Chicken Meat Program

2003-2004 Research Report

Chicken Meat Program

January 2005 RIRDC Publication No 04/076

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© 2005 Rural Industries Research and Development Corporation All rights reserved.

ISBN 0 642 ISSN 1440-6845

2003-2004 Research Report RIRDC Chicken Meat Program” Publication No 04/076 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 Commonwealth of Australia, Rural Industries Research and Development Corporation, 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, RIRDC encourages wide dissemination of its research, providing the Corporation is clearly acknowledged. For any other enquiries concerning reproduction, contact the Communications Manager on phone 02 6272 3186. RIRDC Chicken Meat Program Research Manager Dr Vivien Kite RIRDC PO Box 579 NORTH SYDNEY NSW 2059 Phone: 02 9929 4077 Fax: 02 9925 0627 Email: [email protected] RIRDC Publications Manager Rural Industries Research and Development Corporation Level 1, AMA House 42 Macquarie Street BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6272 3186 Fax: 02 6272 5877 Email: [email protected] Website: http://www.rirdc.gov.au

Published in January 2005 Printed on environmentally friendly paper by Union Offset, Canberra

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Foreword This year RIRDC has produced Research in Progress, June 2004, which contains short summaries of continuing projects as well as those that were completed during 2003-2004 for all of the Corporation’s 22 program areas. The complete report on all the programs is only available in electronic format on our website at www.rirdc.gov.au. The following report is a hardcopy extract covering the Chicken Meat Sub-program. It contains all entries from continuing and completed Chicken Meat research projects funded by RIRDC in 2003-2004. Additional information on other activities funded by the program in 2003-2004 and projects and activities to be funded in 2004-2005 has also been included in this publication. The objective of the Chicken Meat Program is to support increased sustainability and profitability in the chicken meat industry by focussing on research and development on those areas which will enable the industry to become more efficient and globally competitive and which will assist in the development of good industry and product images. Research reported upon herein was funded from industry revenue which is matched by funds provided by the Federal Government. This report is an addition to our extensive catalogue of more than 1200 research report, videos and CD-roms of projects supported by RIRDC. Most of our publications are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/fullreports/Index.htm • purchases at www.rirdc.gov.au/eshop Tony Byrne Acting Managing Director Rural Industries Research and Development Corporation

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CHICKEN MEAT PROGRAM ADVISORY COMMITTEE

Chairperson: Mr Barry Shay Food Safety Consultant 27 Uther Street CARINDALE QLD 4152 Ph: (07) 3398 1766 Fax: (07) 3398 6631 Email: [email protected]

Research Manager: Dr Vivien Kite RIRDC Chicken Meat Program PO Box 579 NORTH SYDNEY NSW 2059 Ph: (02) 9929 4077 Fax: (02) 9929 0627 Email: [email protected]

Committee Members: Mr Ian Farran Agribiz Engineering PO Box 279 GEELONG VIC 3220 Ph: (03) 5229 7300 Fax: (03) 5229 7566 Email: [email protected]

Dr Tom Grimes Grimes Consultancy Pty Ltd 4 Henry Street LEWISHAM NSW 2049 Ph: (02) 9569 7436 Fax: (02) 9569 4183 Email: [email protected]

Dr Ron MacAlpine Inghams Enterprises Pty Ltd PO Box 4 LIVERPOOL NSW 2170 Ph: (02) 9606 5666 Fax: (02) 9606 6640 Email: [email protected]

Dr Margaret MacKenzie Inghams Enterprises Pty Ltd PO Box 1100 BROWNS PLAINS QLD 4118 Ph: (07) 3290 4644 Fax: (07) 3297 0578 Email: [email protected]

Dr Pat Blackall Animal Research Institute Department of Primary Industries Locked Mail Bag No 4 MOOROOKA QLD 4015 Ph: (07) 3362 9498 Fax: (07) 3362 9429 E-mail: [email protected]

Mr Gary Sansom 82 Hawkins Road JIMBOOMBA QLD 4280 Ph: (07) 5546 9235 Fax: (07) 5546 0070 E-mail: [email protected]

Dr Jeff Davis RIRDC PO Box 4776 KINGSTON ACT 2604 Ph: (02) 6272 4152 Fax: (02) 6272 5877 Email: [email protected]

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INDEX TO PROJECT SUMMARIES

COMPLETED PROJECTS

Flock Health ...............................................................................................................1 CSA-18J The effect of Newcastle disease vaccination with strain V4 on the course of infection

with the Peats Ridge strain on Newcastle disease virus...................................................1 RMI-11A The development of vaccination strategies to control necrotic enteritis in poultry............3 RMI-15J Mareks disease research in Australia – a review..............................................................5 UMU-29J Control of intestinal spirochaete infections in chickens.....................................................8

Bird Nutrition and Feed Supply................................................................................9 DAQ-277A Developing a slope ratio assay for amino acid availability ................................................9 DAQ-302A Evaluation of new millet varieties as a poultry feed ingredient .......................................11 UNE-82J The net energy value of Australian feed ingredients for poultry......................................13 US-104A Use of dietary fatty acids to increase protein accretion in broilers..................................14

Food Safety ..............................................................................................................16 UA-61A Growth rate of broiler chickens given condensed tannins extracted from grape seed ...16

Environmental Management...................................................................................18 FSE-2A Meat chicken EMS: transfer to industry ..........................................................................18 JSC-2A Sustainability improvements in the Victorian meat chicken industry (phase 2) ..............19

RESEARCH IN PROGRESS

Flock Health .............................................................................................................21 CSA-20A Postgraduate scholarship - Scott Sheedy: Live vectoring of therapeutic and

prophylactic proteins and pathogenesis in necrotic enteritis...........................................21 CSA-21J Postgraduate scholarship - Manija Asif: Cytokines and innate molecules for enhanced

mucosal immunity in the chicken.....................................................................................22 CSA-24J Rapid identification and pathotyping of virulent IBDV, NDV and AI isolates...................23 CSA-25J Postgraduate scholarship - Kristie Jenkins: Improved therapeutics for Marek's disease

virus infection...................................................................................................................24 CSA-26J Use of cytokines to enhance vaccine efficacy in poultry .................................................25 CSA-29A Biological control of necrotic enteritis in meat chickens..................................................26 CSA-30J Characterisation and modulation of virulence of endemic IBDV strains using reverse

genetics ...........................................................................................................................27 DAQ-316A New diagnostic assays to improve control of coccidiosis in poultry................................28 DAQ-319A Preliminary investigations into the efficacy of novel treatments and application

methods for darkling beetle (Alphitobius diaperinus [Panzer]) control in Australian broiler houses..................................................................................................................29

DAW-113A Advanced clinical diagnostics: use of real-time immuno-PCR and LightUp probes .......30 RMI-12J Molecular evaluation of responses to vaccination and challenge by Marek's disease

viruses .............................................................................................................................31 UJC-10J Molecular techniques for monitoring Marek's viraemias in broilers and layers...............32 UM-68A Avian leukosis subgroup-J (ALV-J): epidemiological studies for control of ALV-J in

Australian broiler breeder flocks......................................................................................33 UNE-83J Systematic pathotyping of Australian Marek's disease (MDV) isolates ..........................34 UQ-100J Typing of Pasteurella multocida ......................................................................................35 UTS-4J Efficacy trials of a maternally-delivered recombinant vaccine against coccidiosis .........36 UTS-6A Postgraduate scholarship - Ms Kelly Mai: The molecular basis for oocyst wall

formation in the apicomplexan parasite, Eimeria maxima ..............................................37

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Bird Nutrition and Feed Supply..............................................................................38 GRD-3J Premium grains for livestock program (stage 2) .............................................................38 UNE-86A Postgraduate scholarship - Mr Nicholas Rodgers: Relationships between grain quality,

intestinal integrity, and performance of broiler chickens .................................................39 UQ-107A Digestible amino acids and improved broiler performance .............................................40 US-127A Establishment of a Chair in Poultry Sciences at The University of Sydney ....................41 UWA-76J Mechanical and enzymatic improvements of dehulled lupins for broiler and layer diets

(AECL-managed project WAU-1A)..................................................................................42 Food Safety ..............................................................................................................43

DAQ-282A On-farm reduction strategies for Campylobacter spp. ....................................................43 IMV-5A Development of a sequence-based bacteriophage typing system for Salmonella .........44 RMI-14A Development and validation of Campylobacter microarrays for virulence detection

and strain differentiation in poultry products ...................................................................45 UG-7A Campylobacter bio-replacement program to control food poisoning organisms in

poultry..............................................................................................................................46 Environmental Management...................................................................................47

DAQ-318A Evaluating risks posed by pathogen emissions from meat chicken sheds .....................47 DAQ-312A Efficacy of windbreak walls for odour reduction ..............................................................48 DAV-213A Trials of odour control technologies for broiler farms......................................................49 FSA-3A Risk assessment on the use of chicken litter and guidelines for its safe use .................50

Other.........................................................................................................................51 CPO-1A CRC for the Australian Poultry Industries funding...........................................................51

OTHER SUPPORTED ACTIVITIES

SCHOLARSHIPS ......................................................................................................52

PROGRAM REVIEW AND DEVELOPMENT............................................................52

TRAVEL/CONFERENCE/WORKSHOPS .................................................................52

RESEARCH TO BE SUPPORTED IN 2004/2005.....................................................53

CHICKEN MEAT PROGRAM – COMPLETED PROJECTS

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COMPLETED PROJECTS

Flock Health Project Title:

The effect of Newcastle disease vaccination with strain V4 on the course of infection with the Peats Ridge strain on Newcastle disease virus

RIRDC Project No

CSA-18J

Researcher: Dr. Paul Selleck, Dr. Hans Heine, Dr. John Bingham, Dr. David Boyle and Dr. Peter Daniels

Organisation: CSIRO Australian Animal Health Laboratory Private Bag 24 GEELONG VIC 3220

Phone: (03) 5227 5000 Fax: (03) 5227 5555 Email: [email protected] Objectives

• To determine whether prior vaccination with V4 vaccine can reduce subsequent

infection with Peats Ridge strain and/or reduce the amount and duration of shedding of Peats Ridge strain Newcastle disease virus.

• To determine whether V4 vaccination during a Peats Ridge infection can modify the course of the Peats Ridge infection.

• To determine the serological response to the Peats Ridge strain alone and to Peats Ridge infection following prior vaccination with V4 vaccine.

Background Newcastle disease may cause extensive losses in the poultry industry, through illness

and death of infected birds and through slaughter-out control policies. In Australia, virulent Newcastle disease virus (NDV) may evolve from avirulent, endemic forms of the virus. Control of virulent and avirulent NDV outbreaks can be carried out through vaccination of flocks, using a live virus vaccine, V4. Vaccination of flocks prior to an NDV outbreak provides substantial immunity. However, it is not known to what extent prior vaccination with V4 reduces the extent of shedding of a subsequently acquired wild NDV virus, or if vaccination of flocks in the face of an outbreak is effective at curtailing the progression of the outbreak.

Research Two separate trial protocols were conducted, using V4 vaccine and Peats Ridge (avirulent) strain challenge. The first trial was a vaccinate-challenge trial, with 21 days between vaccine and challenge virus administration. The second was a challenge-vaccinate trial, with two days between challenge and vaccine administration. Oral-tracheal swabs and cloacal swabs were taken from each bird and tested for the presence of virus by egg inoculation. A selection of virus isolates and original positive swabs were tested by TaqMan PCR technology to determine which of the two virus types – V4, Peats Ridge strain or both – was present in the sample. Serum samples were taken at intervals after challenge and tested for antibody titres.

Outcomes In birds that had received prior immunisation with V4, shedding was almost exclusively confined to the oral-tracheal route and this was in a much lower percentage of the birds compared to unvaccinated control groups. There were no major differences in shedding patterns between unvaccinated controls and birds that were vaccinated two days after Peats Ridge strain challenge. Serology studies


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indicate that there was 100% seroconversion to Peats Ridge strain challenge and seroconversion was rapid with over 90% of birds showing seroconversion by day 7.

Implications The implications of this study are (a) prior vaccination with V4 vaccine is effective at significantly reducing the shedding of virus during subsequent Peats Ridge strain infection; (b) post-challenge vaccination with V4 vaccine during active Peats Ridge strain infection does not appear to significantly affect the number of birds that shed Peats Ridge strain, nor does it shorten the duration of shedding; and (c) Peats Ridge strain infection induces a significant frequency of rapid seroconversion.


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Project Title:

The development of vaccination strategies to control necrotic enteritis in poultry

RIRDC Project No.:

RMI-11A

Researcher: Prof. Peter Coloe Organisation: RMIT University

Plenty Road BUNDOORA VIC 3083


• To lay a foundation for understanding and controlling necrotic enteritis by

developing a model for the reproduction of necrotic enteritis in chickens. • To develop and evaluate a vaccine, based on toxoid, for the control of necrotic

enteritis that can be delivered orally, that is cost effective to manufacture and that can be delivered within established farming practices.

