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Analyse dans les aliments

Est-il nécessaire d’analyser chaque ingrédient de la ration individuellement?

Selon le type de ration servie, une ration totale mélangée (RTM) ou une ration de concentrés et de fourrages distribués séparément, on peut choisir une stratégie d’échantillonnage et d’analyse visant à limiter les efforts et les coûts inutiles. Ainsi, pour la RTM, il peut être avisé de procéder par étape (figure 9). Si le résultat est négatif à la présence de mycotoxines, l’analyse de chacun des ingrédients ne sera pas nécessaire.

Figure 9. Stratégie d’échantillonnage selon le type de ration

Comment s’assurer de prélever un échantillon représentatif ?

Après avoir déterminé la pertinence d’une analyse et ciblé les aliments à soumettre en priorité, l’étape d’échantillonnage doit être réalisée dans les règles de l’art. En effet, la précision et la représentativité des résultats d’analyse en laboratoire dépendent grandement de la qualité de l’échantillonnage. Tel que démontré à la figure 10, la contamination par des mycotoxines n’est pas nécessairement uniforme dans un même lot d’aliments.

Figure 10. 

Pour refléter aussi fidèlement que possible la réalité, l’échantillon doit impérativement être composé de plusieurs sous-échantillons prélevés de plusieurs repas ou de lots différents, selon le cas. Autrement dit, le prélèvement d’une seule « poignée » d’aliments en une seule étape, n’est pas adéquat.

Procédures d’échantillonnage des aliments

Les procédures d’échantillonnage recommandées diffèrent selon le taux d’humidité dans les aliments. Pour assurer une préservation adéquate des échantillons entre le moment du prélèvement jusqu’à l’arrivée au laboratoire, il est important de choisir le protocole approprié pour chaque aliment (tableau 1).

Tableau 1. Choix du protocole d’échantillonnage selon la teneur en humidité de l’aliment à analyser

Protocole 1 : aliments à <12 % d’humidité

ex. grains secs, suppléments protéiques, foin sec et concentrés

  1. Au moment de reprendre les aliments de leur entreposage pour la préparation d’un repas, prélever 8 à 12 sous-échantillons de chaque ingrédient suspect, de façon aléatoire, dans l’ensemble de la masse.* Répéter pour au moins 3 à 5 repas de jours différents.
  2. Pour chaque repas, bien mélanger tous les sous-échantillons prélevés pour créer un échantillon composite de 500 g.
  3. Conserver l’échantillon composite dans un sac en papier propre à double épaisseur broché, au réfrigérateur ;
  4. Combiner tous les échantillons composites de l’étape 2 (au moins 3 à 5) en un seul et unique échantillon composite de 500 g, pour envoyer au laboratoire 
  5. Répéter l’étape 4 afin de prévoir un échantillon supplémentaire pour un éventuel besoin de contre-validation. Conserver au réfrigérateur.

Protocole 2 : aliments à >12 % d’humidité

ex. RTM, ensilages, grains humides

  1. À la reprise* ou juste avant de servir la ration, prélever 8 à 12 sous-échantillons et répéter pour au moins 3 à 5 repas.* Les procédures d’échantillonnage recommandées pour les fourrages sont décrites dans le Guide sur l’interprétation des analyses d’ensilages et présentées sous forme de fiche aide-mémoire et en vidéo pour chaque type d’entreposage.;
  2. Pour chaque repas, combiner et bien mélanger les sous-échantillons prélevés pour créer un échantillon composite de 750 g ;
  3. Conserver l’échantillon composite dans un sac propre en plastique et refermable (style Ziplock), expulser autant d’air que possible et fermer le sac hermétiquement.
  4. Décongeler et mélanger les 3 à 5 échantillons de l’étape 2 pour créer 3 échantillons composites finaux de 1 kg dans un sac de plastique hermétique pour l’analyse des mycotoxines, de la matière sèche (nécessaire pour établir la concentration de mycotoxines sur une base de MS) et un échantillon en réserve pour d’éventuelles analyses supplémentaires.;
  5. Pour éviter le développement de nouvelles moisissures et de levures, conserver les échantillons d’aliments humides au congélateur et expédier dans un sac isotherme avec des sachets réfrigérants.

*Assurer vous de porter des gants propres à chaque échantillonnage.

Comment choisir la méthode d’analyse des aliments?

Différentes méthodes sont utilisées en laboratoire pour l’analyse des mycotoxines dans les aliments (tableau 2). Aucune méthode n’est parfaite en toutes circonstances et les plus complètes sont évidemment les plus coûteuses. Le type d’aliment, la rapidité du test, le coût, la précision des résultats et l’objectif de l’analyse influenceront le choix la méthode.

De manière générale, plus la méthode choisie fournit des résultats fiables et détaillés, plus il sera possible de prendre des décisions éclairées sur les actions à entreprendre pour contrôler et prévenir les problèmes liés aux mycotoxines dans le troupeau.

Et puisque les co-contaminations sont fréquentes et que la concentration de mycotoxines au-delà du seuil préoccupant (tableau 3) influence le risque d’observer des problèmes de santé et de productivité, les méthodes quantitatives et permettant de détecter plus d’une mycotoxine à la fois sont à privilégier.

