Detection of necrotic enteritis risk through non-invasive monitoring of Clostridium perfringens in feces.

Necrotic enteritis (NE), caused by the gram-positive, anaerobic bacterium, Clostridium perfringens, results in an estimated $6 billion in annual economic losses to the global poultry industry. C. perfringens is part of the normal microflora of the poultry gastrointestinal tract, but damage to the intestinal epithelium can lead to increased cell proliferation and production of toxins which gives rise to disease. Conventional methods to diagnose NE are invasive and are typically performed after the disease has manifested. In a pen trial using a necrotic enteritis model, we demonstrate that non-invasive monitoring in feces can detect an increase in average C. perfringens counts that correlates with higher lesion scores and reduced body weight gain in birds with NE. This was achieved using a C. perfringens-specific fluorescence in situ hybridization (FISH) probe and a high throughput platform (PIPER ^TM ) for concentrating, imaging, and automated counting of labeled cells. The assay detects all tested strains of C. perfringens while excluding closely related bacteria, including other Clostridium species commonly found in poultry feces. The counts by the PIPER assay show a linear log-log relationship with the counts obtained by conventional plating of spiked fecal samples. Furthermore, fecal samples can be stored for up to 72 h without a dramatic loss in C. perfringens detection on this platform using the recommended sample collection and storage conditions. This non-invasive assay could open new opportunities for early identification of NE risk as well as for quantifying intervention efficacy.
Competing Interests: Declaration of interests The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Margaret Kiss reports financial support was provided by Ancera. Teja Shidore reports financial support was provided by Ancera. Diane Buhr reports financial support was provided by Ancera. Justin Morano reports financial support was provided by Ancera. Thabani Dhlakama reports financial support was provided by Ancera. James Barton reports financial support was provided by Ancera. Matthew Ritch reports financial support was provided by Ancera. Brian Westgate reports financial support was provided by Ancera. Zoe Moscato reports financial support was provided by Ancera. Mark Fiandaca reports financial support was provided by Ancera. Anastasia Sevostiyanova reports financial support was provided by Ancera. Craig Kiebler reports was provided by Ancera. Matthew Zwilling reports financial support was provided by Ancera. Charles Copley reports financial support was provided by Ancera. Margaret Kiss reports a relationship with Ancera that includes: employment and equity or stocks. Teja Shidore reports a relationship with Ancera that includes: employment and equity or stocks. Diane Buhr reports a relationship with Ancera that includes: employment and equity or stocks. Justin Morano reports a relationship with Ancera that includes: employment and equity or stocks. Thabani Dhlakama reports a relationship with Ancera that includes: employment and equity or stocks. James Barton reports a relationship with Ancera that includes: employment and equity or stocks. Matthew Ritch reports a relationship with Ancera that includes: employment and equity or stocks. Brian Westgate reports a relationship with Ancera that includes: employment and equity or stocks. Zoe Moscato reports a relationship with Ancera that includes: employment and equity or stocks. Mark Fiandaca reports a relationship with Ancera that includes: employment. Anastasia Sevostyanova reports a relationship with Ancera that includes: employment and equity or stocks. Craig Kiebler reports a relationship with Ancera that includes: employment and equity or stocks. Matthew Zwilling reports a relationship with Ancera that includes: employment and equity or stocks. Charles Copley reports a relationship with Ancera that includes: employment and equity or stocks. Anastasia Sevostyanova has patent #Systems, devices, and methods for analysis (2024/0287585A1) pending to Ancera, Inc. Mark Fiandaca has patent #Systems, devices, and methods for analysis (2024/0287585A1) pending to Ancera, Inc. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2025. Published by Elsevier Inc.)