Background A major aim of the poultry industry is to produce high quality product without the

need for supplementation with antimicrobial agents. To achieve this objective, over recent years there has been a strong move towards the use of vaccination strategies to replace long term chemical use for the control of diseases. Effective vaccination of poultry against necrotic enteritis will not only illustrate the industry’s proactive stance with respect to replacing chemical control measures for disease, but should enhance the productivity of flocks. Since an outbreak of necrotic enteritis (which is caused by the pathogen, Clostridium perfringens) can result in between 2% to 10% mortality, as well as causing significant losses in flock productivity, effective vaccination will have a major cost benefit outcome for the industry. Effective vaccination to control C. perfringens alpha toxin will also ensure the development of a healthy, effective gut microflora. This microflora will limit the possibility of other undesirable organisms colonising the chicken and posing a threat to human health after processing.

Research The approach taken in this project was to establish a reproducible model for NE, to produce and test a killed vaccine based on toxoid and, if this proved successful, to clone and deliver a live modified vaccine. A variety of attempts were undertaken to reproduce necrotic enteritis in poultry. These ranged from varying the challenge strain, varying the diet of the birds and varying the dates of challenge as well as co-infecting birds with Eimeria spp and C. perfringens. In addition, birds were also pre-treated with antibiotics to reduce the microbial flora prior to challenge. The C perfringens toxin was cloned in E. coli and purified toxin was used in mouse proof of concept studies to show the toxin was antigenic. A range of truncates of the toxin were also prepared and tested in mice.

Outcomes Unfortunately, a reproducible model for the reproduction of necrotic enteritis in chickens was not successfully established. Necrotic enteritis was achieved at a low level using a variety of approaches, but the incidence of disease was inconsistent and not reproducible enough to be used as a model for vaccine evaluation. Nevertheless, the C. perfringens alpha toxin was cloned into E. coli and a variety of truncates prepared. The purified toxin, when used as a toxoided vaccine in mice, was shown to protect against an intra-muscular challenge of C. perfringens. Truncates of the C. perfringens toxin were less effective than the whole toxin. However, without an effective chicken model to test the purified toxin in this work


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did not proceed further. The availability of cloned alpha toxin will enable evaluation of this as a protective antigen when a model of necrotic enteritis in chickens becomes available.

Implications The development of an efficacious vaccine for necrotic enteritis would be a significant step forward for the chicken industry. A vaccine that is cost effective and can be delivered orally would enhance the productivity of the industry and also enhance disease control and flock welfare. However, it is necessary for an effective disease model to be developed in order that the efficacy of any vaccine or new treatment method can be properly evaluated. Such a model will require a much higher incidence of disease and reproducibility than was achieved in this project, before it can be used to evaluate efficacy against necrotic enteritis. Know-how on production and purification of toxin gained in the course of this project will facilitate future progress towards a toxoid vaccine approach and provide material for alternative vaccine approaches. However, additional work is required on a disease model so that vaccine approaches can be effectively evaluated.


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Project Title:

Mareks disease research in Australia – a review

RIRDC Project No.:

RMI-15J

Researcher: Prof. Greg Tannock Organisation: RMIT University

Department of Biotechnology and Environmental Biology Bundoora West Campus PO Box 71 BUNDOORA VIC 3083


• To undertake and publish a comprehensive review of all relevant background to

and Australian research undertaken on Marek’s disease in response to the problems that were experienced in the field in Australia with Marek’s breaks in the mid 1990s.

Background Marek’s disease (MD) continues to cause major economic losses in all poultry

producing countries of the world. In Australia, control of MD has been achieved with locally produced vaccines, namely HVT (herpes virus of turkeys; serotype 3) and Maravac (serotype 2). However, during the early 1990s, the need for a serotype 1 MD vaccine was apparent due to the frequent isolation of increasingly virulent strains of Marek’s disease virus (MDV). Poultry farms were experiencing excessive losses due to MD, despite vaccination with the serotype 2 and 3 vaccines available at that time. A similar situation in the USA resulted in the introduction of the serotype 1, Rispens CVI988 strain vaccine. The use of a serotype 1 vaccine in the USA appeared to be the solution to many of the problems being experienced there with the newly emerged virulent strains. In Australia, however, the likelihood of importing the Rispens vaccine appeared at the time remote. Consequently, a series of contract trials were funded by the RIRDC to evaluate, under controlled conditions, the efficacy of local, commercially available vaccines against a more recent field strain of MDV. These would evaluate if the problems that were experienced at the time were due to the evolution of new viruses for which current vaccines failed to provide adequate protection or due to problems with vaccine application. The industry also encouraged the development of a serotype 1 vaccine from an Australian MDV strain in the event that the currently available vaccines were not protecting against circulating viruses. The effectiveness of Rispens vaccine was also evaluated in later challenge trials as it became apparent that available local vaccine strains were not going to be sufficient to control MD in Australia, and hence importation of this vaccine was countenanced. Laboratory research associated with MDV can provide new and improved methods for combating the MD problem in Australia. Because of limitations associated with the detection techniques available at the time for MDV serotype 1 virus, it was considered that a MDV1 specific probe that could be used in a rapid identification assay, which is less expensive and more specific than those currently available, would aid field diagnosis and prove important in vaccine evaluation. Rapid tests for the detection of MDV ensure that chicken flocks are protected by vaccination and assists in containment measures to prevent the spread of infection. Adjuvants that are used to improve the immunogenicity of a vaccine without increasing the amount of infectious virus in the vaccine have the potential to add value to the cheap and readily available existing HVT vaccine, and could minimise the need for newer vaccines. The adaptation of MDVs to growth in a continuous cell line could be useful for


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vaccine production, compared with the labour and reagent intensive CEF cultures that are currently used. The availability of new vaccines in Australia increases the need for a reliable assay system to evaluate their effectiveness. Previously, vaccine assays were carried out by individual manufacturers without access to standard reference preparations. Finally, a review of this scope was deemed necessary to place together all the disparate bits of work and to record them in one document why and how they were done.

Research This review begins by describing the development of a live attenuated serotype 1 MDV vaccine from a highly virulent Australian strain, the Woodlands No. 1 strain. Clones of the virus were developed and tested at various passages for attenuation and loss of pathogenicity. In further work described in the review, a potential vaccine candidate, the Woodlands clone and existing MD vaccines, were subjected to a series of challenge trials to evaluate their effectiveness. The trials included the newly imported CR6 vaccine and used SPF and commercial birds (presumably hosting maternal antibodies to other vaccines). Bi- and trivalent vaccine combinations were also evaluated. Two cell culture-prepared MDV challenge viruses, produced from the Woodlands No. 1 and MPF 57 strains were used as challenge strains in the trials. Additional research on MD included many laboratory studies and each was discussed in full throughout the review. These began with the development of a probe labelled with digoxigenin (DIG) for the detection of MDV type 1 by dot-blot hybridisation. The sensitivity and specificity of this procedure was then compared with virus isolation in cell culture and with PCR, which was used as the reference procedure. In order to achieve a representative collection of MDV strains, approximately 300 blood samples were collected from flocks in different parts of the country that had been experiencing losses due to MD. These blood samples were processed so they could be screened for the presence of serotype 1 MDV. HVT and MDV 1 were adapted to the Vero continuous cell line and their yields determined by the microtitre technique in primary CEF cultures. A PCR was used to detect the presence of 132 bp direct repeat sequence specific for serotype 1 MDVs. Commercial broiler chickens were used to assess the possible role of γ-inulin as an adjuvant to improve the efficacy of HVT vaccination against MD. Chickens were administered a cell-associated HVT vaccine with or without γ-inulin using three different vaccination procedures. The Virology Laboratory also set up a vaccine assay facility and was accredited with the National Association of Testing Authorities (NATA). Studies were also carried out in response to industry concerns about possible losses to vaccine potency from the widely used practice of using chilled diluent whilst administering vaccines.

Outcomes The review collateds research that was undertaken in response to the MD problems experienced by Australia in the mid 1990s. It describes the design and results of challenge trials which showed that currently available local vaccines did not adequately protect against MD. The challenge trials also evaluated the efficacy of potential overseas vaccines against the local type 1 viruses and showed that Rispens vaccine did provide protection. The review also provides an exact account of the development of an alternative local vaccine and how attenuation of the Woodlands No. 1 virus was observed at passage 60 in a chicken pathogenicity test, which was confirmed in later pathogenicity experiments on clone No. 2 from the same passage (the 60/2 clone). Large-scale


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safety and protection tests confirmed that the 60/2 clone was both safe and efficacious. No gross tumours were observed in any of the vaccinated birds, although some mild immune organ depletion was evident in a safety test. Research undertaken also showed that the 60/2 clone provided high levels of protection compared with other vaccines, but the Rispens vaccine performed better. On the other hand, the newly imported CR6, when used alone, produced very poor protection. Of work performed in the laboratory, the research affirmed that the highest sensitivity rates for detection of MDV 1 were achieved by dot-blot hybridisation using the 132 bp PCR probe, compared with virus isolation and identification by immunoperoxidase or immunofluorescence, and, despite their much lower sensitivity, higher specificity was obtained by both culture detection methods than for the dot-blot hybridisation. Intact DNA with a size of approximately 180 kb was found in both serotype 1- and HVT-infected Vero cells after isolation and characterisation, indicating that whole copies of both types of DNA were present and providing further evidence for adaptation to growth of the serotype 1 virus. This is the first report of the growth of either virus in a continuous line. The vaccine assay developed at RMIT University proved successful and now provides Australian vaccine manufacturers with a reliable assay facility. During its development, it was found that the use of diluent held at RT substantially decreases virus titre loss (as opposed to 4°C), while the difference in vaccine titre of parallel assays of two operators were non-significant and, therefore, should not affect the assay result. Finally, it is noted that γ-inulin did not appear to function as an adjuvant when administered with the HVT vaccine.

Implications The MD challenge trials facilitated the importation of the Rispens vaccine into Australia and its subsequent availability to the Australian chicken industry. This in turn lessened the need for an Australian serotype 1 MD vaccine. Nevertheless, the RMIT vaccine that was developed may be useful if further change occurs in the evolution of MDVs in Australia. Furthermore, the challenge trials have established a protocol to evaluate vaccine efficacy against emerging Marek’s strains in the future. The dot-blot hybridisation technique using the DIG-labelled probe specific for MDV 1 was shown to be potentially very useful as a rapid and economical test to detect MDV. Also, the adaptation of the growth of MDV 1 to the Vero continuous cell line will enable new and improved vaccine production procedures. RMIT University now provides a national independent assay facility for the standardised assessment of MD vaccines for general use by all Australian manufacturers. In addition to this, RMIT provides and maintains a repository of MDV serotype 1 strains which will allow future trends in the evolution of MDV to be studied.

Publications De Laney, D.B., Morrow, C.J., Read, K.M. and Tannock, G.A. (1998) The development and evaluation of two tissue culture-grown Marek’s disease challenge viruses. Avian Pathology 27: 472-477.

Cipriani, T. L. (2000) The development of two digoxigenin-labelled probes for the detection of Marek’s disease virus by dot-blot hybridisation. B. App.Sc.Honours Thesis, RMIT University.

Jaikumar, D. (1998) Propagation and molecular characterisation of Marek’s disease virus. Master of App.Sc. Thesis, RMIT University.

Jaikumar, D., Gilmore, K.M. and Tannock, G.A. (2001) Adaptation of Marek’s disease virus to the Vero continuous cell line. Veterinary Microbiology 70: 75-82.


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Project Title:

Control of intestinal spirochaete infections in chickens

RIRDC Project No.:

UMU-29J

Researcher: Prof. David Hampson Organisation: Murdoch University

Division of Veterinary and Biomedical Sciences MURDOCH WA 6150


• To develop improved methods to control infection by Brachyspira intermedia and

Brachyspira pilosicoli - bacterial pathogens causing significant economic loss in Australian layer and broiler breeder flocks.

Background Avian intestinal spirochaetosis (AIS) is a condition of layers and broiler breeders

caused by infection of the large intestine with anaerobic intestinal spirochaetal bacteria. Colonisation leads to problems of wet litter and reduced egg production. The spirochaetes Brachyspira intermedia and Brachyspira pilosicoli infect many Australian flocks.

Research The on-farm epidemiology of the infections was examined on a broiler breeder and a layer flock. Birds were monitored and individual isolates identified and typed. Studies were undertaken to examine the survival of the spirochaetes in chicken faeces and in the presence of disinfectants. Experiments were conducted to investigate interactions between different diets and colonisation in experimentally-infected layers.

Outcomes Infection with different spirochaete species and with multiple individual strains was detected in the layer flock. Infection apparently originated from other birds on the site rather than from environmental sources. The organisms survived briefly in the environment and were highly susceptible to common disinfectants. Dietary influences on colonisation were found for both spirochaete species. Diets based on wheat predisposed to colonisation with B. intermedia. Wheats differing in their non-starch polysaccharide (NSP) content predisposed differently to infection with B. intermedia compared to B. pilosicoli. No clear relationship was found between the soluble NSP content of a given diet and the viscosity of the digesta in the ileum. Addition of dietary enzymes did not significantly reduce the digesta viscosity in the ileum. Increased dietary soluble NSP and/or increased viscosity of the ileal digesta did not necessarily lead to an increased colonisation with B. intermedia.

Implications Cycles of infection can be broken by thorough cleaning of sheds between batches of birds and applying effective biosecurity measures to prevent transmission between sheds. Where AIS is a problem, substitution of wheat with other cereals may reduce colonisation. Addition of enzymes to diets for laying hens is unlikely to provide effective control.

