Effet chez le porcelet d'une exposition Ã un rÃ©gime co-contaminÃ© en ...

More documents

Recommendations

Info

These are not the final page numbersMol. Nutr. Food Res. 2011, 55, 1–11 DOI 10.1002/mnfr.2010004021RESEARCH ARTICLEIndividual and combined effects of subclinical dosesof deoxynivalenol and fumonisins in pigletsBertrand Grenier 1,2 , Ana-Paula Loureiro-Bracarense 3 , Joelma Lucioli 3 ,Graziela Drociunas Pacheco 1,3 , Anne-Marie Cossalter 1 , Wulf-Dieter Moll 2 , Gerd Schatzmayr 2and Isabelle P. Oswald 11 INRA, Unité de Pharmacologie-Toxicologie, Toulouse, France2 BIOMIN Research Center, Tulln, Austria3 Universidade Estadual de Londrina, Lab Patologia Animal, Londrina, BrazilScope: Deoxynivalenol (DON) and fumonisins (FB) are the most frequently encounteredmycotoxins produced by Fusarium species and most commonly co-occur in animal diets.These mycotoxins were studied for their toxicity in piglets on several parameters includingplasma biochemistry, organ histopathology and immune response.Methods and results: Twenty-four 5-wk-old animals were randomly assigned to four differentgroups, receiving separate diets for 5 wk, a control diet, a diet contaminated with either DON(3 mg/kg) or FB (6 mg/kg) or both toxins. At days 4 and 16 of the trial, the animals weresubcutaneously immunized with ovalbumin to assess their specific immune response. Thedifferent diets did not affect animal performance and had minimal effect on hematologicaland biochemical blood parameters. By contrast, DON and FB induced histopathologicallesions in the liver, the lungs and the kidneys of exposed animals. The liver was significantlymore affected when the two mycotoxins were present simultaneously. The contaminateddiets also altered the specific immune response upon vaccination as measured by reducedanti-ovalbumin IgG level in the plasma and reduced lymphocyte proliferation upon antigenicstimulation. Because cytokines play a key role in immunity, the expression levels of IL-8, IL-1b, IL-6 and macrophage inflammatory protein-1b were measured by RT-PCR at the end ofthe experiment. The expression of these four cytokines was significantly decreased in thespleen of piglets exposed to multi-contaminated diet.Conclusion: Taken together, our data indicate that ingestion of multi-contaminated dietinduces greater histopathological lesions and higher immune suppression than ingestion ofmono-contaminated diets.Received: August 25, 2010Revised: November 6, 2010Accepted: November 25, 2010Keywords:Co-contamination / Deoxynivalenol / Fumonisins / Immunity / Subclinical doses1 IntroductionMycotoxins are secondary metabolites of fungi that maycontaminate animal feeds and human foods. They arefrequently detected in grains, but also in fruits, vegetables,nuts and silages. The Food and Agricultural Organizationestimated that as much as 25% of the world’s agriculturalcommodities are contaminated with mycotoxins and theeconomic losses due to mycotoxin contamination are estimatedin billions of dollars annually worldwide [1]. Clinicalsigns caused by mycotoxins range from mortality to slowgrowth and reduced reproductive efficiency. ConsumptionCorrespondence: Dr. Isabelle P. Oswald, INRA-Laboratoire dePharmacologie-Toxicologie, 180 chemin de Tournefeuille BP93173, 31027 Toulouse Cedex 3, FranceE-mail: isabelle.oswald@toulouse.inra.frFax: 133-5-61-28-53-10Abbreviations: APC, antigen-presenting cells; BALT, bronchioleassociatedlymphoid tissue; DON, deoxynivalenol; FB,fumonisins; HE, hematoxylin–eosin; MAPK, mitogen-activatedprotein kinase; MIP-1b, macrophage inflammatory protein-1b;OVA, ovalbumin& 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheimwww.mnf-journal.com
