Capability and Capacity of Ohio ADDL Enhanced as Member of the USDA NAHLN
Melanie Prarat, MS, Laboratory Operations Manager
The Ohio ADDL is a key player in the state’s animal disease outbreak response, as highlighted by the ongoing national incident of highly pathogenic avian influenza (HPAI). As the state’s only AAVLD-accredited veterinary diagnostic laboratory, the ADDL is expected to prepare and rapidly respond when there is an animal disease outbreak, particularly if the disease outbreak is of economic consequence to Ohio’s aquaculture, livestock or poultry production. The ADDL’s capability and capacity to respond is possible because the laboratory is part of the National Animal Health Laboratory Network (NAHLN), a network of veterinary diagnostic laboratories that provides ongoing disease surveillance, responds quickly to disease events, communicates diagnostic outcomes to decision makers, and has the capability and capacity to meet diagnostic needs during animal disease outbreaks.
In December 2023, the molecular diagnostics laboratory’s workload increased >1500% in response to multiple outbreaks of HPAI occurring at the same time throughout the state, with more than 9,000 tests conducted to detect avian influenza. The lab’s receiving and molecular staff have also worked 7 days/week to maintain continuity of business for farms impacted by HPAI. The lab’s participation in the NAHLN allowed for lab staff to be proficiency tested to perform the avian influenza testing and funding support provided through USDA National Veterinary Services Laboratory (NVSL) and USDA National Institute of Food and Agriculture (NIFA) allowed the ADDL to purchase the equipment necessary to perform high-throughput testing of hundreds of samples every day. The ADDL works closely with ODA’s Division of Animal Health and USDA staff to coordinate daily testing needs and stockpile reagents for surge testing that occurs during an outbreak.
For more information about the 2022-2024 detections of HPAI: https://www.aphis.usda.gov/aphis/ourfocus/animalhealth/animal-disease-information/avian/avian-influenza/2022-hpai
Porcine Reproductive and Respiratory Syndrome Annual Update and New Lineage Classification System
Jonah Perkins, MPH, Laboratory Scientist 2, Virology
Porcine reproductive and respiratory syndrome (PRRS) is one of the most impactful diseases on the swine industry, causing an estimated economic loss of $664 million per year in the United States alone. Clinical signs include respiratory disease in all ages and reproductive failure and late-term abortion in sows. PRRS is caused by the porcine reproductive and respiratory syndrome virus (PRRSV), of which there are two species: Betaarterivirus suid 1 (PRRSV-1, formerly known as the European genotype) and Betaarterivirus suid 2 (PRRSV-2, formerly known as the North American genotype). Most cases in the United States are caused by PRRSV-2, but small outbreaks of PRRSV-1 do occur.
 Number of samples submitted to the ADDL for PRRSV PCR, by month, from 2019 through 2/21/2024.
The ADDL offers a polymerase chain reaction (PCR) test to identify PRRSV infections as well as sequencing of the ORF5 region of the PRRSV genome. ORF5 is a highly variable region of the genome that encodes the major envelope protein (GP5) of PRRSV. Sequence analysis of this region can provide insight into genetic variation for monitoring distribution, emerging variants, and changes within herds. Restriction length fragment polymorphism (RFLP) typing has been used since 1998 to differentiate between vaccine associated and wild-type viruses. However, RFLP typing relies on very specific sites within ORF5 and can be changed by single nucleotide mutations. In 2022, the ADDL began reporting both RFLP type and lineage classification for PRRSV sequences, allowing comparison of the entire ORF5 region. This system assigned an individual sequence to one of nine lineages (L1-L9). In 2023, a research team at Iowa State University published a new classification system* that expanded the previous system to include two new lineages (L10 and L11) as well as multiple sub-lineages. This new classification system utilizes global PRRSV sequences and allows more specific comparison within individual lineages. The ADDL will now be reporting PRRSV lineage using this new classification system. Descriptions of each lineage can be found here.
 Number of PRRSV ORF5 sequences detected at the ADDL from 2018-2024. (2024 data as of 2/21/2024)
In 2023, the ADDL conducted 6,468 PRRSV PCR tests and sequenced 32 PRRSV isolates. The most common lineage observed at the ADDL was L1A, while SDRS reported L1C.5 as the most prevalent lineage nationwide. In the first few months of 2024, the ADDL has detected mostly L1C.5 lineage viruses.
