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Rapid identification and genotyping of the honeybee pathogen Paenibacillus larvae by combining culturing and
Hannes Beims1,2, Martina Janke1, Werner von der Ohe1
1Lower Saxony State Office for Consumer Protection and Food Safety, Institute of Apiculture, Celle 29221, Germany.
Background:
American Foulbrood (AFB) is a devastating disease of honey bee (Apis mellifera) larvae caused by the spore-forming, Gram-positive bacterium Paenibacillus larvae. In most countries, the law requires mandatory reporting of AFB to the veterinary authority.
Aim And Methods:
To speed up detection and genotyping of P. larvae spores, we compared different culturing protocols on Columbia sheep blood agar and developed a new multiplex quantitative polymerase chain reaction to distinguish between the two relevant P. larvae genotypes enterobacterial repetitive intergenic consensus (ERIC) I and ERIC II.
Results And Conclusion:
As confirmed by P. larvae reference strains and field isolates, the new identification and genotyping protocol halves the time of current workflows, lessens labor-intension, allows a higher throughput of samples for monitoring, and permits a faster intervention to prevent the spread of AFB.
Insights
A new method rapidly detects and genotypes Paenibacillus larvae, the cause of American Foulbrood (AFB) in honey bees. This faster detection aids in preventing the spread of this devastating bee disease.
Area of Science:
- Veterinary Microbiology
- Apiculture Science
- Bacterial Pathogenesis
Background:
- American Foulbrood (AFB) is a severe infectious larval disease in honey bees (Apis mellifera).
- It is caused by the spore-forming bacterium Paenibacillus larvae.
- Mandatory reporting of AFB is required by law in many countries.
Purpose of the Study:
- To accelerate the detection and genotyping of Paenibacillus larvae spores.
- To develop a more efficient diagnostic workflow for AFB management.
- To differentiate between the two main P. larvae genotypes, ERIC I and ERIC II.
Main Methods:
- Comparison of various culturing protocols on Columbia sheep blood agar.
- Development of a novel multiplex quantitative polymerase chain reaction (qPCR) assay.
- Validation using P. larvae reference strains and field isolates.
Main Results:
- The new protocol significantly reduces processing time compared to existing methods.
- It decreases labor intensity and increases sample throughput for monitoring.
- Genotyping of P. larvae strains was achieved efficiently.
Conclusions:
- The developed multiplex qPCR assay offers a faster and more efficient method for AFB diagnosis.
- This rapid detection enables quicker interventions to control disease spread.
- The protocol supports enhanced surveillance and management strategies for American Foulbrood.
