Molecular Detection of Enterococcus faecalis as a Secondary Agent of European Foulbrood Disease in Honey Samples

Document Type : Original Articles

Authors

Department of Honey Bee, Silk Worm and Wildlife Research, Razi Vaccine and Serum Research Institute (RVSRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

10.22092/ari.2026.370975.3893

Abstract

Introduction: European foulbrood (EFB) disease is the most important bee larvae disease in honey bees, caused by Melissococcus plutonius, and subsequently the larvae are infected with secondary bacteria such as Enterococcus faecalis, Brevibacillus laterosporus, and Paenibacillus alvei. Therefore, the presence of these secondary bacteria can be considered as a remarkable sign for EFB disease. Since the preparation of honey bee samples for the assessment of apiary health presents principal problems, this research aimed to use honey samples to track E. faecalis as a secondary agent of EFB disease in apiaries all over Iran. 
Materials & Methods: A total of of 260 apiaries were selected, and honey samples were collected during the autumn and winter of 2023. After preparation of the honey samples according to the standard protocol, genomic DNA was extracted from the samples, and then a pair of specific primers was used to amplify the target fragment of E. faecalis using the polymerase chain reaction (PCR) technique. As the positive and negative controls, the standard bacterium and the distilled water were used, respectively. A sensitivity test was performed using specific concentrations of DNA extracted from the standard E. faecalis under the corresponding optimal PCR reaction conditions, and the appropriate concentration was considered for the PCR reaction of the collected samples.
Results: PCR results showed that 119(46%) of the 260 honey samples were positive for E. faecalis. 
Conclusion: The findings show that the quick and easy use of honey samples for diagnosing E. faecalis can be introduced as an appropriate method to detect this bacterium. Therefore, honey samples can be recommended as a source for the detection of E. faecalis. Also, severe infection of the Iranian apiaries with E. faecalis as an indicator of EFB disease should be recognized as a very significant problem. 

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Archives of Razi Institute
Volume 81, Issue 3
May and June 2026
Pages 715-722

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