*Yim-im W, Anderson TK, Paploski IAD, VanderWaal K, Gauger P, Krueger K, Shi M, Main R, Zhang J.2023.Refining PRRSV-2 genetic classification based on global ORF5 sequences and investigation of their geographic distributions and temporal changes. Microbiol Spectr11:e02916-23. https://doi.org/10.1128/spectrum.02916-23
 Front Row (L to R): Christy Ammons, Anne Parkinson, Dee Davidson; Back Row (L to R): Ervan Rodgers, Katie Esbenshade, Amanda Gillard, Ali Gibson, Rose Blocker
Meet the Section: Serology
Katie Esbenshade, BS, Laboratory Scientist 3, Serology
The Serology Section is responsible for high throughput/high volume regulatory, diagnostic, export, and monitoring/surveillance testing of Ohio’s livestock and poultry production systems using a variety of serological-based test methods. Section staff provide testing of diseases that are of economic significance to producers, that add additional value to their herds and flocks, guard the integrity and safety of Ohio’s food supply, and protect public health in the state from zoonotic diseases.
The section offers required animal health testing for shows and many public and privately managed sales in Ohio and nationally, while additionally supporting organizations and youth in the state during fairs and exhibitions locally and nationally. Regulatory testing is performed on a daily basis for the state’s licensed livestock/poultry auctions and markets to prevent the spread of disease into Ohio’s animal populations.
Serology section staff assist metropolitan and privately-owned zoos and wildlife conservation areas in maintaining the health of their collections and with testing for exports to other zoological/wildlife conservation areas nationally and globally. Section staff also provide regulatory and diagnostic testing services to fellow state agency the Ohio Department of Natural Resources to detect and monitor disease in the state’s fish and wildlife populations.
In all, the section manages a significant catalog of tests and completed 365,186 total tests in 2023. The most requested tests were the MG/MS ELISA, AI ELISA, EIA ELISA, and BT ELISA. Most recently a Q-Fever ELISA has been added and the staff are currently compiling methods for a Small Ruminant testing panel.
The Serology Section is led by Anne Parkinson (MS1) and includes Katie Esbenshade (LS3), Christy Ammons (LS2), Amanda Gillard (LS2), Ali Gibson (LS2), Rose Blocker (LS2), Dee Davidson (LS1), and Ervan Rodgers (LS1). Dee and Ervan are the newest members to the section. Dee joined the team in June of 2023, and holds a B.S. in Biology from Kentucky State University and brings 10 years of laboratory experience to the section. In her free time, she enjoys travelling and completing mind puzzles - sudoku being her favorite. Ervan recently joined the section in early 2024. He graduated in 2023 from The Ohio State University with a B.S. degree in Environmental Science with a concentration in molecular science. In his free time, he enjoys hiking, listening to music, or playing the piano.
The Serology Section is proud to be supporting the state’s diverse animal agriculture populations and protecting the integrity and safety of the food supply in Ohio.
Brucella canis Sample Submission Reminder
Dr. Melissa Simmerman, DVM, Division of Animal Health
Canine brucellosis is an infectious and zoonotic disease caused by the bacterium Brucella canis (B. canis) and has been reported worldwide. In dogs, canine brucellosis manifests with abortion, reproductive failure, enlarged lymph nodes, and can affect the osteoarticular system. The occurrence of asymptomatic infections in dogs is not uncommon. Treatment of infected dogs is associated with a high frequency of relapse and is not curative. Currently there are no commercially available vaccines for prevention of canine brucellosis
In Ohio, canine brucellosis is a reportable disease Rule 901:1-21-02 - Ohio Administrative Code | Ohio Laws and there are certain regulatory requirements that must be followed when testing a dog for brucellosis. These include:
- Any blood samples which are taken to conduct a canis test shall only be taken by an accredited veterinarian, or by a registered veterinary technician who is supervised by an accredited veterinarian
- Information regarding the owner of the dog
- Information regarding the dog
- Official identification of the dog
All regulatory requirements can be found here: Rule 901:1-5-12 - Ohio Administrative Code | Ohio Laws. If any of these requirements are missing, it can slow down the reporting process.
You can find answers to all you B. canis sample submission questions here: Canine Brucellosis Testing | Ohio Department of Agriculture
Additionally, any sample submission to ADDL needs to be accompanied by a submission form. A link to ADDL’s electronic submission form can be found here: General Submission Form (electronic) | Ohio Department of Agriculture. A hard copy of this form must be included with the samples and the electronic copy needs emailed to submissions@agri.ohio.gov
Best Practices for Submitting Swabs for PCR Testing
Ashley Johnson, MS, Molecular Diagnostics Section Head
The following are general guidelines for submitting swabs for PCR testing to the Ohio ADDL. Exceptions to these guidelines include, but are not limited to, avian sampling using either 3 or 5.5 mL BHI tubes and FMD sampling using TBTB media. If you have additional questions, you may call the ADDL at 614-728-6220 or email the Molecular Diagnostics section at AGR-Molecular@agri.ohio.gov.
