Antibacterial activity of green-synthesised silver nanoparticles (AgNPs) on major bacterial fish pathogens with emphasis on their extracellular enzymatic and haemolytic activities

Basanta Kumar Das; Tamasi Senapati; Jyotirmayee Pradhan; Kausalya K. Nayak; Barsha Baisakhi; Debasmita Mohanty; Mala Kumari; Swagatika Sahu

doi:10.21077/ijf.2025.72.4.170069-12

Authors

Basanta Kumar Das ICAR-Central Inland Fisheries Research Institute, Barrackpore, West Bengal, India - 700120. https://orcid.org/0000-0002-6629-8992
Tamasi Senapati ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar, Odisha, India – 751002.
Jyotirmayee Pradhan Kuntala Kumari Sabat Women’s College, Balasore, Odisha, India, 756001 https://orcid.org/0000-0001-6466-0874
Kausalya K. Nayak Kshitrabasi Dayanand Anglo Vedic College, Nirakarpur, Khurdha, Odisha, India - 752019
Barsha Baisakhi Kuntala Kumari Sabat Women’s College, Balasore, Odisha, India, 756001. https://orcid.org/0009-0004-0440-136X
Debasmita Mohanty ICAR-Central Inland Fisheries Research Institute, Barrackpore, West Bengal, India - 700120. https://orcid.org/0000-0003-0730-3955
Mala Kumari ICAR-Central Inland Fisheries Research Institute, Barrackpore, West Bengal, India - 700120.
Swagatika Sahu Krishi Vigyan Kendra, Baliapal, Balasore, Odisha, India – 756027

https://doi.org/10.21077/ijf.2025.72.4.170069-12

Keywords:

Antimicrobial, extracellular enzyme, fish pathogens, silver nanoparticles

Abstract

Silver nanoparticles (AgNPs) were synthesised using a green synthesis approach using silver nitrate, starch and glucose under optimised conditions. Morphometric characterisation of the green-synthesised AgNPs was carried out by Fourier-transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), dynamic light scattering (DLS), and X-ray diffraction (XRD). The antibacterial, extracellular enzymatic and haemolytic activities of AgNPs were evaluated against nine fish pathogenic bacteria, viz., Aeromonas hydrophila (AH1), Edwardsiella tarda (ATCC15947), Pseudomonas putida (ATCC49128), Pseudomonas aeruginosa (ATCC35072), Pseudomonas fluorescens (PF1), Vibrio alginolyticus (ATCC17749), Vibrio parahaemolyticus (ATCC17802), Escherichia coli (EC1) and Staphylococcus aureus (ATCC6538). Bactericidal activity of AgNPs was assessed using both minimum inhibitory concentration (MIC) and disc diffusion assays. In A. hydrophila, starch hydrolysis was completely inhibited from 72 h onwards in cultures exposed to both 50 μl and 100 μl of AgNPs. Similarly, the haemolytic activity of V. alginolyticus showed a marked reduction from 24 h onwards at both AgNP exposure levels (50 μl and 100 μl). Bacterial sensitivity to AgNPs was found to vary among species, indicating pathogen-specific responses to AgNP treatment.

Keywords: Antibacterial activity, Bacterial sensitivity, Minimum inhibitory concentration, Nanomaterials in aquaculture, Virulence factors

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Author Biographies

Basanta Kumar Das, ICAR-Central Inland Fisheries Research Institute, Barrackpore, West Bengal, India - 700120.

Director
Tamasi Senapati, ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar, Odisha, India – 751002.

Research Scholar
Jyotirmayee Pradhan, Kuntala Kumari Sabat Women’s College, Balasore, Odisha, India, 756001

Assistant Professor, P.G. Dept. of Zoology
Kausalya K. Nayak, Kshitrabasi Dayanand Anglo Vedic College, Nirakarpur, Khurdha, Odisha, India - 752019

Assistant Professor, Dept. of Zoology
Barsha Baisakhi, Kuntala Kumari Sabat Women’s College, Balasore, Odisha, India, 756001.

PhD Scholar, P.G. Dept. of Zoology
Debasmita Mohanty, ICAR-Central Inland Fisheries Research Institute, Barrackpore, West Bengal, India - 700120.

PhD Scholar
Mala Kumari, ICAR-Central Inland Fisheries Research Institute, Barrackpore, West Bengal, India - 700120.

