Antimicrobial activity of chitosan and combination with antibiotics against mastitis-causing pathogens

Thi Kieu Oanh Huynh; Thi Minh Khanh Pham; Thuc Quyen Huynh; Van Ty Tran; Quynh Thuong Nguyen; Hong Phuong Ngo; Phuong Thao Nguyen; Thi Thu Hoai Nguyen

doi:10.15625/vjbt-19815

Author affiliations

Authors

Thi Kieu Oanh Huynh \(^1\) School of Biotechnology, International University – Vietnam National University of HCMC, Vietnam https://orcid.org/0009-0006-2543-4528
Thi Minh Khanh Pham \(^1\) School of Biotechnology, International University – Vietnam National University of HCMC, Vietnam https://orcid.org/0009-0003-8923-3387
Thuc Quyen Huynh \(^2\) Research Center for Infectious Diseases, International University, Vietnam National University of HCMC, Vietnam https://orcid.org/0009-0002-7229-0441
Van Ty Tran \(^3\) Vietnam Food Joint Stock Company, Vietnam
Quynh Thuong Nguyen \(^3\) Vietnam Food Joint Stock Company, Vietnam
Hong Phuong Ngo \(^4\) University of Agriculture & Forestry – Ho Chi Minh City, Vietnam
Phuong Thao Nguyen \(^1\) School of Biotechnology, International University – Vietnam National University of HCMC, Vietnam
\(^2\) Research Center for Infectious Diseases, International University, Vietnam National University of HCMC, Vietnam
Thi Thu Hoai Nguyen \(^1\) School of Biotechnology, International University – Vietnam National University of HCMC, Vietnam
\(^2\) Research Center for Infectious Diseases, International University, Vietnam National University of HCMC, Vietnam https://orcid.org/0000-0003-4869-1827

DOI:

https://doi.org/10.15625/vjbt-19815

Keywords:

Antimicrobial activity, bovine mastitis, chitosan, pathogens

Abstract

Bovine mastitis (BM), primarily caused by bacterial pathogens infecting mammary glands, stands as the most prevalent disease in dairy cattle. Traditionally, antibiotics have been the primary choice of treatment, yet their overuse has led to widespread resistance and the presence of antibiotic residues in dairy products. Today, chitosan has emerged as a promising alternative in dairy farming. In this study, we systematically screened and assessed the antibacterial efficacy of five chitosan preparations of different viscosities and components. Additionally, we explored the synergistic antimicrobial potential of the most potent chitosan sample in combination with commonly employed antibiotics, including ampicillin, amoxicillin, oxacillin, and levofloxacin against four prevalent BM-causing pathogens: Staphylococcus epidermidis, Streptococcus agalactiae, Streptococcus uberis and Pseudomonas sp. Agar well diffusion, micro-dilution, and checkerboard techniques were applied to assess the antimicrobial activity and interaction effect. Results indicated that, at a concentration of 1%, low and medium viscosity samples (samples 1, 2, 3) exhibited relatively low activity, compared to very low viscosity ones (samples 4, 5). Notably, sample 5, a combination of chitosan sample 1 with orange and grapefruit essential oils, demonstrated the most potent antibacterial activity with a minimal inhibitory concentration (MIC) of 19.53 mg/L against S. agalactiae, S. uberis and S. epidermidis and 78.13 mg/L against Pseudomonas sp.. Furthermore, the combination of this chitosan sample and antibiotics exhibited some synergistic interactions against BM-causing pathogens, as indicated by the fractional inhibitory concentration (FIC) values ranging from ≥ 0.5 to ≤ 1. While these effects were notable, they did not reach the threshold for strong synergism (FIC < 0.5). In summary, our study highlighted the high antibacterial activity of low viscosity chitosan, particularly in combination with essential oils. Although there were observed synergistic effects with antibiotics against BM-causing pathogens, the strength of these interactions was not robust enough to conclusively categorize them as strongly synergistic. Chitosan, however, emerges as a promising agent in the ongoing exploration of alternatives to antibiotics in the management of BM in dairy farming.

