GD Update april 2013

GD animal health service

Update 1

2013

GD health Courses new oral fluid tests and neW inFlUenZa strain GiQs: association for international cooperation

GD Animal Health Service

Custom made animal health solutions GD Proficiency testing schemes R&D Veterinary laboratory GD Diagnostics Consultancy

w w w. g d d eve n t e r. c o m

| COLUMN

| CONTENT 04 New oral fluid tests and new influenza strain

05 PARR: a PCR-based test for

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unravelling canine lymphoproliferative disorders

06 GIQS: association for international cooperation

07 GD Health Courses 08 News & communications

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Multiple roles, one goal One of the aims of GD Update is to introduce you to the many roles GD plays in the field of animal health.

We are a researcher and developer, but also a service provider, an advisor, a partner and an entrepreneur. In every edition we try to show some of these facets.

This GD Update focuses specifically on GD’s role as developer. In our veterinary-diagnostic laboratory, we not only carry out millions of tests a year, but we also develop and validate many new tests. For example, last year GD introduced several tests for use in the oral fluids of pigs (Happy Bite). So far, PCR and ELISA tests have been validated for PRRS and PCV2.

New tests are coming shortly. Meanwhile a new H1N1 Influenza strain has been discovered in pigs. Will this impede the introduction of new tests? You can read more one page 4. On page 5 you can read about a new PCR-based test GD developed for unravelling canine lymphoproliferative disorders.

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CONTACT information

If you want to know more about what GD can do for you, please contact one of our staff members, responsible for international sales:

Ton de Gee, DVM PhD

Eveline Peereboom – de Haan, BSc

Manager international

Account manager

projects

Pharmacy

Annemiek Kolkman Slothouber, BSc

Madelon Bolderman, MSc

Product sales manager

Account manager

­Diagnostics and PTS

Feed

phone In the Netherlands 0900-1770, Outside the Netherlands +31 (0)570-63 33 91 Fax + 31 (0)570-63 41 04 | E-mail info@gddeventer.com | Website www.gddeventer.com | Mail address GD, P.O. Box 9, 7400 AA Deventer, the Netherlands | Delivery address for samples and post mortem m ­ aterial GD, CDM, Arnsbergstraat 7, 7418 EZ Deventer, the Netherlands.

This GD Update also looks at GD’s role as an international partner. On page 6, we explore the work of GIQS, a major network for scientific research in the agriculture and food sector. GD has been a member for more than 10 years and has participated in at least four international projects.

Finally, on page 7, you can read more about one of the ways in which GD fulfills its role as advisor by organizing Health Courses. Thanks to the many specialists working at GD, we have been providing Health Courses to participants from across the world for many years, much to their satisfaction.

In all of GD’s roles, we always keep one goal in mind, and that is our aim of contributing to improving animal health in the interest of animals, all stakeholders in the food chain and society as a whole.

Jan Jansen, PhD Managing Director

GD Update | April 2013 |

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TESTS | CoPy: THEO GEUDEKE, PHD, AND TOM DUINHOF, MSC

New oral fluid tests and new influenza strain

Happy Bite: the easy way to test for pathogens in pigs by investigating oral fluids.

In 2012, GD Animal Health Service introduced several tests for use in the oral fluids of pigs (Happy Bite).

So far, PCR and ELISA tests have been validated for PRRS and PCV2. New tests are coming shortly. Meanwhile a new H1N1 Influenza strain has been discovered in pigs in the Netherlands. Will this impede the introduction of new tests? oral fluid tests Back in 1986 in the US, a test was developed to detect antibodies against HIV in human saliva. This proved to be very successful, so soon other tests followed for several hepatitis viruses, measles, mumps and rubella. In recent years, tests have been developed for use in pigs.

Although the amount of antibodies in oral fl uids differs from that in blood, and potentially disturbing factors like enzymes and bacteria are present, oral fl uid testing has already proven to be a reliable tool. If samples are stored under cooled conditions, antibodies and pathogens can easily be detected.

However, it is essential to validate antibody tests (e.g.

ELISA tests) and antigen tests (e.g. PCR tests) thoroughly. Tests should be made suitable for use in oral fl uids and the results should be compared to results of serological tests. An important consideration is that, with oral fl uid tests, almost always groups of animals are investigated, whereas serological tests mostly involve individual animals. In fact, collected oral fl uids are pooled samples. So far, GD has established that its PCR and ELISA tests for both PCV2 and PRRSV in oral fl uids are very reliable.

