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New Pathogen Discovery Diagnostic Laboratory Under Construction At UKVDL

The University of Kentucky Veterinary Diagnostic Laboratory is excited to announce the opening of a New Pathogens Discovery Diagnostic Laboratory this fall. The laboratory will serve as an extension to the existing UKVDL Molecular Biology, Virology and Bacteriology laboratories to help identify potential pathogens, both endemic and emerging, that may can cause morbidity and mortality in animals.

Pathogen detection will be done using cutting-edge Next Generation Sequencing (NGS) technology, metagenomics and bioinformatics methods. The Gluck Equine Research Center has plans for a similar laboratory, primarily for infectious disease discovery research.

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Traditionally, the identification of infectious disease agents in specimens/animals submitted to the UKVDL has been done through microscopic observation (morphology and Gram stain), bacterial media culture, viral cell culture, polymerase chain reaction (PCR) and other traditional diagnostic methods.

Reliable cultivation of bacteria was first done by Louis Pasteur and Robert Koch in the late 19th century, and both were named the Fathers of Microbiology for their contributions.

The first virus (vaccinia) was grown in cell culture in 1913. The PCR method was discovered in 1985 by Kary Mullis, an industrial chemist. Unfortunately, conventional bacterial and viral cultures can take days, weeks and even months to lead to pathogen identification and sometimes grow nothing at all. Unlike culture, PCR can only identify specific agents that might be suspected.

In 2015, the UKVDL acquired a MALDI-TOF instrument (Matrix-Assisted Laser Desorption/Ionization-Time of Flight) for identification of culturable bacteria and fungi by using mass spectrometry and laser technology. The instrument can identify most agents within minutes, improving the turn-around time by about 24 hours. However, the organism must first be cultured, which still requires about 24 hours or more depending on the microorganism.

The implementation of NGS and leading-edge bioinformatics systems in the new UKVDL laboratory will assist UKVDL pathologists and microbiologists and Gluck scientists in the discovery of new pathogens and identify known pathogens more precisely than traditional methods. It will also enable innovative disease surveillance methods for endemic and emerging animal diseases.

Metagenomics is a method to study organisms that are difficult or impossible to culture, some of which may be potential pathogens. We are happy to announce that Litty Paul, PhD, an experienced investigational biologist, has been recruited to the UKVDL to design and launch the new NGS laboratory.

Tissue samples from sick or deceased animals are sent to a diagnostic laboratory by a veterinarian or an owner curious about the cause (etiology) of an illness or death loss. Once this new method is validated, the laboratory will extract and sequence the genetic material from the samples and store them in databases. The databases will then be analyzed by state-of-the-art bioinformatics software that can identify and classify the community of microorganisms present in the samples, which can aid in disease diagnosis. Even more powerful is the ability to compare analyses from sick and healthy animals to help identify and characterize new and emerging pathogens.

Several of the 60-member veterinary diagnostics laboratories (VDLs) of the National Animal Health Laboratory Network (NAHLN) are already utilizing this technology.

This past spring, many Bluegrass horse farms were experiencing outbreaks of severe diarrhea in very young foals, and traditional diagnostic tests were not useful in providing the answers. Samples were gathered systematically from many local Thoroughbred farms by Emma Adam, DVM, PhD, DACVIM, DACVS, assistant professor, research and industry liaison at the Gluck Equine Research Center, and brought to the UKVDL for testing in the microbiology laboratory of Erdal Erol, DVM, MSc, PhD, head of diagnostic microbiology and professor at the UKVDL.

He then forwarded select samples to the Texas A&M VDL and the University of Missouri for electron microscopy (EM) for testing. Both labs identified rotavirus on EM. Oddly, all PCR tests for rotavirus group A run at UKVDL had been negative. Feng Li, DVM, PhD, professor and William Robert Mills Chair in Equine Infectious Disease at the Gluck Center, sent fecal specimens to the South Dakota State University VDL for NGS and metagenomics analysis. This collaborative effort identified a novel group B rotavirus in the foal specimens. Until this time, only group A rotavirus was targeted by the UKVDL PCR test.

This is a prime example of the outstanding collaborative diagnostic effort among the UKVDL, Gluck Center and other institutions, and demonstrates how these new methods can rapidly identify a new, emerging pathogen.