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References

Aureus in dairy herds at Hawassa milk shed, South Ethiopia. BMC Vet Res 12: 1-11. https://doi.org/10.1186/s12917-016-0905-3

Ardean C, Davidescu CM, Nemeş NS, Negrea A, Ciopec M, Duteanu N, Negrea P, Duda-Seiman D, Musta V (2021) Factors influencing the antibacterial activity of chitosan and chitosan modified by functionalization. Int J Mol Sci 22: 7449. https://doi.org/10.3390/ijms22147449

Bajaksouzian S, Visalli MA, Jacobs MR, Appelbaum PC (1997) Activities of levofloxacin, ofloxacin, and ciprofloxacin, alone and in combination with amikacin, against acinetobacters as determined by checkerboard and time-kill studies. Antimicrob Agents Chemother 41: 1073-1076. https://doi.org/10.1128/aac.41.5.1073

Bellio P, Fagnani L, Nazzicone L, Celenza G (2021) New and simplified method for drug combination studies by checkerboard assay. MethodsX 8: 101543. https://doi.org/10.1016/j.mex.2021.101543

Yadav P, Yadav AB, Gaur P, Mishra V, Huma ZI, Sharma N, Son YO (2022) Bioengineered ciprofloxacin-loaded chitosan nanoparticles for the treatment of bovine mastitis. Biomed 10: 3282. https://doi.org/10.3390/biomedicines10123282

Breser ML, Felipe V, Bohl LP, Orellano MS, Isaac P, Conesa A, Rivero VE, Correa SG, Bianco ID, Porporatto C (2018) Chitosan and cloxacillin combination improve antibiotic efficacy against different lifestyle of coagulase-negative Staphylococcus isolates from chronic bovine mastitis. Sci Rep 8: 5081. https://doi.org/10.1038/s41598-018-23521-0

Cheng WN, Han SG (2020) Bovine mastitis: risk factors, therapeutic strategies, and alternative treatments - A review. Asian-Australas J Anim Sci 33: 1699-1713. https://doi.org/10.5713/ajas.20.0156

Chi TV, Vy PH, Minh NDN, Lambert P, Hoai NTT (2017) A deletion mutation in nfxB of in vitro-induced moxifloxacin-resistant Pseudomonas aeruginosa confers multidrug resistance. Acta Microbiol Immunol Hung 64: 245-253. https://doi.org/10.1556/030.64.2017.012

Chung YC, Su YP, Chen CC, Jia G, Wang HL, Wu JC, Lin JG (2004) Relationship between antibacterial activity of chitosan and surface characteristics of cell wall. Acta Pharmacol Sin 25: 932-936.PMID: 15210068

CLSI (2021) Performance Standards for Antimicrobial Susceptibility Testing. CLSI supplement M100 31st ed.

Daraghmeh NH, Chowdhry BZ, Leharne SA, Omari MMA, Badwan AA (2011) Chitin. Profiles Drug Subst Excip Relat Methodol 36: 35-102. https://doi.org/10.1016/B978-0-12-387667-6.00002-6

Devlieghere F, Vermeulen A, Debevere J (2004) Chitosan: antimicrobial activity, interactions with food components and applicability as a coating on fruit and vegetables. Food Microbiol 21: 703-714. https://doi.org/10.1016/j.fm.2004.02.008

Fernandez-Saiz P, Lagarón JM, Ocio MJ (2009) Optimization of the film-forming and storage conditions of chitosan as an antimicrobial agent. J Agric Food Chem 57: 3298-3307. https://doi.org/10.1021/jf8037709

Fetrow J, Mann. D, Butcher. K, McDaniel. B (1991) Production losses from mastitis: Carry-over from the previous lactation. J Dairy Sci 74: 833-839. https://doi.org/10.3168/jds.S0022-0302(91)78232-5