- Swabs should be made of synthetic materials such as polyester and nylon. Organic materials such as cotton inhibit PCR and are unacceptable. 6” swabs are standard for most samples.
- Swabs should be placed in sterile tubes with no additives. Snap cap tubes are preferred, but red top and other tubes with a stopper are also acceptable. Tubes with additives such as heparin inhibit PCR and are unacceptable.
- For best collection practices, place the swab in the tube and snap the upper portion of the plastic shaft off. This prevents the portion of the shaft that is being handled from entering and contaminating the sterile tube.
- Swabs should be submerged in ~1 mL of viral transport media. If you do not have viral transport media (preferred), PBS and sterile saline are acceptable alternatives. Bacterial transport media can inhibit PCR and is unacceptable. A minimum volume of 0.2 mL is needed to perform the PCR test.
- Swabs must be kept cold (refrigerated or on ice packs) immediately after collection until they arrive at the ADDL and testing is initiated.
- Swabs must be tested withing 7 days of collection, but sooner is better.
Changes to Serology Testing Schedule
Anne Parkinson, BS, Serology Section Head
Due to the increasing number of tests coming into the ADDL, the Serology Section has made changes to their testing schedule to better accommodate our clients – effective immediately. Changes to the schedule are as follows:
All other testing in the Serology Section not listed here will remain on their regularly scheduled test days. Please note that samples MUST be in the lab the day BEFORE the test day to be reported close of business on the test day. For more information on Serology Section test days, please refer to our Tests and Fees page on our ADDL website.
 Top Left: JoJo Seay; Top Right: Dr. Dixie Burner; Bottom Left: Jada Davenport, Bottom Right: Dr. Daniela Peña
New ADDL Staff
Dominika Jurkovic, PhD, Central Receiving Section Head
Several new team members have joined the ADDL in the past year: Amanda "JoJo" Seay, Laboratory Scientist 3 in Bacteriology; Dr. Dixie Burner, DVM, DACVP, Veterinary Pathologist; Jada Davenport, Laboratory Scientist 2 in Virology; and and Dr. Daniela Peña, DVM, MS, Virology Section Head.
JoJo graduated from Florida Gulf Coast University with my B.A. in Biology and a minor in Interdisciplinary Studies. JoJo believes that to achieve the greatest level of success requires a collection of different approaches from different fields. She spent several years in pharmacology before moving from Florida to Ohio and then into food microbiology. In her free time, she watches an inordinate number of documentaries and enjoys reading scientific literature. JoJo and her husband are huge Columbus Blue Jackets hockey fans and can frequently be found at home games.
Dr. Dixie Burner earned her B.S. in Animal Science and a Doctor of Veterinary Medicine degree from the Ohio State University before dedicating 12 years to private practice as an equine practitioner. As an U.S. Army Veterinary Medical Officer, Dr. Burner managed small clinics, specializing in the care of military dogs and then completed an Anatomic Pathology residency at the Joint Pathology Center. During her time as an Army Veterinarian, Dr. Burner was involved in pathological studies, particularly focusing on trauma and biodefense in various animal species. Before joining the ADDL in October, Dr. Burner was a research pathologist for a medical device research company.
Jada graduated from North Carolina State University in 2023 with a Bachelor’s in Animal Sciences. Prior to that, she was engaged in research at the NCSU Comparative Biomedical Sciences Research Program and Veterinary School, focusing on developing therapeutic inhalant treatments for managing symptoms of pulmonary and lung diseases, notably COPD and Cystic Fibrosis. During her leisure time, she finds enjoyment in hiking, weightlifting, gardening, bird watching, and taking care of my plethora of animals from Chickens to Oscars to a Bearded Dragon.
Dr. Daniela Peña earned her DVM degree at the National University of Colombia in 2019. She completed a clinical microbiology residency at Purdue University and graduated in 2023 with a Master’s in Science in comparative pathobiology. During her residency at the Indiana Animal Disease Diagnostic Laboratory, Dr. Peña led a one-year comparison study to determine the most sensitive and rapid method for detecting Salmonella spp. from environmental samples. During the same period, she collaborated with the small-animal hospital in two studies characterizing the in vitro antimicrobial activity of different formulations used in small animal surgery. Dr. Peña is an inventor contributor in a nonprovisional patent dealing with the use of a carrier for sustained release levering antimicrobial properties of silver nanoparticles. Dr. Peña enjoys traveling to Colombia with her husband to visit their family while enjoying the tropical weather.
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