GD Update 1 april 2015

Teaming up for animal health

Update

Intestinal integrity:

Testing the effect of nutrition and additives New test available for

genotyping BVD

Introducing our specialists

The importance of diagnosing

M. gallisepticum

01 APRIL 2015

Teaming up for animal health

www.gdanimalhealth.com: our new website website

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Diagnostics ProďŹ ciency testing schemes z Lab services (e.g. laboratory tests per animal species) z Consultancy (e.g. setting up lab units and contract research) z Courses & trainings z GD

GD, P.O. Box 9, 7400 AA Deventer, the Netherlands, T. +31 (0)570-63 33 91, F. +31 (0)570-63 41 04 www.gdanimalhealth.com, info@gdanimalhealth.com

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GD Health hasAnimal a new international TheAnimal website of GD Health offerswebsite: you

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04

Intestinal integrity

06

Our specialists: passionate and skilled

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GD applies for GLP

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New BVD-genotyping test

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Diagnosis of M.g.: Accuracy is crucial

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| COLUMN

| CONTENT

Special time

News & communications Every day, we are working to improve our services and the opportunities for collaboration with our partners in animal health. This is a bit of a special time, though, since we are currently going through some major changes.

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A big change for GD is the completion of the Good Laboratory Practice (GLP) certification. One of the key staff members involved in implementing GLP in our organisation would like to tell you the story behind the story.

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All the stories here are about our people and their enthusiasm for their work. The team of specialists at GD dedicated to small ruminants for example, is internationally known for their knowledge and expertise.

We are proud to tell you a bit more about their work, their dedication and their stories of animals and their owners.

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

Annet Velthuis, BSc Head International and Special Accounts Ad Interim

Eveline Peereboom, BSc Senior account manager Pharmacy

Annemieke Spitse, BSc

Madelon Bolderman, MSc

International Services Coordinator

Senior account manager Feed

Annemiek Kolkman, BSc

Fanny Nieuwenhuis

Senior account manager Laboratories and Test kit manufacturers

These stories show once again that everything we do is with and through people. In that sense it is business as usual: dedicated staff doing high-quality work and working on our mission to team up for animal health in the interest of animals, their owners and society at large.

YNTE SCHUKKEN, DVM, PHD MANAGER ANIMAL HEALTH

Senior account manager Pharmacy

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@gdanimalhealth.com | WEBSITE www.gdanimalhealth.com | MAIL ADDRESS GD, P.O. Box 9, 7400 AA Deventer, the Netherlands | DELIVERY ADDRESS FOR SAMPLES AND POST MORTEM MATERIAL GD, CMD, Arnsbergstraat 7, 7418 EZ Deventer, the Netherlands.

GD Update | April 2015 |

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INTESTINAL HEALTH | COPY: MADELON BOLDERMAN, MSC & NAOMI DE BRUIJN, DVM, DIPL. ECVP

Intestinal integrity GD Animal Health supports you in the The intestinal tract is a very dynamic system, it supplies the body with nutrients for growth and performance, but it also plays an essential role in the immune system. Therefore optimal intestinal health is essential for maximum productivity and efficiency in the poultry industry.

GD Animal Health’s vast repository of expertise in the field of intestinal health ensures you receive relevant advice on the current intestinal health of your animals and on the effect of nutrition and additives on intestinal integrity.

Furthermore, our team of specialists will support you on the study design of your research.

Histological analyses In both layers and broilers intestinal health is determined by the genetic background, feed composition and intestinal integrity. Maximal productivity requires an intestinal tract that is able to absorb available nutrients, and forms a functional barrier that offers protection against potential harmful substances within the intestinal lumen.

Intestinal health problems that arise at early age are usually caused by impaired

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intestinal maturation. In this critical period, the intestinal cells (both enterocytes and goblet cells) develop into the functional cells responsible for absorption of nutrients and protection of the delicate intestinal barrier. Intestinal health problems that occur at later age both have a direct relationship with the integrity and functionality of intestinal cells, as with the presence of inflammatory cells within the intestinal wall.

GD Animal Health offers various histological analyses that are useful to evaluate the effect of your products or concepts at poultry farms. For evaluating feed and additives, we can support you in the selection of appropriate tests that generate useful insights on your products. The extent of the support varies from a basic level to more advanced support that may involve the conduct of the entire experiment.

infectious diseases and feed components.

The current state of the mucus can be estimated by a goblet cell analysis, which measures the number of goblet cells per villus surface area, and the size of these cells. This test measures the effect of your product on intestinal mucus.