Publications Phillips, N.D., La, T. and Hampson, D. J. (2003) Survival of intestinal spirochaete strains from chickens in the presence of disinfectants and in faeces held at different temperatures. Avian Path. 32: 639-643.


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Bird Nutrition and Feed Supply

Project Title:

Developing a slope ratio assay for amino acid availability

RIRDC Project No.:

DAQ-277A

Researcher: Dr. Rider Perez-Maldonado Organisation: Department of Primary Industries & Fisheries (Qld)

QPRDC PO Box 327 CLEVELAND QLD 4163


• To establish and validate a slope ratio chick assay for determining the availability

of lysine in selected Australian canola meals (CM) and cottonseed meals (CSM). • To compare lysine availability values determined using this assay with ileal

apparent digestibility values determined in the same samples of CM and CSM.

Background The levels of undigested amino acids (AA) appearing at the terminal ileum or in excreta are used to determine the AA digestibility of feed ingredients for poultry. It is assumed that, if an AA is not recovered at the terminal ileum or in excreta, then it has been absorbed and used for maintenance and production. This is not always true, a fact which has led to the concept of amino acid ‘availability’. Availability differs from digestibility in that it involves a measure of utilisation or potency of the absorbed AA. Available AAs are those which are actually supplied at the appropriate sites for protein synthesis for incorporation into body protein or as metabolites for other body uses.

Research A pilot assay and two slope ratio assay trials were conducted to evaluate the availability of lysine in CM from Boree (extruded meal), Riverland and Melbourne, and one CSM from Narrabri. The pilot study established the dose levels to ensure a linear response to standard lysine and the test proteins. The slope ratio assay developed an intersecting lines multiple linear regression model to estimate the lysine bioavailability of the four test proteins meals. The criteria analysed for lysine availability were live weight gain (LWG) and feed conversion ratio (FCR).

Outcomes The methodology for a chick slope ratio assay for determining the AA availability was established. The availability estimates when using FCR were in close agreement with the LWG figures. For CSM, the availability of lysine (at 0.56) was similar to lysine digestibility values obtained in previous trials. These low lysine digestibility and availability values are almost certainly due to anti-nutritional factors present in the CSM, such as condensed tannins, and to the effects of heat during plant processing. The lysine availability of CM from Boree, Riverland and Melbourne of 1.11, 0.91, and 0.83, respectively, were 49-100% higher than the lysine availability in CSM. Processing conditions are the primary cause of these lysine availability differences within the CM samples. The bioavailability of lysine in extruded meals was higher than for solvent extracted meals. In CM, the ileal digestibility method underestimated the CM lysine availability by 20-51%.

Implications The combined results from previous work and this slope ratio bioassay for CM and CSM suggest that, when ingredients display high lysine bioavailability (>0.80), generally there is no need to formulate diets on a digestible basis as no gain in terms of broiler performance would be obtained. However, when lysine bioavailability is less than 0.65 (as in the case of CSM) substantial gain would be obtained from


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formulating diets on a digestible amino acid basis. The duration and amount of temperature used during oil extraction, and differences in the type and amounts of condensed tannins which are present in CM and CSM, are likely to be responsible for the observed differences in lysine availability in these two feed ingredients and further trials are needed to confirm this.


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Project Title:

Evaluation of new millet varieties as a poultry feed ingredient

RIRDC Project No

DAQ-302A

Researcher: Mr. Danny Singh Organisation: Department of Primary Industries & Fisheries (Qld)

QPRDC PO Box 327 CLEVELAND QLD 4163


• To evaluate the nutritional value of Australian pearl millet varieties as a high

quality feed for poultry, by determining their gross chemical composition as well as the metabolisable energy content and digestible amino acid values of the selected millet varieties when fed to broilers.

• To generate data to assist in the promotion of the benefits of pearl millet varieties as a feed grain to both grain growers and poultry producers.

Background Pearl millet has been identified as a suitable alternative crop to sorghum in low

rainfall and sandy areas in Queensland. In the United States of America, pearl millet is being heralded as the new feed grain. New pearl millet hybrids that are now being developed in Australia have never been evaluated as a feed grain for poultry. The potential benefits from further research into the nutritional value of pearl millet for the development of this feed grain crop in warm-temperate agriculture are substantial.

Research The pearl millet hybrids were grown at Biloela Research Station, Biloela, Queensland and clean grain from three selected elite hybrids was transported to the Department of Primary Industries & Fisheries’ Poultry Research and Development Centre, Alexandra Hills, for evaluation. Chemical analyses were performed on these three grains to measure their protein, fat, fibre, starch, phosphorus, calcium and amino acid content. Feeding experiments were conducted using broiler chickens to measure the metabolisable energy content and the digestibility of amino acids in the three pearl millet hybrids.

Outcomes In comparison with sorghum, the millet hybrids had higher protein, fat and neutral-detergent fibre contents. Pearl millet hybrids contained nearly 2% more protein than sorghum. The fat content of the millet hybrids was more than twice that of sorghum. The fibre content of hybrid PM31 was lower than that of PM3 and PM4 but was higher than sorghum. The starch content and gross energy (GE) content of the three pearl millet hybrids were the same. Condensed tannin was not detected in any of the three hybrids. Linolenic acid ((ClS:3n-3) comprised 3% of the total fatty acid in the pearl millet hybrids, which is higher than other cereal grains. The ratios of n-6:n-3 fatty acids in the three hybrids were 13.72, 12.68 and 14.13 for PM31, PM3 and PM4 respectively, compared to sorghum’s 28.43. The pearl millet hybrids had at least a 0.6 MJ/kg higher energy content than sorghum. The total amino acid profile of the three elite pearl millet hybrids is much more suitable for poultry diets than sorghum. Pearl millet hybrids contained 50% more lysine, methionine, threonine and tryptophan than sorghum. Except for the protein content, where PM31 had slightly less protein, the three hybrids had very similar chemical compositions. The fat content was 6.4% and fibre content 2.0%. The amino acid profile of PM31 was poorer than that of PM3 and PM4. The lysine content of PM31 was 2.83 g/kg compared to 3.18 g/kg and 3.09 g/kg for PM3 and PM4 respectively. The digestibilities of cystine, lysine and threonine in the pearl millets were higher than that of sorghum, whereas the digestibilities of the other amino acids in the elite pearl millet hybrids were very similar to sorghum.


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Implications There is little doubt that the demand for cereal grains for poultry feed will increase

substantially in the future. It is essential that detailed knowledge of any potential alternative feed ingredient, such as pearl millet, be established to allow it to be used effectively and efficiently. The experiments conducted within this project have demonstrated the nutritive value of the millets, and provided details of their nutritional profile which is essential if they are to be formulated into least cost poultry diets. The results suggest that the three pearl millet hybrids could be used as feed ingredients in broiler diets. However, further research on the upper inclusion levels of pearl millet in broiler diets, together with the effects of feeding pearl millet on carcass quality and yield, and on the interaction of enzymes on the utilisation of nutrients in the millet hybrids, is warranted. Three elite parent lines were identified in previous yield trials (conducted under a GRDC project) and only small quantities of these parent lines are currently held in stock. Any subsequent evaluation of pearl millet hybrids is likely to be on a larger scale than the trials reported upon herein and will require several kilograms of seed. Since production of F1 hybrid seed is labour intensive and can take several cycles of regeneration, it would seem prudent to begin increasing seed stocks of the three identified elite parent lines.

Publications Singh, D.N. and Perez-Maldonado, R.A. (2003) Pearl millet as an alternative feed grain for pigs and poultry. Proceedings of the Nutrition Society of Australia, Vo1 27. S40.

Singh, D.N., Trappett, P.C., Nagle, T.A. and Perez-Maldonado, R. (2004) Digestibility of pearl millet in broiler diets. Proceedings of the Nutrition Society of Australia (in press).


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Project Title:

The net energy value of Australian feed ingredients for poultry

RIRDC Project No.:

UNE-82J

Researcher: Prof. Mingan Choct Organisation: The University of New England

School of Rural Science and Agriculture ARMIDALE NSW 2351

Phone: (02) 6773 5121 Fax: (02) 6773 3050 Email: [email protected] Objectives:

• To measure the net energy (NE) values of the common Australian feed

ingredients for broilers and layers. • To compare the performance of broilers fed diets formulated on either a net

energy or apparent metabolisable energy (AME) basis.

Background Energy is by far the most expensive part of a poultry diet, and the potential for improvement in production efficiency is large if dietary energy can be accurately measured. The NE bioassay is one system for measuring the energy value of feed ingredients that reflects the true availability of energy to the bird.

Research A series of experiments were conducted to test the NE value of commonly used energy and protein sources for both broilers and layers using a closed circuit calorimetric system.

Outcomes The measured NE values (MJ/kg) of wheat, barley, sorghum, millrun, sweet lupin, soybean meal (48% crude protein), canola meal and meat meal for broilers were: 11.89, 10.64, 13.18, 8.75, 3.87, 7.74, 5.28 and 7.44 respectively. The measured NE values (MJ/kg) of wheat, barley, sorghum, millrun, sweet lupin, soybean meal (48% crude protein), canola meal, meat meal and oats for layers were: 10.31, 10.26, 12.25, 6.73, 12.90, 11.71, 9.27, 15.75 and 11.44 respectively. Broiler diets formulated on NE values gave a clear advantage in feed conversion efficiency over those formulated using their AME values, resulting in savings of over 80 grams of feed per kg liveweight gain over a 35-day growth period.

Implications The formulation of poultry diets using NE values has clear advantages. However, due to the tedious nature of the NE system, the establishment of a NE database for practical feed formulation requires much more work.


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Project Title:

Use of dietary fatty acids to increase protein accretion in broilers

RIRDC Project No.:

US-104A

Researcher: Dr. Ron Newman Organisation: The University of Sydney

Faculty of Veterinary Science 405 Werombi Rd CAMDEN NSW 2570


• To develop a simple feed technology to reduce fat deposition and increase muscle

protein accretion in broilers. • To achieve this by manipulating dietary fatty acid intake to alter tissue sensitivity

to the metabolic hormones involved in lipid and protein metabolism. • To improve nutrient utilisation and reduce feed costs through the adoption of this

technology.

Background It had previously been established in RIRDC project US-44A that dietary fatty acids from the n-3 and n-6 series alter the carcass composition of the broiler, albeit by different mechanisms. In these earlier studies, diets contained 80 g/kg of edible tallow, fish oil or sunflower oil (giving diets high in saturated fatty acids, n-3 polyunsaturated fatty acids (PUFA) or n-6 PUFA, respectively) were fed to broilers. Feeding PUFA gave a significant reduction in the abdominal fat pad mass and an increase in feed efficiency of 5.4% for the fish oil and 7.7% for the sunflower oil fed broilers compared to tallow feeding. Furthermore, the results from this earlier project also indicated that dietary n-3 and n-6 fatty acids are assimilated into the phospholipids of all cell membranes with subsequent changes in the pattern of energy utilisation of broiler chickens, resulting in increased lipid oxidation and decreased glucose oxidation. This fatty acid-induced alteration in energy utilisation was proposed as a mechanism by which decreased lipogenesis could increase both protein synthesis and improve nutrient utilisation. The current project was initiated to build upon this information in order to develop a simple dietary strategy for reducing fat deposition and increasing muscle protein accretion in broilers.

Research A series of studies were conducted at the Camden farms, University of Sydney, which determined both the feeding period and the level of inclusion of dietary n-3 and n-6 PUFA that were required to orchestrate desirable changes in carcass composition and broiler performance. In addition, studies also examined the effectiveness of other dietary fatty acid sources and the dietary n-3:n-6 PUFA and n-3:saturated fatty acid ratios that optimised broiler performance, enhanced breast muscle mass and reduced abdominal fat pad mass.

Outcomes Studies conducted in the course of this project identified a positive correlation between n-6 PUFA and protein accretion, n-6 PUFA and improved feed conversion and a positive correlation between n-3 PUFA and a reduction in fat deposition. Feeding broilers increasing levels of n-6 PUFA increases breast muscle mass in a dose dependent manner. This result is attributed to both an increase in the n-6 fatty acid subtype, linoleic acid, and the reduction in the n-3 fatty acid subtypes, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. In contrast, feeding broilers increasing levels of n-3 PUFA results in a linear decrease in fat deposition. This outcome is attributed to the presence of the n-3 PUFA EPA and DHA. Further, these studies have shown that to achieve significant responses in both carcass composition and feed efficiency, birds need to be fed these fatty acid subtypes for a period of six weeks. Furthermore, other dietary n-3 and n-6 fatty acids sources can be


15

substituted for both fish oil and sunflower oil to achieve these outcomes. Studies undertaken demonstrate that linseed oil (n-3 PUFA) can be substituted for fish oil and that canola oil (n-6 PUFA) can be substituted for sunflower oil, as these n-3 and n-6 fatty acid sources gave similar production responses to those of fish and sunflower oil. In addition, feeding broilers pelleted diets containing a fatty acid ratios of 3% n-3 PUFA and 3% n-6 PUFA reduces carcass fatness and maintains breast muscle mass.