This data enabled Erol and Li to swiftly develop and validate a new PCR that can now identify the new group B rotavirus in about four hours. This test is now offered by UKVDL. The good news is the UKVDL is now prepared to detect the new virus in upcoming foaling seasons.

Once NGS technology is in place at the UKVDL and the Gluck Center, new and emerging pathogens in horses and other animals will be detected faster and easier than ever before.

Erdal Erol, DVM, MSc, PhD, head of diagnostic microbiology and professor, and Litty Paul, PhD, both from the UK Veterinary Diagnostic Laboratory, provided this information.

New Strain Of Hendra Virus Discovered In Australia

Australian researchers have determined a new strain of Hendra virus was responsible for the unexplained death of a horse in 2015. Hendra virus has a mortality rate of nearly 80 percent in horses. It's also lethal to humans, with a 60 percent mortality rate. Since its discovery in 1994, Hendra virus has killed over 100 horses and four humans in Australia.

The new strain of Hendra virus had not been seen in routine equine biosecurity testing, but has been detected in grey-headed flying fox samples from 2013. The samples collected in Adelaide share 99 percent of their sequence identity with the virus infecting the 2015 horse sample.

Prior to this discovery, Hendra virus was only known to occur in areas where black flying foxes and spectacled flying foxes live. The research team has developed diagnostic lab techniques to identify the new strain. They expect the currently available Hendra virus vaccine will be effective against the newly discovered strain.

The research team, led by Dr. Peter Reid, suggests that unwell horses living in areas where flying foxes are present should be treated with the same precautions as horses that have had a positive Hendra diagnosis, even if their initial Hendra test comes back negative.

The scientists suggest that preventative measures to curtail the spread of Hendra virus be put in place, including vaccination, good hygiene, the use of personal protective equipment and putting biosecurity measures in place.

EHV Outbreaks And Equine Insurance

Equine insurance is utilized by many horse owners, whether the horse is kept for competition, breeding, or as a family pet.

With the EHV-1 outbreak across the United States and Europe, many horse owners are considering insuring their horses. But would insurance cover a horse that may have been exposed to a contagious disease? Horse Network recently delved into that question.

Each insurance carrier has its own set of rules regarding insuring a horse during a contagious disease outbreak; some may have a heightened sense of awareness about where the horse is located and what it has been exposed to, while others proceed with business as usual.

Insurance agencies and carriers deal with viral disease outbreaks similarly to how they handle weather events: they cover exactly what they say they will cover for current customers – nothing changes. However, they may limit their risk by not accepting new clients in the storm's path.

Insuring a horse that has been imported from Europe, where the EHV outbreak is intensifying, may not be possible, so contacting an equine insurance company before the import is planned is imperative.

Atypical Myopathy: Why Are Only Some Horses Affected?

Equine atypical myopathy is an unusual, deadly disease which is poorly-understood. If a group of horses is turned out together, only some will be affected by the debilitating disease, caused by the ingestion of the seeds or saplings of specific trees in the Acer family.

Equine atypical myopathy typically affects horses that live on pasture in the spring and fall. It has been seen in Europe since the mid-1990s. Some 2,500 cases have been reported in the last decade.

Researchers believe intestinal bacteria may play a role in why some horses get the disease while herdmates don't. Drs. Christina Wimmer-Scherr, Bernard Taminiau, Benoît Renaud, Gunther van Loon, Katrien Palmers, Dominique Votion, Hélène Amory, Georges Daube and Carla Cesarini hypothesized that fecal microbiota may play a role in whether or not horses became affected by the disease.

The study team used fecal samples from 59 horses with atypical myopathy that were referred to Belgian clinics: 29 horses that survived and 30 that didn't. The team also used 26 healthy horses that shared pastures with the affected horses during the outbreak periods.

The team concluded that horses suffering from atypical myopathy have different fecal microbiota than their healthy counterparts. The changes were more severe in horses that didn't survive the disease.

Horses impacted by atypical myopathy will suddenly become stiff and weak; they will have a rapid heart rate and dark urine. As the disease progresses, the horse may be unable to rise, experience breathing issues and eventually die. There is no cure and treatment of symptoms is often unsuccessful. The mortality rate is between 43 and 97 percent.

Read the article here.