INTERNATIONAL On-going report by Jean Llewellyn and Claartje van Andel: In last month’s issue of BN, we published a ‘Chronology’ of WFFS that has now been updated, as promised, for this issue. However, given the volume of news, the update is fairly brief – including only the most important announcements. Since last month, BN has been in correspondence with Dr. Nena Winand PhD of Cornell University in Ithaca, NY, who was responsible for developing the genetic test for WFFS Type 1, which became commercially available in 2013.

Nena Winand has worked at Cornell since 1986, after completing several years of veterinary pathology preceptorships at a number of highly respected schools of veterinary medicine. Winand’s impressive professional resumé lists her research interests as ‘medical genetics and comparative genetics, emphasis: equine genetics.’ She currently holds five patents, including the genetic test for WFFS Type 1. DNA testing On May 20, the Dutch Horses.nl website published an article that appears to question the validity of the DNA testing carried out, so far, in the Netherlands by the Van Haeringen Laboratory (VHL) on behalf of a number of major stud farms. Winand reported that in Europe only Laboklin has the license to test. The Van Haeringen Laboratory (VHL), which does all the tests for KWPN and NRPS, does not. “This is not credible science, but business.” Winand continued by saying, “The laboratories that test without a license have not had access to the data on this condition, only the patents issued by Cornell and the patent applications. That is not professional and certainly not science”.

The editors of Horses.nl contacted the VHL laboratory, whose spokesperson, Leanne van de Goor, said, “We have had the patent status checked by a lawyer. The patent is not valid in the Netherlands. It is completely legal that we carry out the test in the Netherlands”. Van de Goor continued by saying, “The mutation for WFFS is published in a scientific article. Based on this information we have developed and validated the test”. The publication to which VHL refers is the case report by Mounthoux et al., published in BMC Veterinary Research.

Winand responded to VHL’s claim by saying, “The mutation is not in that publication, the mutation is in the Cornell patent application. You can never set up a valid test without the proper controls, for which you need markers of homozygous WFFS cases and carriers. What VHL and other labs have done is to copy a published DNA test protocol without the controls to validate it.” Winand reports that VHL and a lab in Ireland (where horses from KWPN partner WSI are being tested) would be wise to get a sub-license from Laboklin. “I hope that these laboratories can continue to test, but then through a sub-license of Laboklin. I do not want to interfere with studbooks that use VHL to test, but testing without proper controls is not ideal.” Horses.nl has asked the Van Haeringen Laboratory for a reaction, but has not received one, to date. (End of Horses.nl item.)

Breeding News has been in direct communication with Nena Winand since the Horse.nl item was published, and she confirmed that, “At this time the patent covers testing only in Germany, France and the United Kingdom. It does not apply to testing in The Netherlands and Ireland. It does mean that Animal Genetics in Wales must sub-license from Laboklin to test legitimately. I don’t know where their EU lab is, but the same would apply if it was in Germany or France. As for testing at any other labs, these are commercial labs and not per se research labs. “The only labs that have worked with me to validate testing are Laboklin, and more recently UC Davis. It is best when establishing testing to validate results for a sample set of tests with a lab that has access to the original material.

It isn’t required, as I am unaware of any regulation of the animal DNA testing industry, but it is an issue of best practice. “I cannot comment on the accuracy of testing at these other labs but I hope it will be high, for the sake of breeders. It is, of course, ideal for KWPN and WSI to funnel testing through VHL, as that is the lab that provides their parentage verification services. “Cornell patented the test mainly to control where testing was done to ensure that quality services were being provided. There is no required external oversight of the animal DNA testing industry, as there is for human genetic disease diagnostic testing (although admittedly the landscape for regulation may differ between countries).”

Solutions

During BN’s correspondence with Nena Winand, I asked her about the best measures studbooks and breeders can take in terms of both short-and long-term management of WFFS: In terms of what registries and breeders can do to manage the trait: “I am an advocate for compulsory testing for known genetic defects through registries, and public disclosure. This gives breeders complete control over their own programs and it gives registries the data and power that they may need going forward, in the event it is desirable to make policy changes. I am also an advocate for coupling testing for defects to parentage verification as this really helps prevent fraudulent sample submission. Putting this strategy in place now will ensure transparency and foster ethical business practices. It’s important to remember that breeding is a commercial enterprise, and as such is subject to relevant statutes and law.

“Data from broadly testing by many registries and large-scale population surveys will demonstrate accurately what the carrier frequency is in today’s horses and may fine-tune the amount of testing needed in the future. Whether carriers should be removed from breeding programs is presently an individual decision of the owner, or by policy of individual stallion stations [and studbooks].

