OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Avian influenza
Australian Animal Health Laboratory
CSIRO Livestock Industries
Private Bag 24, Geelong, Victoria, 3220, Australia
Tel.:
(61-3) 52.27.50.00
Fax:
(61-3) 52.27.55.55
e-mail address:
website:
Name of Head of Laboratory
(Responsible Official):
Name of OIE Reference Expert:
Name of writer of this report
(if different from above):
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Paul.Selleck@csiro.au
www.csiro.au
Dr Martyn Jeggo
Dr Paul Selleck
Dr Peter Daniels
1
Avian influenza
Part I: Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease/topic at your laboratory
AAHL maintains a broad range of diagnostic tests and supporting laboratory technologies for avian influenza
which include virus isolation in embryonated eggs, virus characterization by haemagglutination and neuramidase
inhibition (HAI and NI), molecular diagnosis by TaqMan assay for H5, H7 and Influenza A, rapid diagnosis by
antigen detection tests such as indirect fluorescent antibody tests on tissue impression smears,
immunohistochemistry on formalin fixed tissues, electron microscopy and immuno-EM. Isolates may be
pathotyped by pathogenicity tests in chickens and molecularly by sequence analysis. Further characterization is
undertaken by sequence analysis for molecular epidemiology and detection of other nucleotide changes of interest
in specific situations. Serology is conducted primarily by Type A group specific C-ELISA and HAI tests, and the
agar gel diffusion test (group antigen) can be offered. Specialized HI testing in support of antigenic cartography is
conducted. Isolates are tested phentypically for susceptibility or resistance to antiviral agents.
During 2010 the following tests were performed:
2.
Test
For
Specificity
2010 Total
C-ELISA (Flu A)
Antibody
Group
2635
HI (H5) N1 & N3
Antibody
H Type
573
HI (H7) N3
Antibody
H Type
202
HI (other)
Antibody
H Type
85
VN (Avian Haemagglutinin typing)
Antibody
Type
6
AGID
Antibody
Group
1512
Real time PCR (Flu A)
Molecular test
Group
669
Real time PCR (H5 & H7)
Molecular test
H Type
288
Embryonated chicken egg culture
Virus isolation
Group
499
IFA on Impression smears
Antigen Detection
Group
0
H typing
Virus characterization
H Type
161
Immunohistochemistry
Antigen Detection
Group
42
N typing
Virus characterization
N Type
3
Pathotyping - molecular
Virus characterization
HA cleavage site
12
HA gene sequencing and phylogenetic
analysis
Virus characterization
113
Production and distribution of diagnostic reagents
Reagents produced for Avian Influenza
H1 – H16 reference antigens
H1 – H16 reference antisera in chickens
PCR Positive controls
Antigenic cartography serum for OFFlu
Antigenic cartography antigens for OFFlu
2
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Avian influenza
OIE Reference Antiserum for the avian influenza agar gel immunodiffusion test
Diagnostic reagents supplied nationally (including own laboratory)
H5N1 HI antigens
50 ml
PCR Proficiency Testing panels
2 x 10 samples (0.25 ml each)
AI ELISA Proficiency Testing panels
2 x 10 samples (0.3 ml each)
Miscellaneous AIV antigen
100 ml
Miscellaneous AIV antisera
20 ml
AIV C-ELISA reagents
25,000 tests
AGID Reagents
500 tests
Diagnostic reagents supplied to other OIE Members
H5N1 HI antigens
500 ml
H5N1 reference antisera
250 ml
PCR Proficiency Testing panels
2 x 10 samples (0.25 ml each) to each of 17 laboratories
H5N1 HI Proficiency Testing panels
2x 10 samples (0.3 ml each) to each of 17 laboratories
Miscellaneous AIV antigen
200 ml
Miscellaneous AIV antisera
100 ml
Live viruses supplied to other laboratories
10 viruses (1 ml each)
AIV NP Monoclonal antibody
30 ml
AIV C-ELISA reagents
10,000 tests
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
EQA programs for both serology and real time PCR are supported in Indonesia (9 laboratories) and more broadly
in Southeast Asia, in eight ASEAN countries national laboratories. Each proficiency testing panel contains 10
samples for PCR diagnosis and 10 samples for serology (haemagglutination inhibition). Two rounds were
distributed in Indonesia in 2010 and 2 rounds to the ASEAN national laboratories.
In 2010 AAHL participated in two rounds of the GC Deventer ring trial for avian influenza serology, both
haemagglutination inhibition and ELISA.