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Currently GD’s R&D department is putting a great deal of effort into validating ELISA tests on several other pathogens for respiratory diseases like Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae and Infl uenza virus, and also PCR tests for Infl uenza (general and subtypes).

Preliminary results are very promising, so GD expects to be able to offer oral fl uid tests for these pathogens later this year.

new influenza h1n1 – new tests?

Sometimes unexpected but interesting challenges can occur. Last autumn GD discovered a new H1N1 Infl uenza strain in the Netherlands, a finding with potential consequences for test panels. On some Dutch farms, finisher pigs suffered from Infl uenza-like respiratory symptoms.

However, serological tests with the available haemagglutination inhibition tests (HI) on the three known strains H1N1, H1N2 and H3N2 all revealed negative results. On post mortem investigation, the pigs showed pathological lesions characteristic of an Infl uenza infection, and PCR tests were positive. This Infl uenza virus was then characterised as a thus far unknown H1N1 strain in the Netherlands. The strain is quite different from the one that is used in the regular HI test at the GD laboratory.

However, the newly discovered strain still belongs to the classic European H1N1 cluster and not to the pandemic H1N1-2009 cluster or the H1N2 group.

GD has subsequently developed a new HI test and the above-mentioned diseased animals showed significant antibody titres and seroconversion with this new test. By now the new HI test is incorporated in the test panel.

Copy: EVERT VAN GARDEREN, DVM PHD DIPL. ECPHM | TESTS

PARR: a PCR-based test for unravelling canine lymphoproliferative disorders Lymphoproliferative disorders re­present a significant clinical problem in dogs. It is crucial to differentiate neoplastic lymphoid disease; i.e. the lymphomas and leukaemias from reactive, hyper­ plastic lymphoid conditions.

Recently, GD developed a new PARR test which makes this differentiation possible.

Unravelling nodal or extranodal lymphoproliferative conditions frequently represents a diagnostic challenge for veterinary pathologists. Until now, they had to rely on cytology, immunocytology, histology and immunohistochemistry to solve this differential diagnosis. Whereas the combination of histopathology and immunohistochemistry is a powerful tool to accurately assess the neoplastic or reactive nature of a lymphoproliferative condition in a significant percentage of such cases, the essence of lymphoid neoplasia, i.e. the clonal expansion of neoplastic lymphoid cells, cannot be proved by this microscopic approach.

Lymphoproliferative disorders represent a significant clinical problem in dogs.

GD’s new PARR test for formalin-fixed and paraffin-embedded tissues will supply additional information about the clonal (i.e. neoplastic) nature of lymphoid cells, or the polyclonal (i.e. reactive) nature of lesional lymphoid cells.

Basics of PARR PARR is the abbreviation of PCR for antigen receptor rearrangement. Antigen receptor rearrangement represents a crucial phase in the biology of B-cells and T-cells, since this process enables the generation of the antigen receptor diversity that is a hallmark of the lymphoid system. This diversity is generated through recombinations in the DNA that encode the immunoglobulins in B-cells and the T-cell receptor (TCR) in T-cells. PARR for B-cell probing is based upon recombinations that occur in the V, D and J genes that encode the antigen-binding sites of the immunoglobulin heavy chains (IgH), whereas in PARR for T-cell probing the DNA segments that are a result of V and J genes recombinations for the gamma-TCR gene are amplified.

The resulting PCR products are best visualized by capillary electrophoresis.

Sensitivity and specificity of PARR GD’s PARR is based upon the latest information on primer sequences. The PARR has been tested and validated and the overall specificity is 90%, and sensitivity 92%. It will generate results even when small lymph node Tru-Cut biopsies are used as DNA source. Inherent to these sensitivity and specificity data, it is therefore always prudent to interpret PARR data in combination with other findings, such as history, histopathology and immunohistochemistry.

Additional information about our PARR test Please feel free to contact us via email if you need additional information about the test, price and lead time: Jeanine Brussee, Doctor of Veterinary Medicine j.brussee@gddeventer.com Judith Hofland, Market manager companion animals j.hofland@gddeventer.com

GD Update | April 2013 |

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INTERNATIONAL PROJECTS | Copy: PETER VAN DER WOLF, PHD

GIQS:

association for international cooperation GIQS is an international association that aims to stimulate, initiate and coordinate international projects in the field of quality improvement in agriculture. GD Animal Health Service has been a member for more than 10 years.