Inflammatory cells The presence of macrophages and CD3, CD4 and CD8 inflammatory T lymphocytes can be specifically investigated. These cell types play a key role in the development of the immune system and regulate the immune-system response to infections.

The presence of these cells and their number determine to a large degree the health status of the intestines. GD Animal Health has a specific histological test to demonstrate these cells.

Presence of intestinal viruses and coccidiosis

selection of tests We give recommendations regarding the necessary random-sample size, the sampling procedures and the time of sampling.

If required, GD staff can assist in sample collection and shipment.

Chronic enteritis score (CE score) This score gives a reliable impression of the intestinal health of laying hens and pullets. The CE score runs from 0 (healthy intestines) to 5 (chronic enteritis). Since this score shows dynamic changes in time, repetition of this score visualizes the effect of a product on intestinal health after a period of exposure.

Villus: Crypt ratio This ratio indicates the functional intestinal surface area available for digestion.

The villus length determines the capacity for digestion and absorption of nutrients.

The crypt depth determines the degree re-

generation of the intestinal tract. Various infectious and non-infectious elements can lead to damage and healing of the intestinal epithelium, which can disrupt the normal ratio between villi and crypts.

Feed composition and additives are able to influence this ratio.

Intestinal viruses and coccidiosis have a direct effect on the integrity of the intestines, especially in chickens. The functional intestinal surface area is negatively influenced by these infections, and furthermore an enteritis induces increased numbers of inflammatory cells. These intestinal changes can even lead to abnormalities in the bacterial balance of the intestines, which in turn can lead to an exacerbation of the negative effects. GD Animal Health has a PCR that can detect intestinal viruses and coccidiosis in quantitative terms.

Study design Goblet cell test The intestinal epithelium is covered by mucus that forms a barrier between the intestinal wall and the intestinal lumen. In addition to the protective function, the mucus is also important in the transportation of nutrients through the intestinal wall and local immunity in the intestines. Mucus comprises an acid and a neutral component that can be visualized by histological staining.

The production and loss of mucus during digestion is a dynamic process, influenced by the bacterial microbiota, but also by

The feed industry is continually improving and developing new products or concepts to positively impact poultry health in relation to intestinal health. Due to their vast experience of such tests, the GD specialist will assist you with your study design.

More information Want to know more about what GD can do for you? Then contact our veterinary pathologist with specific knowledge in this field of expertise, Naomi de Bruijn via +31(0)570 660385 or at n.d.bruijn@gdanimalhealth.com.

GD Update | April 2015 |

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OUR SPECIALISTS | COPY: YONI PASMAN, MA

Passionate and skilled The specialists of GD Animal Health focus their daily work on gathering and spreading their extensive knowledge of animal diseases. How did they obtain their passion and what are their focus areas? Every edition of the GD Update, we emphasise on a specific sector. This time: the specialists of the department of Small Ruminants.

Piet Vellema, DVM, PhD, dip. ECSRHM European specialist small ruminants Head of the Department of Small Ruminant Health

“After studying Veterinary Sciences, I started practicing at a veterinarian’s, but soon I discovered that the routine there wasn’t for me. I gained my doctorate in 1996 with a study into vitamin B12-deficiency in grazing lambs. The results of my research still apply.”

I have also been involved in crisis meetings for foot-and-mouth disease, blue-tongue and Q-fever. And that is very demanding, but also gives you lots of energy, which I guess comes from my drive to solve things.”

Leading the pack Sampling “I’m particularly interested in zoonotic agents. But also, when we begin a study or launch a new product, When we tested the Q-fever vaccine, I monitored the temperature of fifty goats on the first day.

This means you gain a keen insight into what goes on at the farm. I try to sample a little bit of every step, so I can tell the manufacturer a few days later what’s going on in the field.

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“At GD our motivation is to always be the first to obtain the latest knowledge. Last year, for instance, we were the first to discover resistance to Monepantel, and we have a great deal of experience on zoonosis. We might be a small country, and have only relatively few animals, but researchers in neighbouring countries see that we are doing things right.”

Piet Vellema

At home All our specialist enjoy breeding sheep on a small scale. On Holland’s famous green meadows, Piet keeps Friesian milk sheep, Rene Texel and Karianne Charollais sheep.

René van den Brom, DVM, PhD, dip. ECSRHM European specialist small ruminants Veterinarian, specialised in monitoring animal health, Q-fever and aborted foetuses “I was interested in small ruminants already at a young age, especially sheep.