Australia has coordinated its government laboratories into a functional network for responses to emergency animal
diseases – Laboratories for Emergency Animal Disease Diagnosis and Response, LEADDR – with the mission to
coordinate a national laboratory network operating on a partnership approach to detect and manage outbreaks of
selected EADs. The network will coordinate an expanding diagnostic and surge capacity for EAD outbreaks
nationwide, including those which have the potential to threaten human health. Through its role in LEADDR, and
specifically the AI working group in LEADDR, AAHL has helped coordinate adoption of an agreed national real
time PCR test for AI detection that detects all strain and particularly detects AI viruses that occur in Asia.
Proficiency Testing in support of this test is being provided.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
An Agar Gel Immunodiffusion Reference Serum was prepared and tested for the OIE. A total volume of 2.5 litres
was prepared. This has been lyophilized and will be distributed during 2011 after appropriate labelling.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
3
Avian influenza
5.
Research and development of new procedures for diagnosis and control
AAHL is undertaking various projects relating to AI pathogenesis and transmission as well as studies of AI
vaccines and vaccination. There is significant ongoing activity in H5N1 in both domestic poultry and mammalian
infection models with particular emphasis on the impact of route of exposure on infection outcomes, virulence
determinants associated with host adaptation, and mechanisms of protection afforded by vaccines. Additional
projects have been developed in response to pandemic H1N1 including evaluation of the efficacy of novel antiviral
therapies in suppressing virus replication in the lung and of new vaccine delivery systems.
Two major studies with an international poultry breeding company are continuing; one using transgenic techniques
to develop poultry with an enhanced immune system that will be substantially resistant to AI infection and another
to develop anti-viral therapeutics.
Evaluation of the anti-viral properties of commercial chemical compounds is also being conducted. The relative
susceptibility of AI isolates from various Southeast Asian countries to the major influenza antivirals is being tested
in an ongoing study.
Regarding diagnostic tests, work has been conducted to update the Influenza type A TaqMan assay by including a
primer to increase sensitivity for the pandemic 2009 strain, and for HPAI (H5 subtype) by inclusion of an
additional target region to minimise risk of assay failure due to sequence changes in the target region in the virus.
Other test development has addressed Pandemic H1N1 influenza diagnosis through design of an N1-Taqman
Assay for pH1N1 and also the fine tuning of a pandemic H1 subtype Taqman assay and an NP gene Taqman
assay. In addition, an inactivated pH1N1-like antigen preparation has been tested against a range of reference
antisera and has exhibited exceptional utility for the diagnosis of pH1N1.
A molecular diagnostic test for rapid identification and differentiation of important poultry viruses is under
development which can detect and differentiate influenza type A, H5, H7, pandemic H1N1 and NDV field and V4
vaccine strains.
For agent characterization’ algorithms have been established for high-throughput whole genome capillary
sequencing for HPAI and pH1N1 2009 together with phylogenetics workflow for virus characterization and
compared with the Roche 454 whole genome pyrosequencing for H5N1 HPAI. In particular
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The AAHL workflow is similar to the algorithm for HPAI phylogenetic analysis suggested by Richard
Webby at the September 2010 Vaccine Composition Meeting. This will be important for inter-laboratory data
sharing and results comparison. AAHL supports participation any further development and consultations on
the harmonization of recommended methods by participating laboratories.
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AAHL collaborated with the WHO Collaborating Centre in Melbourne for using genome segment-marker
specific primers for rapid screening of pH1N1 2009 swine influenza viruses for possible genomic
reassortments using the QIAGEN pyrosequencer. Swine isolates from 2009 disease outbreaks in Australian
commercial piggeries were characterized with this assay.
Under an OFFLU project antigenic and genetic characterisation and evolution of H5N1 HPAI viruses in Indonesia
and SE Asia is being studied (see also Section 11). The work included
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genetic and antigenic characterization of clade 2.3 H5N1 viruses from Indonesian poultry sectors under the
OFFLU Indonesia project; this work will be continuing in 2011.
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identification and characterization of clade 1, clade 2.3.2, and clade 2.3.4 H5N1 viruses from Lao PDR,
Myanmar, and Vietnam over 2010. Clade 2.3.4 viruses were predominant in these samples.
Wild bird isolates of avian influenza viruses from Australia and Indonesia are being sequenced and analysed.
4
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A limited number of Australian avian influenza viruses were genetically characterized including H6, H10 and
low path H5 LPAI’s (eg. H5N9).
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Capability has been established for molecular identification of non-H5 avian influenza subtypes, including H
and N sub-typing by panHA and NA-subtype specific sequencing assays respectively.
