In vitro antimicrobial efficacy of laser-synthesized silver nanoparticles against antibiotic-resistant Escherichia coli isolated from dairy cattle wastewater

Vet World Vol.19 February-2026 Article - 4

Research Article

Veterinary World, 19(2): 511-522

https://doi.org/10.14202/vetworld.2026.511-522

Sheila Marty Yanestria¹, Freshinta Jellia Wibisono¹, Mustofa Helmi Effendi^2,3,4, Tri Untari⁵, Aswin Rafif Khairullah⁶, Fidi Nur Aini Eka Puji Dameanti⁷, John Yew Huat Tang⁸, Saifur Rehman⁹, Wasito Wasito⁶, and Riza Zainuddin Ahmad⁶

1. Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Wijaya Kusuma Surabaya, Jl. Dukuh Kupang XXV No.54, Dukuh Kupang, Dukuh Pakis, Surabaya, 60225, East Java, Indonesia.

2. Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Airlangga, Jl. Dr. Ir. H. Soekarno, Kampus C Mulyorejo, Surabaya 60115, East Java, Indonesia.

3. Research Group of Antimicrobial Resistance, Faculty of Veterinary Medicine, Universitas Airlangga Surabaya, 60115, East Java, Indonesia.

4. School of Food Industry, Faculty of Bioresources, and Food Industry, Universiti Sultan Zainal Abidin (Besut Campus), Besut 22200, Malaysia. .

5. Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Bulaksumur, Caturtunggal, Depok, Sleman 55281, Daerah Istimewa Yogyakarta, Indonesia.

6. Research Center for Veterinary Science, National Research and Innovation Agency (BRIN), Jl. Raya Bogor Km. 46 Cibinong, Bogor 16911, West Java, Indonesia.

7. Laboratory of Veterinary Microbiology and Immunology, Faculty of Veterinary Medicine, Universitas Brawijaya, Jl. Puncak Dieng, Kunci, Kalisongo, Dau, Malang 65151, East Java, Indonesia.

8. School of Food Industry, Faculty of Bioresources, and Food Industry, Universiti Sultan Zainal Abidin (Besut Campus), Besut 22200, Malaysia.

9. Department of Pathobiology, Faculty of Veterinary and Animal Sciences, Gomal University, RV9W+GVJ, Indus HWY, Dera Ismail Khan 27000, Pakistan.

Background and Aim: Poorly managed dairy farm wastewater is a significant reservoir of antibiotic-resistant bacteria, particularly Escherichia coli, contributing to the environmental spread of antimicrobial resistance (AMR) and posing risks to animal and public health. Conventional wastewater treatment systems are often insufficient to inactivate these resistant organisms. Silver nanoparticles (AgNPs), especially those synthesized by pulsed laser ablation (PLA) in liquid, offer a high-purity, chemical-free nanomaterial with promising antimicrobial properties. This study aimed to evaluate the in vitro antimicrobial efficacy of laser-synthesized AgNPs against antibiotic-resistant E. coli isolated from dairy cattle wastewater within a One Health framework.

Materials and Methods: Wastewater samples were collected aseptically from 50 smallholder dairy farms in East Java, Indonesia. E. coli isolates were identified using standard cultural, morphological, Gram staining, and biochemical (Indole, methyl red, Voges–Proskauer, citrate) methods. Antibiotic resistance was screened using the Kirby–Bauer disk diffusion method against streptomycin, erythromycin, penicillin, and tetracycline. AgNPs were synthesized via PLA in polyvinylpyrrolidone medium and characterized using transmission electron microscopy, ultraviolet–visible spectroscopy, and Fourier transform infrared spectroscopy. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AgNPs were determined by broth microdilution and agar subculture methods, respectively, across concentrations ranging from 0.195 to 100 ppm. Statistical analysis was performed using one-way analysis of variance followed by Tukey’s post hoc test at a significance level of p < 0.05.

Results: PLA successfully produced monodisperse AgNPs with a mean diameter of 11.62 ± 1.8 nm and a characteristic surface plasmon resonance peak at 418 nm, confirming high-purity and stability. Twenty antibiotic-resistant E. coli isolates were evaluated. MIC values ranged from 37.5 to 100 ppm, with erythromycin-resistant isolates showing the lowest MICs (45.0 ± 10.5 ppm) and streptomycin-resistant isolates the highest (75.0 ± 33.3 ppm). Most isolates (75%) exhibited MBC values >100 ppm, indicating predominantly bacteriostatic activity at the tested concentrations. No statistically significant differences in MIC values were observed among resistance groups (p > 0.05). A concentration of 62.5 ppm was identified as the most effective inhibitory dose across resistance profiles.

Conclusion: Laser-synthesized AgNPs demonstrated consistent in vitro inhibitory activity against antibiotic-resistant E. coli from dairy wastewater, with an optimal MIC of approximately 62.5 ppm. These findings highlight the potential application of AgNPs as a supplementary control strategy in dairy waste management and AMR mitigation, supporting an integrated One Health approach.

Keywords: antimicrobial resistance, dairy farm wastewater, Escherichia coli, laser ablation, One Health, silver nanoparticles, wastewater management, zoonotic bacteria.

How to cite this article: Yanestria SM, Wibisono FJ, Effendi MH, Untari T, Khairullah AR, Dameanti FNAEP, et al. In vitro antimicrobial efficacy of laser-synthesized silver nanoparticles against antibiotic-resistant Escherichia coli isolated from dairy cattle wastewater. Vet World. 2026;19(2):511–522.

Received: 22-09-2025 Accepted: 05-01-2026 Published online: 10-02-2026

Corresponding author: Mustofa Helmi Effendi E-mail: mhelmieffendi@gmail.com

DOI: 10.14202/vetworld.2026.511-522

Copyright: Yanestria, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.