“Because this trait segregates in the bloodlines that have been popular in recent decades and that have produced the level of international performance that we see in showjumping and dressage, I cannot advocate for completely removing carriers from the breeding population. Basing selection solely against a single mutation may ultimately relieve selection on performance parameters and change the level at which the horses perform. I am rather inclined to recommend selecting outstanding non-carrier bloodstock over a longer period (decades) going forward. One hopes this would gradually reduce the allele frequency in the population. One cannot eliminate recessive traits from bloodstock unless all carriers are removed from the breeding population. Ultimately it isn’t for me to decide what each registry should do-that is their decision, but genotype knowledge is power for both individual breeders and for registries.”

Updated chronology

Retroactively added: 2007: A French veterinarian who was practicing in northern Germany at the time, states that this was when she saw her first cases of WFFS in Hanoverian and Oldenburg foals. However, as there was no DNA test for WFFS commercially available until 2013, it would be impossible to prove retroactively without direct familial DNA samples.

March 2014: Suspected case of WFFS at Spy Coast Farm (notified by Lisa Lourie), but it went undiagnosed at the time.

2018 April 28: (Learned after the May issue went to print.) Gert-Jan Van Olst announced on Paardenkrant-Horses.nl that Van Olst Horses will assume the cost of mare testing for any owner who wishes to use Everdale (a known carrier stallion) in 2018.

May 4: Two stallion sons of Everdale confirmed as WFFS carriers: Inclusive (2013/ KWPN out of Doutzen x Uptown x Kennedy) and Inspire (2013/KWPN out of Don Furstia C-Two x Don Romantic x Don Schufro).

May 4: NRPS studbook announces mandatory WFFS DNA testing for all their approved horses and ponies. According to NRPS Chairman Ronald Funke Küpper, “We assume that it is only a few isolated cases, but if we do not take action now, we will never find out, and the disorder will have the opportunity to spread further.” It is also the NRPS’s intention to give mare owners the opportunity to have their mare or foal tested soon. (NRPS are using VHL for their testing.)

May 4: The Warmblood Studbook of Ireland (WSI) issues a statement saying that WSI has “never been informed of a case of WFFS in our population, and we are unaware of any Irish Warmblood foals that have died with symptoms similar to WFFS.” They are, however, proactively seeking Department of Agriculture support for an on-going investigation regarding the prevalence of the defective gene in the Irish Warmblood population, and to deliver an education program to breeders.

May 7: The Oldenburg Horse Breeders’ Society and German Oldenburg Verband publish a statement encouraging members to “stay informed and to test breeding stock. […] We encourage stallion owners to test and publish the results of their stallions. Owners of WFFS carrier stallions should require mare owners to produce a negative WFFS genetic test result in order to fulfill a breeding contract.”

May 9: Team Nijhof and Stal Brouwer in the Netherlands confirm that for reasons of transparency they will be testing all their stallions (KWPN registered as well as non-KWPN).

May 20: An announcement that three Blue Hors stallions, out of 10, have tested positive for WFFS and have been temporarily taken out of breeding: Blue Hors Veneziano (2010/DWB: Vivaldi - Dorina x Blue Hors Don Schufro) – in breeding since 2015 with four foal crops: Blue Hors Emilio (2014/Westf: Escolar - La Fraviata x Lauries Crusador xx) – one foal crop: Blue Hors Londoner (2015/ Hann: Londontime - Duffy x Danone I) – 2018 has been his first covering season. Stud farm manager Esben Møller expressed his shock at this result, and Blue Hors veterinarian Jesper Møller Nielsen has advised breeders and owners with offspring from these three carrier stallions to have their offspring tested.

May 23: A representative from the Trakehner Verband confirmed that the German studbooks are on the eve of setting up a study to evaluate, among other parameters, the prevalence of WFFS, which will be orchestrated by the German Federation.

May 24: The Glock Horse Performance Centre announces the DNA test results for all their stallions, saying that only Glock’s Total US (2012: Totilas [KWPN] - Sondra [Hann] x Sir Donnerhall I [Oldbg]) is a carrier of the WFFS gene. Their other Glock-prefixed stallions have been listed as non-carriers: Dream Boy, Flavio, Johnson TN, Romanov, Toto Jr, Trafalgar, Voice and Zonik. Glock have been quoted as saying that Total will continue to be available for breeding, but they will only be accepting mares with non-carrier status.