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Genetic characterization assays for wildtype H9 viruses, specifically H9N2, are being established and tested.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Avian influenza
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
Influenza virus isolates have been received from Myanmar, Laos and Vietnam during the reporting period, as
listed in Section 9 of this report.
AAHL is collaborating in an international partnership coordinated by OFFLU to address issues of antigenic
mismatch between some circulating field strains and H5 antigens in available vaccines in Indonesia. On the basis
of sequence analyses, HI studies and antigenic cartography done in collaboration with Erasmus University
recommendations have been made to Indonesian animal health authorities (see also Sections 9 and 11 below).
A program of assistance to the national veterinary laboratories in Indonesia includes a laboratory outreach
component with the objective of offering support in epidemiological investigations to develop international best
practices in that country for outbreak investigations and the conduct of active surveillance. Under this
collaboration studies regarding H5N1 epidemiology in Indonesia are being conducted.
AAHL is providing experts to FAO in Indonesia to work on the Government of Indonesia program of detection,
reporting and control of H5N1 infections in village and commercial poultry through the Participatory Disease
Surveillance and Response (PDSR) program.
Epidemiological studies in Indonesia and Vietnam are investigating AI transmission in markets, AI infection in
ducks, and the impact of vaccination on AI transmission in both commercial and village sectors. This work
includes investigation of whether long term persistence and shedding of HPAI H5N1 viruses occurs in ducks.
7.
Provision of consultant expertise to OIE or to OIE Members
With respect to avian influenza, AAHL has contributed to the following OIE and FAO activities:
Dr Emma Watkins and Dr Kerry Mulqueen were based in Makassar in South Sulawesi province of Indonesia
contributing to the Indonesian national government program for the control of H5N1 HPAI in poultry in that
country January – October 2010.
Dr Peter Daniels attended a joint OFFLU Steering and Executive Committee meeting at OIE Headquarters in
Paris, 25 January 2010
Dr Martyn Jeggo and Dr Linfa Wang attended the Second FAO-OIE-WHO Joint Scientific Consultation:
Influenza and other Emerging Zoonotic Diseases at the Human Animal Interface in Verona, Italy on April 27-29
2010.
Dr John Allen attended the Annual Meeting of the EU FLUTrain Project in Legnaro, Italy, 15-17 June 2010.
Dr Peter Daniels attended an OFFLU Steering Committee and Executive Committee meeting at IAEA in Vienna,
14-15 July 2010.
Dr Peter Daniels and Mr Chris Morrissy participated in the FAO/OIE Consultative Meeting of Regional
Laboratory Network Partners for HPAI Diagnosis in Southeast Asia in Bangkok 20-21 July 2010.
Mr Chris Morrissy, Ms Gemma Carlile, Mr Paul Selleck and Mr John Allen attended the Regional FAO LabNet
meeting at Veterinary Research Institute in Ipoh, Malaysia, 3-6 October 2010
Dr John Allen, Dr Peter Daniels, Mr Paul Selleck and Dr Frank Wong attended the OFFLU Technical Review
Meeting in Jakarta, 28-29 October 2010, to specifically address the scientific issues surrounding an international
response to the situation that has emerged in Indonesia wherein strains of H5N1 resistant to currently available
vaccines have emerged in the commercial poultry sector.
Dr Peter Daniels and Mr Paul Selleck participated in an OFFLU Technical Group Meeting held at FAO
Headquarters, Rome 16-17 November 2010.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
5
Avian influenza
8.
Provision of scientific and technical training to personnel from other OIE Members
AAHL has entered a formal OIE Twinning agreement with the Malaysia Veterinary Research Institute (VRI) at
Ipoh for Avian Influenza and Newcastle Disease,
The Indonesia Veterinary Laboratory Capacity Building Project (AusAID), part of the Support of the Pandemic
Influenza and Emerging Infectious Diseases Prevention and Preparedness Program in Indonesia, developed
laboratory capacity for HPAI in particular, supporting the network of national government veterinary laboratories
throughout Indonesia. This work included an outreach component that enhanced epidemiological capacity for the
better utilization of the laboratory capacity.
Another project that developed surveillance capability was the project: Support of the Pandemic Influenza and
Emerging Infectious Diseases Prevention and Preparedness Program in Indonesia through the Animal Health
Support for South Sulawesi and Papua Provinces (AusAID), operated in conjunction with the FAO.
AAHL delivered regional country capacity building and capability transfer (in-country training) for genetic (HA
sequence analysis) and antigenic (Ag cartography) analysis of H5N1 viruses in SE Asia under various programs
over 2009/2010, including for OFFLU and OIE. This included a major project developing laboratory capacity for
bioinformatics of HPAI and the development of antigenic cartography for monitoring of the antigenic match of
vaccines to circulating field strains with the Indonesian laboratory network and international collaborators.