Tableau 2. Comparaison des méthodes d’analyse des mycotoxines disponibles en laboratoire.

MéthodeType d’alimentAvantagesDésavantagesEstimation des coûts
ELISA1Grains uniquement

Rapide

Abordable

Ne détecte qu’une seule mycotoxine à la fois ;

Ne peut être utilisé pour les ensilages et le foin

Ne fournit pas de valeur quantitative

35-50 $/mycotoxine
HPLC2

Grains

Ensilage

Foin

Fourni des valeurs quantitatives ;

Peut détecter plus d’une mycotoxine ;

Coût élevé100-150 $/1-10 mycotoxines
LC/MS/MS3

Grains

Ensilage

Foin

Fourni des valeurs quantitatives ;

Détecte plusieurs mycotoxines par analyse

Coût élevé140-350 $/10-50 mycotoxines
HPLC dans le sérum ou plasmasérum ou plasmaDétecte le déoxynivalénol (DON) et son métabolite (DOM-1)Variations métaboliques de l’animal75 $/DON + DOM-1

1ELISA : méthode immunoenzymatique à double détermination d’anticorps; 2HPLC : chromatographie liquide à haute performance ; 3LC/MS/MS : chromatographie en phase liquide combinée à la spectrométrie de masse

La méthode ELISA

Pour détecter la présence des mycotoxines les plus fréquentes dans les grains – par exemple le DON et la ZÉA – la méthode ELISA est l’option la plus rapide et la plus abordable. Cependant, si on souhaite connaître la concentration précise des mycotoxines dans l’aliment, il faudra alors opter pour l’une des deux autres méthodes. Étant donné leur composition hétérogène et riche en acides organiques, la méthode ELISA ne convient pas pour l’analyse dans les ensilages.

Les méthodes HPLC et LC/MS/MS

Bien que plus dispendieuse, l’analyse par chromatographie en phase liquide à haute performance (HPLC) a l’avantage de pouvoir être utilisée pour tous les types d’aliments. De plus, elle permet une mesure quantitative des mycotoxines, mais seulement pour une dizaine d’entre elles.

Pour une analyse plus complète, on devra opter pour la méthode HPLC combinée à la spectrométrie de masse (LC/MS/MS). Si cette option s’avère la plus coûteuse, elle se démarque fort avantageusement par les résultats les plus précis et exhaustifs.

Au-delà du choix de la méthode d’analyse, il est important de se rappeler que la fiabilité et la validité des résultats dépend d’abord grandement de la qualité de l’échantillonnage

Quels sont les seuils de contamination préoccupants dans les aliments?

La présence de mycotoxines dans les aliments est fréquente, mais les effets toxiques ne s’observent généralement qu’à partir d’un certain seuil de concentration dans la ration.

Le tableau 3 indique les concentrations auxquelles on peut s’attendre à observer des signes perceptibles, selon le type de mycotoxine et le stade de développement de l’animal.

Tableau 3. Seuils de concentration préoccupants (PPM) sur une base de matière sèche (MS) des principales mycotoxines présentes dans l’alimentation des bovins laitiers et de boucherie

MycotoxinesStade de développement Dose maximale (PPM) sur une base MSRéférence
Bovins laitiersBovins de boucherie
Déoxynivalénol (DON) et ses dérivés
Lactation15FDA, CE, ACIA
Veau < 3 mois22FDA, CE, ACIA
Veau > 3 mois55FDA, CE, ACIA
Fumonisines (FUM)
Lactation3030FDA
Veau < 3 mois1010FDA
Veau > 3 mois3060FDA
Zéaralénone (ZÉA)
Lactation2-45-10MO
Veau < 3 mois0,50,5CE
Veau > 3 mois0,50,5CE
T2/HT-2
Lactation0,10,1
Veau < 3 mois0,0250,1ACIA
Veau > 3 mois0,0250,1ACIA

PPM : Partie par million ; ACIA : Agence canadienne d’inspection des aliments ; FDA: Agence américaine de l’alimentation et des médicaments ; CE : Commission européenne ; MO : Laboratoire de diagnostic en médecine vétérinaire de l’Université du Missouri à Columbia ; ND : Laboratoire de diagnostic vétérinaire de l’Université du Dakota du Nord.

Ces seuils de concentration préoccupants permettent de situer l’importance de la contamination d’un aliment, d’évaluer le risque qu’il présente pour la santé et la productivité des animaux et ainsi de décider des actions à entreprendre, s’il y a lieu.

Voilà pourquoi il est pertinent de choisir une méthode d’analyse qui permettent non seulement d’identifier la ou les mycotoxines en cause, mais aussi d’évaluer l’ampleur de leur présence dans l’aliment.

Puisque plusieurs facteurs influencent l’apparition et l’intensité des effets toxiques, il est possible d’observer des signes à des concentrations moindre. De même, l’absence de signes est possible même lorsque la quantité mesurée est au-delà du seuil préoccupant.

Ce projet est financé par l’entremise du programme Innov’Action agroalimentaire, en vertu du Partenariat canadien pour l’agriculture, entente conclue entre les gouvernements du Canada et du Québec.

 
 

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Par Younès Chorfi, Maxime Leduc et Julie Baillargeon