Performing Department
Biomedical & Diagnostic Sciences
Non Technical Summary
Meningeal worm (Parelaphostrongylus tenuis) infection is one of the most important infectious diseases plaguing the camelid industry today. Infected animals present with a head tilt, arching of the neck, incoordination, difficulty getting up, and/or gradual weight loss. Treatment for camelids and ungulates with moderate to severe clinical disease is often expensive, but unrewarding, despite use of numerous anthelmintics and supportive care. Recently a nested-PCR (nPCR) protocol was developed for the detection of P. tenuis DNAfrom fresh, frozen, or formalin-fixed, paraffin-embedded tissues. Although the nPCR assay for P. tenuis is successful at identifying the presence of the parasite in post-mortem samples, there is no consistently accurate live animal diagnostic test available. We propose to use the nPCR on cerebrospinal fluid (CSF) of camelids and other ungulates where P. tenuis infection is present or suspected. If validated, the nPCR would represent a live-animal assay for meningeal worm allowing veterinarians to make swift and accurate diagnosis of infected animals. The nPCR CSF test could additionally be used to accurately track the spread and outbreak of P. tenuis infections in different domestic animal and wildlife populations via phylogenetic analysis of P. tenuis DNA sequences. Because animal safety is our greatest concern, the samples used in this study will only be from animals presented for necropsy or from cases seen at UTCVM in which a CSF tap was part of the client-elected diagnostic workup. These samples will be considered surplus samples and according to the University of Tennessee's attending veterinarian this research project will not require IACUC approval.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
50%
Developmental
50%
Goals / Objectives
UsenPCR on cerebrospinal fluid of camelids and other ungulates where P. tenuis infection is present or suspected to develop alive-animal diagnostic test for P. tenuisto beused byveterinarians, diagnosticians, and researchers, allowingthem to makeswift and accurate diagnoses of infected farm animals.
Project Methods
Validate the nPCR assay in camelids and wild ungulates by testingarchivedcerebral spinal fluid (CSF) from camelids and other ungulate where P. tenuis is present or suspected. To date we have collected two camelid and five moose CSF samples. We will request surplus camelid CSF samples from other veterinary colleges and diagnostic laboratories to add to the sample queue, and we expect to acquire at least 30 samples for testing. If animals are euthanized and a necropsy performed, histological findings for each animal will be compared to the PCR results to correlate necropsy findings with molecular testing. In all DNA extractions, we will perform a negative extraction control, and negative and positive PCR controls will also be used. Amplicons from PCR-positive samples will be bidirectionally sequenced and resultant sequences will be phylogenetically analyzed and subjected to a GenBank BLAST analysis. CSF from camelids with CNS disorders that do not involve P. tenuis will also be tested to determine specificity of the test. It is our hope that our research will enhance the ability of veterinarians to diagnose P. tenuis in live camelids and ungulates. Following this study, we expect to advertise the results and availability of the P. tenuis PCR testing, and increase diagnostic revenue as an added benefit. Because animal safety is our greatest concern, the samples used in this study will only be from animals presented for necropsy or from cases seenin which a CSF tap was part of the client-elected clinical diagnostic workup. Thus these samples will be considered surplus samples and do not require IACUC approval. In addition to the assays and analysis,we will also review medical records and slides of tissues from all included cases. After we have compiled and analyzed all of our data,we will write a manuscript.We also plan to review numerous recently published articles on P. tenuis and on neurological diseases in camelids and ungulates so that we may better understand the various factors associated meningeal infections.