Research Scholar
Swagatika Sahu, Krishi Vigyan Kendra, Baliapal, Balasore, Odisha, India – 756027

Senior Scientist

References

Alderman, D. J., & Smith, P. 2001. Development of draft protocols of standard reference methods for antimicrobial agent susceptibility testing of bacteria associated with fish diseases. Aquac., 196 (3-4): 211-243.https://doi.org/10.1016/S0044-8486(01)00535-X DOI: https://doi.org/10.1016/S0044-8486(01)00535-X

Ansari, M. A., & Alzohairy, M. A. 2018. One‐pot facile green synthesis of silver nanoparticles using seed extract of Phoenix dactylifera and their bactericidal potential against MRSA. eCAM., 2018 (1) 1860280. https://doi.org/10.1155/2018/1860280 DOI: https://doi.org/10.1155/2018/1860280

Balantrapu, K., & Goia, D. V. 2009. Silver nanoparticles for printable electronics and biological applications. J. Mater. Res., 24 (9): 2828-2836. https://doi.org/10.1557/jmr.2009.0336 DOI: https://doi.org/10.1557/jmr.2009.0336

Chabbert, Y. A. 1963. The antibiogram: sensitivity and resistance of bacteria to antibiotics. Editions de la Tourelle. 257.

Feng, Q. L., Wu, J., Chen, G. Q., Cui, F. Z., Kim, T. N., & Kim, J. O. 2000. A mechanistic study of the antibacterial effect of silver ions on Escherichia coli and Staphylococcus aureus. J. Biomed. Mater., 52 (4): 662–8. https://doi.org/10.1002/1097-4636(20001215)52 DOI: https://doi.org/10.1002/1097-4636(20001215)52:4<662::AID-JBM10>3.0.CO;2-3

Gupta, A., & Silver, S. 1998. Molecular genetics: silver as a biocide: will resistance become a problem? Nat. Biotechnol., 16 (10): 888. https://doi.org/10.1038/nbt1098-888 DOI: https://doi.org/10.1038/nbt1098-888

Hatchett, D. W., & White, H. S. 1996. Electrochemistry of sulfur adlayers on the low-index faces of silver. J. Phys. Chem., 100 (23): 9854–9859. https://doi.org/10.1021/jp953757z DOI: https://doi.org/10.1021/jp953757z

Kawata, K., Osawa, M., & Okabe, S. 2009. In vitro toxicity of silver nanoparticles at noncytotoxic doses to HepG2 human hepatoma cells. Environ. Sci. Technol., 43 (15): 6046–6051. https://doi.org/10.1021/es900754q DOI: https://doi.org/10.1021/es900754q

Kheybari, S., Samadi, N., Hosseini, S.V., Fazeli, A., Fazeli, M.R. 2010. Synthesis and antimicrobial effects of silver nanoparticles produced by chemical reduction method. Daru., 18 (3):168-72. PMID: 22615613; PMCID: PMC3304363.

Kim, J.S., Kuk, E., Yu, K.N., Kim, J.H., Park, S.J., Lee, H.J., Kim, S.H., Park, Y.K., Park, Y.H., Hwang, C.Y. and Kim, Y.K. 2007. Antimicrobial effects of silver nanoparticles. NBM, 3 (1): 95-101. https://doi.org/10.1016/j.nano.2006.12.001 DOI: https://doi.org/10.1016/j.nano.2006.12.001

Kvítek, L., Panáček, A., Soukupova, J., Kolář, M., Večeřová, R., Prucek, R., Holecová, M. and Zbořil, R. 2008. Effect of surfactants and polymers on stability and antibacterial activity of silver nanoparticles (NPs). J. Phys. Chem. C., 112 (15): 5825–34. https://doi.org/10.1021/jp711616v DOI: https://doi.org/10.1021/jp711616v

Lee, H. J., Yeo, S. Y., & Jeong, S. H. 2003. Antibacterial effect of nanosized silver colloidal solution on textile fabrics. J. Mater. Sci., 38 (10): 2199–204. https://doi.org/10.1023/A:1023736416361 DOI: https://doi.org/10.1023/A:1023736416361

Liau, S. Y., Read, D. C., Pugh, W. J., Furr, J. R., & Russell, A. D. 1997. Interaction of silver nitrate with readily identifiable groups: relationship to the antibacterial action of silver ions. Lett. Appl. Microbiol., 25 (4): 279–283. https://doi.org/10.1046/j.1472-765X.1997.00219.x DOI: https://doi.org/10.1046/j.1472-765X.1997.00219.x

Lok, C.N., Ho, C.M., Chen, R., He, Q.Y., Yu, W.Y., Sun, H., Tam, P.K.H., Chiu, J.F. and Che, C.M. 2007. Silver nanoparticles: partial oxidation and antibacterial activities. J. Biol. Inorg. Chem., 12 (4): 527-534. https://doi.org/10.1007/s00775-007-0208-z DOI: https://doi.org/10.1007/s00775-007-0208-z

Matsumura, Y., Yoshikata, K., Kunisaki, S. I., & Tsuchido, T. 2003. Mode of bactericidal action of silver zeolite and its comparison with that of silver nitrate. Appl. Environ. Microbiol., 69 (7): 4278-4281. https://doi.org/10.1128/AEM.69.7.4278-4281.2003 DOI: https://doi.org/10.1128/AEM.69.7.4278-4281.2003

