Download Real-time PCR

OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
White spot disease
Aquaculture Pathology Laboratory
Department of Veterinary Science and Microbiology
University of Arizona
Building 90, Room 202 Pharmacy/Microbiology
Tucson, AZ 85721
UNITED STATES OF AMERICA
Tel.:
(1.520) 621.84.14
Fax:
(1.520) 621.48.99
e-mail address:
website:
[email protected]
http://microvet.arizona.edu/research/aquapath/index.htm
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Dr Donald V. Lightner
Professor
Name(including Title and
Position) of OIE Reference
Expert:
Dr Donald V. Lightner
Professor
Name (including Title and
Position) of writer of this report
(if different from above):
Annual reports of OIE Reference Centres, 2011
Dr Donald V. Lightner
&
Linda Nunan, Assistant Staff Scientist
1
White spot disease
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
PCR
Surveillance & SPF verification
All known WSD viral
genotypes
1294
Histology
Initial diagnosis & surveillance
Lesions
2385
ISH
Confirmatory diagnosis
All known WSD viral
genotypes
0
Real-time PCR
Confirmatory diagnosis &
quantification of viral load
All known WSD viral
genotypes
68
Production and distribution of diagnostic reagents
Date
Country
Material provided
Quantity sent
02/23/2011
USA
WSSV infected tissue
1vial
03/29/2011
Saudi Arabia
WSSV PCR kit
1 Kit
05/26/2011
Mexico
WSSV infected tissue
1vial
05/31/2011
Argentina
WSSV infected tissue
1vial
06/01/2011
Saudi Arabia
WSSV infected tissue
1vial
06/11/2011
India
WSSV infected tissue
1vial
07/14/2011
Malaysia
WSSV infected tissue
1vial
09/29/2011
Vietnam
WSSV infected tissue
1vial
10/06/2011
Singapore
WSSV plasmid DNA
1 vial
11/23/2011
Saudi Arabia
WSSV infected tissue
1vial
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
2
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
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
Annual reports of OIE Reference Centres, 2011
White spot disease
Ring trials: White spot syndrome virus detection by PCR
Country
Month & Year
No. Labs participating
Correct results
Ecuador
August 2011
1
YES
Madagascar
February 2011
1
YES
August 2011
1
YES
Saudi Arabia
February 2011
1
YES
Mexico
February 2011
7
Lab 1: YES Lab 2: YES Lab
3: YES Lab 4: YES Lab 5:
YES Lab 6: YES
Lab 7: YES
August 2011
5
Lab 1: YES Lab 2: YES Lab
3: YES Lab 4: YES
Lab 5: YES
Panama
February 2011
1
YES
India
August 2011
1
YES
Vietnam
February 2011
2
Lab 1: YES
Lab 2: YES
August 2011
1
YES
N-An = not analyzed
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
See section Part I-2
.5.

White spot syndrome virus infected shrimp tissue

Plasmid DNA for use as positive control in PCR reactions

White spot syndrome virus kits
Research and development of new procedures for diagnosis and control
UAZ developed and published an improved, one-step PCR method for detection of WSSV, which is based on the
OIE recommended method published by LO et al. (1996).
Nunan, L.M. and D.V. Lightner. 2011. Optimized PCR assay for detection of white spot syndrome virus (WSSV).
Journal of Virological Methods 171: 318-321.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
None
Annual reports of OIE Reference Centres, 2011
3
White spot disease
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned


8.
Quality assurance of sample material provided is achieved by pre-testing the sample material for the pathogen
of interest, by PCR, prior to shipment.
Biosafety and biosecurity is achieved by following federal regulations for disposal of infectious materials and
through the inland location of the laboratory, as no marine pathogens would be able to survive in the natural
environment of the Sonora Desert.
Provision of consultant expertise to OIE or to OIE Member Countries
Chapters for the OIE Aquatic Animal Health Code – assisted in preparation
Lightner DV. 2011. Aquatic Animal Health Code, 14th Edition. World Organization for Animal Health (Office
International des Epizooties = OIE), Paris:
- Infectious hypodermal and haematopoietic necrosis, Chapter 9.2.
- Infectious myonecrosis, Chapter 9.3.
- Necrotizing Hepatopancreatitis, Chapter 9.4.
- Taura syndrome, Chapter 9.5.
- White spot disease, Chapter 9.6.
- White tail disease, Chapter 9.7.
- Yellow head disease, Chapter 9.8.
Chapters for the OIE Manual of Diagnostic Tests for Aquatic Animals
Lightner DV. In press (2012). Diagnostic Manual for Aquatic Animal Diseases, 7th Edition. World Organization
for Animal Health (Office International des Epizooties = OIE), Paris:
- Disinfection of Crustacean Farms, Section C of Methods for Disinfection of Aquaculture Establishments.
- Diseases of Crustaceans, General Information.
- Infectious Hypodermal and Hematopoietic Necrosis.
- Infectious Myonecrosis.
- Taura Syndrome.
- Tetrahedral Baculoviruosis (Baculovirus penaei).
- Spherical Baculovirosis (Penaeus monodon-type baculovirus).
Advisor to the Aquatic Animal Health Standards Commission, Office International Des Epizooties, Paris. Term
2009-2011, Advisor (~10 days/year). I was appointed to a third 3-year term by the OIE Central Bureau, beginning
in May 2009, on the OIE Aquatic Animal Health Standards Commission (AAHSC) which meets 2 times per year
to consider new diagnostic methods for OIE listed and notifiable aquatic animal diseases; to consider changes to
the OIE Aquatic Animal Code; and to make recommendations to the Director General and to the OIE World
Assembly of Member Country Delegates. In 2011, I participated in the AAHSC meeting of October 3-7 at the OIE
Headquarters in Paris.
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
• Hosted Borlaug scholar Mr Bambang Hanggono, of the Indonesian Government’s Situbondo Laboratory.
Mr Hanggono visited the Aquaculture Pathology Laboratory for 12 weeks (17 September to 3 December, 2011).
Mr Hanggono’s training emphasized histopathology and molecular diagnostic tests for major shrimp diseases
including white spot syndrome virus.
10. Provision of diagnostic testing facilities to other OIE Member Countries
We received “official” samples for confirmation of WSSV infection from the Kingdom of Saudi Arabia and
Mozambique.
4
Annual reports of OIE Reference Centres, 2011
White spot disease
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
None
12. Participation in international scientific collaborative studies
• Development of qPCR kits with Life Technologies of Singapore & USA.
Provided samples of frozen shrimp tissue infected with IMNV and genome sequence information to Life
Technologies (formerly Applied Biosystems) of Singapore & USA for development of qPCR/RT-PCR test kits.
In late 2011, we received a real time thermocycler for use in validating shrimp virus kits for Life Technologies.
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
Invited speaker OECD Symposium, Halifax, Canada. Given by OECD Co-operative Research Program, Paris,
8/7/2011.
Invited keynote speaker, Honduras. Given by V Simposio Nicovita, Honduras, 11/24/2011. (100%).
Invited keynote speaker, Ecuador. Given by XIII Congreso Ecuatoriano de Acuicultura and Aquaexpo,
10/20/2011.
Invited speaker - OIE Global Conference on Aqiuatic Animal Health, Panama. Given by OIE , 6/29/2011.
Invited keynote speaker, Panama City, Panama. Given by El Comite Organizador del X Simposio International de
Camaron de Cultivo y Exhibicion Comercial, 5/13/2011.

Scientific publications in peer-reviewed journals
Lightner D.V. and R.M. Redman. 2010. The global status of significant infectious diseases of farmed shrimp.
Asian Fisheries Society Journal 23(4): 383-426.
Lightner D.V. 2011. Virus diseases of farmed shrimp in the Western Hemisphere (the Americas): a review.
Journal of Invertebrate Pathology 106: 110-130.
Lightner, D.V. 2011. Status of shrimp diseases and advances in shrimp health management. pp. 121-134, in M.G.
Bondad-Reantaso, J.B. Jones, F. Corsin and T. Aoki (editors), Diseases in Asian Aquaculture VII. Fish Health
Section, Asian Fisheries Society, Selangor, Malaysia. 385 p.
Nunan, L.M. and D.V. Lightner. 2011. Optimized PCR assay for detection of white spot syndrome virus (WSSV).
Journal of Virological Methods 171: 318-321.
Stentiford, G.D. and D.V. Lightner. 2011. Cases of White Spot Disease (WSD) in European shrimp farms.
Aquaculture 319: 302-306.
Lightner, D.V. and R.M. Redman. "In press". Development of specific pathogen-free (SPF) shrimp stocks and
their application to sustainable shrimp farming. Woodhead Publishing, LTD., Cambridge. UK.
Aoki, T., J.R. Bonami, D.V. Lightner, C.F. Lo, G.D. Stentiford, J.H. Leu, J. M. Vlak, K. Soderholl, P.J. Walker,
T.W. Flegel, H.C. Wang, X. Xu, and H. Yang. 2012. Nimaviridae. In A.M.Q. King, M.J. Adams, E.B. Carstens
and E J. Lefkowitz, editors: Virus Taxonomy. 9th Report of the International Committee on Taxonomy of Viruses,
Oxford, Elsevier, pp. 229 - 234.
Annual reports of OIE Reference Centres, 2011
5
White spot disease

Other communications
Popular Press
Lightner, D.V. 2011. Enfermedades virales en camarones cultivados en el hemisferio oeste: una reseña. .
Enfermedades virales en camarones cultivados en el hemisferio oeste: una reseña. Panorama Acuícola 16(4): 8-20.
Arce, S.M., S.M. Moss and D.V. Lightner. 2011. Biosecurity principles for sustainable production using SPF
shrimp. Global Aquaculture Advocate. May/June 2011, 14-16.
Cuéllar-Anjel, J., R. Chamorro, B.White-Noble, P.Schofield and D.V. Lightner. 2011. Testing finds resistance to
WSSV in shrimp from Panamanian breeding program. Global Aquaculture Advocate July/August 2011, 65-66.
Cuéllar-Anjel, J., R. Chamorro, B.White-Noble, P.Schofield and D.V. Lightner. 2011. White shrimp Litopenaeus
vannamei significantly resistant against WSSV. Panorama Acuícola 16(5): 46-48.
Cuéllar-Anjel, J., , B.White-Noble, P.Schofield R. Chamorro and D.V. Lightner. 2011. Selected families of white
shrimp Litopenaeus vannamei from Panama breeding program show important resistance to WSSV.
November/December AQUA Culture Asia Pacific Magazine, pp. 36-37
_______________
6
Annual reports of OIE Reference Centres, 2011