In these 10 years, GD has participated in at least 4 large international GIQS projects, including the development of a system to collect swine herd management data which was up till then only gathered on paper. By developing an OCR (Optical Character Reading) based form, these data could be put in a database to be analysed. Within this project, standard operating procedures were also developed to advise farmers on certain illnesses.

Early warning system for swine fever A second project involved developing an early warning system for Classical Swine Fever based on fallen stock numbers. Using these numbers a response time could be obtained that was comparable to reporting by swine farmers and veterinarians. The advantage of this system was that it was independent of the willingness to report suspected outbreaks. One drawback was the operational costs of such a system.

MRSA In a large international study (called ‘SafeGuard’) involving many partners, multiple animal species and humans,

More about GIQS GIQS stands for “Grenzüberschreitende Integrierte Qualitätssicherung”, which is German for Cross-boarder Integrated Quality Assurance. Members are the Universities of Bonn, Wageningen, Warsaw, Göttingen and “Hochschule Rhein-Waal”, public agricultural authorities like “Landwirtschaftskammer NRW and Niedersachsen”, several swine production integrations like EGO, EZG and EGR in Germany, animal health services in Germany and the Netherlands, VION food group, and several SMEs, both in Germany and the Netherlands.

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Peter van der Wolf, veterinarian at GD Animal Health Service, has been a member of the GIQS board for more than 10 years.

GD participated in a longitudinal study on MRSA in pig farms, at both animal and herd level.

Monitoring weaned pigs In another part of the same SafeGuard project, GD participated in the development of a quality-assured monitoring system for weaned pigs. The aim of this project was to establish advice on the uniform health status of weaned pigs destined to be sold to fattening farms. This was done in collaboration with the animal health services in North Rhine-Westphalia and Lower Saxony and with the help of several veterinary laboratories.

A spin-off of the monitoring project was the development of proficiency testing schemes (ring trials) for PCRs for Circovirus (PCV2), Brachyspira hyodysenteriae and Lawsonia intracellularis. The first two where developed by GD, the last of these was developed by BioScreen in Hanover.

Checklists for Salmonella risk factors Finally, GD was involved in the development of web-based checklists for Salmonella risk factors in both finishing farms and slaughterhouses as well as in cutting plants.

A weighing system for the questions enabled the evaluation of companies both over time (repeated checks) and between companies within the same category.

Financing and coordination Many of these projects were financed by the EU or its EUregion offices. Co-financing came from national governments, the German federal government and provinces in the Netherlands. GIQS coordinated all of these projects, both technically and financially. More information, and its many international projects, can be found at www.giqs.org.

Copy: EVELINE PEEREBOOM, BSC | EDUCATION

GD Health Courses Globally, there is a huge demand for animal health courses. For many years, GD Animal Health Service has been a valued partner in providing courses on pig, poultry and cattle health.

On site demonstration by Klaas Peperkamp (veterinary pathologist at GD) of organs from pigs with and without pathological changes.

The aim of the GD Health Courses is to provide attendees with the most recent scientific insights and practical expertise. The training sessions are organized by GD’s Pharmacy Department, which focuses on the development and execution of test methods and studies compliant with internationally accepted regulations. Also, the Pharmacy Department is specialised in providing advice in this field.

GD: expert partner Thanks to the team of specialists working at GD, the GD Health Courses have been organized for years with great satisfaction of attendees. GD has veterinarians, scientific researchers and specialised staff members. All expertise acquired is utilised when studies are set up and executed, scientific articles and reports are written and when GD is participating in international conferences and lectures.

GD has modern state-of-the-art facilities for carrying out tests and studies.

Wide range of GD Health Courses Courses can be compiled at any desired level. The subject of a course can be broad, treating a number of diseases associated with one particular animal species, or more specific, with only one organ system of a certain animal species being studied. Each course is supplemented with expertise from different professional fields (for instance bacteriology, immunology and histology). Cases provide participants with useful tips and hints to apply in their profession. At completion of the course all attendees receive a certificate of participation.

For groups of 10 persons or more, GD can prepare a customized course in the field of certain animal diseases for one or more species. The training sessions take place at GD, however on-site courses are also possible.

More information: Eveline Peereboom - de Haan, Key Account Manager Pharmacy, pharma@gddeventer.com

Training session at GD.