Morones, J.R., Elechiguerra, J.L., Camacho, A., Holt, K., Kouri, J.B., Ramírez, J.T. and Yacaman, M.J. 2005. The bactericidal effect of silver nanoparticles. Nanotechnology, 16 (10): 2346. 10.10880957-4484/16/10/059 DOI: https://doi.org/10.1088/0957-4484/16/10/059

Oluwafemi, O. S. 2009. A novel “green” synthesis of starch-capped CdSe nanostructures. Colloids Surf. B. Bio., 73 (2): 382-386. https://doi.org/10.1016/j.colsurfb.2009.06.011 DOI: https://doi.org/10.1016/j.colsurfb.2009.06.011

Pirtarighat, S., Ghannadnia, M., & Baghshahi, S. 2019. Green synthesis of silver nanoparticles using the plant extract of Salvia spinosa grown in vitro and their antibacterial activity assessment. J. nanostructure chem. 9 (1): 1-9. https://doi.org/10.1007/s40097-018-0291-4 DOI: https://doi.org/10.1007/s40097-018-0291-4

Prakash, P., Gnanaprakasam, P., Emmanuel, R., Arokiyaraj, S., & Saravanan, M. 2013. Green synthesis of silver nanoparticles from leaf extract of Mimusops elengi, Linn. for enhanced antibacterial activity against multi drug resistant clinical isolates. Colloids Surf. B Biointerfaces , 108: 255-259. https://doi.org/10.1016/j.colsurfb.2013.03.017 DOI: https://doi.org/10.1016/j.colsurfb.2013.03.017

Raveendran, P., Fu, J., & Wallen, S. L. 2003. Completely “green” synthesis and stabilization of metal nanoparticles. J. Amer. Chem. Soc., 125 (46): 13940–13941. https://doi.org/10.1021/ja029267j DOI: https://doi.org/10.1021/ja029267j

Saxena, A., Tripathi, R. M., & Singh, R. P. 2010. Biological synthesis of silver nanoparticles by using onion (Allium cepa) extract and their antibacterial activity. Dig. J. Nanomater. Biostruct., 5(2): 427–432.

Sharma, V. K., Yngard, R. A., & Lin, Y. 2009. Silver nanoparticles: green synthesis and their antimicrobial activities. Adv. Colloid. Interface. Sci., 145 (1-2): 83–96. https://doi.org/10.1016/j.cis.2008.09.002 DOI: https://doi.org/10.1016/j.cis.2008.09.002

Shrivastava, S., Bera, T., Roy, A., Singh, G., Ramachandrarao, P., & Dash, D. 2007. Retracted: Characterization of enhanced antibacterial effects of novel silver nanoparticles. Nanotechnology, 18 (22): 225103. 10.1088/0957-4484/18/22/225103 DOI: https://doi.org/10.1088/0957-4484/18/22/225103

Singh, M., Sinha, I., & Mandal, R. K. 2009. Role of pH in the green synthesis of silver nanoparticles. Mater. Lett., 63 (3-4): 425–427. https://doi.org/10.1016/j.matlet.2008.10.067 DOI: https://doi.org/10.1016/j.matlet.2008.10.067

Sondi, I., & Salopek-Sondi, B. 2004. Silver nanoparticles as antimicrobial agent: a case study on E. coli as a model for Gram-negative bacteria. J. Colloid. Interface. Sci., 275 (1): 177–182. https://doi.org/10.1016/j.jcis.2004.02.012 DOI: https://doi.org/10.1016/j.jcis.2004.02.012

Sondi, I., Goia, D. V., & Matijević, E. 2003. Preparation of highly concentrated stable dispersions of uniform silver nanoparticles. J. Colloid. Interface. Sci., 260 (1): 75-81. https://doi.org/10.1016/S0021-9797(02)00205-9 DOI: https://doi.org/10.1016/S0021-9797(02)00205-9

Vasileva, P., Donkova, B., Karadjova, I., & Dushkin, C. 2011. Synthesis of starch-stabilized silver nanoparticles and their application as a surface plasmon resonance-based sensor of hydrogen peroxide. Col. and Sur. A: Physicochem. Eng. Asp., 382 (1-3): 203-210. https://doi.org/10.1016/j.colsurfa.2010.11.060 DOI: https://doi.org/10.1016/j.colsurfa.2010.11.060

Yakout, S. M., & Mostafa, A. A. 2015. A novel green synthesis of silver nanoparticles using soluble starch and its antibacterial activity. Inter. J. Cli. Exp. Med., 8(3): 3538-3544. PMCID: PMC4443080 PMID: 26064246