DIVA metabolomics: Differentiating vaccination status following viral challenge using metabolomic profiles

Abstract

Bovine Respiratory Disease (BRD) is a major source of economic loss within the agricultural industry. Vaccination against BRD-associated viruses does not offer complete immune protection and vaccine failure animals present potential routes for disease spread. Serological differentiation of infected from vaccinated animals (DIVA) is possible using antigen-deleted vaccines, but during virus outbreaks DIVA responses are masked by wild-type virus preventing accurate serodiagnosis. Previous work by the authors has established the potential for metabolomic profiling to reveal metabolites associated with systemic immune responses to vaccination. The current study builds on this work by demonstrating for the first time the potential to use plasma metabolite profiling to differentiate between vaccinated and non-vaccinated animals following infection-challenge. Male Holstein Friesian calves were intranasally vaccinated (Pfizer RISPOVAL®PI3+RSV) and subsequently challenged with Bovine Parainfluenza Virus type-3 (BPI3V) via nasal inoculation. Metabolomic plasma profiling revealed that viral challenge led to a shift in acquired plasma metabolite profiles from day 2 to 20 p.i., with 26 metabolites identified whose peak intensities were significantly different following viral challenge depending on vaccination status. Elevated levels of biliverdin and bilirubin and decreased 3-indolepropionic acid in non-vaccinated animals at day 6 p.i. may be associated with increased oxidative stress and reactive oxygen scavenging at periods of peak virus titre. During latter stages of infection, increased levels of N-[(3α,5β,12α)-3,12-dihydroxy-7,24-dioxocholan-24-yl]glycine and lysophosphatidycholine and decreased enterolactone in non-vaccinated animals may reflect suppression of innate immune response mechanisms and progression to adaptive immune responses. Levels of hexahydrohippurate were also shown to be significantly elevated in non-vaccinated animals from days 6 to 20 p.i. These findings demonstrate the potential of metabolomic profiling to identify plasma markers that can be employed in disease diagnostic applications to both differentially identify infected non-vaccinated animals during disease outbreaks and provide greater information on the health status of infected animals.

Technological Tools for the Early Detection of Bovine Respiratory Disease in Farms

“…Another promising combination would be the use of BAS with pathogens or biomarker detection. Metabolomics has been used as a DIVA approach to discriminate between BRD-infected and vaccinated animals [ 28 ], for classifying animals as BRD or non-BRD [ 98 ] and even for predicting the outcome of BRD cases [ 29 ]. Results could be integrated into a big data system and, through AI, the software could learn and integrate behavior differences between sick and healthy animals or vaccinated and infected animals, or could provide information about the severity of outcomes.…”

“…The accuracy of the BRD diagnosis and treatment through PLF may be increased when laboratory diagnosis is integrated. Thus, metabolomics may be able to differentiate BRD-infected from vaccinated animals [ 28 ] and predict disease outcomes after secondary bacterial infection [ 29 ]. Moreover, antimicrobial susceptibility testing could also be integrated into big data analysis to improve treatment for BRD.…”

Review on Discriminatory Tests of Immune Reaction due to Infection and Vaccination (DIVA Test)

“…The DIVA immune response can be detected by companion diagnostic tests such as enzyme linked immunosorbent assay. Companion serology based tests rely on seroconversion, and upon exposure to wild type virus the antibody response to DIVA vaccines will be masked by that of the wild type virus [5]. More recently, the use of genetic DIVA has been introduced, which is based on identifying genetic differences between live vaccines and the field viruses.…”

“…Successful differentiation of vaccinated from nonvaccinated animals using this technique requires concurrent vaccination and infection, with a narrow diagnostic window post-infection for detection of DIVA vaccine and viral genetic material. Furthermore, detection of vaccine genetic material only demonstrates exposure to the vaccine and not the successful generation of immune protection, limiting functionality in assessment of herd level immunity [5].…”

Profiling of local disease-sparing responses to bovine respiratory syncytial virus in intranasally vaccinated and challenged calves

“…In the post-genomic era, the rapid growth in 'omic' technologies has led to a greater understanding of physiological processes within the body, allowing researchers to understand how a particular perturbation in a biological system affects up-stream and downstream biological processes A C C E P T E D M A N U S C R I P T [10,11,12]. Proteomics, in contrast to transcriptomics, has the capacity to highlight actual changes in functional protein levels and provide information not only on proteins produced within tissue of interest but also on those effector proteins originating from the circulatory system.…”