Abortion has my special interest, specifically infectious cases, as these are very frustrating. Researching aborted foetuses is essential for farmers, not just for the animals themselves, but also for his own health and that of his family and visitors, and sometimes even the community.”

including the GD. Because the start of the study coincided with me commencing employment at GD, I was involved from the start. Which is why my thesis focused on the veterinary aspects of a Q-fever outbreak in the Netherlands between 2005 and 2012. On February 5th 2015 I obtained my doctorate.”

Humans and animals Q-fever “When I joined GD in 2008, a large-scale study was initiated in the Netherlands into the Q-fever outbreak in 2007 and 2008. During the outbreak, hundreds of people in the Netherlands were infected with Q-fever. The most probable cause was dairy goat herds with problems with abortion due to C. burnetii. A study was initiated with people from various institutes,

“GD works on animal health in the interests of both humans and animals. This goal really appeals to me and I am keen to contribute to it. There are still patients suffering the after effects of Q-fever, and the disease is still a sensitive subject for many goat and sheep farmers. In social and political terms, Q-fever is still a live issue.”

René van den Brom

Karianne Lievaart-Peterson, DVM, PhD, Grad. Cert. In UL&T, dip. ECSRHM European specialist small ruminants Veterinarian, specialised in paratuberculosis, Maedi Visna and CAE and udder health “During my Veterinary Sciences studies I was sure that farm animals were my primary interest. When I was asked after my study to help research a project about artificial insemination (A.I.) in goats, I was certain – this is for me. I gained my doctorate in small ruminants shortly after that, focusing on Maedi Visna, CAE and sperm. I worked at the faculty till 2007 as a tutor and put small ruminants on the curriculum there with elective subjects that are still being given. After that, I worked at an Australian university to help to set up a school of Veterinary Sciences.

Here I focused primarily on the large sheep industry: wool as well as meat.”

Paratuberculosis

Karianne Lievaart-Peterson

“At GD I was involved from the start (2011, ed.) in the paratuberculosis project. This disease is linked to CAE and Maedi Visna, and relates to the same white blood cells. So paratuberculosis was not such a mystery to me, specifically how the

disease behaves. Together with dairy goat farmers, I studied how we could manage the disease. Norway is doing particularly well here, but they have a totally different structure. We can learn much from them, but their system cannot be implemented point-to-point, unfortunately.”

Unique “The Netherlands is a specialist in the field of goats. The Dutch goat industry and the size of goat farms here is unique in global terms. People who want to get involved in the intensive dairy-goat industry could learn a great deal from this.

And in France, Belgium and the United Kingdom too, we are seeing more and more dairy-goat farms. That’s both exciting and interesting. GD possesses a great deal of in-house expertise, but in the end it’s our lab that is our core business, and really is the hub of our most productive operations.”

GD Update | April 2015 |

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GLP | COPY: GERT JAN BOELM, PHD

applies GD for GLP Currently GD Animal Health is in the process of applying for Good Laboratory Practice (GLP).

The aim is to conduct trials and/or tests for which GLP can be claimed from 2016 onwards.

GLP will broaden the portfolio of GD’s services for our industrial partners, hence GD Animal Health wants to become GLP-compliant. The GD laboratory has already been ISO-17025 accredited by the Dutch Accreditation Board (RvA) for a large number of laboratory tests (registration number L120) and this system is used as a scaffold to enrol GLP over its entire site. As a veterinary multidisciplinary laboratory, GLP implementation will make GD Animal Health unique, even in global terms.

Trials During implementation of GLP, GD Animal Health uses the expertise of a well-known consultant in this area. The focus for the first half of 2015 is the installation of

Soon, our laboratory will be GLP-certified.

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a QA-department and finalisation of procedures required for GLP. In the second half of 2015 GD Animal Health will opt for conducting trials for which GLP can be claimed.

These trials will be necessary for governmental recognition of GD Animal Health as a GLP-compliant laboratory, which we hope to receive in 2016.

Outsource entire projects Principally, the portfolio of GD Animal Health always will contain non-GLP trials and tests, which can be performed simultaneously with GLP trials. Becoming GLP-compliant will make it easier for industrial partners to outsource entire projects to GD Animal Health.

COPY: KEES VAN MAANEN, PHD | BVD GENOTYPING

GD introduces

genotyping test

The GD Animal Health laboratory has a comprehensive array of tests for the detection of Bovine Virus Diarrhoea (BVD) or BVD antibodies. A real-time PCR is now being added that can distinguish between BVD genotypes 1 and 2.