GD Update | April 2013 |

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news & communications GD AT THE WAVLD 2013

TUrNKEY LAB PrOJECT

GD Animal Health Service is one of the sponsors of the 16th International Symposium of the World Association of Veterinary Laboratory Diagnosticians (WAVLD) that will take place in Berlin from June 5-8, 2013. Experts from all over the world will meet in the Estrel Hotel & Convention Center. We look forward to meeting you there. If you would like to make an appointment to meet us in Berlin or if you would like to receive more information, please send an email to annemiek Kolkman, a.kolkman@gddeventer.com.

GD has developed a new product for companies and organizations that want to establish a veterinary diagnostic laboratory. Together with our partners we offer a turnkey project for the establishment of a laboratory, including training laboratory personnel and field support staff, such as veterinarians. We assist with the design of the laboratory and supply high-quality furniture and equipment. The project is adaptable to a client’s specific requirements.

The project may be limited to a small laboratory with limited tests, but might also consist of a large laboratory with several disciplines, automated processes and the requirement to obtain quality accreditation.

We are proud that the WAVLD Board of Directors has voted unanimously for Dr. Gerard Wellenberg to succeed Dr. Gerrit Borst on the Board. Gerard Wellenberg is a senior scientist in Molecular Biology & Virology at GD Animal Health Service. He is also active as veterinary consultant in national and international projects.

Dr. Gerard Wellenberg, senior scientist at GD, will be a member of the WAVLD Board of Directors.

MOrE iNfOrMATiON? on our website www.gddeventer.com you will find information about our products, services and various GD projects as well as the latest information about submitting samples to GD. Furthermore, we have an e-letter to keep you informed about new tests, recent publications by GD employees, proficiency testing schemes and much more. you can register for our e-letter on the website.

GD, Po box 9, 7400 aa Deventer, the netherlands, t. +31 570 63 33 91, F. +31 570 63 41 04 www.gddeventer.com info@gddeventer.com

Upgrading and renovating existing laboratories are also options.

Please contact Gerard Wellenberg (g.wellenberg@gddeventer.com) if you want a partner for the establishment of a veterinary laboratory equipped to meet all your requirements.

PTS PrOGrAMME, JUNE–OCTOBEr 2013 In addition to the regular use of internal controls, laboratories should participate in inter-laboratory comparison or Proficiency Testing Schemes (PTS). Participation is one of the requirements of the ISO 17025 guideline for laboratory quality systems. GD Animal Health Service is formally accredited Cat. no

GD PTS

according to ISO 17043:2010 for the organization of PTS for antibody detection.

Below you will find our PTS programme from June–October.

Please go to www.gddeventer.com for the online PTS application form. Feel free to contact us via email: pts@gddeventer.com Closing date for registration

Shipment of the samples

Submission of test results

Final report + certificate

11 July 2013

Week 30 2013

30 August 2013

Week 40 2013

15 August 2013

Week 35 2013

04 October 2013

Week 45 2013

19 September 2013

Week 40 2013

08 November 2013

Week 50 2013

24 October 2013

Week 46 2013

13 December 2013

Week 4, 2014

JUNE VLDIA172*

PTS for Infectious Bursal Disease Virus (IBDV)/ Gumboro antibody detection in serum

VLDIA219*

PTS for Newcastle Disease Virus (NDV) antibody detection in serum

VLDIA235*

PTS for Small Ruminant Lentiviruses (SRLVs)/ Maedi Visna Virus (MVV)/Caprine Arthritis Encephalitis Virus (CAEV) antibody detection in serum

JULY VLDIA255*

PTS for Avian Metapneumovirus (aMPV)/Turkey Rhinotracheitis (TRT) antibody detection in serum

VLDIA277*

PTS for Avian REO Virus (ARV/REO) antibody detection in serum

VLDIA292

PTS for Infectious Laryngotracheitis (ILT) antibody detection in serum

AUGUST VLDIA232*

PTS for Avian Infl uenza Virus (AI) antibody detection in serum

VLDIA233*

PTS for Salmonella (SA) antibody detection in chicken serum

VLDIA171*

PTS for Salmonella (SA) antibody detection in porcine serum

OCTOBEr VLDIA291

PTS for Brachyspira hyodysenteriae detection in porcine feces via Nucleic acid Amplification Techniques (NAT) e.g. PCR

VLDIA295

PTS for Swine Infl uenza Virus (SIV) antibody detection in serum

* officially accredited according to the international guideline ISO 17043:2010