Abstract
Honey bees are vital pollinators essential for ecosystems and global agriculture, but their populations are declining due to pressures caused by pests. This study reports the presence of Escherichia coli, Klebsiella pneumoniae, Proteus terrae, and Pseudomonas spp. in four different honey bee species in Pakistan for the first time, uncovering a previously neglected aspect of honey bee microbiology. From January 2023 to June 2024; 1190 honey bees were collected from four districts in Khyber Pakhtunkhwa and they were identified. Molecular screening based on PCR targeting the microbial 16 S rDNA resulted in the detection of potential bacterial pathogens across different districts. In Buner, K. pneumoniae and E. coli were detected in Apis mellifera and Apis cerana individuals while Apis florea was positive only for K. pneumoniae. In Swat, Pseudomonas spp. was identified in A. mellifera, Apis dorsata and A. florea, while in A. cerana no pathogen was detected. In Lower Dir, A. mellifera and A. florea were positive for Pseudomonas spp., and A. cerana for K. pneumoniae, with A. dorsata being pathogen-free. In Malakand, K. pneumoniae and E. coli was found in A. mellifera and A. cerana, while A. florea harbored only Pseudomonas spp., and A. dorsata had no pathogens. Finally, P. terrae was detected exclusively in A. mellifera from Buner and Swat. Morphological analysis revealed significant interspecies variation, with A. dorsata exhibiting the largest values in tongue length (4.27 ± 0.21 mm), forewing size (12.8 ± 0.19 mm), and the highest cubital index (2.71 ± 0.04 mm). A. mellifera was classified in second place, followed by A cerana and A. florea, which characterized by the lowest measurement values. In the present study significant bacterial pathogens were identified representing emerging threat to honey bees in Pakistan while data regarding their phenotypic characterization and phylogenetic position were provided for the first time in the region.
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Data availability
Sequences are submitted to GenBank with the accession numbers given by GenBank are as followed: Escherichia coli (PQ223672, PQ223673), Proteus terrae (PQ223674), Klebsiella pneumoniae HB4 (PQ223670, PQ223671), and Pseudomonas spp. (PQ223667, PQ223668, PQ223669).
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Acknowledgements
The authors extend their appreciation to the Deanship of Scientific Research and Graduate Studies at King Khalid University for funding this work through the Large Research Project under grant number RGP2/715/46.
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This work is financially supported by the Deanship of Scientific Research and Graduate Studies at King Khalid University for funding this work through the Large Research Project under grant number RGP2/715/46.
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Author Contributions: Shazia Amin: Methodology, Formal Analysis, Investigation, Writing – original draft. Shakir Ullah: Writing – original draft, Writing – review & editing, Validation. Nasreen Nasreen: Methodology, Formal Analysis, Data curation, Validation. Ioannis A. Giantsis: Writing – review & editing, Validation. Maria V. Alavanou, Esmael M. Alyami, and Youssouf Ali Younous: Data curation, Software, Formal Analysis, Writing – original draft. Sadaf Niaz: Investigation, Validation, Resources. Adil Khan, and Turki M. Dawoud: Conceptualization, Supervision, Project administration.
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The Institutional Research Ethics Committee of Abdul Wali Khan University Mardan Pakistan approved the study. Notably, all animal experimentation was conducted in accordance with applicable laws, regulations, and guidelines, prioritizing animal welfare and minimizing any potential harm.
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Amin, S., Ullah, S., Nasreen, N. et al. Molecular characterization of emerging bacterial communities associated with honey bees. Sci Rep (2026). https://doi.org/10.1038/s41598-026-42610-z
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DOI: https://doi.org/10.1038/s41598-026-42610-z
