<abstract xmlns="http://www.w3.org/1999/xhtml">

<sec>
<h3>Introduction</h3>
<p><italic>Mycobacterium avium</italic> subsp. <italic>paratuberculosis</italic> (MAP) is the causative agent of paratuberculosis, a chronic infectious intestinal disease occurring in domestic and wild ruminants. Early diagnosis of infected herds enabling timely adoption of control measures is tremendously important in view of the fact that the disease has a significant economic impact on farmers. The aim of this study was to evaluate the possibility of rapid detection of viable MAP on small ruminant farms based on environmental sample examination using a novel phage-based test named Actiphage.</p>
</sec>
<sec>
<h3>Material and Methods</h3>
<p>A total of 9 fresh and 28 frozen (8 or 11 years at -70°C) environmental samples originating from paratuberculosis-affected farms were analysed for the presence of MAP by four different diagnostic methods: Actiphage combined with real-time PCR targeting insertion sequence <italic>900</italic> (IS<italic>900</italic> qPCR), conventional phage amplification assay, culture (frozen samples only), and direct <italic>ĪS900</italic> qPCR.</p>
</sec>
<sec>
<h3>Results</h3>
<p>Viable MAP was detected in one fresh environmental sample using Actiphage–IS<italic>900</italic> qPCR. None of the frozen samples tested positive using this diagnostic approach, which was consistent with the results of culture examination also providing information on viability.</p>
</sec>
<sec>
<h3>Conclusion</h3>
<p>This study describes other possible and innovative uses of phage-based methods in paratuberculosis control strategies. The Actiphage-qPCR was found to be less laborious than culture and provided results within six hours, suggesting that it may be a valuable tool for rapid initial determination of the infectious status of farmed animals based on environmental sample examination.</p>
</sec>
</abstract>

Introduction
Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis, a chronic infectious intestinal disease occurring in domestic and wild ruminants. Early diagnosis of infected herds enabling timely adoption of control measures is tremendously important in view of the fact that the disease has a significant economic impact on farmers. The aim of this study was to evaluate the possibility of rapid detection of viable MAP on small ruminant farms based on environmental sample examination using a novel phage-based test named Actiphage.


Material and Methods
A total of 9 fresh and 28 frozen (8 or 11 years at -70°C) environmental samples originating from paratuberculosis-affected farms were analysed for the presence of MAP by four different diagnostic methods: Actiphage combined with real-time PCR targeting insertion sequence 900 (IS900 qPCR), conventional phage amplification assay, culture (frozen samples only), and direct ĪS900 qPCR.


Results
Viable MAP was detected in one fresh environmental sample using Actiphage–IS900 qPCR. None of the frozen samples tested positive using this diagnostic approach, which was consistent with the results of culture examination also providing information on viability.


Conclusion
This study describes other possible and innovative uses of phage-based methods in paratuberculosis control strategies. The Actiphage-qPCR was found to be less laborious than culture and provided results within six hours, suggesting that it may be a valuable tool for rapid initial determination of the infectious status of farmed animals based on environmental sample examination.

Utilisation of Actiphage in combination with IS<italic>900</italic> qPCR as a diagnostic tool for rapid determination of paratuberculosis infection status in small ruminant herds

Journal of Veterinary Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Utilisation of Actiphage in combination with IS qPCR as a diagnostic tool for rapid determination of paratuberculosis infection status in small ruminant herds

Introduction
Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis, a chronic infectious intestinal disease occurring in...

Utilisation of Actiphage in combination with IS900 qPCR as a diagnostic tool for rapid determination of paratuberculosis infection status in small ruminant herds

Published Online: Sep 20, 2023

Page range: 347 - 352

Received: Feb 20, 2023

Accepted: Jul 04, 2023

DOI: https://doi.org/10.2478/jvetres-2023-0041

Keywords
Actiphage-qPCR, environmental samples, subsp., phage amplification assay, small ruminants.

© 2023 Monika Beinhauerova et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Utilisation of Actiphage in combination with IS900 qPCR as a diagnostic tool for rapid determination of paratuberculosis infection status in small ruminant herds

Published Online: Sep 20, 2023

Page range: 347 - 352

Received: Feb 20, 2023

Accepted: Jul 04, 2023

DOI: https://doi.org/10.2478/jvetres-2023-0041

KeywordsActiphage-qPCR, environmental samples, subsp., phage amplification assay, small ruminants.

© 2023 Monika Beinhauerova et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
Actiphage-qPCR, environmental samples, subsp., phage amplification assay, small ruminants.