These are not the final page numbers2 B. Grenier et al. Mol. Nutr. Food Res. 2011, 55, 1–11& 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheimof mycotoxins may also result in impaired immunity anddecreased resistance to infectious diseases [2].Worldwide surveys on the occurrence and contaminationlevels of mycotoxins in raw materials indicate that toxinsproduced by Fusarium mold species are of concern [3–5].Among the fusariotoxins, deoxynivalenol (DON) and fumonisins(FB) are frequently detected with concentrations up to927 mg DON/kg and 300 mg FB/kg [4]. Among cerealsamples collected from European countries, 54% were cocontaminatedwith DON and FB [6]. Similarly, in France, 65%of the maize kernels harvested during 2004–2006 were cocontaminatedwith DON and FB (Arvalis-Institut du végétal,unpublished data). These two mycotoxins are of majorconcern not only in terms of their ubiquitous distribution butalso because of their effects on human and animal health.At high concentrations, FB cause equine leukoencephalomalaciaand porcine pulmonary edema, and it is nephroandhepatotoxic and carcinogenic in rats and mice. FB1 hasbeen classified as a potential human carcinogen (class 2B)by the International Agency for Research on Cancer. Inhumans, consumption of FB-contaminated food has beenlinked with human esophageal cancer and neural tubedefects [7]. Disruption of sphingolipid biosynthesis appearsto be one mechanism involved in FB toxicity, with inhibitionof ceramide synthase [7] leading to accumulation of sphingoidbases (sphinganine and sphingosine). The effects ofingestioning low doses of FB are less documented butrevealed pathological alterations of the lungs and anincrease in intestinal colonization by opportunistic pathogenicbacteria in piglets [8–10].Acute exposure to high doses of DON induces diarrhea,vomiting, leukocytosis and gastrointestinal hemorrhage.Anorexia, growth retardation and immunotoxicity occur inrodents and pigs following chronic DON ingestion [11]. At thecellular level, DON interferes with the active site of peptidyltransferase on ribosomes, and inhibits protein synthesis [11].Further, binding of DON to the ribosome in eukaryotic cellstriggers a ‘‘ribotoxic stress response’’, which involves phosphorylationof the mitogen-activated protein kinases (MAPKs)[12]. MAPK activation modulates the expression of genesassociated with the immune response, chemotaxis, inflammationand apoptosis. The cellular and molecular mechanismsof the immunomodulating action of DON have beendescribed in numerous studies using mice and murine celllines [13]. Depending on the dose and frequency of exposure,DON can be either immunosuppressive or immunostimulatory[11, 14]. Prolonged ingestion of DON produces elevationof immunoglobulin A in plasma [13–15] while increasing thesusceptibility to infectious diseases [11].The toxicity of combinations of mycotoxins cannot alwaysbe predicted based upon their individual toxicities [1]. Interactionsbetween concomitantly occurring mycotoxins can beantagonistic, additive