Lymphocyte subpopulations in blood and duodenal epithelium of broilers fed diets contaminated with deoxynivalenol and zearalenone
Introduction
Mycotoxins are secondary metabolites of mould that may contaminate food and feed materials. Consumption of mycotoxins may result in impaired immunity and decreased resistance to infectious diseases (Li et al., 2005, Awad et al., 2008).
According to chemical structure, deoxynivalenol (DON) is a member of trichothecene mycotoxin group and belongs to most commonly detected mycotoxins in feedstuffs produced in EU countries. The mode of toxic action of DON is inhibition of protein synthesis, thus affecting rapidly dividing cells, such as those of the gastrointestinal tract and the immune system. DON is absorbed through the intestinal epithelium by simple diffusion and inhibits intestinal cell proliferation (Sergent et al., 2006).
Numerous studies demonstrated trichothecene effects as immunostimulatory or immunosuppressive depending on dose, exposure frequency and timing of functional immune assay (Bondy and Pestka, 2000, Bouhet and Oswald, 2005). High trichothecene exposure injures actively dividing tissues including bone marrow, lymph nodes, spleen, thymus and intestinal mucosa and leads to reduction in circulating blood leukocytes (Pestka et al., 2004). In contrast, low trichothecene exposure may elevate serum IgA levels due to enhanced premature differentiation of IgA secreting cells in Peyer's patches (Pestka, 2007, Pinton et al., 2008). DON was shown as the agent altering claudin expression and thus reducing the barrier function of the intestinal epithelium in pigs (Pinton et al., 2009). In poultry, DON disrupts gut glucose and amino acid transport and reduces the height and width of villi in the duodenum (Awad et al., 2006, Awad et al., 2008). Poultry is known to be relatively resistant to dietary DON in comparison to pigs but more sensitive than ruminants. Diet contaminated with DON at 10 mg/kg had no effect on the growth performance of broilers (Awad et al., 2004). A lowest observed adverse effect level (LOAEL) 9 mg DON/kg feed was suggested for poultry (Eriksen and Pettersson, 2004). Finding of median lethal oral dose (LD50) of 140 mg DON/kg b.w. in 1-day-old broiler chickens also indicates a relative insensitivity of broilers to DON toxicity (Huff et al., 1981).
Zearalenone (ZEA) is a worldwide contaminant of cereals and grains, including maize and soybean. Despite its non-steroidal structure, ZEA activates estrogen receptors resulting in functional and morphological alteration in reproductive organs (Dänicke et al., 2003, Fink-Gremmels and Malekinejad, 2007). The immune system is a potential target for estrogenic endocrine disruptors considering that its cells express estrogen receptors (Igarashi et al., 2001). Contrary to other species, poultry appear to be fairly resistant to ZEA effects. No adverse effects on zootechnical performance of broilers and laying hens were observed at dietary ZEA level up to 800 mg/kg (Chi et al., 1980, Allen et al., 1981).
In practice the food and feed commodities are frequently contaminated with more than one mycotoxin (Speijers and Speijers, 2004); therefore the experimental approach using diets significantly contaminated with both DON and ZEA may yield new information. It is well known that some Fusarium toxins can exert additive and/or synergistic effects. In this study we investigated the effects of feed contaminated with lower and higher DON and ZEA levels on poultry lymphocyte subpopulations by flow cytometry examination of peripheral blood and duodenum epithelium.
Section snippets
Experimental design, animals and housing
Sixty chickens of broiler Ross 308 hybrid of both sexes were obtained from a single commercial source LP-Parovske Haje, Slovakia at day of hatching. After delivery, birds were randomly divided into three groups with twenty birds in each and all were fed an identical uncontaminated starter diet for two weeks. After this pre-treatment, an experimental period of further 14 days followed using a completely randomised design: chickens from Groups 2 and 3 were fed contaminated diets differing only in
Results
Broilers did not show any visible clinical signs of toxicosis during the whole of our experiment. No mortality appeared in birds.
Discussion
Numerous in vitro studies on trichothecenes demonstrated that such compounds may decrease chemotaxy, microbicidal and phagocytic activity of murine neutrophils or macrophages (Corrier et al., 1987, Ayral et al., 1992). Results of present study show reduced phagocytic activity after exposure of chickens to diets contaminated with both lower (DON and ZEA, both being 3.4 mg/kg) and higher (DON and ZEA at 8.2 and 8.3 mg/kg, respectively) levels of Fusarium mycotoxins. On the other hand, flow
Acknowledgements
The study was supported by the Grant Agency for Science (VEGA) of the Slovak Republic, Grant Nos. 1/0044/08, 1/0580/08, 1/0609/09, 2/0006/09, and by the Slovak Research and Development Agency, Slovak Republic, Grants Nos. APVT-51-004804, LPP-0213-06, and LPP-0219-09. This work was also supported by the project INFEKTZOON – Centrum of excellence for infections of animals and zoonoses, on the basis of support of operational program Research and Development financed by European fund of regional
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