The following specific training in avian influenza was conducted in member countries:
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Mr Trevor Taylor visited the Maros Disease Investigation Centre (DIC) in Indonesia to assist with training in
PCR of HPAI samples from market surveys and to assist with live bird sampling, 7-23 January 2010
Dr Patrick Kluver visited Disease Investigation Centres in Indonesia, Wates and Maros, to conduct
epidemiological field training exercises for Avian Influenza, 14-25 January 2010
Mr Paul Selleck visited the Malaysia Veterinary Research Institute under the OIE Twinning project for Avian
Influenza and Newcastle Disease, 25-31 January 2010
Dr John Bingham provided training as part of the Vietnam AusAID Partnership Program 15-25 January 2010
Ms Wendy Ha, Mr Chris Morrissy, Mr Paul Selleck and Dr Frank Wong conducted a Sub-regional training
workshop in PCR, virus isolation, biosafety, HI serology and sequencing as part of the FAO project
‘Strengthening Laboratory Capacity in Molecular Characterization of H5N1 and Pandemic H1N1 2009
Diagnosis’ at the Veterinary Research Institute Ipoh, Malaysia, 1-3 February 2010. Participants included
laboratory staff from participating countries in Asia including Cambodia, Indonesia, Lao PDR, Malaysia,
Myanmar, Philippines, Thailand and Vietnam
Ms Wendy Ha, Mr Selleck and Dr Frank Wong conducted follow up in-country training at the Veterinary
Research Institute Ipoh, Malaysia as part of the FAO project ‘Strengthening Laboratory Capacity in Molecular
Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 4-6 February 2010
Ms Wendy Ha and Dr Frank Wong conducted follow up in-country training for molecular diagnosis at the
National Institute of Animal Health (NIAH), Bangkok, Thailand as part of the FAO project ‘Strengthening
Laboratory Capacity in Molecular Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 8-10
February 2010
Mr Trevor Taylor provided training and assistance to Balitvet and two Disease Investigation Centres in
Indonesia, Denpasar and Jogjakarta, for the molecular diagnosis of Avian Influenza, 8-19 February 2010.
Mr Paul Selleck provided training and assistance to Balitevet and two Disease Investigation Centres, Wates &
Subang, in Indonesia, in serology techniques for the diagnosis of Avian Influenza, 8-19 February 2010.
Mr Ross Lunt conducted follow up in-country training for virology/serology at the National Institute of Animal
Health (NIAH), Bangkok, Thailand as part of the FAO project ‘Strengthening Laboratory Capacity in
Molecular Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 15-17 February 2010
Dr Patrick Kluver visited four Disease Investigation Centres in Indonesia, Denpasar, Subang, Wates and
Maros, to conduct epidemiological field training exercises for Avian Influenza, 15-25 February 2010
Mr Nick Gudkovs provided training and assistance to Disease Investigation Centres in Indonesia, Bukittinggi
and Subang, for the molecular diagnosis of Avian Influenza, 15-27 February 2010.
Ms Wendy Ha and Ms Kim Newberry conducted follow up in-country training at the National Veterinary
Research Institute (NaVRI), Phnom Penh, Cambodia as part of the FAO project ‘Strengthening Laboratory
Capacity in Molecular Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 3-5 March 2010
Ms Wendy Ha and Ms Kim Newberry conducted follow up in-country training at the National Animal Health
Centre, Vientiane, Lao PDR as part of the FAO project ‘Strengthening Laboratory Capacity in Molecular
Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 8-10 March 2010
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Avian influenza
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Dr Patrick Kluver visited two Disease Investigation Centres in Indonesia, Medan and Bukittinggi, to conduct
epidemiological field training exercises for Avian Influenza, 8-19 March 2010
Mr Paul Selleck conducted follow up in-country training at the Yangon Veterinary Diagnostic Laboratory,
Myanmar as part of the FAO project ‘Strengthening Laboratory Capacity in Molecular Characterization of
H5N1 and Pandemic H1N1 2009 Diagnosis’, 11-13 March 2010
Mr Eric Hansson and Mr Paul Selleck conducted follow up in-country training at the Phillippines Animal
Health Centre, Philippines as part of the FAO project ‘Strengthening Laboratory Capacity in Molecular
Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 17-19 March 2010
Mr Chris Morrissy and Dr Frank Wong conducted follow up in-country training at the National Centre for
Veterinary Diagnosis (NCVD), Hanoi, Vietnam as part of the FAO project ‘Strengthening Laboratory Capacity
in Molecular Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 23-25 March 2010