or synergistic. The data on combinedtoxic effects of mycotoxins are limited and, therefore, theactual combined health risk from exposure to mycotoxins isunknown. Assessment of the interaction of Fusarium mycotoxinshas been investigated in vitro on immune cells andintestinal epithelial cells [16, 17]. In vivo experiments have alsobeen done on mice, pigs and poultry using high doses oftoxins in which the authors mainly looked for differences inanimal performance. Among them, few studies wereconcerned with the interaction between DON and FB [18, 19].The purpose of this study was to compare the effects of lowdoses of DON and FB in pigs when fed individually and incombination with particular emphasis on their effects on theimmune response. The experimental design was a factorialassay including control feed and feed contaminated with 3and 6 mg/kg DON and FB individually and in combination,respectively. These contamination levels correspond to levelsthat frequently occur naturally in cereals [1]. Results havebeen reported in terms of both general toxicological parametersincluding weight gain, hematology, plasma biochemistryand organ histology as well as specific parametersdescribing immune system responses (total and specificantibody, lymphocyte proliferation, cytokine expression).2 Materials and methods2.1 AnimalsAll animal experimentation procedures were carried out inaccordance with the European Guidelines for the Care andUse of Animals for Research Purposes (Directive 86/609/EEC). Twenty-four 4-wk-old weaned castrated male pigs(Pietrain/Duroc/Large-white) were obtained locally. Malepigs were used in this protocol as it was previouslydemonstrated that a greater effect of DON and FB occurs inmale pigs compared to female pigs [20]. Animals wereacclimatized for 1 wk in the animal facility of the INRALaboratory of Pharmacology and Toxicology (Toulouse,France), prior to being used in experimental protocols. Sixpigs were allocated to each treatment on the basis of bodyweight. During the 35-day experimental period each treatmentgroup was given free access to water and the assigneddiet. The pigs were observed daily and weighed weekly.2.2 Experimental dietsDiets were manufactured at INRA facilities in Rennes(France), and formulated according to the energy and aminoacid requirements for piglets. Feed composition is detailedin Table 1. Four different batches were prepared, onecontrol batch and three batches artificially contaminatedwith the mycotoxins. Two strains of Fusarium, F. graminearumDSM-4528 and F. verticillioides M-3125 were used toproduce DON and FB, respectively. These strains weregrown separately on rice. FB were produced as previouslydescribed [21]. DON was extracted with ethyl acetate, andthe extract dried on silica gel 60 (Merck, Darmstadt,Germany). The homogenized extracts contained 24 andwww.mnf-journal.com
Page 2 and 3:
AUTEUR : Bertrand GRENIERTITRE : Ef
Page 4 and 5:
REMERCIEMENTSLe travail ici présen
Page 6 and 7:
J’ai une pensée particulière po
Page 8 and 9:
Grenier, B., Loureiro-Bracarense, A
Page 10 and 11:
TABLE DES MATIERES ................
Page 12 and 13:
LISTE DES ABREVIATIONSAFAflatoxineD
Page 14 and 15:
FIGURES ET TABLEAUXFigure 1 : Mycot
Page 16 and 17:
INTRODUCTION8
Page 18 and 19:
INTRODUCTIONCONTEXTE DE L’ETUDELe
Page 20 and 21:
INTRODUCTIONElle se présentait sou
Page 22 and 23:
