional Autonoma De Mexico
item MERINO-GUZMAN, RUBEN - Universidad Nacional Autonoma De Mexico
item VICENTE, JOSE - University Of Arkansas
item Donoghue, Ann - Annie
item CROSS, DAVID - University Of Arkansas
item HARGIS, BILLY - University Of Arkansas
item TELLEZ, GUILLERMO - University Of Arkansas

Submitted to: Frontiers in Veterinary Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/1/2016
Publication Date: 2/15/2016
Citation: Galarza-Seeber, R., Latorre, J.D., Bielke, L., Kuttappan, V.A., Wolfenden, A., Hernandez-Velasco, X., Merino-Guzman, R., Vicente, J.L., Donoghue, A.M., Cross, D., Hargis, B.M., Tellez, G. 2016. Leaky gut and mycotoxins: Aflatoxin B1 does not increase gut permeability in broiler chickens. Frontiers in Veterinary Science. 3:10. doi: 10.3389/fvets.2016.00010.

Interpretive Summary: Previous studies conducted in our laboratory have demonstrated that intestinal barrier function can be adversely affected by diet ingredients or feed restriction, resulting in increased intestinal inflammation-associated permeability. Two experiments were conducted in broilers to evaluate the effect of 3 concentrations of Aflatoxin B1 (AFB1; 2, 1.5 or 1 ppm) on serum fluorescein isothiocyanate-dextran (FITC-d; 3–5 kDa) levelsgastrointestinal leakage and liver bacterial translocation (BT). In Exp 1, 240 day-of-hatch male broilers were allocated in two groups, each group had six replicates of 20 chickens (n = 120/group): Control feed or feed + 2 ppm AFB1. In Exp 2, 240 day-of-hatch male broilers were allocated in three groups, each group had 5 replicates of 16 chickens (n = 80/group): Control feed; feed + 1 ppm AFB1; or feed + 1.5 ppm AFB1. In both experiments, chickens were fed starter (d1-d7) or grower diets (d8-d21) ad libitum and performance parameters were evaluated every week. At day 21, all chicks received an oral gavage dose of FITC-d (4.16 mg/kg) 2.5h before collecting blood samples to evaluate gastrointestinal leakage of FITC-d, and in Exp 2 a hematologic analysis was also performed. Liver sections were aseptically collected and cultured using TSA plates to determine BT on TSA plates. Cecal contents were collected to determine total CFUcfu/g of gram-negative bacteria; lactic acid bacteria (LAB) or anaerobes by plating on selective media. In Exp 2, liver, spleen and bursa of fFabricious were removed to determine organ weight ratio, and also intestinal samples were obtained for morphometric analysis. Performance parameters, organ weight ratio and morphometric measurements were significantly different between control and AFB1 groups in both experiments. Gut leakage of FITC-d was not affected by the three concentrations of AFB1 evaluated (P > 0.05). Interestingly, a significant reduction in BT was observed in chickens that received 2 and 1 ppm AFB1. An increase (P < 0.05) in total aerobic bacteria, total gram negatives, and total LAB were observed in chickens fed 2 and 1.5 ppm of AFB1 when compared with control and 1 ppm chickens. The integrity of gut epithelial barrier was not compromised after exposure to the mycotoxin.

Technical Abstract: Previous studies conducted in our laboratory have demonstrated that intestinal barrier function can be adversely affected by diet ingredients or feed restriction, resulting in increased intestinal inflammation-associated permeability. Two experiments were conducted in broilers to evaluate the effect of 3 concentrations of Aflatoxin B1 (AFB1; 2, 1.5 or 1 ppm) on serum fluorescein isothiocyanate-dextran (FITC-d; 3–5 kDa) levelsgastrointestinal leakage and liver bacterial translocation (BT). In Exp 1, 240 day-of-hatch male broilers were allocated in two groups, each group had six replicates of 20 chickens (n = 120/group): Control feed or feed + 2 ppm AFB1. In Exp 2, 240 day-of-hatch male broilers were allocated in three groups, each group had 5 replicates of 16 chickens (n = 80/group): Control feed; feed + 1 ppm AFB1; or feed + 1.5 ppm AFB1. In both experiments, chickens were fed starter (d1-d7) or grower diets (d8-d21) ad libitum and performance parameters were evaluated every week. At day 21, all chicks received an oral gavage dose of FITC-d (4.16 mg/kg) 2.5h before collecting blood samples to evaluate gastrointestinal leakage of FITC-d, and in Exp 2 a hematologic analysis was also performed. Liver sections were aseptically collected and cultured using TSA plates to determine BT on TSA plates. Cecal contents were collected to determine total CFUcfu/g of gram-negative bacteria; lactic acid bacteria (LAB) or anaerobes by plating on selective media. In Exp 2, liver, spleen and bursa of fFabricious were removed to determine organ weight ratio, and also intestinal samples were obtained for morphometric analysis. Performance parameters, organ weight ratio and morphometric measurements were significantly different between control and AFB1 groups in both experiments. Gut leakage of FITC-d was not affected by the three concentrations of AFB1 evaluated (P > 0.05). Interestingly, a significant reduction in BT was observed in chickens that received 2 and 1 ppm AFB1. An increase (P < 0.05) in total aerobic bacteria, total gram negatives, and total LAB were observed in chickens fed 2 and 1.5 ppm of AFB1 when compared with control and 1 ppm chickens. The integrity of gut epithelial barrier was not compromised after exposure to the mycotoxin.