Implications The implications of this research for the selective use of dietary n-3 and n-6 fatty acids on broiler production are significant. Incorporation of either n-3 PUFA or n-6 PUFA into broiler diets will selectively produce birds with specific carcass qualities ie one with less fat or one with a greater muscle mass. In addition, improvements in nutrient utilisation can be achieved by selective incorporation of n-6 PUFA into broiler diets.

Publications Newman R.E., Wilkinson S.J., Downing J.A. and Bryden, W.L. (2002) N-3 and n-6 fatty acids: dietary levels, growth performance and carcass composition of broilers Proceedings of the Australian Poultry Science Symposium 14: 103.


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Food Safety Project Title:

Growth rate of broiler chickens given condensed tannins extracted from grape seed

RIRDC Project No.:

UA-61A

Researcher: Dr. Bob Hughes, Assoc. Prof. John Brooker and Dr. Christopher Smyl Organisation: Livestock Systems Alliance

The University of Adelaide and SARDI Pig and Poultry Production Institute Roseworthy Campus ROSEWORTHY SA 5371


• To determine whether condensed tannins extracted from grape seed retard the

growth rate of broiler chickens.

Background Food safety, disease control and the search for alternatives to antibiotics are major issues facing the chicken meat industries worldwide. Condensed tannins are secondary plant products that are known to bind to proteins and inhibit microbial function, possibly by preventing attachment of microorganisms to the gut lining. Tannins have also been demonstrated to have anthelmintic properties. A hypothesis arising from these studies is that tannins, if incorporated into the feed of chickens, may inhibit attachment of microbial pathogens in the gut and reducing bacterial growth without adversely effecting bird growth rates. This would reduce or eliminate pathogen colonisation of broiler chickens, and reduce the spread of pathogens that can occur through faecal contamination of feed and water.

Research A simple feeding trial was conducted with broiler chickens to determine whether they would grow at a rate close to their genetic potential when fed a nutritionally adequate diet containing tannins extracted from grape seed.

Outcomes Growth rate, feed efficiency and mortality were not significantly affected by inclusion of up to 10 g/kg of active ingredient (condensed tannins extracted from grape seed) or by the dietary addition of antibiotic (Tylosin) in the positive control diet. The reduced feed intake, without any change in feed efficiency, recorded for chickens given the diet with the highest concentration of condensed tannins, indicates that depression of growth rate was due mainly to reduced feed intake rather than to more subtle effects on digestion and absorption of essential nutrients. There was no difference between the negative and positive control groups, suggesting that the dietary addition of antibiotic product provided no detectable advantage in this particular experiment. Any antimicrobial effects of condensed tannins were unlikely to be demonstrable under these benign conditions. On the other hand, dietary condensed tannin concentrations of 10 g/kg or less were not detrimental to growth and feed efficiency. The results of this preliminary investigation tend to suggest that tannin extract has some potential as a non-antibiotic growth promotant in commercial broiler feed.

Implications Grape seed from the wine industry is currently a waste product and can be difficult to dispose of in an environmentally friendly manner. If tannins from this material are suitable for feeding to chickens then two major industries stand to benefit from this type of research. Antibacterial and anthelmintic properties of grape seed extract in dietary concentrations less than 30 g/kg active ingredient need to be evaluated more


17

fully. Future studies should determine the nature and extent of specific components of condensed tannins with potential antimicrobial properties.

Publications Hughes, R.J., Brooker, J.D. and Smyl, C. (2005) Growth rate of broiler chickens given condensed tannins extracted from grape seed. Australian Poultry Science Symposium, 17, in press.


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Environmental Management

Project Title:

Meat chicken EMS: transfer to industry

RIRDC Project No.:

FSE-2A

Researcher: Mr. Eugene McGahan Organisation: FSA Environmental

National Centre for Engineering in Agriculture (NCEA) PO Box 2175 TOOWOOMBA QLD 4350


• To develop a practical, competency based training and assessment workshop package in “Environmental Management Planning and Training for Meat Chicken Producers”.

• To recommend methods for facilitating on-site audit processes for meat chicken farms.

Background Recently completed RIRDC Meat Chicken Program projects produced valuable

information for producers on environmental issues relevant to their industry. There is now a need to take the information gathered in these recently completed projects and deliver it to producers through training and assessment.

Research The first step undertaken as part of this project was to investigate, plan and develop a training package which covers general environmental issues associated with chicken meat production, environmental management plans, environmental risk assessment and any monitoring, interpreting and reporting requirements. The package that was developed was trialled with producers and identified facilitators from at least three states. Workbook “master copies” for facilitators and producers were then edited and refined. Recommends were also developed with regards to best-bet methods for facilitating on-site audit processes for meat chicken farms.

Outcomes A training package was developed that will assist meat chicken producers by increasing their knowledge of environmental issues, improving their ability to minimise or manage environmental impacts, enhancing their ability to interpret basic environmental monitoring results and enabling them to produce a meaningful draft environmental management plan.

Implications The training package provides an excellent structure for developing farm-based environmental management systems. Resources incorporated in the training package in that has been prepared are a Facilitators Training Manual, Participants Training Manual, Example Environmental Management Plan, and the national Environmental Mangement Plan. By creating draft environmental management plans, producers are made aware of industry and site specific environmental issues. Based on the resources prepared during the project, the training package should now be implemented throughout the meat chicken industry.


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Project Title:

Sustainability improvements in the Victorian meat chicken industry (phase 2)

RIRDC Project No.:

JSC-2A

Researcher: Mr. James Smith Organisation: James Smith Consulting Pty Ltd

55 Sims Street SANDRINGHAM VIC 3191


• To achieve broader uptake of Chicken Care best practices by Victorian broiler

growers. • To identify the extent of Chicken Care implementation by growers and address

any barriers to its further adoption. • To identify and provide improvements to Chicken Care information transfer,

processes and tools as required by the community, regulators and growers. • To incorporate the RIRDC National Environment Management System, the

national Biosecurity Manual requirements and Animal Welfare Audit programs into the Chicken Care best practices model.

Background The industry Chicken Care initiative was launched in Victoria in 2000 in order to

improve its environmental performance and to address broad community concerns. This was in response to the increased broiler farm performance standards expected by the community and reflected in tough prescriptive regulatory controls adopted in 2001 in the State. Good initial understanding of and participation in the initiative by growers (40%) was achieved by early 2003 and the program was reconfirmed as needed, effective and valuable by its industry steering committee. However, it was felt that broader uptake of the program would be beneficial and should be sought.

Research Research was undertaken to survey the extent of implementation of the Chicken Care best practices by approximately 50 growers and to analyse areas of common needs. A process for external verification of grower self-assessment was investigated, developed and trialled. The existing Chicken Care best practices model was reviewed and updated to reflect recently adopted national industry codes and State regulations. At least twelve grower training workshops, industry briefings and community meetings were held as a means to identify barriers and improvements and to brief growers on new guidance materials developed and adopted.

Outcomes A total of thirty barriers to adoption of the Chicken Care best practices were identified. Changes addressing seven of them were incorporated into the program and ten will be recommended to the program steering committee as areas of priority future work. The Chicken Care best practices model was updated with 200 changes made to fully align it with the RIRDC’s Environmental Management code, the RIRDC Animal Welfare audit program, the requirements of the industry’s Biosecurity Manual and with format improvements suggested by growers and the community. Four new guidance notes covering Landscaping, Broiler Farm OHS Hazards, Regular Maintenance and Farm Training and Skills required were also developed. Fifty-five growers were trained in four Chicken Care workshops on the updated model. Grower implementation of the Chicken Care best practices was increased to 115 farms (60% of the Victorian industry) by this training, by appointment of a Chicken Care officer in each grower branch and by thirteen other arranged briefings for growers during the project.


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An external verification process was developed and trialled at five farms. It confirmed that 96% of grower self-assessments were accurate or understated. This added credibility to grower survey results that indicated that implementation of best practices by participating growers had risen by 11% (now up to 69% of the practices overall) in the two years since the earlier (end-2001) survey. Several public acknowledgements by regulators and the Community Advisory Panel that the environmental performance and cooperative attitudes of the industry had improved were identified and documented.

Implications Community and regulator comments, changes made to the Chicken Care program and surveys of growers confirm that the target of near-full participation and implementation by end-2005 (five years after program launch) as adopted by the industry steering committee is achievable. The remaining twenty potential improvements to the program to assist its adoption should be considered by the Steering Committee for incorporation in future years. Chicken Care processes and outcomes should be an integral part of the rationale and performance assurance cited by industry in support of its advocacy aimed at resolving anomalies in the Victorian rural odour standard, the code for broiler farms and other planning regulations.

Publications Smith, J. and Durkin, P. (2002) Workshop Outcomes Summary Proceedings of EPA Victoria, VFF and VCMC Odour Controls Workshop.

Smith, J. (2002) Chicken Care – Improving Sustainability in the Victorian Chicken Meat Industry. Proceedings of the 12th Australian Poultry and Feed Convention pp 517-523.

Smith, J. (2003) Sustainability improvements in the Victorian chicken meat industry (phase 1) RIRDC Publication No 03/035

Smith, J. (2004) Chicken Care Implementation Workshop VFF Manual.

CHICKEN MEAT PROGRAM – RESEARCH IN PROGRESS 21

RESEARCH IN PROGRESS

Flock Health

Project Title

Postgraduate scholarship - Scott Sheedy: Live vectoring of therapeutic and prophylactic proteins and pathogenesis in necrotic enteritis

RIRDC Project No.:

CSA-20A

Start Date: 01/03/2002 Finish Date: 28/02/2005 Researcher: Dr. Robert Moore Organisation: CSIRO Livestock Industries

Private Bag 24 GEELONG VIC 3220


• To use live bacterial vectors to deliver therapeutic proteins (eg. cytokines;

vaccine antigens) to the gut of chickens. Such delivery may have applications in the treatment of gut diseases such as necrotic enteritis.

• To investigate the importance of the alpha toxin produced by Clostridium perfringens in the pathogenesis of necrotic enteritis.

Current Progress Earlier experiments identified three marked E. coli strains that were able to

persist in the gastro-intestinal tract of chickens for up to 42 days. A trial is currently underway studying the in vivo stability of plasmids incorporated into these persisting strains, and how they compete against other non-persisting strains. Furthermore, this trial investigates the proportion of marked strains against the total resident E. coli population in each bird, where they reside, and colony numbers throughout the various regions of the gut. In all trials to date, inoculation of chickens with these strains has not caused any signs of ill-health in the birds. Cytokines such as IL-6 and IFN-γ, in expression plasmids, will be transformed into these strains. A particular bacteriocin that has specific activity against Clostridium perfringens is currently being tested for activity in the E. coli strains, and will then be ready for delivery. The project will focus on the delivery of biological agents from the above using the bacterial vectors to help control necrotic enteritis in chickens. Delivery of an attenuated alpha-toxin will investigate if the alpha-toxin from Clostridium perfringens is important in disease pathogenesis of necrotic enteritis in chickens.


Project Title

Postgraduate scholarship - Manija Asif: Cytokines and innate molecules for enhanced mucosal immunity in the chicken

RIRDC Project No.:

CSA-21J

Start Date: 01/04/2002 Finish Date: 31/03/2005 Researcher: Dr. Andrew Bean Organisation: CSIRO Livestock Industries



• To reduce disease by enhancing mucosal immunity in chickens with the aid

of cytokine therapy.

Current Progress Many livestock diseases take place at the mucosa, as these surfaces are in intimate contact with the external environment. There is an increasing need for non-viable vaccines that are capable of promoting effective mucosal and systemic immunity to pathogens. Oral vaccination with protein antigen alone generally induces poor immunity; therefore, the development of a potent mucosal adjuvant is essential. Adjuvants such as cytokines have the potential to mediate immune responses and offer an alternative to currently used adjuvants. Newly discovered chicken cytokines such as chicken IL-6 (ChIL-6) have the potential to control the outcome of mucosal responses. This highlights the potential for the use of cytokines as adjuvants to augment mucosal immunity. To date, ChIL-6 has been expressed in both prokaryotic and eukaryotic expression systems and the recombinant protein has been shown to be biologically active. To investigate the immunological role of ChIL-6 in chickens, anti-ChIL-6 monoclonal antibodies have been established to allow the development of an ELISA and have been used as a tool to detect ChIL-6. In vivo trials involving administration of recombinant ChIL-6 protein in chickens have been carried out to elucidate its effect on immune response. Currently, the role of ChIL-6 during infectious bronchitis (IB), a respiratory disease of chickens, is being investigated.


Project Title

Rapid identification and pathotyping of virulent IBDV, NDV and AI isolates

RIRDC Project No.:

CSA-24J

Start Date: 01/07/2002 Finish Date: 30/11/2005 Researcher: Dr. Hans Heine Organisation: CSIRO Livestock Industries



• To provide fast, sensitive and specific molecular diagnostic tests for the

identification of very virulent isolates of infectious bursal disease virus (IBDV), highly pathogenic Newcastle disease virus (NDV) and avian influenza virus (AIV), by developing and implementing new real-time PCR assays (Sequence Detection System based on TaqMan) that will give reliable results in less than five hours.

• To develop a common assay format, enabling simultaneous testing for IBDV, NDV and AI as well as the differentiation of very virulent/highly pathogenic strains from circulating endemic non/low-pathogenic strains.

• To implement the new tests at the Australian Animal Health Laboratory (AAHL) to ISO17025 standards by June 2004.