INTRODUCTIONl’utilisation de ce m
Page 24 and 25:
INTRODUCTION1. Les mycotoxines : g
Page 26 and 27:
INTRODUCTIONc) La zéaralénone (vo
Page 28 and 29:
INTRODUCTIONleucoencéphalomalacies
Page 30 and 31:
INTRODUCTIONMycotoxins co-contamina
Page 32 and 33:
INTRODUCTIONINTRODUCTIONFood safety
Page 34 and 35:
INTRODUCTIONCHARACTERIZATION OF THE
Page 36 and 37:
Quail(140 d)AF-FBRabbit(21 d)AF-FBR
Page 38 and 39:
INTRODUCTIONdue to the ingestion of
Page 40 and 41:
Table 2 : Interaction between Aflat
Page 42 and 43:
(49 d) - RW-L ↗- Ab SRBC ↘- RW-
Page 44 and 45:
INTRODUCTIONb) effects AF and OTA o
Page 46 and 47:
INTRODUCTIONThe depletion of lympho
Page 48 and 49:
AF-T2Chicken 0.3 - 3.0(35 d)AF-T2Ra
Page 50 and 51:
INTRODUCTIONonly seen in birds give
Page 52 and 53:
(30 d) 2AF-RUBGuinea pig 0.02 bw -(
Page 54 and 55:
INTRODUCTIONbetween AF and CPA on t
Page 56 and 57:
(21 day) - RBC, hemoglobin ↗- AST
Page 58 and 59:
INTRODUCTIONII. INTERACTIONS BETWEE
Page 60 and 61:
INTRODUCTIONwhereas in the experime
Page 62 and 63:
INTRODUCTION2.2) TCT type A and BTw
Page 64 and 65:
Table 6 : Interaction between Ochra
Page 66 and 67:
T2-CPAChicken(28 d)T2-CPAChicken(21
Page 68 and 69:
INTRODUCTIONexposed group (Brown et
Page 70 and 71:
INTRODUCTION2.3) Interaction betwee
Page 72 and 73:
INTRODUCTIONeffects in comparison t
Page 74 and 75:
INTRODUCTION3. Procédés de décon
Page 76 and 77:
INTRODUCTIONPhysical and chemical m
Page 78 and 79: INTRODUCTIONINTRODUCTIONConsumption
Page 80 and 81: INTRODUCTIONmm screen shows that fr
Page 82 and 83: INTRODUCTIONseparate the grain into
Page 84 and 85: Table 7 : toxicological evaluation
Page 86 and 87: INTRODUCTIONextrusion parameters ra
Page 88 and 89: INTRODUCTION- an important loss of
Page 90 and 91: INTRODUCTIONal., 2005). The fate of
Page 92 and 93: INTRODUCTIONby a steeping period. S
Page 94 and 95: INTRODUCTIONon cell tissue cultures
Page 96 and 97: INTRODUCTIONCONCLUSIONDetoxificatio
Page 98 and 99: INTRODUCTIONMycotoxin Reduction in
Page 100 and 101: INTRODUCTIONINTRODUCTIONMycotoxin-p
Page 102 and 103: INTRODUCTIONI. ADSORPTION OF MYCOTO
Page 104 and 105: Rats3000 ppb 0.5 %2500 ppb 0.5 %250
Page 106 and 107: INTRODUCTIONIn studies of the poten
Page 108 and 109: INTRODUCTIONSubstances investigated
Page 110 and 111: Table 9 : Outcome of yeast cell wal
Page 112 and 113: INTRODUCTIONFurthermore, cholestyra
Page 114 and 115: INTRODUCTIONwall peptidoglycans and
Page 116 and 117: INTRODUCTIONII. BIOLOGICAL DETOXIFI
Page 118 and 119: INTRODUCTIONdecrease in toxin-conce
Page 120 and 121: INTRODUCTIONCONCLUSIONPrevention an
Page 122 and 123: TRAVAILEXPERIMENTAL90
Page 124 and 125: TRAVAIL EXPERIMENTALtoxicité gén
Page 126 and 127: TRAVAIL EXPERIMENTALRESUME DES ETUD
Page 130 and 131: These are not the final page number
Page 140 and 141: TRAVAIL EXPERIMENTALChronic ingesti
Page 142 and 143: TRAVAIL EXPERIMENTALINTRODUCTIONMyc
Page 144 and 145: TRAVAIL EXPERIMENTALMATERIAL & METH
Page 146 and 147: TRAVAIL EXPERIMENTALthrough a grade
Page 148 and 149: Figure 5ABCDVilli height (µm)5432a
Page 150 and 151: TRAVAIL EXPERIMENTALRESULTS1) Histo
Page 152 and 153: Figure 8ILEUM2.01.6TNF-αbbb2.52.0I
Page 154 and 155: TRAVAIL EXPERIMENTAL3) Intestinal i
Page 156 and 157: TRAVAIL EXPERIMENTALtreated group t
Page 158 and 159: TRAVAIL EXPERIMENTALThe potencial e
Page 160 and 161: TRAVAIL EXPERIMENTALQuantification
