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Green Synthesized Silver Nanoparticles Using Lactobacillus Acidophilus as an Antioxidant, Antimicrobial, and Antibiofilm Agent Against Multi-drug Resistant Enteroaggregative Escherichia Coli

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Abstract

The present study was envisaged to employ the green synthesis and characterization of silver nanoparticles (AgNPs) using the potential probiotic strain Lactobacillus acidophilus, to assess its antibacterial as well as antibiofilm activity against multi-drug-resistant enteroaggregative Escherichia coli (MDR-EAEC) strains and to investigate their antioxidant activity. In this study, AgNPs were successfully synthesized through an eco-friendly protocol, which was then confirmed by its X-ray diffraction (XRD) pattern. A weight loss of 15% up to 182 °C with a narrow exothermic peak between 170 °C and 205 °C was observed in thermogravimetric analysis-differential thermal analysis (TGA-DTA), while aggregated nanoclusters were observed in scanning electron microscopy (SEM). Moreover, the transmission electron microscopy (TEM) imaging of AgNPs revealed a spherical morphology and crystalline nature with an optimum size ranging from 10 to 20 nm. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of green synthesized AgNPs against the MDR-EAEC strains were found to be 7.80 mg/L and 15.60 mg/L, respectively. In vitro time-kill kinetic assay revealed a complete elimination of the MDR-EAEC strains after 180 min on co-incubation with the AgNPs. Moreover, the green synthesized AgNPs were found safe by in vitro haemolytic assay. Besides, the green synthesized AgNPs exhibited significant biofilm inhibition (P < 0.001) formed by MDR-EAEC strains. Additionally, a concentration-dependent antioxidant activity was observed in 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Hence, this study demonstrated potential antibacterial as well as antibiofilm activity of green synthesized AgNPs against MDR-EAEC strains with antioxidant properties and warrants further in-depth studies to explore it as an effective antimicrobial agent against MDR infections.

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All data generated or analysed during this study are included in this published article [and its supplementary information files].

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Acknowledgements

The authors thank Director, ICAR-National Research Centre on Meat, Hyderabad; Vice-Chancellors and Directors of Research of KVASU and MAFSU; and Deans and respective Heads of Department of CVAS, Pookode, and NVC, Nagpur, for providing facilities for this research.

Funding

This work was supported by a financial grant from National Agricultural Science Fund (ICAR-NASF; NASF/ABA-8007) to SBB, DBR, JV, and NVK.

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Authors and Affiliations

  1. Department of Veterinary Public Health, College of Veterinary and Animal Sciences, Pookode, Kerala Veterinary and Animal Sciences University, Lakkidi Post, Wayanad, 673 576, India

    Padikkamannil Abishad, Jess Vergis, Varsha Unni & Prejit Nambiar

  2. ICAR- National Research Centre On Meat, Chengicherla, Hyderabad, 500 092, India

    Vemula Prasastha Ram, Pollumahanti Niveditha, Jyothsana Yasur, Sukhadeo Baliram Barbuddhe & Deepak Bhiwa Rawool

  3. Department of Veterinary Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Pookode, Kerala Veterinary and Animal Sciences University, Lakkidi Post, Wayanad, 673 576, India

    Sanis Juliet

  4. Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Pookode, Kerala Veterinary and Animal Sciences University, Lakkidi Post, Wayanad, 673 576, India

    Lijo John

  5. Centre for Research and Innovations, BGSIT, Adichunchanagiri University, B.G. Mandya, 571 418, India

    Kullaiya Byrappa

  6. Department of Veterinary Pathology, Nagpur Veterinary College, Maharashtra Animal and Fishery Sciences University, Nagpur, 440 006, India

    Nitin Vasantrao Kurkure

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  1. Padikkamannil Abishad
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Contributions

Deepak Bhiwa Rawool; Jess Vergis; Nitin V Kurkure; Sukhadeo Baliram Barbuddhe took part in conceptualization; Abishad Padikkamannil; Varsha Unni; Jess Vergis; Sanis Juliet; Lijo John involved in methodology; Abishad Padikkamannil; Varsha Unni; Sanis Juliet; Lijo John; Kullaiya Byrappa; Prejit Nambiar; Jess Vergis took part in formal analysis, visualization, and investigation; Abishad Padikkamannil; Varsha Unni; Vemula Prasastha Ram; Niveditha Pollumahanti; Jyothsana Yasur; Kullaiya Byrappa; Nitin V Kurkure took part in validation; Varsha Unni; Abishad Padikkamannil; Vemula Prasastha Ram; Niveditha Pollumahanti; Jyothsana Yasur involved in writing—original draft preparation; Jess Vergis; Deepak Bhiwa Rawool; Sukhadeo Baliram Barbuddhe; Nitin V Kurkure; Kullaiya Byrappa took part in writing—review and editing; Deepak Bhiwa Rawool; Jess Vergis; Nitin V Kurkure; Sukhadeo B Barbuddhe involved in funding acquisition; Jess Vergis; Deepak Bhiwa Rawool took part in resources; Jess Vergis; Deepak Bhiwa Rawool involved in supervision.

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Abishad, P., Vergis, J., Unni, V. et al. Green Synthesized Silver Nanoparticles Using Lactobacillus Acidophilus as an Antioxidant, Antimicrobial, and Antibiofilm Agent Against Multi-drug Resistant Enteroaggregative Escherichia Coli. Probiotics & Antimicro. Prot. 14, 904–914 (2022). https://doi.org/10.1007/s12602-022-09961-1

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