Current Progress Very virulent IBDV, virulent NDV and highly pathogenic AI virus strains are a

major threat to the Australian poultry Industry and have enormous trade implications. The rapid diagnosis of index cases following the incursion or outbreak of any of these viruses is crucial to support the timely activation of disease control measures. Fast and accurate diagnostic methods are also essential for epidemiological surveys to confirm freedom from disease post outbreak, in order to lift trade restrictions and restore consumer confidence. Rapid and highly sensitive real-time PCR tests (TaqMan based Sequence Detection System) have been developed for detection of IBDV, NDV and AIV and to discriminate very virulent IBDV from classical strains and NDV V4 vaccine from field strains. The development of further type or subtype specific tests is in progress. These tests have been evaluated with a number of representative strains and have the capacity to detect and differentiate important pathogens under uniform reaction conditions that allow multiplex assays which provide results in less than five hours. The assays will be further validated on clinical material from experimentally infected chickens and the assay procedures implemented to ISO 17025 standard to facilitate the transfer of these tests to other laboratories or to industry if needed.


Project Title

Postgraduate scholarship - Kristie Jenkins: Improved therapeutics for Marek's disease virus infection

RIRDC Project No.:

CSA-25J




• To examine the early immune response to Marek's disease virus infection

and apply this information to enhance the development of therapeutic strategies.

Current Progress The continual evolution of hyper-virulent Marek’s disease virus (MDV) is

considered to pose a major future threat to the Australian poultry industries. Understanding the interactions of this herpes virus and the host immune system is fundamental to understanding the disease mechanisms. In additional, utilising and augmenting the immune response may provide protection to this economically significant disease. This project will focus on assessing the early responses to MDV infection, focusing on innate immunity. Work has been carried out to try and identify which Toll-like receptors may be involved in the innate response to MDV. Characterisation of these cell surface receptors may help in targeting vaccine adjuvants to improve control of MDV infection. Additionally, preliminary work has been performed to identify innate immune molecules such as specific cytokines that are produced during MDV infection as these may have potential as adjuvants in MDV vaccines.


Project Title

Use of cytokines to enhance vaccine efficacy in poultry

RIRDC Project No.:

CSA-26J




• To develop novel vaccine formulations and therapeutics that will enhance

the efficacy of vaccines currently used to control Marek's disease in poultry. This will involve the evaluation of various cytokines and an assessment of their ability to enhance vaccine efficacy and immune competence.

Current Progress Marek’s disease (MD) is an economically important disease in chickens caused by MD herpes virus (MDV). The disease is characterised by early

immunosuppression and development of T lymphomas. However, resistance to disease can be acquired after vaccination. The mechanisms underlying resistance to MDV are not understood and little is known about the responses involved in disease control. Understanding the interactions of this virus and the host immune system is fundamental to understanding the disease mechanisms. Currently, chickens are protected from MD by vaccination with multiple nononcogenic serotypes of MDV-related strains, but recently, increased vaccination breaks have been observed. It may be expected that this could continue and that the current vaccine strains may become less able to protect chickens against MD, thereby posing a threat to the Australian poultry industries. The continuing emergence of new hypervirulent strains of MDV

necessitates improving vaccination efficiency and the development of new prophylactic strategies. Many studies have identified cell-mediated immunity as important in early protective mechanisms against viral replication. With this in mind, work has been carried out to identify immune molecules in chickens that are of importance during MDV infection. Additionally, utilising and augmenting these molecules and the associated immune response may provide alternative strategies for the protection of Australian chicken flocks against this economically significant disease.


Project Title

Biological control of necrotic enteritis in meat chickens

RIRDC Project No.:

CSA-29A

Start Date: 08/07/2003 Finish Date: 07/07/2004 Researcher: Dr. Robert Moore Organisation: CSIRO Livestock Industries



• To complete the development of a reproducible necrotic enteritis (NE)

disease model that can be implemented in PC2 containment conditions, thus allowing the testing of disease treatments, such as bacteriocin products and recombinant vaccines developed in previous projects.

Current Progress Six trials have been completed as part of the development of a necrotic enteritis

disease induction model. During the course of the trials, a number of factors potentially affecting the course of disease development have been evaluated. A series of local field isolates of Clostridium perfringens have been screened for their ability to induce disease in birds and a number of pathogenic strains have been identified. It has been found that the exact composition of the media in which the C. perfringens is grown before infecting the birds has a large impact on the degree of disease subsequently seen. Other key aspects of the current disease model are (a) the use of a high protein ration, (b) withdrawal of feed during the bacterial challenge, and (c) the use of multiple bacterial doses during the challenge. Housing of the birds at a lower than normal temperature may marginally aggravate the disease. In this laboratory, a light challenge with a mixture of Eimeria strains did little to exacerbate disease. In trials using the current optimal challenge method, 50 % of the challenged birds are consistently being found to have some sign of necrotic enteritis, ranging from single small lesions to extensive patches of necrotic tissue.


Project Title

Characterisation and modulation of virulence of endemic IBDV strains using reverse genetics

RIRDC Project No.:

CSA-30J

Start Date: 01-Jul-03 Finish Date: 31-Jul-06 Researcher: Dr. Sandra Sapats Organisation: CSIRO Livestock Industries

Private Bag No 24 GEELONG VIC 3220


• To monitor genetic changes in circulating IBDV strains (Molecular

Epidemiology) in order to determine if field strains are changing significantly.

• To optimise the already developed Reverse Genetic system for IBDV for the genetic modification and the molecular characterisation of biological properties of IBDV.

• To apply the Reverse Genetics system for the construction of recombinant IBDV to ascertain the importance of specific mutations in VP2 gene related to virulence and antigenicity and in particular how these effect virulence of endemic strains.

Current Progress Samples collected in 2003 from seven broiler farms in NSW were tested in

ELISA for the presence of IBDV antigen. All seven farms tested positive for IBDV, with broilers ranging in age from 21-35 days of age. Representative isolates from each of the seven farms were sequenced and their deduced amino acid sequences compared to previously isolated strains. All seven isolates were genetically similar to Australian IBDV vaccine strains. A number of unique amino acid substitutions were identified in four isolates, however none of these amino acid substitutions have been associated with increased virulence of IBDV. IBDV contains an RNA genome consisting of two segments, "Annie Robertson" <[email protected]> A and B. In order to manipulate the viral sequence a DNA copy of the genome must first be cloned. Clones containing DNA copies of segments A and B of Australian IBDV strain 002/73 were previously generated, however these clones failed to produce infectious virus. Nucleotide sequencing identified three incorrect amino acid substitutions within segment B. These three incorrect amino acid substitutions have now been corrected by PCR and the nucleotide sequence confirmed. The system is currently being optimized to facilitate the production of viral RNA which is identical to that produced in the native virus.


Project Title

New diagnostic assays to improve control of coccidiosis in poultry

RIRDC Project No.:

DAQ-316A

Start Date: 01/01/2004 Finish Date: 31/12/2006 Researcher: Dr. Glenn Anderson Organisation: Department of Primary Industries & Fisheries (Qld)

Animal Research Institute Locked Mail Bag No. 4 MOOROOKA QLD 4105


• To develop and validate new DNA-based quantitative assays for the

diagnosis of poultry coccidiosis. To develop and validate new serological assays for the diagnosis of poultry coccidiosis.

Current Progress PCR primers and real-time PCR probes have been designed for a region of

small subunit ribosomal DNA that exhibits consistent interspecific variation for the seven species of Eimeria in chickens. Evaluation of the primers to ensure they work for a range of different strains within each species is underway. A bank of pre and post inoculation reference sera has been assembled from chickens experimentally infected with each of the seven species of Eimeria. Methods have been developed for preparing parasite antigens from gut scrapings and egg culture. The two sources of antigen contain a different range of contaminant proteins, allowing the development of mouse immunization protocols that maximize the immune response to parasite-specific antigens and enhance the prospects for obtaining useful monoclonal antibodies (Mabs). A high throughput western blot method suitable for screening hybridoma supernatants for the presence of parasite-specific Mabs has been developed. One antigen that is apparently unique to E. tenella and a second antigen that is apparently common to all species of Eimeria have been identified. Mice have been inoculated with whole antigen preparations of either the E. tenella–specific antigen or the common antigen, and Western blots have shown that all mice are producing high titre antibodies.


Project Title

Preliminary investigations into the efficacy of novel treatments and application methods for darkling beetle (Alphitobius diaperinus [Panzer]) control in Australian broiler houses

RIRDC Project No.:

DAQ-319A

Start Date: 01/07/2003 Finish Date: 31/12/2004 Researcher: Mr. Trevor Lambkin Organisation: Department of Primary Industries & Fisheries (Qld)

Entomology Building, Indooroopilly Research Centre 80 Meiers Road INDOOROOPILLY QLD 4068


• To undertake preliminary, mainly laboratory based investigations into the

efficacy of a number of novel litter treatments in controlling darkling beetle (Alphitobius diaperinus).

Current Progress Previous research has identified where control strategies should be targeted for the management of A. diaperinus in Australian broiler houses. Results from a range of investigations conducted in previous RIRDC funded research projects on darkling beetle control, indicated that the highest populations of beetles and their immature stages occur in the litter of broiler houses, and predominately within the first three weeks of each batch. Results from this research also strongly suggested that floor applications of insecticides, in particular pyrethroids, do little in controlling pest numbers. Therefore, a number of novel compounds were trialed as litter treatments in the laboratory. To date, four different formulations have been trialed: (a) imidacloprid, a Bayer® product (liquid [SC 20%] and bait granule applications), (b) beta cyfluthrin, a Bayer® product (liquid [SC 2.5%] application), (c) spinosad, an Elanco® product (liquid [25g/L] application), and (d) a Queensland diatomaceous earth product (powder application), from Grow Choice® and Grose “N” Grows Silica Products®. The most effective formulation in reducing progeny in trials undertaken to date has been the imidacloprid bait granule. It was quite surprising how little of this compound was required in laboratory test boxes to produce a very high level of progeny reduction, while imidacloprid SC 20% and beta cyfluthrin SC 2.5% did not give as high progeny reductions. The application of diatomaceous earth to litter in the laboratory also gave very good progeny reductions. Preliminary laboratory results using spinosad found it to be ineffective when trialled this way, so field trials will now be undertaken to further evaluate the efficacy of this compound. These trials are planned to commence in July 2004.


Project Title

Advanced clinical diagnostics: use of real-time immuno-PCR and LightUp probes

RIRDC Project No.:

DAW-113A

Start Date: 01/02/04 Finish Date: 30/12/2004 Researcher: Dr. Debby Cousins Organisation: Department of Agriculture (WA)

Animal Health Laboratories Locked Bag 4 BENTLEY DELIVERY CENTRE WA 6983


• To evaluate new molecular biology protocols for use as a variety of real-

time and 'on site' diagnostic tests. • To gain an understanding of the science involved in the production of

LightUp probes and the use of real-time immuno-PCR. • To conduct preliminary studies into the production of LightUp probes and

their use in routine diagnosis. • To assess the feasibility and availability of this technology in Australia .

Current Progress Researcher Carmel Hancock has been resident at Chalmers University, Gotenburg, Sweden, since 19 March 2004 where she has been developing an understanding of the technologies (conjugated peptide nucleic acid RT PCR and immunoPCR) developed by Prof Mikael Kubista and his group as a means of determining the feasibility of using such techniques for use in improved diagnostic tests for the chicken meat industry. Training has been received in the design of probes, quantification and RT-PCR methods and experience gained in the use of various types of real time equipment (Lightcycler, Corbett Rotorgene and the iCycler). Optimisation of the Campylobacter tests developed has been progressing, with some specificity issues still to finalise. Once optimisation is complete, the synthesized Lightup probes will be applied to samples of DNA prepared in Perth prior to travel. Opportunities for multiplexing (testing for different organisms in a single test using different dyes for each probe) appear promising, since the same programs can be used for detection of C. jejuni and C. coli. Experience in ImmunoPCR will be gained during the month of June. An industry survey to gain feedback on opportunities for diagnostic technologies was designed and sent to key industry contact in February. Results from this survey are being collated.


Project Title

Molecular evaluation of responses to vaccination and challenge by Marek's disease viruses

RIRDC Project No.:

RMI-12J

Start Date: 19/12/2002 Finish Date: 30/12/2005 Researcher: Prof. Greg Tannock Organisation: Royal Melbourne Institute of Technology

Department of Biotechnology and Environmental Biology PO Box 71 BUNDOORA VIC 3083


• To develop and refine a QPCR test that will be suitable for estimating

Marek's disease virus DNA copy number in cell cultures infected with virulent and vaccine viruses.

• To extend the use of QPCR to measure viral loads in various organs of infected chickens after infection with virulent and vaccine viruses.

• To determine similar loads in organs of the developing chick embryo in an attempt to determine alternative markers for virulence that may not involve inoculation of chickens.

• To measure the distribution of serotype 1 and -3 vaccine viruses after inoculation to chick embryos under conditions used for in ovo vaccination.

• To carry out protection experiments after in ovo and day 1 vaccination in which the endpoints will be measured from the challenge virus load in critical target organs.

