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Validation of an Analytical Method for the Determination of the Activity of Protease in Animal Feed Additives and in Animal Feedingstuffs

Published online by Cambridge University Press:  23 January 2015

G.P. Dillon*
Affiliation:
Alltech Ireland, Sarney, Summerhill Road, Dunboyne, Co Meath, Ireland
K. Filer
Affiliation:
Alltech Inc, 3031 Catnip Hill Pike, Nicholasville, 40356 KY, USA
C.A. Moran
Affiliation:
Alltech France SARL, ZI Belle Etoile, 25 allée des Sapins, 44483 Carquefou, France
*
*Corresponding author:gdillon@alltech.com

Summary

The nutrient availability in animal feeds can be improved by including exogenous enzymes to the feed, either by helping breakdown anti-nutritional factors or by increasing digestibility of complex ingredients thereby releasing more nutrients for utilisation. This process can improve the efficiency of meat and egg production, increase animal health, decrease feeding costs and reduce nutrients in animal waste. Proteases are protein-digesting enzymes that are used in animal nutrition to break down storage proteins in various plant materials and proteinaceous anti-nutrients in vegetable proteins. The analysis of exogenous proteases in feed additives and after they have been added to feed has proven technically challenging. Accordingly, the purpose of this work was to validate a method for the determination of the activity of protease in animal feed additives and supplemented animal feed. The approach used for the assay was to adapt an assay based on the hydrolysis of haemoglobin. The method validations examined a range of parameters including; linearity & range; uncertainty, sensitivity, accuracy and studies designed to highlight any possible matrix effects on various types of supplemented feed. The assay method described herein is convenient and inexpensive and could be applied to the analysis of proteases in animal feeds during quality control and in investigating fraudulent adulteration of feed to ensure the authenticity and traceability of the product.

Type
Original Research
Copyright
Copyright © Cambridge University Press and Journal of Applied Animal Nutrition Ltd. 2015 

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References

Association Française de Normalisation (AFNOR) (1999) AFNOR Standard Method Decembre 1999. Protocole d’évaluation d'une méthode alternative d'analyse physico-chimique quantitative par rapport à une méthode de reference. Norme NF XPT 90-210. AFNOR, La Plaine Saint-Denise, Paris, France. pp. 158Google Scholar
Barletta, A. (2010) Introduction: Current Market and Expected Developments; in: Partridge, G.G.; Bedford, M.R. (Eds.), In Enzymes in Farm Animal Nutrition, 2nd edition. Publisher: CABI North American Office, Cambridge, MA 02139, USA, pp. 111Google Scholar
Bedford, M.R. (2000) Exogenous enzymes in monogastic nutrition – their current value and future benefits. Animal Feed Science and Technology 86, 113.Google Scholar
Chiou, P.W.S., B. Yu, B., Wu, S.T., Liu, C.C. and Gauthier, R. (2007) Effects of enzyme inclusion in a maize–soybean diet on broiler performance. Animal Feed Science and Technology 134 (3–4), 283294.Google Scholar
Doherty, J.V.O., O'Shea, C.J., Mc Alpine, P.O., Solan, P., Curran, T., Varley, P.F. and Walsh, A.M. (2014) The effect of protease and xylanase enzymes on growth performance, nutrient digestibility, and manure odour ingrower–finisher pigs. Animal Feed Science and Technology 189, 8897Google Scholar
Fidelis Fru-Nji, F. (2011) Use of a protease in poultry feed offers promising environmental benefits. International Journal of Poultry Science 10 (11), 842848.Google Scholar
Glenney, P. and Filer, K. (2005) Development of an analytical method for the analysis of acid proteases in feed samples. Poster Presentation at ADSA/ASAS Joint Annual Meeting.Google Scholar
Hajati, H. (2010) Effects of enzyme supplementation on performance, carcass characteristics, carcass composition and some blood parameters of broiler chicken. American Journal of Animal and Veterinary Sciences 5 (3), 221227.Google Scholar
Hayashi, K., Saleh, F., Ohtsuka, A. and Tanaka, T. (2004) Carbohydrates are Digested by Proteases present in enzyme preparations during in vitro digestion. Journal of Poultry Science 41, 229235.Google Scholar
Isaksen, M.F., Cowieson, A.J. and Kragh, K.M. (2010) Starch- and Protein-degrading Enzymes: Biochemistry, Enzymology and Characteristics Relevant to Animal Feed Use, In: Partridge, G.G.; Bedford, M.R. (Eds.), Farm Animal Nutrition, 2nd edition.; Publisher: CABI North American Office, Cambridge, MA 02139, USA, 8595.Google Scholar
Johnson, F.N. (1996) General tests and Assays, in: Food Chemicals Codex. Publisher: National Academy Press, Washington, USA, 812813.Google Scholar
Kalmendal, R. and Tauson, R. (2012) Effects of a xylanase and protease, individually or in combination, and an ionophore coccidiostat on performance, nutrient utilisation, and intestinal morphology in broiler chickens fed a wheat-soybean meal-based diet. Poultry Science 91 (6), 1387–93Google Scholar
Ljubojević, D.B., Milošević, N., Bjedov, S. and Stanaćev, V. (2011) The nutritive value of extruded corn in nutrition of broiler chickens. Biotechnology in Animal Husbandry 27 (4), 17331740.Google Scholar
McCleary, B.V. (2001) Analysis in Feed Enzymes; in: Partridge, G.G.; Bedford, M.R. (Eds.), In Enzymes in Farm Animal Nutrition; Publisher: CABI Publishing, Wallingford, UK85108Google Scholar
Mehri, M. and Shirmohammad, F. (2011) Effects of dietary supplementation of multi-enzyme complex on the energy utilisation in rooster and performance of broiler chicks. African Journal of Biotechnology 10(38), 75417547Google Scholar
Sheehan, N. (2010) Analysis of Enzymes, principles and Problems: Developments in Enzyme Analysis in: Partridge, G.G.; Bedford, M.R. (Eds.), In Enzymes in Farm Animal Nutrition, 2nd edition.; Publisher: CABI North American Office, Cambridge, MA 02139, USA, 260272Google Scholar
Simone, G. and Robouch, P. (2014) EURL-FA Guide: Protocol for verification studies of single laboratory/in-house validated methods. Working document v3.00. European Commission, Directorate-General Joint Research Centre. https://ec.europa.eu/jrc/sites/default/files/EURLFA-technical%20guide%20for%20validation%20and%20verification%20v2014.pdfGoogle Scholar
Thompson, M., Ellison, S.L.R., Wood, R. (2002) Harmonised Guidelines For Single Laboratory Validation Of Methods Of Analysis (IUPAC Technical Report) Pure and Applied Chemistry, 74, 835855.Google Scholar
Südekum, K.H., Edmunds, B., Spiekers, H. and Schwarz, F.J. (2012) Estimating ruminal crude protein degradation of forages using in situ and in vitro techniques. Animal Feed Science and Technology 175, 95105.Google Scholar
Vieira, S.L., Freitas, D.M., Angel, C.R., Favero, A. and Maiorka, A. (2011) Performance and nutrient utilisation of broilers fed diets supplemented with a novel mono-component protease. Journal of Applied Poultry Research 20, 322334Google Scholar
Zakaria, H.A.H., Jalal, M.A.R. and Abu Ishmais, M.A. (2010) The influence of supplemental multi-enzyme feed additive on the performance, carcass characteristics and meat quality traits of broiler chickens. International Journal of Poultry Science 9 (2), 126133.Google Scholar