OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated
OIE Reference Laboratory:
Address of laboratory:
Contagious agalactia
Animal Health and Veterinary Laboratories Agency
Woodham Lane,
Addlestone,
Surrey, KT15 3NB,
UNITED KINGDOM
Tel.:
Tel.: (+44-1932) 35.73.79
Fax:
Fax:(+44-1932) 35.74.23
e-mail address:
website:
robin.nicholas@ahvla.gsi.gov.uk
http://www.defra.gov.uk/vla/diseases/dis_mycoplasma.htm
Name (including Title and
Position) of Head of
Laboratory (Responsible
Official):
Miss Catherine Brown
Name(including Title and
Position) of OIE Reference
Expert:
Professor Robin A.J. Nicholas
Name (including Title and
Position) of writer of this
report
(if different from above):
Annual reports of OIE Reference Centres, 2011
1
Contagious agalactia
Part I: Summary of general activities related to the disease
The Mycoplasma Group at the AHVLA performs diagnosis and research into all aspects of animal
mycoplasmology including as a contagious agalactia-free country monitoring small ruminant herds for diseases
suggestive of CA. We also perform fundamental studies into the behaviour of M agalactiae and related CA
mycoplasmas including the effects of biofilm formation on survival and pathogenicity. We have created a
universal molecular typing scheme to enable the classification of isolates and help disease tracing. We
routinely perform antibiotic sensitivity tests on mycoplasma isolates
1.
Test(s) in use/or available for the specified disease/topic at your laboratory
Total
Test
For
Animal
Surveillance
Diagnosis
ELISA
Antibody
Sheep
2
0
CFT
Antibody
Goat
0
0
Immunoblotting
Antibody
Sheep and goat
0
0
Mycoplasma isolation
Mycoplasma organisms
Sheep and goat
0
0
PCR/DGGE*
Mycoplasma DNA
Sheep and goat
49
* Denaturing gradient gel electrophoresis (McAuliffe et al., 2005)
2.
Production and distribution of diagnostic reagents
Antigen for the ELISA for antibodies to M agalactiae was provided to our laboratory testing division at
AHVLA
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
None
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
None
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
None
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Annual reports of OIE Reference Centres, 2011
Contagious agalactia
5.
Research and development of new procedures for diagnosis and control
In an effort to develop improved diagnostic tests for contagious agalactia, representative strains of the
causative mycoplasmas: M agalactiae, M mycoides subsp capri and M capricolum subsp. capricolum were
analysed by 2-D electrophoresis. Unique proteins were identified by MudPIT analysis and immunogenic
proteins were matched by mass spectrometry. A number of proteins were identified from each mycoplasma
species that could be used as antigens in a diagnostic test. Real time PCRs have also been assessed for their
ability to detect causative mycoplasmas in clinical material. They were found to be 100 time more sensitive
than conventional PCRs and easier to standardise and automate. Further validation is required before they can
be used in routine diagnosis
6.
Collection, analysis and dissemination of epizootiological data relevant to international
disease control
Sheep flocks from three prefectures of Japan, Hokkaido, Iwate and Aumori, were screened for antibodies to
Mycoplasma agalactiae by indirect ELISA. None of the 246 sera were serologically positive suggesting that
this region is free of contagious agalactia
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
ISO 17025 and ISO9001-2008 audited by Lloyds Registry Quality Assessors on behalf of United Kingdom
Accreditation System. We follow Advisory Committee on Dangerous Pathogens for handling live mycoplasma
in containment level 2 facility using class II microbiological safety cabinets and follow stipulations set by the
Specified Animal Pathogens Order..
8.
Provision of consultant expertise to OIE or to OIE Member Countries
RN chaired a group of experts to analyse information gaps in our knowledge of contagious agalactia under the
EU DISCONTOOLS initiative. A report was submitted
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
Staff from Italy (1) and Czech Republic (1) were trained in diagnostic and research methods concerning
ruminant mycoplasmas including those causing contagious aglactia
We began discussions with the CVDRL in Kabul, Afghanistan under the twinning initiative. This process will
begin in early 2012 and will cover mycoplasma diseases of small ruminants including those causing contagious
agalactia
10. Provision of diagnostic testing facilities to other OIE Member Countries
None
11. Organisation of international scientific meetings on behalf of OIE or other international
bodies
We organised an international meeting on mycoplasmas at AHVLA in June 2011 entitled “Whats new in
Mycoplasmology” which attreacted nearly 100 delegates from Europe, Middle East, Africa and the USA. This
included talks on contagious agalactia, diagnostics, vaccines and molecular epidemiology
Annual reports of OIE Reference Centres, 2011
3
Contagious agalactia
12. Participation in international scientific collaborative studies
None
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
Nicholas, R.A.J. Molecular typing –a guide. Immunologia e benessere animale: strumente e indicatori
della sanita pubblica veterinaria, IZS Sicilia Feb 2011

Scientific publications in peer-reviewed journals
La Manna, M.P. Agnone, A. Villari, S. Puleio, R. Vitale, M. Nicholas, R. Sireci , G., Dieli F. and Loria
G.R. (2011)Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae
infection in sheep. Research in Veterinary Science doi: 10.1016/j.rvsc 2011.01.029
Giangaspero, M., Nicholas, RAJ., Hlusek, M., Orusa, R. & Ayling, RD (2011)Seroepidemiological
survey of sheep flocks from Northern Japan for Mycoplasma ovipneumniae and M. agalactiae. Tropical
Animal Health and Production DOI 10.1007/s11250-011-9952-8
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Annual reports of OIE Reference Centres, 2011