Current Progress Over the past year, tests for estimating virus loads by serotype 1 and 3 strains of

Marek’s by quantitative polymerase chain reaction (QPCR) have been fully standardised using specific PCR products cloned into vectors as internal standards. The standardized test has been used to measure the distribution of serotype 3 herpes virus of turkeys (HVT) in various organs of infected chick embryos. Results have shown that the highest number was obtained in the spleen. Copy number at day 20 was measured in embryos inoculated at different times of embryogenesis. Yields in embryos inoculated at days 6 and 11 were significantly higher than in embryos inoculated at day 17. However, significant mortality occurred in embryos inoculated at day 6 and, therefore, day 11 may be a more satisfactory age for in ovo vaccination. The growth of HVT in the organs of chickens was measured by QPCR between one and eight weeks after infection with HVT. Highest yields were obtained in the spleen and lower yields were present in the kidney, liver and thymus and brain. Peak yields in the brain were present one to two weeks after infection; peaks for the kidney, liver and thymus and heart occurred after the peak was observed in the spleen. The lowest copy number for HVT was observed in the thymus. It is concluded that the spleen is the key target organ for measuring the presence of HVT in vaccinated birds.


Project Title

Molecular techniques for monitoring Marek's viraemias in broilers and layers

RIRDC Project No.:

UJC-10J

Start Date: 01/07/2002 Finish Date: 31/07/2005 Researcher: Dr. Graham Burgess Organisation: James Cook University

Department of Microbiology and Immunology TOWNSVILLE QLD 4811


• To develop and transfer to the industry molecular techniques for monitoring

blood samples for the presence of Marek's disease virus (MDV); • To use a panel of simple, cost-effective molecular diagnostic techniques to

describe the effects of various management strategies on Marek's infection; • To develop and validate quantitative assays for Marek's disease viral DNA

based on real-time PCR; • To assemble a comprehensive collection of DNA for Australian wild type

Marek's disease viruses.

Current Progress Due to inconsistencies in the results obtained from filter paper testing, one new material (glass fibre sheet) and method (treated capillary tubes) are being trialed for adaptation to quantitative PCR. PCR testing of unvaccinated broiler flocks indicated high levels of MDV in feather follicle samples. Following the initiation of a HVT vaccination program, subsequent broiler flocks were routinely tested over a six month period and no type 1 MDV was found during the vaccination period. Following the six month vaccination trial, the use of HVT vaccine was discontinued and MDV Type-1infection returned to high levels (>10% of birds tested) within two batches. A small number of samples from unvaccinated birds reacted in the type 2 PCR, indicating the presence of type 2 MDV in commercial flocks. The laboratory has recently taken delivery of a Corbett Rotorgene 3000 for real time quantitative PCR and all diagnostic assays are currently being converted to suit this amplification platform. SYBR Green assays for serotype 1 MDV have now been established in quantitative and end point format. A genotyping assay has been successfully trialed on Australian cell culture and field strains in parallel with samples from CVI 988. This assay is capable of differentiating between CVI 988 and other strains at very low levels of template.


Project Title

Avian leukosis subgroup-J (ALV-J): epidemiological studies for control of ALV-J in Australian broiler breeder flocks

RIRDC Project No.:

UM-68A

Start Date: 01/07/2003 Finish Date: 31/07/2005 Researcher: Dr. Trevor Bagust Organisation: The University of Melbourne

Faculty of Veterinary Science, Building 404 Cnr Park Drive & Flemington Road PARKVILLE VIC 3010


• To undertake a structured epidemiological survey to determine the current

prevalence and distribution of ALV-J in Australian broiler breeder stocks at Great Grandparent (GGP) and GP levels.

• To undertake molecular characterisation and comparative pathotyping of recent Australian strains of ALV-J isolated from large commercial poultry breeding organisations.

• To provide the tools that would allow industry to control ALV-J in Australian broiler breeder flocks.

Current Progress Avian leucosis virus subgroup-J (ALV-J), which produces tumours and

decreased production in broiler breeders, was inadvertently imported into Australia with genetic stocks in the last decade. Using the PCR identification technology developed at these laboratories, an epidemiological survey is in progress to determine the current prevalence and distribution of ALV-J in Australia. During July 2003 to January 2004, some 3364 samples from five states have been screened. Only three isolates of ALV-J have been obtained in the present cycle of sampling, and these were from the one parent breeder flock sampled in June-2003 and then again in August-2003. While widespread sampling has revealed largely negative results for ALV-J, a significant number (37) of ALV-A isolations have been obtained and confirmed. The number of ALV-J isolates being made is heavily reduced compared to previous work (RIRDC project UM-49A) and appears to indicate that the prevalence of ALV-J is successfully being reduced as known positive flocks are eliminated by the commercial poultry industry. Targeted sampling of Australian great grandparent, grandparent and parent flocks will be continues over the next twelve months in order to definitively establish freedom from ALV-J. Scientific liaison with commercial collaborators is continuing in order to also test newly imported broiler stocks while they are being held in quarantine. Samples from this genetic stock are tested for both ALV-J and ALV-A prior to release into the Australian industry. Some further insights may also be obtained from planned future work into the undesirable ongoing circulation of ALV-A in Australian chicken flocks.


Project Title

Systematic pathotyping of Australian Marek's disease (MDV) isolates

RIRDC Project No.:

UNE-83J

Start Date: 01/07/2002 Finish Date: 30/11/2005 Researcher: A/Prof. Stephen Walkden-Brown Organisation: University of New England

Animal Science School of Rural Science and Natural Resources ARMIDALE NSW 2351


• To pathotype nine current and three older Australian isolates of Marek's

disease virus (MDV) using internationally recognised protocols. • To undertake molecular characterisation of these isolates to facilitate

identification and relationships between isolates. • To test the pathogenicity of two isolates in current Australian strains of meat

and layer bird. • To determine the extent of protection provided by HVT and bivalent

serotype 2/HVT vaccines against recent isolates. • To determine whether recent isolates are more pathogenic than older

isolates. • To develop an improved knowledge base for rational decision making

regarding MDV vaccines and vaccination procedures.

Current Progress Difficulties with the isolation, decontamination and bulking up of new isolates of Marek’s disease virus (MDV) have been experienced on this project over the past twelve months. Contamination of samples with vaccinal MDV strains and the failure of isolates to grow strongly in chicken fibroblast and chicken embryo kidney cell culture have been the main problems encountered. The first pathotyping experiment, conducted in August/September 2003, proved disappointing, with no evidence of infection with the sole new MDV isolate tested, and very low levels of infection and pathogenicity in the reference MDV strain. In response to these problems, and in close consultation with RIRDC and AECL, significant efforts have been expended on improving virus isolation and bulking up procedures. Chick kidney cells from two week-old SPF chicks have proved to be the best culture medium for growing MDV and it is planned that MDV isolates will be grown and titrated in this medium from now on. In order to limit contamination of isolates with vaccinal strains and other viruses, it is intended that MDV will be isolated from feather tip or dander material, initially in SPF chickens. Only MDV isolates that cause tumours or significant pathology in these birds will be further developed for use in pathotyping experiments. The first attempt at this approach will commence on June 8, 2004 with six positive dander samples from the field under test.


Project Title

Typing of Pasteurella multocida

RIRDC Project No.:

UQ-100J

Start Date: 01/01/2002 Finish Date: 31/12/2004 Researcher: Prof. Linda Blackall Organisation: The University of Queensland

Department of Microbiology & Parasitology ST LUCIA QLD 4072


• To establish a Multi-locus Sequence Typing (MLST) system for

Pasteurella multocida. • To facilitate the rapid, accurate typing of P. multocida isolates that will

allow any isolate to be directly compared with any previous isolate already typed and allow an understanding of the epidemiology of fowl cholera outbreaks.

• To provide typing tools which will support improved prevention and control programs for fowl cholera.

Current Progress

The project has been undertaken as a Masters of Science project by Ms. Sounthi Subaaharan. Ms. Subaaharan is supervised by Prof. Linda Blackall (University of Queensland) and Dr.Pat Blackall (DPI&F). A literature review has been produced that outlines the broad details of this work and the relevant published literature. A total of 56 isolates of Pasteurella mulltocida have been revived and their identity confirmed by biochemical tests and a species-specific PCR. DNA was extracted from these isolates using a DNA extraction kit. A total of seven house-keeping enzymes were selected for this study based on a prior MLEE study. Great difficulty was experienced in matching the enzyme names with the names used by the researchers who annotated the genome. After considerable searching, five enzymes were confidently located on the genome. Primers were designed and amplification by these primers was proven successful. To date, DNA of 30 isolates has been amplified for all five enzymes and sequencing of the resultant PCR products completed. Currently, these sequences are being cleaned up and prepared for the MLST analysis.


Project Title

Efficacy trials of a maternally-delivered recombinant vaccine against coccidiosis

RIRDC Project No.:

UTS-4J

Start Date: 01/05/02 Finish Date: 30/04/04 Researcher: A/Prof. Nicholas Smith Organisation: University of Technology, Sydney

Institute for the Biotechnology of Infectious Diseases Westbourne Street GORE HILL NSW 2065


• To test the immunogenicity of recombinant versions of gametocyte antigens

of Eimeria maxima. • To test the efficacy of recombinant versions of Eimeria maxima gametocyte

antigens as a subunit, maternally-delivered vaccine against coccidiosis.

Current Progress

The genes encoding two immunodominant antigens of Eimeria maxima, gam56 and gam82, have been cloned into a bacterial expression vector and the proteins expressed and purified. Both recombinant proteins are recognised by protective chicken antibodies that were raised to the native 56 and 82 kDa gametocyte proteins, by immunoblotting. In a competitive ELISA, a combination of the recombinant proteins inhibits the binding of protective antibodies to the native proteins by 76%, which is comparable to the inhibition of 98% observed when the native antigens themselves are used as the competing protein in the assay. In two breeds of chicken (Australorp and Cobb500), the recombinant proteins alone, or in combination, elicited a dose-dependent, antibody response that recognises the native proteins by ELISA, and gametocytes by immunoblotting. Together, the results suggested that the development of a recombinant subunit vaccine that maintains the antigenic and immunogenic properties of the native protein vaccine is feasible. Efficacy trials of the recombinant proteins are now underway.


Project Title

Postgraduate scholarship - Ms Kelly Mai: The molecular basis for oocyst wall formation in the apicomplexan parasite, Eimeria maxima

RIRDC Project No.:

UTS-6A

Start Date: 01/03/2003 Finish Date: 28/02/2006 Researcher: A/Prof. Nicholas Smith Organisation: University of Technology, Sydney

Institute for the Biotechnology of Infectious Diseases Westbourne Street GORE HILL NSW 2065


• To identify and characterise structural proteins and enzymes involved in

oocyst wall formation in Eimeria maxima.

Current Progress Thus far, truncated recombinant versions of a 56kDa, tyrosine-rich, gametocyte protein of E.maxima have been produced, one truncated protein being rich in tyrosines and the other possessing only a small number of tyrosine residues in its sequence. The 56kDa protein is known to be processed by the parasite and incorporated into the oocyst wall, allowing it to harden and thereby protecting the parasite as it is passed in the chicken's faeces. The hypothesis being investigated is that the formation of strong bonds between tyrosine residues (ie dityrosine bonds) is critical for this process. This hypothesis has been tested this experimentally. Thus, in the presence of exogenous peroxidase (commercially available horseradish peroxidase) and t-butylhydroperoxide, the tyrosine-rich protein appears to polymerase into high molecular weight aggregates whereas the truncated protein that lacks many tyrosine residues polymerases to a much smaller degree. This indicates that peroxidase(s) may be critically involved in catalysing dityrosine bond formation in the oocyst wall of E. maxima. It will now be important to confirm these results and identify the endogenous peroxidase(s) responsible for cross-linking reactions.


Bird Nutrition and Feed Supply

Project Title

Premium grains for livestock program (stage 2)

RIRDC Project No.:

GRD-3J

Start Date: 01/07/00 Finish Date: 30/06/03 Researcher: Dr. John Black Organisation: John L Black Consulting

Locked Bag 21 WARRIMOO NSW 2774


• To develop rapid and objective analytical tests for assessing the quality of

feed grains. • To enhance grain nutritional value through breeding and processing. • To develop a model(s) to predict feed grain quality for certain livestock

species. • To provide an integrated and coordinated framework in which the above

objectives can be achieved.

Current Progress During the Premium Grains for livestock Program, 136 cereal grains have been fed to broiler chickens and 116 fed to laying hens. The grains, including wheat, barley, triticale, sorghum oats and rice, were selected for wide variation in energy availability within plant species. There were relatively small differences between Apparent Metabolisable Energy (AME) values for broilers and layers. AME values for layers tended to be higher for grains with high concentrations of non-starch polysaccharides and lipids. Rice had the highest AME which exceeded 17 MJ/kg DM, with little variation between samples. The second highest was for sorghum with a range in AME from 15.1 to 16.8 MJ/kg DM. AME values for wheat, barley and triticale ranges over 20-25% and differences exceeded 3.6 MJ/kg DM for triticale. Voluntary intake of all grain species examined except rice also varied widely, with up to 28% difference between samples within a grain species. Frost affected grains had significantly lower energy values than un-frosted samples, whereas germination increased AME for most grains. Extrusion of a barley sample reduced AME by almost 3 MJ/kg DM. Preliminary NIR calibrations indicate that both AME and intake for broilers can be predicted with acceptable accuracy. The Program has been extended for two years to fully exploit the results obtained and assist adoption of findings by industry.