May 24: The KWPN studbook published a list of DNA test results with the message: “The KWPN decided to test the active KWPN approved stallions for WFFS. The breeding season is in full swing and breeders are now making the choice of a partner for their mares. By publishing the WFFS status of the KWPN-approved stallions, we are informing breeders in a timely manner.” The results show that 96% of KWPN stallions are non carriers. WFFS-positive stallions are the already announced Everdale, as well as his sons, Inclusive and Inspire, alongside the aforementioned Glock’s Total US, plus Guardian S (2011: Bodyguard Moorland - Amadin S x Trento B); Habanna (2012: Vivaldo - Dimare x Van Gogh); Indian Rock (2013: Apache - Crisjena x Vivaldi); Jack (2014: Fernando-H - Narobi x Colino [Holst]); Jubel ES (2014: Harley VDL - Maxime O x Lauriston [Oldbg]); Regino (1998: Animo - Regina Adelheid Z [Zangersheide] x Rebel II Z [Hann]).

May 24: Katrinelund – another major Danish stud farm confirmed that all their stallions have been tested but, so far, only two results are known: Hesseljøj Donkey Boy (2012/DWB Era Dancing Hit - Søbakkehus Kamilja x Milan) and Romeo (2012/Hann: Romanov/ Blue Hors Romanov - Wolke JM x Wolkentanz), of which the latter has tested positive as a WFFS carrier: Quoted as saying that they believe the whole issue is a ‘storm in a teacup’ as no cases have yet been reported in Denmark!

May 25: An email to BN from Jeannette Nijhof confirmed that all the Team Nijhof stallions have been DNA tested for WFFS, and all are non carriers.

May 25: A press release is received from the Swedish Warmblood Association... In light of the recent events regarding Warmblood Fragile Foal Syndrome (WFFS) the Board of the Swedish Warmblood Association has made the following decision on May 24, 2018: All stallions active or intending to be active in SWB breeding will be tested for the WFFS carrier status. This also applies to stallions activated with the one-mare license. During 2018 the information on the WFFS status of each stallion tested will posted on the SWB website under the heading ‘Hingstlicenser’ as soon as the information becomes available.

Starting in 2019, a stallion's WFFS status is mandatory before he can be activated. This also applies to the one-mare licenses. WFFS will be added to the SWB list of defects that has to be officially reported. SWB will initiate a research project to map out the frequency of carriers in the SWB population. The study will begin with this year's foal crop in order to identify the current status of carriers. The study will be finalized this coming fall/winter, in due time before the breeding season of 2019 is to begin. Samples from this year's foal crop will be available as registrations of the foals begin. The study will not generate any individual results for the foals, but will give a clear indication of the carrier frequency in the population of foals born 2018.

Conclusion

Published on May 27 on the WFFS Awareness Group Facebook page, extremely helpful words of wisdom written by Tom Reed of Warmblood Studbook of Ireland....

Warmblood Studbook of Ireland (WSI) has acted quickly and responsibly in response to WFFS1, as have many other studbooks. But there are many reasons why some studbooks may appear slow to respond that is nothing to do with cover-ups, selective sampling, putting heads in sand, etc., reasons that have been attributed on social media. Some examples of these reasons are:

1) Each studbook has its own governance structure, which varies in layers and complexity. A flat and simple structure can produce quick decisions; a vertical and complex structure that requires consultations with or approvals by committees, members councils, etc. will slow down the decision-making process.

2) Each studbook has to make a response plan while taking into account legal liability. National law, the studbook’s constitution, and the stallion inspection contract all will determine whether the studbook is free to analyse a DNA sample and report the result without the owner’s permission or if informed consent must be obtained from the owner.

3) Studbooks have varying degrees of technical and managerial competence in their top management teams. Some studbooks will be required to consult extensively with external experts to develop a response strategy.

4) Many studbooks began DNA analyses (for parentage testing, for example) only in recent years. These studbooks face the logistical challenge of collecting DNA samples from stallions while ensuring the integrity of the sample collection process and chain of custody.

5) Other studbooks that have conducted parentage testing for many years will face the following problem: many of the DNA profiles they use have been supplied by other studbooks, and cooperation will be required from the source studbooks to have the WFFS1 tests performed. DNA profiles supplied on a piece of paper cannot be used to perform the WFFS1 test.

6) Stallions that have died may not have DNA samples or data on file with a laboratory that can be used for WFFS1 testing, even if parentage testing has been done for many years using the stallion’s original DNA profile. Breeders can rightly demand transparency and communication from their breed associations and studbooks. But breeders have to give their organisations time to manage appropriate responses to this challenge.

BN will continue to publish appropriate WFFS news and updates for as long as necessary.