Antimicrobial activities of chitosan nanoparticles against pathogenic microorganisms in Nile tilapia, Oreochromis niloticus
- 84 Downloads
Nanotechnology is a recent unique technique generally used for nutrition and therapy purposes among others. In this respect, the present study was carried out to evaluate the antimicrobial activity of chitosan nanoparticles (CNP) against various microorganisms (fungi and bacteria) isolated from diseased or health Nile tilapia, Oreochromis niloticus. The CNP was prepared based on the ionic gelation of chitosan with tripolyphosphate anion and its mean size was 35 nm with a narrow size distribution and zeta potential of 61.2 mV. The lethal dose of pathogenic bacterial isolates for Nile tilapia was successfully standardized. Clinical signs including weakness, slower movement, swimming closer to the surface, fin hemorrhages, and red patches at the gut regions were observed. Enlargement of spleen followed by tissue necrosis along with signs of hemorrhagic septicemia was also seen in infected fish. Fungal and bacterial isolates were exposed to different CNP doses and it is noticed that CNP inhibited all examined fungal and bacterial isolates in a dose-dependent manner. However, high CNP doses (80 μg/ml) gave highest inhibition zones where Aspergillus flavus, Mucor sp., and Candida sp. were more susceptible, whereas Aspergillus niger, A. fumigatus, and Fusarium sp. were more resistant. Similarly, largest inhibition zones of tested bacteria were obtained at high CNP dose (20 μg/ml). And Aeromonas sobria, A. hydrophila, and Pseudomonas aeruginosa were the most susceptible bacterial strain; meanwhile, Staphylococcus aureus and Pseudomonas fluorescens were the most resistant ones. The minimal inhibitory concentration of CNP against the examined bacteria ranged from 0.156 to 2.5 μg/ml causing their minimal counts. The transmission electron microscope images revealed that CNP showed antagonistic action against A. hydrophila causing disruption of cell membranes and the leakage of cytoplasm. In a practical experiment, Nile tilapia fed dietary CNP at levels of 0.0 and 1.0 g/kg diet for 3 weeks and post-challenged with different pathogenic bacteria via intraperitoneal injection. It is noticed that fish fed a CNP-enriched diet showed less mortality with all bacterial strains (6.7–20%), while, fish fed a CNP-free diet showed highest mortality (66.7–100%). The dietary CNP protected Nile tilapia efficiently against A. hydrophila, A. sobria, and Streptococcus agalactiae infections with relative level of protection (RLP) value of 93.3%, while the RLP against Staphylococcus aureus was 70.0%.
KeywordsChitosan nanoparticles Antimicrobial activity Pathogenic fungi Pathogenic bacteria Nile tilapia
This study was funded and supported by the Central Laboratory for Aquaculture Research (CLAR), Abbassa, Abu-Hammad, Sharkia, Egypt.
Compliance with ethical standards
Conflict of interest
The author declares that she has no conflict of interest.
The author declares that she followed all guidelines for the care and use of fish in the present study.