Project Title

Postgraduate scholarship - Mr Nicholas Rodgers: Relationships between grain quality, intestinal integrity, and performance of broiler chickens

RIRDC Project No.:

UNE-86A

Start Date: 01/02/2003 Finish Date: 31/01/2006 Researcher: Prof. Mingan Choct Organisation: University of New England

School of Rural Science and Agriculture ARMIDALE NSW 2351


• To elucidate the mechanisms by which grain processing and feed

constituents affect gut physiology of birds from early life, which in turn affects life-long productivity.

Current Progress Two trials have been completed, and another is in progress. The first trial,

completed in late 2003, examined the effect of sorghum particle size on growth and performance of male and female broiler chickens in group-cages. The commercially formulated experimental diets were pelleted and contained sorghum (50%) of three different particle sizes (<0.5mm, 0.5mm to >2mm, and >2mm) prepared at UNE and one diet, which was obtained from a commercial feed manufacturer (~0.5mm to 1.6mm). Results indicate that the intermediate (0.5mm to >2mm) particle size increased growth relative to the fine (<0.5mm) particle size with no effect on feed efficiency. The second experiment (completed in early 2004) was conducted to study the effects of sorghum grain particle size, fed as mash, in a complex commercial diet (the same formulation used in experiment 1), on the performance and health of male broiler chickens to six week of age, under a Clostridium perfringens challenge, and housed in floor pens. Preliminary results show that an intermediate to coarse particle size is beneficial to growth and reduces the incidence of mortalities due to necrotic enteritis. An experiment is currently being undertaken in conjunction with the Australian Poultry CRC and the Agricultural University of Norway. The aim of the study is to determine the effects of sorghum particle size and milling method on male broiler growth, gastrointestinal development, duodenal particle size, digesta transit time, starch digestibility, metabolisable energy, and pancreatic enzyme activity. The experimental diets are based on sorghum (75% of the diet) and soy isolate, and include the following particle sizes (before conditioning and pelleting): (1) whole sorghum; (2) 3mm screen, hammermilled; (3) equivalent particle size to diet 2, using a rollermill; and (4) 1mm screen, hammermilled.


Project Title

Digestible amino acids and improved broiler performance

RIRDC Project No.:

UQ-107A

Start Date: 01/04/2003 Finish Date: 31/03/2006 Researcher: Prof. Wayne Bryden Organisation: The University of Queensland

School of Animal Studies GATTON QLD 4343


• To delineate the digestible amino acid supply from the grain portion of the

diet. • To estimate the ileal digestible amino acid requirements of broilers fed

wheat/sorghum based diets. • To estimate the availability and requirements of digestible amino acids such

as lysine and methionine for lean tissue deposition. • To improve broiler performance by integration of the above information into

feed formulation practices.

Current Progress An initial study examined the performance of male and female broilers fed diets formulated on a digestible amino acid basis throughout the growing cycle (1-42 days). Feed intake, weight gain, feed conversion efficiency and body protein proportions were significantly increased when diets were formulated on a digestible amino acid basis. There was a significant diet x sex interaction for weight gain which was due to a greater response in males to diets formulated using digestible amino acid values. Studies have commenced to examine the influence of different wheat cultivars and the protein content of wheat on the relationship to digestible amino acid values. These experiments are being run in parallel with studies that seek to determine the digestible amino acid requirements in the starter phase for lysine, methionine and threonine.


Project Title

Establishment of a Chair in Poultry Sciences at The University of Sydney

RIRDC Project No.:

US-127A

Start Date: 01/01/2004 Finish Date: 30/12/2007 Researcher: Prof. Tom Scott Organisation: The University of Sydney

Chair of Poultry Science Faculty of Veterinary Science PMB 3 University of Sydney Poultry Research Unit CAMDEN NSW 2570


• To establish a Chair in Poultry Science at the University of Sydney in order

to maintain a high level research capability for the industry at that facility. • To assist in the development of a centre of excellence in poultry science at

the University of Sydney and to develop a targeted research program in poultry nutrition, health and safety focused on the needs of the Australian poultry industries.

Current Progress Tom Scott assumed his new role as Chair of Poultry Science at the University of Sydney on July 14, 2003. Immediate challenges he faced were: to assume the role of chair and editor for the Australian Poultry Science Symposium and work with committee members to identify symposium themes and invited speakers; call for paper submissions, prepare and facilitate the 2004 meeting; develop the Poultry Production course for fourth year Animal Science (Faculty of Agriculture, Food and Natural Resources) students and specific poultry course work for other undergraduates; and to assume the responsibility of Director for the Poultry Research Foundation. Professor Scott has obtained university support to install 96 new bioassay cages to facilitate future broiler studies. Furthermore, a number of changes to infrastructure have also been made to develop the program’s ability to conduct experiments with poultry and facilitate a higher quality of worker safety and productivity. Applications for research support (from RIRDC, AECL, Autralian Poultry CRC) have been made in collaboration with other research associates and industry members. Currently, Professor Scott has three 4th year students conducting thesis projects in poultry, one of whom was awarded a CRC Honours scholarship. Postgraduate student involvement is being actively developed, but is dependent on industry and University support.


Project Title

Mechanical and enzymatic improvements of dehulled lupins for broiler and layer diets (AECL-managed project WAU-1A)

RIRDC Project No.:

UWA-76J

Start Date: 01/05/2003 Finish Date: 21/03/2005 Researcher: Dr. Ian Williams Organisation: University of Western Australia

Animal Biology 35 Stirling Highway CRAWLEY WA 6009


• To improve the nutritional value of whole and dehulled lupins so that they

can replace soybean meal in diets for broilers and layers, with major savings in feed costs.

• To destroy the thick cell walls and their main antinutritional factor, pectic substances, by expansion and enzymatic treatment.

• To increase the inclusion rates of whole and dehulled lupins up to 20% in broiler and layer diets by the above treatments without significant losses in productivity.

Current Progress There are several nutritional advantages of breaking down pectin in lupins.

First, viscosity of digesta can be reduced allowing a more complete digestion of nutrients. Second, more nutrients will be released from cells. Third, water-holding capacity will be reduced which may reduce wet droppings. Whole and dehulled lupins have been treated with a pectinase, polygalacturonase (PG), and fed to layers. Results obtained show that water intake and wet droppings decrease, metabolisable energy increases, and egg shells become thicker with pectinase treatment of lupins. PG only breaks down about 10% of the bonds between the galacturonic acid units because they are protected by methyl ester radicles. Another pectinase, pectin methyl esterase (PME), can remove these radicles allowing PG to break down much more of the pectin. Dehulled lupins were treated in vitro with a combination of PG + PME and it was found that 47% of the pectin was broken down, cell-walls were reduced by 27%, viscosity by 18%, and water-holding capacity by 15% compared with either enzyme used on its own. If this greatly improved breakdown of pectin can be demonstrated in vivo it may allow substantial improvements to be made in the nutritive value of lupins for poultry and in reducing the incidence of wet droppings in flocks fed on lupins.


Food Safety

Project Title

On-farm reduction strategies for Campylobacter spp.

RIRDC Project No.:

DAQ-282A

Start Date: 01/01/2002 Finish Date: 31/12/2004 Researcher: Ms. Jillian Templeton Organisation: Department of Primary Industries (Qld)

Agency for Food and Fibre Sciences Locked Mail Bag No 4 MOOROOKA QLD 4105


• To develop targeted control strategies, based on knowledge of sources of the

organism, to prevent the entry of Campylobacter spp. into broiler flocks.

Current Progress In-depth field and laboratory-based experiments have been completed to investigate if darkling beetles (Alphitobius diaperinus) play a role in the transmission of Campylobacter spp. in broiler flocks. Results from studies undertaken suggest that darkling beetles and larvae can carry Campylobacter spp., indicating that they are potential vectors. However, no evidence has been obtained from longitudinal field studies to indicate that they are the primary source of introduction of the organism to the broiler flocks. Experimental studies on naturally and artificially infected beetles confirm that, unless very short turnaround times are employed (e.g. <72 hours), darkling beetles colonised in one cycle are unlikely to still be colonised at placement of the next cycle of chickens. During 2004, the focus of the project has been on determining the appropriate level of biosecurity required on commercial broiler farms to prevent the entry and spread of Campylobacter spp..


Project Title

Development of a sequence-based bacteriophage typing system for Salmonella

RIRDC Project No.:

IMV-5A

Start Date: 01/07/2003 Finish Date: 01/07/2006 Researcher: Dr. Michael Heuzenroeder Organisation: Institute of Medical & Veterinary Science

Infectious Diseases Laboratories PO Box 14 Rundle Mall PO ADELAIDE SA 5000


• To continue to provide conventional typing (serotyping and phage typing) to

the industry on an ongoing basis for the purposes of organism tracing and the monitoring of Salmonella serovars for public health and research objectives.

• To use AFLP and PFGE typing (where appropriate) for typing industry Salmonella strains when conventional typing methods fail to offer sufficient discriminatory power.

• To establish an objective sequence-based molecular typing system that would enhance and extend the current classical phage typing system.

• To characterise genes in endogenous phages in Salmonella to determine the predicted impact of these genes on current typing methods.

Current Progress Specimens have continued to be received from the industry in comparable

numbers to previous years. In 2003, Salmonella Sofia remained the most common chicken isolate (39%), followed by serovar Typhimurium (30%). In humans, Typhimurium is the most common isolate (26.1%). Genomic regions of prophage genes found in the Salmonella chromosome are being targeted for MLST (Multilocus Sequence Typing) analysis. Salmonella housekeeping genes are also being tested for comparative purposes. Seventy-eight isolates of S. Typhimurium are currently being tested to evaluate MLST. Forty-one isolates include the closely related (by phenotypic lysis patterns) phage-types DT12, 108 and 170. The remaining isolates are DT126 isolates from two separate outbreaks as well as epidemiologically unrelated strains. Results show that S. Typhimurium isolates can be distinguished from each other based on the presence or absence of prophage genes in the chromosome as determined by PCR. This first level discrimination is equal to or exceeds pulsed-field gel electrophoresis (PFGE) the current ‘gold-standard’ for molecular typing. This result can be achieved in 24 hours, in contrast to PFGE, which can take up to five days. A higher level discrimination between individual isolates can be achieved if necessary by DNA sequencing of the prophage genes. These are more variable than Salmonella housekeeping genes, which are the usual targets for MLST for other species. These results indicate that the prophage genes are superior to housekeeping genes as a discriminatory tool for Salmonella isolates.


Project Title

Development and validation of Campylobacter microarrays for virulence detection and strain differentiation in poultry products

RIRDC Project No.:

RMI-14A

Start Date: 01/06/2002 Finish Date: 31/05/2005 Researcher: Prof. Peter Coloe Organisation: Royal Melbourne Institute of Technology

Department of Biotechnology & Environmental Biology PO Box 71 BUNDOORA VIC 3083


• To utilise existing knowledge on Campylobacter spp. to select the most

appropriate virulence factors for use in the development of a Campylobacter microarray.

• To design and select appropriate oligonucleotides for use in a Campylobacter array through the use of bacterial genomic techniques and alignment of gene sequences from known virulence factors to the Campylobacter genome.

• To investigate the design of microchips for the immobilisation of oligonucleotides and hybridsation conditions for simultaneous detection of a variety of selected virulence factors.

Current Progress An extensive search of international databases containing Campylobacter species sequences has been carried out and, based on this, virulence factors selected for the initial investigations. Five virulence factors (flagella, cytolethal distending toxin, capsule, sialic acid synthesis and adhesion and invasion) and one Campylobacter specific gene, the 16S rRNA, were selected and extensive gene alignments were performed enabling the design and synthesis of PCR oligonucleotides and molecular probes for microarray fabrication. The PCR oligonucleotides were tested on selected Campylobacter isolates from the RMIT Campylobacter culture collection and the resulting amplicons sequenced and realigned against known genomic sequences, such as the sequenced C. jejuni NCTC11168 and other Campylobacter sequences lodged with various international databases. A number of species specific sequences were identified and are currently undergoing further analysis, including Southern hybridisation. Over the next six months, the project team will array both PCR amplicons from the selected virulence genes and molecular probes onto amino-saline coated glass slides. The arrayed slides will enable the detection and virulence characterisation of wild type Campylobacter strains isolated from breeder farms or processing environments and provide for a better understanding of Campylobacter pathogenesis. Other applications could include the use of the microarrays for monitoring expression of or changes in bacterial virulence as a result of changed processing conditions and for monitoring for the emergence of new Campylobacter strains and other pathogens that may be of importance to public health, thus enhancing the quality and safety of poultry products.


Project Title

Campylobacter bio-replacement program to control food poisoning organisms in poultry

RIRDC Project No.:

UG-7A

Start Date: 01/07/2003 Finish Date: 31/07/2007 Researcher: Dr. Victoria Korolik Organisation: Griffith University

School of Health Sciences, Gold Coast Campus PMB 50 GOLD COAST MAIL CENTRE QLD 9726


• To develop methods for controlling Campylobacter spp in poultry by

utilising a collection of non pathogenic Campylobacter strains as bio-replacement organisms to exclude pathogenic Campylobacter spp from commercial chicken flocks.