Page 162 and 163: Figure 10 :(a) Réaction de deesté
Page 164 and 165: TRAVAIL EXPERIMENTALrévélé de l
Page 166 and 167: TRAVAIL EXPERIMENTALABSTRACTFumonis
Page 168 and 169: TRAVAIL EXPERIMENTALintestinal leve
Page 170 and 171: TRAVAIL EXPERIMENTAL3) Experimental
Page 172 and 173: TRAVAIL EXPERIMENTALthe importance
Page 174 and 175: Figure 11 : Effects of FB1 and HFB1
Page 176 and 177: Table 15 : Effects of FB1 and HFB1
Page 178 and 179:
Table 17 : Effects of FB1 and HFB1
Page 180 and 181:
TRAVAIL EXPERIMENTALDISCUSSIONThe a
Page 182 and 183:
TRAVAIL EXPERIMENTALassociated lymp
Page 184 and 185:
TRAVAIL EXPERIMENTALHartinger et al
Page 186 and 187:
Figure 13 :(a) Transformation par v
Page 188 and 189:
Figure 14 :Illustrations de la proc
Page 190 and 191:
Figure 15 :Plan expérimental de la
Page 192 and 193:
TRAVAIL EXPERIMENTAL4) Analyses du
Page 194 and 195:
TRAVAIL EXPERIMENTAL9) Analyses sta
Page 196 and 197:
TRAVAIL EXPERIMENTALRESULTATS1) Eff
Page 198 and 199:
TRAVAIL EXPERIMENTAL2) Effet des ag
Page 200 and 201:
TRAVAIL EXPERIMENTALb,cca,ba,b,da,d
Page 202 and 203:
TRAVAIL EXPERIMENTALL’exposition
Page 204 and 205:
Figure 21 : Effet de l’exposition
Page 206 and 207:
1,41,21,00,80,60,40,20,0IL‐8a,cc
Page 208 and 209:
TRAVAIL EXPERIMENTALDISCUSSIONLes o
Page 210 and 211:
TRAVAIL EXPERIMENTALœdèmes pulmon
Page 212 and 213:
DISCUSSIONGENERALE160
Page 214 and 215:
DISCUSSION GENERALEDans ce travail
Page 216 and 217:
DISCUSSION GENERALEdes vaches laiti
Page 218 and 219:
Tableau 24 : Analyse de denrées ag
Page 220 and 221:
DISCUSSION GENERALEet al., 2008b).
Page 222 and 223:
DISCUSSION GENERALE• une réactio
Page 224 and 225:
DISCUSSION GENERALE2. Les systèmes
Page 226 and 227:
DISCUSSION GENERALEMycoplasma agala
Page 228 and 229:
DISCUSSION GENERALEoptimale. Les au
Page 230 and 231:
DISCUSSION GENERALEsphingomyéline
Page 232 and 233:
DISCUSSION GENERALEsignalisation ce
Page 234 and 235:
DISCUSSION GENERALEL’IMMUNITÉ IN
Page 236 and 237:
DISCUSSION GENERALEplus spécifique
Page 238 and 239:
DISCUSSION GENERALEsouligner que le
Page 240 and 241:
DISCUSSION GENERALELA CARBOXYLESTER
Page 242 and 243:
DISCUSSION GENERALEde HFB1. Toutefo
Page 244 and 245:
CONCLUSIONS184
Page 246 and 247:
CONCLUSIONSLes effets d’expositio
Page 248 and 249:
CONCLUSIONSest utilisée lorsque la
Page 250 and 251:
REFERENCES BIBLIOGRAPHIQUESAABDEL-W
Page 252 and 253:
REFERENCES BIBLIOGRAPHIQUESBHANDARI
Page 254 and 255:
REFERENCES BIBLIOGRAPHIQUESCASADO,
Page 256 and 257:
REFERENCES BIBLIOGRAPHIQUESDEGIRMEN
Page 258 and 259:
REFERENCES BIBLIOGRAPHIQUESETIENNE,
Page 260 and 261:
REFERENCES BIBLIOGRAPHIQUESGRENIER,
Page 262 and 263:
REFERENCES BIBLIOGRAPHIQUESHERZALLA
Page 264 and 265:
REFERENCES BIBLIOGRAPHIQUESKERKADI,
Page 266 and 267:
REFERENCES BIBLIOGRAPHIQUESKUMAR, M
Page 268 and 269:
REFERENCES BIBLIOGRAPHIQUESMARZOCCO
Page 270 and 271:
REFERENCES BIBLIOGRAPHIQUESODHAV, B
Page 272 and 273:
REFERENCES BIBLIOGRAPHIQUESPFOHL-LE
Page 274 and 275:
REFERENCES BIBLIOGRAPHIQUESREFAI, M
Page 276 and 277:
REFERENCES BIBLIOGRAPHIQUESSCHWARTZ
Page 278 and 279:
REFERENCES BIBLIOGRAPHIQUESSWANSON,
Page 280 and 281:
REFERENCES BIBLIOGRAPHIQUESVEKIRU,
Page 282 and 283:
REFERENCES BIBLIOGRAPHIQUESYAN, D.,
show all

Effet chez le porcelet d'une exposition Ã un rÃ©gime co-contaminÃ© en ...

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?