Dr Patrick Kluver visited two Disease Investigation Centres in Indonesia, Banjarbaru and Maros, to conduct
epidemiological field training exercises for Avian Influenza, 19-29 April 2010
Mr Chris Morrissy provided training and assistance to the Regional Animal Health Office (RAHO-6) in Ho
Chi Minh City, Vietnam under the OIE/AAHL Project on Strengthening Veterinary Laboratory Services
(PSVS) in Southeast Asia, 6-7 May 2010
Ms Julie Cooke provided training and assistance to three Disease Investigation Centres, Banjarbaru,
Bukittinggi & Maros, in Indonesia, in serology techniques for the diagnosis of Avian Influenza, 10-27 May
2010
Mr Trevor Taylor provided training and assistance to three Disease Investigation Centres in Indonesia,
Banjarbaru, Denpasar & Maros for the molecular diagnosis of Avian Influenza, 10-25 May 2010
Mr Chris Morrissy provided training and assistance National Centre for Veterinary Diagnosis (NCVD) in
Hanoi, Vietnam under the OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in
Southeast Asia 18-19 May 2010
Dr Patrick Kluver visited three Disease Investigation Centres in Indonesia, Bukittinggi, Denpasar & Medan, to
conduct epidemiological field training exercises for Avian Influenza, 24 May – 12 June 2010
Mr Paul Selleck visited the Malaysia Veterinary Research Institute to conduct training in serology and standard
laboratory QA procedures under the OIE Twinning project for Avian Influenza and Newcastle Disease, 31
May – 5 June 2010
Mr Paul Selleck provided training and assistance to two Disease Investigation Centres, Medan & Wates, in
Indonesia, in serology techniques for the diagnosis of Avian Influenza, 7-10 June 2010.
Mr Paul Selleck and Mr Eric Hansson conducted sequencing training on behalf of OFFLU at PUSVETMA in
Surabaya, Indonesia, 14-19 June 2010
Mr Eric Hansson and Ms Kim Newberry conducted follow up in-country training at the Bandar Lampung
Disease Investigation Centre (DIC), Indonesia as part of the FAO project ‘Strengthening Laboratory Capacity
in Molecular Characterization of H5N1 and Pandemic H1N1 2009 Diagnosis’, 22-24 June 2010
Dr John Allen provided training and assistance to the Bbalitvet Research Institute in Indonesia under the
OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast Asia, 20-21 July
2010
Mr Paul Selleck provided training and assistance to the national Avian Influenza laboratories in Myanmar,
Cambodia, Thailand and Lao PDR under the OIE/AAHL Project on Strengthening Veterinary Laboratory
Services (PSVS) in Southeast Asia, 25-31 July 2010
Dr John Allen provided training and assistance to the National Animal Health Centre in Lao PDR under the
OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast Asia, 26-27 July
2010
Dr Frank Wong visited the Malaysia Veterinary Research Institute to conduct training for Molecular
Diagnostics & Phylogenic Sequencing and Sequence Analysis training under the OIE Twinning project for
Avian Influenza and Newcastle Disease, 9-15 August 2010
Mr Paul Selleck provided training and assistance to the Philippine Animal Health Center under the OIE/AAHL
Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast Asia, 29 August – 3 September
2010
Ms Julie Cooke conducted Antigen Cartography for Avian Influenza training as part of the OFFLU Project at
the Wates Disease Investigation Centre, Indonesia, 19-25 September 2010
Dr John Allen, Ms Gemma Carlile and Mr Paul Selleck conducted training on the production of test reagents
and positive and negative controls and distributing panels, compiling, conducting, analyzing and reporting on
the results of Proficiency Test rounds under the OIE Twinning project for Avian Influenza and Newcastle
Disease, 6-9 October 2010
Dr Frank Wong conducted Avian Influenza sequence analysis training as part of the OFFLU project in Jakarta,
Indonesia, 25-27 October 2010
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
7
Avian influenza
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Mr Paul Selleck conducted update training for pre-screening for Antigenic Cartography as part of the OFFLU
project at the Subang Disease Investigation Centre, Indonesia, 1-3 November 2010
Mr Paul Selleck provided training and assistance to the Bbalitvet Research Institute in Indonesia under the
OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast Asia, 4 November
2010
Mr Paul Selleck provided training and assistance to the National Institute of Animal Health in Thailand under
the OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast Asia, 5
November 2010
Mr Paul Selleck provided training and assistance to the Livestock Breeding and Veterinary Department in
Myanmar under the OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast
Asia, 8-10 November 2010
Dr John Allen provided training and assistance to the National Animal Health Centre in Lao PDR under the
OIE/AAHL Project on Strengthening Veterinary Laboratory Services (PSVS) in Southeast Asia, 29 November
2010
Mr Trevor Taylor provided training and assistance to the Philippine Animal Health Center & the National
Veterinary Research Institute in Cambodia under the OIE/AAHL Project on Strengthening Veterinary
Laboratory Services (PSVS) in Southeast Asia, 29 November - 10 December 2010
The following training in Avian Influenza was conducted at AAHL:
 Five Nepalese trainees attended training in virus isolation, biosafety, PCR and serology for HPAI diagnosis at
AAHL as part of the Avian Influenza Control Project supported by FAO, 8-27 February 2010
 A colleague from Botswana undertook training in virus isolation and biosafety at AAHL 12-23 April 2010
 Two trainees from the Malaysia VRI undertook training in Biosecurity & Biosafety, decontamination &
calibration of laboratory equipment under the OIE Twinning project for Avian Influenza and Newcastle
Disease at AAHL 18-25 April 2010
 One trainee from Lao PDR undertook a combination of training in Biosafety, Biosecurity QA and PCR and
phylogenic analysis with a focus on Avian Influenza at AAHL as part of the Avian Influenza Control Project
supported by FAO, 19-30 April 2010
 A colleague from the Bbalitvet Research Institute in Indonesia attended training in HI testing of Indonesian
post vaccination sera against a range of AI Antigens, 26 April – 16 May 2010
 One trainee from EpiCentre Massey University in New Zealand attended training in Avian Influenza
sequencing as part of a NZAID project ‘Development of epidemiological skills and technical information to
support the management and control of Highly Pathogenic Avian Influenza in Vietnam’, 2-29 May 2010.
 One trainee from the Malaysia VRI attended training in Molecular Diagnostics & Phylogenic Sequencing and
Sequence Analysis under the OIE Twinning project for Avian Influenza and Newcastle Disease at AAHL 9-30
May 2010
 Six Indonesian trainees visited AAHL to undertake Molecular Sequencing Training for Avian Influenza as part
of the OFFLU project, 8-12 November 2010
 An Indonesian colleague from the Disease Investigation Centre Wates, Yogyakarta, is undertaking a 3 year
PhD project on Pathogenesis of HPAI in ducks at AAHL.
9.
Provision of diagnostic testing facilities to other OIE Members
Country
Test
Number of Samples
Myanmar
Virus Isolation
6
HA Typing
4
Pathotyping – molecular
4
HA gene sequencing and
phylogenetic analysis
Virus Isolation
4
HA Typing
3
Pathotyping – molecular
3
Laos
8
3
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Avian influenza
Vietnam
HA gene sequencing and
phylogenetic analysis
Virus Isolation
3
20
HA Typing
20
Pathotyping – molecular
20
HA gene sequencing and
phylogenetic analysis
20
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
No major AI meetings were organized this period. Delivery of training as outlined in Section 8 above frequently
requires organization of meetings in countries.;
11. Participation in international scientific collaborative studies
AAHL has continued involvement in the OFFLU coordinated project managed by FAO “Monitoring AI virus
variants in Indonesian poultry and defining an effective and sustainable vaccination strategy”. Isolates of HPAI
from laboratories in Indonesia servicing the agricultural sector have been received for analysis as outlined in
Section 9 and capacity building as outlined in Section 8.
Sequence analyses continues to confirm that all H5N1 isolates studied to date cluster in Clade 2.1, consistent with
an interpretation that Indonesia is not being subject to multiple introductions of virus from other countries in the
region. There are some isolates from recent years that do no cluster with the 3 recognized subclades in Clade 2.1
and the description of a fourth subclade in Clade 2.1 is being investigated. The presentation of antigenic
cartography results to date are consistent with genetic analyses and field experience. The antigenic cartography on
available isolates has identified a general pattern of antigenic drift in Indonesian Clade 2.1 isolates together with a
number of outlying strains. For a comprehensive understanding of the national situation and to allow
recommendations relevant for both commercial and village poultry sectors structured prospective programs of
isolation and analyses of AI viruses from all poultry industry sectors are needed. Recommendations have been
developed regarding suitable strains as potential sources of vaccine antigens and for use as challenge viruses on
the basis of the information available.
12. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)
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Presentations at international conferences and meetings
Arzey, E., Kirkland, P., Arzey, A., Selleck, P. et al. 2010. Infection of commercial poultry and people with an H10N7
influenza virus. 6th Annual Meeting of the Australian Association of Veterinary Laboratory Diagnosticians. Biosecurity
Queensland, Coopers Plains, Brisbane 25 – 26 November 2010.
Bean, A. G. 2010. The Immune responses to influenza virus in the chicken. 9th International Veterinary
Immunology Symposium (9th IVIS). The symposium.
Butler, J., Brown, L., Middleton, D., Rockman, S. & Sapats, S. 2010. Mammalian adaptation of H5N1 avian
influenza: The importance of a good polymerase. 4th European Congress of Virology. Villa Erbe Congress Centre
- Cernobbio, Lake Como, Italy: European Society for Virology.
Butler, J., Brown, L., Middleton, D., Rockman, S. & Sapats, S. 2010. Pathogenic consequences of H5N1
polymerase changes in the mammalian host. Society for General Microbiology Spring 2010 meeting. Edinburgh
international conference centre, Edinburgh Scotland: Society for General Microbiology.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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Avian influenza
Butler, J., Brown, L., Middleton, D., Rockman, S. & Sapats, S. 2010. Rapid Adaptation of H5N1 Avian Influenza
to Mammals: The Importance of the Viral Polymerase. Australian Society for Microbiology 2010 Annual Meeting.
Sydney Convention and Exhibition Centre: Australian Society for Microbiology.
Daniels, P., Allen, J., Wong, F., Morrissy, C., & Selleck, P. 2010. Quality Assurance for Laboratories in a
Network. OFFLU Technical Meeting 16-17 November 2010 FAO Headquarters, Rome, Italy.
Daniels, P. 2010. H1N1 overview of the global pandemic. Pan Pacific Veterinary Conference AVA & NZVA
Embracing challenges. The Conference.
Deng, Y.-M., Iannello, P., Smith, I., Watson, J., Daniels, P., Komadina, N., Barr, I., Harrower, B. & Wong, F.
2010. No Reassortment in A(H1N1) 2009 Pandemic Viruses isolated from Swine in Australia. Options for the
Control of Influenza VII. Hong Kong: Wiley Blackwell.
McKimm-Breschkin, J. & Selleck, P. 2010. Surveillance of neuraminidase inhibitor sensitivity of avian influenza
isolates in South East Asia 2005-2008. OFFLU Technical Review Meeting: Monitoring AI virus variants in
Indonesian poultry and defining an effective and sustainable vaccination strategy. Jakarta, 28-29 October 2010.
McKimm-Breschkin, J., Barrett, S., Mohr, P., Bruce, K., Cooke, J. & Selleck, P. 2010. Surveillance of
neuraminidase inhibitor sensitivity of avian influenza isolates in South East Asia 2005-2008. OFFLU Technical
Meeting 16-17 November 2010 FAO Headquarters, Rome, Italy.
McLachlan, K. 2010. Improved processing of short hairpin RNA molecules targeting H5N1 avian influenza.
Antiviral applications of RNA Interference. European Science Foundation.
Selleck, P. 2010. Repeatable and Reproducible Haemagglutination Tests and AAHL’s HI Protocol. FAO SubRegional Training on Animal Influenza Viruses – Diagnosis and Molecular Characterisation, Veterinary Research
Institute, Ipoh, Malaysia. 1-3 February, 2010.
Selleck, P. 2010. Microbiological Security – Biosafety and Biosecurity. FAO Sub-Regional Training on Animal
Influenza Viruses – Diagnosis and Molecular Characterisation, Veterinary Research Institute, Ipoh, Malaysia. 1-3
February, 2010.
Selleck, P. 2010. Laboratory Safety and H5N1. FAO Sub-Regional Training on Animal Influenza Viruses –
Diagnosis and Molecular Characterisation, Veterinary Research Institute, Ipoh, Malaysia. 1-3 February, 2010.
Selleck, P. & Colling, A. 2010. Internal and External Quality Control. FAO Sub-Regional Training on Animal
Influenza Viruses – Diagnosis and Molecular Characterisation, Veterinary Research Institute, Ipoh, Malaysia. 1-3
February, 2010.
Selleck, P. 2010. Gap Analyses. FAO/OIE Sub-Regional Laboratory Network Meeting. VRI, Ipoh 4-6 October,
2010
Selleck, P., Wong, F., Morrissy, C., Allen, J. & Daniels, P. 2010. OFFlu Activities at AAHL. OFFLU Technical
Review Meeting: Monitoring AI virus variants in Indonesian poultry and defining an effective and sustainable
vaccination strategy. Jakarta, 28-29 October 2010.
