In-vitro cytotoxicity of aflatoxin B1 to broiler lymphocytes of broiler chickens

CEP Zimmermann,JM Santurio,RA Zanette,KB Schlemmer,FC Cadoná,DBR Leal,JAS Jaques,IBM Cruz,C Lautert,AK Machado

doi:10.1590/1516-635x1603307-312

Abstract

The aim of the present work was to study the in-vitro cytotoxic effects of different concentrations of aflatoxin B1 (AFB1) on broiler lymphocytes. Lymphocyte-rich mononuclear cells were separated by Ficoll-Histopaque density and cultured in 96-wellplates containing the evaluated AFB1 concentrations in 5% CO2 atmosphere at 39°C. Thereafter, MTT, PicoGreen, and reactive oxygen species assays were performed. Cell viability decreased in the presence of 10 µg/mL AFB1 at 48 h (p < 0.05) and of 10 and 20 µg/mL AFB1 at 72 h (p < 0.01 and p < 0.001, respectively) when compared to the control (0 µg/mL). However, a dose-dependent increase in the cell-free DNA at 24 h was observed at 1, 10 and 20 µg/mL (p < 0.001). ROS formation significantly increased at 24 h at all concentrations (p < 0.001). The in-vitro results demonstrate that AFB1 is cytotoxic and causes biomolecular oxidative damage in broiler lymphocytes.

Highlights

Aflatoxins are secondary metabolites of Aspergillus fungi, including A. flavus, A. parasiticus, and A. nomius (Hamid et al, 2013; Kalpana et al, 2012)
The lymphocyte viability of broiler chickens was assessed in the presence of aflatoxin B1 (AFB1) using the MTT assay at 24, 48, and 72 h (Table 1)
When compared with the control (0 μg/mL), an increase in reactive oxygen species (ROS) concentrations was observed at 24 h of exposure to AFB1 at the concentrations of 0.1, 1, 10, and 20 μg/mL (p < 0.001) (Fig. 2)

Summary

Introduction

Aflatoxins are secondary metabolites of Aspergillus fungi, including A. flavus, A. parasiticus, and A. nomius (Hamid et al, 2013; Kalpana et al, 2012). The toxigenic potential of aflatoxins dependends on their dose and duration of intake, as well as on animal species, age, and nutritional status (Marai & Asker, 2008). It is directly associated with its rapid absorption in the gastrointestinal tract and immediate binding to serum proteins, such as albumin (Santurio, 2000). AFB1 is recognized as one of the most potent known liver carcinogens, and has genotoxic, immunotoxic, and other adverse effects on several animal species, including poultry (Kalpana et al, 2012). DNA damage occurs when ROS synthesis exceeds the capacity of Zimmermann CEP, Machado AK, Cadoná FC, Jaques JAS, Schlemmer KB Lautert C, Cruz IBM, Zanette RA, Leal DBR, Santurio JM body antioxidant defenses to eliminate them

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