Effet chez le porcelet d'une exposition à un régime co-contaminé en ...

These are not the final page numbersMol. Nutr. Food Res. 2011, 55, 1–11 5France). Besides this inspection, a 200 ng sample of RNA wasanalyzed by electrophoresis. The reverse transcription and realtimePCR steps were performed as already described [26]. RNAnon-reverse transcripted was used as the non-template controlfor verification of a no genomic DNA amplification signal.Specificity of PCR products was checked out at the end of thereaction by analyzing the curve of dissociation. In addition, thesize of amplicons was verified by electrophoresis. The sequencesof the primers used are detailed in Table 3. Primers for macrophageinflammatory protein-1b (MIP-1b), IL-8 and IL-6 detectionwere designed using Primer Express s software (AppliedBiosystems, Courtaboeuf, France). Primers were purchasedfrom Invitrogen (Cergy Pontoise, France). Amplification efficiencyand initial fluorescence were determined by DataAnalysis for Real Time-PCR method; then values obtained werenormalized by both house-keeping genes, b2-microglobulin andribosomal protein L32, and finally, gene expression wasexpressed relative to the control group as already described [27].2.10 StatisticsFollowing the Fisher test on equality of variances, one-wayANOVA was used to analyze the differences between thedifferent groups of animals at each time point. p-Values of0.05 were considered significant.Table 4. Individual or combined effects of DON and FB on weightgain a)Bodyweightgain/day(kg)Days1–14Days14–35Animal dietsControl DON FB DON1FB0.3670.05 a 0.3570.03 a 0.4370.05 a 0.3270.07 a0.7670.05 a 0.6570.03 a 0.7470.06 a 0.6870.03 aa) Results are expressed as mean7SEM for five animals.3 Results3.1 Individual or combined effects of DON and FB onweight gain, hematological and biochemicalparametersDuring the experiment, piglets were weighed weekly and asreported in Table 4, ingestion of individual or combinedDON- and FB-contaminated diets did not significantlyimpair animal growth.At the end of the experiment, blood samples were takenfrom all piglets to investigate the effects of mycotoxins onhematological and biochemical variables (Tables 5 and 6).Piglets fed either FB- or FB1DON-contaminated dietsdisplayed a significant decrease in neutrophil number(Table 5). An increase in creatinin concentration (p 5 0.047)and a decrease in albumin concentration (p 5 0.015) wasalso observed in the animal groups fed with FB- or DONcontaminateddiets, respectively. These alterations were notobserved in animals fed the diet contaminated with bothtoxins (Table 6).3.2 Individual or combined effects of DON and FB onorgan histopathologyLiver, lungs and kidneys were collected at the end of thetrial for histopathological analysis. The lesions observedin these three organs were mild to moderate for animal fedany of the three contaminated diets (DON, FB, DON1FB)(Fig. 1).The main histological lesions observed in the livers werea disorganization of hepatic cords, cytoplasmatic andnuclear vacuolization of hepatocytes and megalocytosis(Figs. 1A and B). Piglets fed either DON- or FB-contaminateddiets displayed significant liver lesions whencompared to animals fed the control diet. The lesion scorewas further increased for animals fed diet contaminatedwith both toxins. The proliferation of hepatocytes wasassessed by counting Ki-67-positive cells in liver sections.Table 5. Individual or combined effects of DON and FB on hematological parameters a)Hematological parameters Animal diets (wk 6)Control DON FB DON7FBWhite blood cells (thousands/mL) 21.271.9 a 19.672.3 a 20.372.8 a 18.271.6 aLymphocytes (thousands/mL) 12.471.9 a 11.471.4 a 14.772.1 a 12.671.0 aNeutrophils (thousands/mL) 7.371.1 a 7.071.1 a,b 4.570.9 b 4.670.6 bRed blood cells (thousands/mL) 6.270.3 a 5.770.2 a 6.170.4 a 5.970.4 aMean corpuscular volume (fL) 47.670.8 a 47.170.7 a 46.270.5 a 50.471.9 aHematocrit (%) 29.871.6 a 27.070.5 a 28.071.9 a 29.571.6 aHemoglobin (g/dL) 9.670.5 a 9.070.2 a 9.470.5 a 9.770.5 aMean corpuscular hemoglobin (pg) 15.470.2 a 15.670.2 a 15.670.2 a 16.570.8 aMean corpuscular hemoglobin concentration (%) 32.470.4 a 33.270.3 a 33.870.6 a 32.870.4 aa) Results are expressed as mean7SEM for six animals. Values in rows with different letters are significantly different.& 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheimwww.mnf-journal.com