• To determine the overall effect of colonisation by super-colonising Campylobacter strains on the general health of chickens.

Current Progress C. jejuni and C. coli are the most common cause of food poisoning in humans worldwide, with poultry derived campylobacteriosis playing a major role in human infections. This project aims at minimising risk of human exposure to pathogenic campylobacters and involves ‘bio-replacement’ of potentially virulent Campylobacter strains by a non-virulent C. jejuni strain. Persistence of ‘bio-replacement’ campylobacteria C. jejuni strain 331 in the gastrointestinal tract of chickens in experimental flocks of 100 birds in a simulated farm environment was tested. Live bacteria were delivered, mixed in chicken feed, to experimental flocks on days 1, 7 and 14 post-hatch. Cloacal secretions were collected by swabbing 15 randomly chosen birds from each flock on days 3, 7, 14, 21, 28, 35, 42 and 56. The identity of resident C. jejuni strains was established by molecular testing by multiplex PCR for each sample taken. All experimental flocks were found to secrete C. jejuni by 28 days of age and continued to secrete live campylobacteria until the termination of the experiment on day 56. Molecular identification of the C. jejuni strain secreted by the chickens throughout this trial showed that the secreted strain was identical to C. jejuni strain 331 used to inoculate the chickens on days 1, 7 and 14. This trial demonstrated the ability of the proposed ‘bio-replacement’ strain to effectively colonise and persist in the chicken flocks in the simulated farm environment.


Environmental Management

Project Title

Evaluating risks posed by pathogen emissions from meat chicken sheds

RIRDC Project No.:

DAQ-318A

Start Date: 01/01/2004 Finish Date: 31/12/2006 Researcher: Dr. Pat Blackall Organisation: Department of Primary Industries & Fisheries (Qld)

Animal Research Institute Locked Mail Bag No 4 MOOROOKA QLD 4105


• To quantify emission rates for pathogenic bacteria (Salmonella spp,

Escherichia coli, Campylobacter jejuni/coli, Staphylococcus) from typical tunnel ventilated meat chicken sheds.

• To evaluate the potential for human health effects by these emissions evaluated by combining measured emissions, gaussian air dispersion models, bacterial survival models and applying quantitative risk assessment methodology.

Current Progress This project, which commenced in January 2004, has concentrated on

establishing and validating methods for the enumeration of pathogens and dust particles in the emissions from chicken meat sheds. Biological aerosol samplers that use three different collection techniques – impingement, impaction and gel filtration – are now being validated. A list of the potential bacteria to be examined in the study has been assembled. As part of the validation of the media to be used to enumerate the target bacteria, a laboratory-based dust generator has been developed. This generator will allow the generation of dust spiked with known amounts of relevant pathogens. This spiked dust will then be used to validate the various bacteriological media (for their ability to revive bacteria captured from aerosols) and the various biological samples (for their ability to capture bacteria from aerosols). Dust sampling in a typical chicken meat shed has commenced. This work involves use of an automatic dust sampling device which will quantify dust levels over a period of time.


Project Title

Efficacy of windbreak walls for odour reduction

RIRDC Project No.:

DAQ-321A

Start Date: 01/05/04 Finish Date: 30/07/05 Researcher: Mr Geordie Galvin Organisation: Department of Primary Industries & Fisheries (Qld) Phone: (07) 4688 1118 Fax: (07) 4688 1192 Email: [email protected] Objectives

• To identify the value of windbreak walls for improving dispersion of

exhaust air from tunnel-ventilated chicken sheds. • To evaluate the use of windbreak walls as an odour reduction strategy for

meat chicken sheds.

Current Progress The project has progressed well since its inception. A portable weather station has been operating on the farm under study since May 2004. The data from the weather station has been used to calculate dispersion parameters (eg. stability class). Suitable sampling sites for the field work have been identified using the dispersion model Ausplume. The modelling has highlighted the differences in dispersion related to air temperature inside the shed, showing that the plume would travel further when the exhauster air is warmer. Sampling will be undertaken between midnight and 7am on the sampling days. This is based on the meteorological data from the site that shows a dominant south-westerly wind under early morning stable conditions. Work has also been undertaken to ensure that the gas analysis method will detect the concentrations required by trapping known concentrations on the sampling media for analysis. Preliminary sampling in a destocked shed will be undertaken on Tuesday 13 July to ensure that all methods are complete before the more intensive sampling regime begins. This sampling involves releasing a known amount of the tracer gas and measuring concentrations downwind. This data will then be compared to the modelling. In addition to the agreed method, a real time sampler has been hired by the QDPI&F for real time plume monitoring.


Project Title

Trials of odour control technologies for broiler farms

RIRDC Project No.:

DAV-213A

Start Date: 01/07/2003 Finish Date: 31/07/2006 Researcher: Dr. Julie Simons Organisation: Department of Primary Industries (Vic)

Primary Industries Research Victoria, Attwood Site 475 Mickleham Road ATTWOOD VIC 3049


• To identify, assess and quantify the performance of technology options to

control odour from broiler sheds. • To define the conditions and application methods in which they work. • To monitor the extent of odour emissions as they occur on a number of

farms. • To develop test protocols for the testing the efficacy of future odour control

technologies as they become available.

Current Progress The first stage of the project was to conduct an on-farm survey of farm and litter conditions. The response for participation in this survey was overwhelming. Chicken meat grower networks from the Victorian Farmers Federation and the Department of Primary Industries secured 19 willing participants, representing growers from most areas of Victoria and five major processing companies. As this number was in excess of the original target of ten producers, not all producers could be included in the survey. The survey involved at least one farm visit, whereby an extensive on-farm interview was conducted with the grower and basic shed conditions were measured, including temperature and humidity. In addition, litter samples were collected from at least one shed and tested for moisture, carbon: nitrogen ratio, protein, fat and starch content. The on-farm survey is in its final stages of completion. Four farms will be selected for more intensive research. The next stage of the research will be to develop the method protocols for the assessment of odour technologies. This project has provided a unique opportunity for industry and government agencies to work together towards a critically important goal and will provide much needed data for understanding odour and opportunities for its control.


Project Title

Risk assessment on the use of chicken litter and guidelines for its safe use

RIRDC Project No.:

FSA-3A

Start Date: 15/04/04 Finish Date: 15/04/05 Researcher: Mr. Peter Nicholas Organisation: FSE Environmental Phone: (07) 4632 8230 Fax: (07) 4632 8057 Email: [email protected] Objectives

• To undertake a risk assessment on the use of raw and treated litter products

for a range of different end-use practices. • To develop a set of guidelines for the safe and sustainable use of litter and

treated or composted litter products based this risk assessment.

Current Progress The main activities undertaken since the recent start of the project have been the commencement of the literature review and planning for the collection and analysis of farm litter samples. The literature review involves two main components – the collation of information relevant to chicken litter characteristics and reuse and the review of relevant Standards, Codes and Guidelines for the reuse of organic manures. The review of chicken litter characteristics and reuse includes research from Australia and overseas. The review of Standards, Codes and Guidelines is primarily focussed on the current situation in Australia and likely future directions that may influence Guidelines for the safe use of litter. There are major gaps in our knowledge that must be addressed before any substantive guidelines can be developed. The project therefore provides for the collection of litter samples from ten farms per state in Victoria, New South Wales and Queensland. These samples will be analysed to determine typical concentrations of key pathogens and heavy metals - areas where there are major gaps in our knowledge. Planning has commenced to determine the factors to be considered for selecting farms for sampling, such as diet and management used (different integrators; one use litter, recycled litter etc). Consideration has also been given to the selection of sample sites within the sheds to obtain representative material and to the co-ordination of sample collection and transport between states.


Other

Project Title

CRC for the Australian Poultry Industries funding

RIRDC Project No.:

CPO-1A

Start Date: 01/07/2003 Finish Date: 30/06/2010 Researcher: Prof. Mingan Choct Organisation: CRC for the Australian Poultry Industries

W21, University of New England ARMIDALE NSW 2351


• To provide funding to the CRC for Australian Poultry, the mission of which

is to enhance the competitiveness of the Australian egg and chicken meat industries and supporting industries through the application of strategic programs delivering cost-effective and socially responsible production of safe, quality poultry products for domestic consumption and for emerging export markets.

Current Progress The CRC was established 1 July 2003. All agreements for projects to be funded

in its first year of operation have been signed and significant progress has been made on some of the projects. The earliest signing took place on 20 October 2003 and the latest was on 8 June 2004. Nineteen project agreements have been executed, with a total investment of $2,717,006 for 2003/2004. Some aspects of the Poultry CRC project agreements are unique in that they contain considerable amounts of information on commercialisation pathways and licensing arrangements.


52


SCHOLARSHIPS

CSA-20A Postgraduate scholarship - Scott Sheedy: Live vectoring of therapeutic and prophylactic proteins and

pathogenesis in necrotic enteritis Refer to report page 21 CSA-21J Postgraduate scholarship - Manija Asif: Cytokines and innate molecules for enhanced mucosal

immunity in the chicken Refer to report page 22 CSA-25J Postgraduate scholarship - Kristie Jenkins: Improved therapeutics for Marek's disease virus infection

Refer to report page 24 UTS-6A Postgraduate scholarship - Ms Kelly Mai: The molecular basis for oocyst wall formation in the

apicomplexan parasite, Eimeria maxima Refer to report page 37 UNE-86A Postgraduate scholarship - Mr Nicholas Rodgers: Relationships between grain quality, intestinal

integrity, and performance of broiler chickens Refer to report page 39 PROGRAM REVIEW AND DEVELOPMENT MS023-28 Steering committee meetings with Marek's disease researchers MS034-02 Campylobacter research groups coordination MS034-30 Contribution towards 2004 Grain Stock Survey TRAVEL/CONFERENCE/WORKSHOPS TA034-04 Recent Advances in Animal Nutrition in Australia 2003 Conference, Armidale, July 2003 – Dr David

Sklan TA034-08 12th International Workshop on Campylobacter, Helicobacter and Related Organisms, September 2003

– Dr Margaret MacKenzie TA034-09 12th International Workshop on Campylobacter, Helicobacter and Related Organisms, September 2003

– Dr Pat Blackall TA034-28 Western Poultry Disease Conference, Sacramento – Tom Grimes TA034-37 7th International Marek’s Disease Symposium – Ms Julie Cooke TA034-38 World’s Poultry Congress 2004 – Rider Perez-Maldonado TA034-45 Enviro04 Odour Conference – Larry Jacobson TA034-46 5th Asia Pacific Poultry Health Conference, Gold Coast – Drs Donoghue & Cavanagh TA034-49 Poultry Information Exchange, April 2004 WS023-09 National Biosecurity Workshops WS023-22 Strategic Planning Workshop, Sydney, 10 July 2003 WS034-08 Support for the 2004 Australian Poultry Science Symposium

RE

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AR

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BE

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PP

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003/

2004

53

R

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Phon

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and p

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is in

necro

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CS

A-20

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Rob

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CS

IRO

Lives

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tries

(03)

5227

5760

Postg

radu

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- Man

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olecu

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d mu

cosa

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chick

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viru

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(03)

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prov

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(03)

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of ne

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mea

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CS

A-29

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Rob

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CS

IRO

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tock I

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tries

(03)

5227

5760

Ch

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(03)

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is in

poult

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Q-31

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ustrie

s and

Fis

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(07)

3362

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the e

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meth

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disea

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rus

RMI-1

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nolog

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3) 99

25 30

88

Molec

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for m

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as in

broil

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UJC-

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(07)

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for c

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in Au

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an

broil

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UM-6

8A

Dr T

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ust

Unive

rsity

of Me

lbour

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(03)

9344

9676

Effec

ts of

orga

nic ac

ids, p

rebio

tics,

and e

nzym

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contr

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necro

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teritis

and

perfo

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ce of

broil

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icken

s UN

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ct Un

iversi

ty of

New

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(02)

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matic

patho

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arek

's dis

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S-4J

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olas S

mith

Unive

rsity

of Te

chno

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Sydn

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(02)

9514

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stgra

duate

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larsh

ip - M

s Kell

y Mai:

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mole

cular

basis

for o

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form

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niver

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(07)

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3081

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or liv

estoc

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(stag

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lack

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(0

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- Mr N

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s Rod

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in qu

ality,

int

estin

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egrity

, and

perfo

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ce of

broil

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icken

s UN

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A Pr

of Mi

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Cho

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ty of

New

Engla

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(02)

6773

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feed

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preb

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on de

velop

ment

of the

dige

stive

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m an

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of

broil

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an C

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of Ne

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Th

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anica

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enzy

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impr

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AECL

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Peter

J Co

loe

Roya

l Melb

ourn

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titute

of Te

chno

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to co

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ganis

ms in

poult

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Gr

iffith

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rsity

(0

7) 55

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at Bl

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partm

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ndus

tries a

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Fishe

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Qld)

(0

7) 33

62 94

98

Effic

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lvin

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Fis

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(07)

4688

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Fis

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farm

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3A

Dr Ju

lie S

imon

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pt of

Prim

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ndus

tries (

Vic)

(03)

9217

4358

Ri

sk as

sess

ment

on th

e use

of ch

icken

litter

and g

uideli

nes f

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safe

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A Co

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(07)

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Documents

Chicken Meat Program