Two BVD genotypes are recognised in western Europe, and various subtypes have been defined within each genotype. GD’s laboratory has now introduced a real-time PCR which can distinguish between the two genotypes. Both genotypes can present a wide range of pathologies, varying from sub-clinical infections, respiratory complaints, abortion, mild or serious enteritis, through to secondary infections caused by suppressed immunity. Moreover, BVD type 2 is associated with thrombocytopenia in calves, which can result in extensive haemorrhaging.

Validation The identifier can be applied for detailed testing in order to genotype individual BVD-positive samples (blood samples, necropsy material or organs). In order to validate this test, the results were compared with those of external reference institutes, and proved perfectly comparable.

Until now, this test has been deployed on a project basis in order to monitor the prevalence of BVD genotype 2 in the Netherlands. The prevalence of this genotype has been extremely low in recent years, despite the presence

of the German BVD type-2 strain, which was recently responsible for high morbidity and mortality rates in the veal sector.

BVD causes major economic losses, and the Netherlands, with its large bovine population, has a reasonably high prevalence of BVD infections. 71 percent of the farms not certified to be BVD-free has BVD antibodies in the bulk milk. Furthermore antibody monitoring among young cattle showed BVD to have recently circulated at 14 percent of the dairy farms and at 19 percent of all other farms. On the other hand, 34 percent of the dairy farmers have now achieved BVD-free/negative status.

Application Should you wish to apply for BVD genotyping, this can be done via re-registration of the sample(s) in question. Please send an e-mail to klantenservicelab@gddiergezondheid.nl, with reference to ‘10937 BVD-genotyping’, the relevant dispatch number and sample identification number(s). The test costs 57.50 euros per sample (excluding VAT and basic costs).

GD Update | April 2015 |

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M.G. | COPY: ANNEKE FEBERWEE, PHD & WIL LANDMAN, PHD

Diagnosis of Mycoplasma gallisepticum

Accuracy

is crucial

Although Mycoplasma gallisepticum (M.g.) occurs worldwide, its prevalence has decreased significantly in reproduction stock due to the implementation of compulsory eradication programmes. The success of any disease control programme is highly dependent on the accurate identification of M.g. infected and free flocks.

M.g. infections can be detected routinely by both serology and PCR. Serology is mostly used for large-scale monitoring as it is cheaper than PCR. The latter is often used as a confirmatory test. Besides test

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sensitivity and specificity, other factors that may affect the accuracy of the diagnosis are sample frequency and sample size as well as flock sensitivity and specificity. Flock diagnosis is never based on

one sample: a sufficient number of birds has to be sampled in order to also enable the detection of the disease in a flock whereas relative small number of chickens are infected. For example, to detect a prevalence of M.g. antibodies of 5 or 10 percent in a flock size of thousand birds or more sixty or thirty blood samples have to be taken, respectively (Table 1).

False positive and false negative test results may occur due to the test itself (low sensitivity and low specificity, respectively) or other factors. Regarding the latter for instance, false positive serological test results have been obtained after using serum of bad quality, after analyzing sera from flocks that had been

Percentage prevalence 50

40

30

25

20

15

10

5

2

1

0.5

0.1

1000

5

6

9

11

14

19

29

57

138

258

450

950

5000

5

6

9

11

14

19

29

59

147

290

564 2253

10000

5

6

9

11

14

19

29

59

148

294

581 2588

> 10000

5

6

9

11

14

19

29

59

149

299

596 2995

Flock size

Table 1. Sample sizes required at a certain flock size to detect different M. gallisepticum sero prevalences with 95% confidence.

Sample

Material

Article No.

Mg RPA

Blood

Serum

10386

Mg/Ms combination

Blood

Serum

10389

ELISA

Blood

Serum

10388

Mg ELISA

Trachea swab / FTA

DNA

10677

Mg PCR

card *Prices available on request (a.spitse@gdanimalhealth.com)

Table 2. M.gallisepticum diagnostic tests at GD Animal Health

treated with inactivated vaccines three to four weeks prior to sampling, in flocks infected with antigenic related mycoplasmas (e.g. M. synoviae), etcetera. Poor immunological seroresponse to mycoplasmas (described in turkeys) and antibiotic treatment may result in a false negative serological test outcome. Regarding PCR, contamination may induce a false positive result, while antibiotic treatment or the presence of PCR inhibitors may give a false negative result.

