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ORIGINAL RESEARCH ARTICLE

Antibacterial activity of green-synthesized silver nanoparticles against Gram-negative bacteria and insights into potential resistance mechanisms

Akamu J. Ewunkem1* ,  Bliss Daodu1 ,  Zahirah J. Williams2 ,  Lydia Merrills2 ,  Brittany L. Justice1 ,  Felicia Simpson3 ,  David Holland3 ,  Tatyana Bowers3 ,  Uchenna Iloghalu1
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1 Department of Biological Sciences, Faculty of Natural and Physical Sciences, Winston-Salem State University, Winston-Salem, North Carolina, United States of America
2 Department of Nursing, Faculty of Natural and Physical Sciences, Winston-Salem State University, Winston-Salem, North Carolina, United States of America
3 Department of Mathematics, Faculty of Natural and Physical Sciences, Winston-Salem State University, Winston-Salem, North Carolina, United States of America
INNOSC Theranostics and Pharmacological Sciences, 025080007 https://doi.org/10.36922/ITPS025080007
Received: 17 February 2025 | Revised: 3 June 2025 | Accepted: 11 June 2025 | Published online: 1 July 2025
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Gram-negative bacterial infections pose a serious public health challenge due to their high global mortality rates and potential to cause severe complications. Antibiotics – one of the most impactful medical innovations of the 20th century – remain vital in treating life-threatening bacterial infections. However, the increasing prevalence of antibiotic resistance has made it progressively harder to treat Gram-negative bacterial infections effectively. Therefore, nanoparticles have gained attention as a promising alternative treatment owing to their targeted antibacterial properties. Among the various synthesis methods, green synthesis is considered one of the most effective approaches for nanoparticle production. In this study, silver nanoparticles were synthesized using a green approach that utilized silver nitrate salt and an extract derived from carpenter bee wings (CBWs). The synthesized nanoparticles were characterized using spectroscopic techniques and scanning electron microscopy. Their antibacterial activity was tested against two pathogenic Gram-negative bacteria using the broth dilution method. Furthermore, whole genome sequencing was conducted to assess the mutagenic effects of the biosynthesized silver nanoparticles on the two bacterial strains. The results demonstrated that the green-synthesized silver nanoparticles exhibit notable antibacterial activity, likely through electrostatic interactions that promote cell binding and induce significant morphological alterations. Genomic analysis revealed mutations associated with efflux pump regulation, neutralization, transport, energy metabolism, cell division, biosynthetic pathways, adaptation, and invasion in the tested strains. These findings demonstrate the potential of CBWs as a novel biological resource for the green synthesis of silver nanoparticles with antibacterial properties. However, the study also raises concerns regarding the potential for bacteria to develop resistance to nanoparticles over time.

Keywords
Carpenter bee wing extracts
Genomics
Gram-negative bacteria
Green synthesis
Nanoparticles
Funding
The study was supported by the Professional Development Committee Research Grant (grant number: 211444), Winston-Salem State University, and The Genomic Research and Data Science Center for Computation and Cloud Computing (grant number: 211512).
Conflict of interest
The authors declare that they have no competing interests.
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