AUSTRALIAN POULTRY CRC FINAL REPORT Program 1 Project No: 03-04: UNE PROJECT LEADER: Dr Paul A. Iji DATE OF COMPLETION: 31 July 2007 Control over life-long productivity by dietary manipulation immediately post hatch © 2008 Australian Poultry CRC Pty Ltd All rights reserved. ISBN 1 921010 14 2 Control over life-long productivity by dietary manipulation immediately post hatch Project No. 03-04 The information contained in this publication is intended for general use to assist public knowledge and discussion and to help improve the development of sustainable industries. The information should not be relied upon for the purpose of a particular matter. Specialist and/or appropriate legal advice should be obtained before any action or decision is taken on the basis of any material in this document. The Australian Poultry CRC, the authors or contributors do not assume liability of any kind whatsoever resulting from any person's use or reliance upon the content of this document. This publication is copyright. However, Australian Poultry CRC encourages wide dissemination of its research, providing the Centre is clearly acknowledged. For any other enquiries concerning reproduction, contact the Communications Officer on phone 02 6773 3767. Researcher Contact Details2 (Name): Dr Paul A. Iji (Address): School of Environmental & Rural Science, The University of New England, Armidale NSW 2351. Phone: (02) 6773 2082 Fax: (02) 6773 3922 Email: [email protected] In submitting this report, the researcher has agreed to the Australian Poultry CRC publishing this material in its edited form. Australian Poultry CRC Contact Details PO Box U242 University of New England ARMIDALE NSW 2351 Phone: 02 6773 3767 Fax: 02 6773 3050 Email: [email protected] Website: http://www.poultrycrc.com.au Published in August 2008 Contents Executive Summary .............................................................................................................................. 3 1. Introduction ....................................................................................................................................... 4 1.1 Objectives ..................................................................................................................................... 5 2. Methodology ...................................................................................................................................... 6 2.1 Strategy A ..................................................................................................................................... 6 2.1.1 Isolation of water-soluble carbohydrates ........................................................................... 6 2.2 Analytical techniques and measurements ................................................................................. 6 2.2.1 Constituent sugar analysis ................................................................................................... 6 2.2.2 High performance size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS) ............................................................................................................. 6 2.2.3 Determination of fructan contents and thin-layer chromatographic (TLC) analyses of fructans ........................................................................................................................................... 7 2.2.4 Extraction of genomic DNA for PCR amplification .......................................................... 7 2.3.5 Isolation and characterization of bacteria ......................................................................... 7 2.2.6 Sequencing of 16S rRNA gene ............................................................................................. 7 2.2.7 Fermentation characteristics of lactobacilli ....................................................................... 8 2.2.8 Enzyme-linked immunosorbent assay (ELISA) ................................................................ 8 2.2.9 Bird husbandry ..................................................................................................................... 8 2.3.1 Treatments ............................................................................................................................ 9 2.3.2 Feed preparation .................................................................................................................. 9 2.3.3 Measurements ..................................................................................................................... 10 2.3.4 Marker preparation and administration .......................................................................... 10 2.4 Experiment 2 .............................................................................................................................. 10 2.4.1 Experimental facilities........................................................................................................ 11 2.4.2 Treatments .......................................................................................................................... 11 2.4.3 Feed preparation ................................................................................................................ 11 2.5 Experiment Three...................................................................................................................... 11 2.5.1 Hypotheses and aims .......................................................................................................... 12 2.5.2 Bird husbandry ................................................................................................................... 12 2.5.3 Feed preparation ................................................................................................................ 13 2.5.4 Analytical procedures ................................................................................................... 13 Particle sizes of feed, milled grain and duodenal digesta ............................................. 13 2.5.5 Intestinal membrane-bound and pancreatic enzyme activity ............................. 14 2.5.7 Digesta flow marker analyses ..................................................................................... 14 2.6 General methods applied to both strategies ............................................................................ 14 2.6.1 Experimental diets .............................................................................................................. 14 2.6.2 Collection and processing of samples ............................................................................... 15 2.6.3 Organ weights ..................................................................................................................... 15 2.6.4 Acid-insoluble ash ............................................................................................................... 15 2.6.5 Apparent metabolisable energy (AME) bioassay ............................................................ 15 2.6.6 Digestibility of nutrients .................................................................................................... 16 2.6.7 Proximate analysis .............................................................................................................. 16 2.6.8 Gut histomorphology ......................................................................................................... 16 2.6.9 Enumeration of intestinal bacteria ................................................................................... 16 2.6.10 Measurement of organic acids ......................................................................................... 17 2.6.11 Necrotic enteritis challenge model .................................................................................. 18 2.7.12 Statistical analysis ............................................................................................................. 18 2.7.13 Animal ethics ..................................................................................................................... 18 3 Results ............................................................................................................................................... 18 3.1 Strategy A ................................................................................................................................... 18 1 3.1.1 Water-soluble prebiotic compounds from Australian and New Zealand plants: isolation and characterisation .................................................................................................... 18 3.1.2 Fructans from Rengarenga lily (Arthropodium cirratum) extract and frutafit as prebiotics for broilers .................................................................................................................. 20 3.1.3 Prebiotic plant extracts for broilers: their effects on growth performance, intestinal morphology, microbial composition and activity ..................................................................... 22 3.1.4 Molecular and biochemical characterisation of lactobacilli isolated from ileal and caecal digesta of broilers fed prebiotic plant extracts .............................................................. 23 3.1.5 Natural plant extracts and prebiotic compounds as alternatives to antibiotics in broilers in a necrotic enteritis challenge model ........................................................................ 24 3.2 Strategy B ..................................................................................................................................