OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Bovine viral diarrhoea
Virology Laboratory
Elizabeth Macarthur Agriculture Institute,
Woodbridge Rd
Menangle
NSW 2568
AUSTRALIA.
Postal address:
PMB 4008,
Narellan,
NSW 2567
AUSTRALIA
Tel.:
+61-2-46406331
Fax:
+61-2-46406429
e-mail address:
website:
peter.kirkland@dpi.nsw.gov.au
www.dpi.nsw.gov.au/
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Dr P.D. Kirkland
Name (including Title and
Position) of OIE Reference
Expert:
Dr P.D. Kirkland
Name (including Title and
Position) of writer of this report
(if different from above):
Annual reports of OIE Reference Centres, 2011
1
Bovine viral diarrhoea
Part I: Summary of general activities related to the disease
1.
2.
Test(s) in use/or available for the specified disease/topic at your laboratory
Test
For
Specificity
Total
AGID
Antibody
Group
9,550
ELISA
Antigen
Group
36,500
ELISA
Antibody (tank milk)
Group
170
VN
Antibody
Type
1,250
Cell culture
Virus isolation
Group
700
Real time PCR
Viral RNA detection
Group
4,100
Production and distribution of diagnostic reagents
Reagent
Supply within Australia
Supply to other countries
BVDV AGID antigen and reference serum
30,000 tests
4,000
BVDV Free serum for cell culture
50 × 500 mL
5 × 500 mL
Pan-pestivirus monoclonal antibodies for
immunoperoxidase staining of cell cultures
40,000 tests
5,000
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
a)
Establishment and maintenance of a network with other OIE Reference Laboratories
designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results
To date there have not been any formal interlaboratory comparisons of BVDV assays by the OIE reference
laboratories. The reliance on reference laboratories for BVDV activities has been less than for some other
diseases because of the availability of highly sensitive, well standardised commercial diagnostic test kits.
However, as more countries make progress towards eradication, the need for reference laboratory support for
difficult cases will inevitably increase.
b)
Organisation of inter-laboratory proficiency testing with laboratories other than OIE
Reference Laboratories for the same pathogens and diseases to ensure equivalence of
results
Although organised by an independent group, the laboratory routinely participates in ANQAP, the Australian
proficiency testing program/ring trial that is conducted each year. This program involves laboratories from
Australia, New Zealand and the USA. As well as being a participant in the AGID, VNT, antigen ELISA,
virus isolation and PCR elements, the laboratory is called upon to undertake preliminary screening of
samples for suitability for inclusion in test panels.
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Annual reports of OIE Reference Centres, 2011
Bovine viral diarrhoea
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
There are no formally recognised international reference standards for BVDV.
5.
Research and development of new procedures for diagnosis and control
For the rapid diagnosis of BVDV, the antigen capture ELISA is now in widespread, almost universal use for the
diagnosis of persistent infections of cattle with BVDV. The laboratory continues to be involved in a project that is
examining the suitability of alternative specimens for the detection of BVDV antigen. In the past, white blood
cells were considered the optimal sample for BVDV diagnosis in the live animal. More recently, with the
availability of commercial assays for the detection of the E rns antigen, skin (“ear notch”) samples have become the
sample of choice for primary screening as this circumvents the problem of maternal antibodies blocking antigen
detection in young or other seropositive animals. This project is focussing on the testing of hair samples. There is
extensive testing of other samples (buffy coat for antigen, serum for antibodies), including repeat sampling of
animals, to confirm that any animals that test positive are actually persistently infected. The results obtained have
confirmed that hair samples can be used as an alternative sample for BVDV diagnosis. In the last 12 months
studies have been completed to test samples from cattle infected with different subtypes of BVDV1 and with
strains of BVDV Type 2. These studies have now included persistently and acutely infected animals which have
been tested by both antigen ELISA and qRT-PCR. These studies have shown that PI animals can be detected by
both assays whereas acutely infected animals are not detected by the antigen ELISA. These data indicate that this
assay will reliably distinguish between acutely and persistently infected animals. During the next 12 months this
research will be extended to the validation of a commercial kit for the detection of BVDV antigens by ELISA.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
There are no recent epidemiological events of interest that would suggest changes in disease transmission patterns.
There has been no recognition of new strains of BVDV or the emergence of new pestiviruses affecting cattle in
Australia.
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
A large amount of time is spent on activities related to quality assurance, biosafety and biosecurity. The laboratory
facilities that we currently occupy are new (have been occupied for less than 6 months) and are fully compliant
with all national and international guidelines to both an enhanced PC2 and PC3 (PSL3 Ag) level.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
None provided
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
There was no formal training provided during this period.
10. Provision of diagnostic testing facilities to other OIE Member Countries
During the latter part of the year, the laboratory tested a number of serum samples by VNT for New Zealand. This
was routine screening testing for preparation of biological materials, rather than disease investigation and none of
the results were provided to OIE. See also comments under Item 12.
Annual reports of OIE Reference Centres, 2011
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Bovine viral diarrhoea
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
No meetings were attended in a formal capacity to represent OIE
12. Participation in international scientific collaborative studies
During the year, investigations were undertaken with colleagues in the USA to undertake follow-up testing of
batches of semen from a bull that is thought to have a persistent testicular infection. Semen from this bull had been
exported to Australia and had been subsequently shown to contain BVDV. Testing in this laboratory and in a
research laboratory in the USA has identified BVDV in most batches of semen. However, the virus titres are very
low and difficult to detect by virus isolation. It is unlikely that such low doses of virus would be infective when
inseminated into susceptible cows. Virus was most consistently detected by qRT-PCR but still required special
extraction methods to consistently identify virus. These investigations identified several shortcomings in the OIE
Manual, including the absence of any suitable qRT-PCR methods.
13. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
Dr Kirkland attended the 9th ESVV pestivirus symposium in Hannover Germany. Since that meeting there have
been ongoing international discussions on changes in pestivirus classification and new criteria for systematic
descriptions of viruses, especially for new strains/isolates that could represent new species.

Scientific publications in peer-reviewed journals
None specifically on BVDV; 2 on Bungowannah virus – a recently discovered novel pestivirus

Other communications
None
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Annual reports of OIE Reference Centres, 2011