- Abdel-Ghany HM, Salem ME-S (2019) Effects of dietary chitosan supplementation on farmed fish; a review. Rev Aquac. https://doi.org/10.1111/raq.12326
- Akmaz S, Adjgüzel ED, Yasar M, Erguven O (2013) The effect of Ag content of the chitosan-silver nanoparticle composite material on the structure and antibacterial activity. Adv Mater Sci Eng. https://doi.org/10.1155/2013/690918
- Amer MSMI (2002) Antimicrobial activity of some species of blue green algae (Cyanobacteria). M.Sc., Botany Dep., Fac. Sci., Tanta Univ, Egypt.Google Scholar
- Avadi MR, Sadeghi AMM, Tahzibi A, Bayati KH, Pouladzadeh M, Zohuriaan-Mehr MJ, Rafiee (2004) Optimized synthesis and characterization of N-Triethyl chitosan Tehrani, M. Eur Polym J 40: 1355–1361.Google Scholar
- Florio D, Gustinelli A, Caوٴara M, Turci F, Quaglio F et al (2009) Veterinary and public health aspects in tilapia (Oreochromis niloticus) aquaculture in Kenya, Uganda and Ethiopia. Ittiopatologia 6:51–93Google Scholar
- Hadwiger LA, Kendra DG, Fristensky BW, Wagoner W (1981) Chitin in nature and technology. In: Muzzarelli RAA, Jeuniaux C, Gooday GW (eds) Chitosan both activated genes in plants and inhibits RNA synthesis in fungi. Plenum, New York, p 584Google Scholar
- Hernandez-Lauzardo AN, Bautista-Banos S, Velazquez-del Valle MG, Mendez-Montealvo MG, Sanchez-Rivera MM, Bello-Perez LA (2008) Antifungal effects of chitosan with different molecular weights on in vitro development of Rhizopus stolonifer (Ehrenb.:Fr.) Vuill. Carbohydr Polym 73:541–547CrossRefGoogle Scholar
- Iqbal Z, Saleemi S (2013) Isolation of pathogenic fungi from a freshwater commercial fish Catla catla. Sci Int 25:851–855Google Scholar
- Kaplan SL, Assaâd Silaa B, Baha Eddine A, Rihab BA, Semia EC, Ali B, Rafik B (2016) Chitin and chitosan from the Norway lobster by-products: antimicrobial and anti-proliferative activities. Int eJ Biological Macromolecules 47:341–345Google Scholar
- Kinner NE, Balkwill DL, Bishop PL (1983) Light and electron microscopic studies of microorganisms growing in rotating biological contactor bio films. Appl Environ Microbiol 45(5):1659–1669Google Scholar
- Lam TD, Hoang VD, Lien LN, Thinh NN, Dien PG (2006) Synthesis and characterization of chitosan nanoparticles used as drug. J Chem 44:105–109Google Scholar
- Larone DH (1987) Medically important fungi: a guide to identification. American Society for Microbiology, Medical, pp 230Google Scholar
- Másson M, Holappa J, Hjalmarsdottir MR, Unarsson OV, Nevalainen T, Jarvinen T (2008) Antimicrobial activity of piperazine derivatives of chitosan. Carbohydrate Polymers 74: 566–571Google Scholar
- Melaku H, Lakew M, Alemayehu E, Wubie A, Chane M (2017) Isolation and identification of pathogenic fungus from African catfish (Clarias gariepinus) eggs and adults in national fishery and aquatic life research center hatchery, Ethiopia. Fish Aqua J 8:213. https://doi.org/10.4172/2150-3508.1000213 Google Scholar
- Mohy-Eldin SM, Soliman EA, Hashem AI, Tamer TM (2008) Antibacterial activity of chitosan chemically modified with new technique. Trends Biomater Atrif Organs 22:121–133Google Scholar
- Nester E W, Anderson D G, Roberts E, Pearshall, N N, Nester M T (2004) Microbiology a human prospect. Published by Boston McGraw-Hill 2e004 https://trove.nla.gov.au/version/49174664. Accessed 30 July 2004
- Refai M (1987) Isolation and identification of fungi. Fac. Vet. Mid., Cairo University, Cairo, EgyptGoogle Scholar
- Refai M, Abdel Halim MM, Afify MMH, Youssef H, Marzouk M (1987) Studies on aspergillomycosis in catfish (Clarias lasera). Allgemeine Pathologic and pathologische Anatomic. Tagung der Deutachen Veterinar—Medizinischen Gesellschaft. der Europeischen Gesellschaft fur Vet. Pathol. 63: 1–12Google Scholar
- Refai MK, Laila AM, Amany KM, Shimaa E-SMA (2010) The assessment of mycotic settlement of freshwater fishes in Egypt. J Am Sci 6(11):594–602Google Scholar
- Shaheen, AA (1986) Mycoflora of some freshwater fish. M.V.Sc. Thesis, Fac. Vet. Med., Zagazic Univ, Zagazig, EgyptGoogle Scholar
- Srivastava RC (2009) Fish mycopathology. Today and Tomorrow’s Printers and Publishers, New Dehli, p 103Google Scholar