Selleck, P. 2010. Evolution of H5N1 virus and the changing threat to Australia. Host, pathogens and their
interactions at the mucosal barrier: evolution, epidemiology, mechanisms and interventions. Sansom Institute
Research Symposium University of South Australia, Adelaide 13th December 2010.
Wibawa, H., Henning, J., Bingham, J., Wong, F., Lowther, S., Davies, K., Stevens, V., Pye, T., Payne, J., Rookes,
J., Nuradji, H., Ha, W., Selleck, P., Johnson, D., Junaidi, A., Middleton, D., Daniels, P. & Meers, J. 2010.
Characterization of Indonesian H5N1 isolates to understand the prolonged infection and spread of H5N1 among
domestic poultry. Global Biosecurity 2010. The Conference.
Wibawa, H., Wong, F., Bingham, J., Davies, K., Stevens, V., Pye, T., Ha, W., Selleck, P., Henning, J., Junaidi, A.,
Middleton, D., Daniels, P., Meers, J. 2010. Diversity of the Hemagglutinin and Neuraminidase Genes of
Indonesian H5N1 Highly Pathogenic Avian Influenza (HPAI) Virus Isolates. Australian Virology Group Meeting,
Lorne, 13-17 December, 2009.
10
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Avian influenza
Wong, F., Selleck, P., Smith, I., Allen, J., Daniels, P. & Kim, M. 2010. OFFLU Update on genetic characterisation
of H5N1 viruses in Indonesia. OFFLU Final Technical Review. Jakarta, Indonesia: OFFLU.

Scientific publications in peer-reviewed journals
Bryant, N. A., Paillot, R., Rash, A. S., Medcalf, E., Montesso, F., Ross, J., Watson, J., Jeggo, M., Lewis, N. S.,
Newton, J. R. & Elton, D. M. 2010. Comparison of two modern vaccines and previous influenza infection against
challenge with an equine influenza virus from the Australian 2007 outbreak. Veterinary Research, 41, 19.
Bulach, D., Halpin, R., Spiro, D., Pomeroy, L., Janies, D. & Boyle, D. 2010. Molecular analysis of H7 avian
influenza viruses from Australia and New Zealand: Genetic diversity and relationships 1976 to 2007. Journal of
Virology, 84, 9957-9966.
Kitelberger, R., McFadden, A. M. J., Hannah, M. J., Jenner, J., Bueno, R., Wait, J., Kirkland, P. D., Delbridge, G.,
Heine, H. G., Selleck, P. W., Pearce, T. W., Piggott, C. J. & O'Keefe, J. S. 2010. Comparative evaluation of four
competitive/blocking ELISAs for the detection of influenza A antibodies in horses. Veterinary Microbiology,
Online.
Oh, S., Selleck, P., Temperon, N.J., Chan, P.K., Capecchi, B., Manavis, J., Higgins, G., Burrell, C.J., Kok, T.
2009. Neutralizing monoclonal antibodies to different clades of influenza A H5N1 viruses. Journal of Virological
Methods 157:161-167.
Paillot, R., Prowse, I., Donald, C., Medcalf, E., Montesso, F., Bryant, N., Watson, J., Jeggo, M., Elton, D.,
Newton, R., Trail, P. & Barnes, H. 2010. Efficacy of a whole inactivated EI vaccine against a recent EIV outbreak
isolate and comparative detection of virus shedding. Veterinary Immunology and Immunopathology, 136, 272-283.
Sullivan, J.S., Selleck, P.W., Downton, T., Boehm, I., Axell, A., Ayob, A., Kapitza, N.M., Dyer, W., Fitzgerald,
A., Walsh, B. & Lynch, G.W. 2009. Heterosubtypic anti-avian H5N1 influenza antibodies in intravenous
immunoglobulins from globally separate populations protect against H5N1 infection in cell culture. Journal of
Molecular and Genetic Medicine 3(2), 217-224.

Other communications
Selleck, P. & Colling, A. 2010. Internal and External Quality Control. 28 January 2010, Veterinary Research
Institute, Ipoh Malaysia
Selleck, P. 2010. Microbiological Security – Biosafety and Biosecurity. 9th February 2010, DIC IV Wates, Jogjakarta,
Indonesia
Selleck, P. 2010. Microbiological Security – Biosafety and Biosecurity. 16th February 2010, DIC VIII Subang,
Indonesia
13. Inscription of diagnostic kits on the OIE Register
i)
Did you participate in expert panels for the validation of candidate kits for inscription on the
OIE Register? If yes, for which kits?
ii)
Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for
which kits?
_______________
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
11