Development of quantitative PCR method for the rapid quantification of Paenibacillus larvae spores in pooled, extracted honey

Tuesday, November 15, 2022

1:54 PM – 2:06 PM PT

Location: Vancouver Convention Centre, Meeting Room 206

Presenting Author(s)

Muhammad Fahim Raza

postdoctoral fellow
University of Saskatchewan
Saskatoon, Saskatchewan, Canada

Co-Author(s)

MZ

Michael Zabrodski

University of Saskatchewan
Saskatoon, Saskatchewan, Canada
OO

Oleksii Obshta

University of Saskatchewan
Saskatoon, Saskatchewan, Canada
GW

Geoff Wilson

Government of Saskatchewan
Saskatoon, Saskatchewan, Canada
Midhun Sebastian Jose

Postdoctoral Fellow
University of Saskatchewan
Saskatoon, Saskatchewan, Canada
Sarah Biganski

Postdoctoral Fellow
University of Saskatchewan
Saskatoon, Saskatchewan, Canada
Jenna thebeau

Graduate Student
University of Saskatchewan
Saskatoon, Saskatchewan, Canada
IK

Ivanna Kozii

University of Saskatchewan
Saskatoon, Saskatchewan, Canada
Sarah Wood

Research Chair in Pollinator Health and Associate Professor
University of Saskatchewan
Saskatoon, Saskatchewan, Canada
ES

Elemir Simko

University of Saskatchewan
Saskatoon, Saskatchewan, Canada

American foulbrood (AFB) is a devastating bacterial disease of honeybees caused by Paenibacillus larvae. The North American beekeeping industry is highly dependent on antimicrobial metaphylaxis to prevent and control outbreaks of AFB, however, infectious P. larvae spores are extremely resilient to environmental factors and are impervious to antimicrobials licensed against AFB. We previously developed a logistically feasible AFB risk assessment tool for large-scale, commercial beekeeping operations through the culture of P. larvae spores in pooled, extracted honey at the yard (i.e., herd) level. By shifting AFB risk assessment from an individual hive to a herd-level perspective, beekeepers will have an improved ability to make evidence-based decisions regarding antimicrobial use, thus reducing the threat of antimicrobial resistance. Nevertheless, quantification of P. larvae spores by this culture-based strategy is time-consuming (~7-14 days). To improve the diagnostic turnaround time for herd-level AFB risk assessment, we will develop a TaqMan-based qPCR assay targeting a single-copy chromosomal metalloproteinase gene that will allow for reliable quantification of P. larvae. We anticipate that this qPCR-based strategy will provide a prognostic assessment of P. larvae spore concentrations in pooled, extracted honey samples within 2 hours. The use of this methodology will advance AFB risk assessment at the herd level through the rapid detection of P. larvae spores, consequently aiding beekeepers in making timely and improved evidence-based decisions regarding antimicrobial use against AFB at the herd level.