Vaccination Live or inactivated vaccines may interfere with the detection of M.g. field strains by serology (live and inactivated vaccines) or by PCR (live vaccines). If inactivated vaccines are used, monitoring may be performed using the more expensive PCR test instead of serology. However, if live vaccines are used, monitoring of these vaccinated flocks is not possible as long as a differentiating M.g. PCR is lacking.

Importance of M.g.

Considering the complexity of infection dynamics and all factors affecting the accuracy of the flock diagnosis of M.g., it is highly recommended to confirm the alleged infection status of flocks using a second test.

M.g. is the most important mycoplasma species in commercial chickens. It may cause chronic respiratory disease, downgrading of carcasses, decreased growth and egg production, and reduced hatch-

ability rates. M.g. can be transmitted vertically (from parent to offspring) and horizontally (by direct and indirect contact). Although M.g. occurs worldwide, its prevalence has decreased significantly in reproduction stock due to the implementation of compulsory eradication programmes. In the Netherlands for instance, its seroincidence has decreased significantly over the past 15 years. The percentage of reproduction farms infected with M.g. decreased significantly from 1.4 to less than 0.5 percent, in layer farms from 10.5 to 2.4 percent and in meat turkey farms from 12.5 to 0.0 percent. This success has been possible amongst others due to the accurate identification of M.g. infected and free flocks.

GD Update | April 2015 |

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Commercial document needed when submitting samples from EUmember states

Jantijn Swinkels earns PhD

In the battle against Q-fever, the key is to use a broad range of measures. That’s what GD’s veterinarian and European specialist René van den Brom concluded in his thesis about the veterinarian aspects of a Q-fever outbreak. He gained his doctorate on February 5th with his study ‘Veterinarian aspects of a Q-fever outbreak in the Netherlands between 2005 and 2012’.

We would like to kindly inform you that we need a commercial document with each sample submission from EU-member states. Therefore we kindly ask you to enclose the commercial document with each submission. You can find the commercial document on our website www.gdanimalhealth.com in the menu ‘Forms’.

For more information about the document or any other questions related to submitting samples, please do not hesitate to contact Annemieke Spitse via a.spitse@gdanimalhealth.com.

WVPA, Cape Town, South Africa GD Animal Health will be present with a booth at the WVPA, the International Congress of the World Veterinary Poultry Association. The WVPA will be held from 7 to 11 September 2015 in Cape Town, South Africa.

MORE INFORMATION? On www.gdanimalhealth.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 eletter 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.gdanimalhealth.com info@gdanimalhealth.com

René van den Brom obtains doctorate

It’s not always a good idea to treat dairy cows with antibiotics for prolonged periods to cure mastitis. GD veterinarian Jantijn Swinkels discovered this with his doctoral research paper entitled ‘Extended antibiotic treatment of persistent bovine mastitis during lactation - efficacy, economics and social infl uences’. On December 9th, he defended his thesis, in which he compared the bacteriological cure, visible recovery, the cell-count cure and the economic impact of prolonged treatment with those of a standard treatment.

This study demonstrates that prolonged treatment is only useful where the mastitis is caused by S. aureus and udderhealth management is suboptimal. In other cases the end result does not justify the costs of prolonged treatment.

Jantijn Swinkels carried out this study alongside his work at GD Animal Health.

In the past he has worked as a practicing veterinarian carrying out research at Boehringer Ingelheim and Intervet (later MSD).

Although the aim of the vaccination (to reduce aborted foetuses and discharge) is realised, the separate effects of other measures are difficult to assess, as they are implemented more or less at the same time. The total range of measures was effective. Read more about René van den Brom on page 6 & 7 of this GD Update.

GD present at WAVLD GD Animal Health will be present as one of the exhibitors at the 17th International Symposium of the Word Association of Veterinary Laboratory Diagnosticians (WAVLD) held from 15-18 June 2015 in Saskatoon, Canada.

The symposium brings together senior level executives, veterinarians and scientists from across a varied range of sectors to present and learn about advancements in science and laboratory techniques related to Veterinary Laboratory Diagnostics. The 2015 symposium will cover a broad range of themes, including New Technologies, Control & Management of Endemic Disease, Detection, Management & Control of New & Emerging Disease, Wildlife & One Health & Turning Laboratory Data into Intelligence. We look forward to meeting you there. If you would like to make an appointment, please send an email to Annemiek Kolkman via a.kolkman@gdanimalhealth.com. For more information about the symposium go to www.wavld2015.com.