Goy RC, Britto Dd, Assis OBG (2009) A review of the antimicrobial activity of chitosan. Polímeros 3: 241-247. https://doi.org/10.1590/S0104-14282009000300013

Hosseinnejad M, Jafari SM (2016) Evaluation of different factors affecting antimicrobial properties of chitosan. Int J Biol Macromol 85: 467-475. https://doi.org/10.1016/j.ijbiomac.2016.01.022

Jovanović GD, Klaus AS, Nikšić MP (2016) Antimicrobial activity of chitosan coatings and films against Listeria monocytogenes on black radish. Rev Argent Microbiol 48: 128-136. https://doi.org/10.1016/j.ram.2016.02.003

Linh DNT, Ngan TQ, Thuc NT, Chau DNP, Hoai NTT (2020) Effects of culture conditions on the antimicrobial activity of Streptomyces spp. Springer Singapore 69: 677-780. https://doi.org/10.1007/978-981-13-5859-3_114

Lorian V (2005) Antibiotics in Laboratory Medicine (5th ed). Clin Infect Dis 1–9: 365–441. ISBN: 0781749832

Liu XF, Guan YL, Yang DZ, Li Z (2001) Antibacterial action of chitosan and carboxymethylated chitosan. J Appl Polym Sci 79: 1324-1335. https://doi.org/10.1002/1097-4628(20010214)79:7<1324::AID-APP210>3.0.CO;2-L

Nadia MC, Lichtfouse E, Torri G, Crini G (2019) Applications of chitosan in food, pharmaceuticals, medicine, cosmetics, agriculture, textiles, pulp and paper, biotechnology, and environmental chemistry Environ Chem Lett 17: 1667-1692. https://doi.org/10.1007/s10311-019-00904-x

No HK, Park NY, Lee SH, Meyers SP (2002) Antibacterial activity of chitosans and chitosan oligomers with different molecular weights. Int J Food Microbiol 74: 65-72. https://doi.org/10.1016/S0168-1605(01)00717-6

Pangprasit N, Srithanasuwan A, Suriyasathaporn W, Pikulkaew S, Bernard JK, Chaisri W (2020) Antibacterial activities of acetic acid against major and minor pathogens isolated from mastitis in dairy cows. Pathog 9: 961. https://doi.org/10.3390/pathogens9110961

Phượng NH, Thương NQ, Ty TV (2022) Antibacterial effects of chitosan from shimp waste on Bacteria strains causing bovine mastitis. Khoa học kỹ thuật chăn nuôi 274: 51-57.

Qin C, Li H, Xiao Q, Liu Y, Zhu J, Du Y (2006) Water-solubility of chitosan and its antimicrobial activity. Carbohydr Polym 63: 367-374. https://doi.org/10.1016/j.carbpol.2005.09.023

Rivera AP, Bruna LT, Alarcón GC, Cayupe RB, González-Casanova J, Rojas-Gómez D, Caro FN (2020) Antimicrobial and antibiofilm capacity of chitosan nanoparticles against wild type strain of Pseudomonas sp. isolated from milk of cows diagnosed with bovine mastitis. Antibiot (Basel) 9: 9-551. https://doi.org/10.3390/antibiotics9090551

Sudarshan NR, Hoover DG, Knorr D (1992) Antibacterial action of chitosan. Food Biotechnol 6: 257-272. http://dx.doi.org/10.1080/08905439209549838

Thuong PLH, Trung TT, Hoai NTT (2015) Antimicrobial activity of Senna alata (l.), Rhinacanthus nasutus and Chromolaena odorata (l.) collected in Southern Vietnam. IFMBE Proceedings 46: 362-366. https://doi.org/10.1007/978-3-319-11776-8_89

Tung DN, Nhi TV, Hoai NTT (2024) Silver Nanoparticle inhibited levofloxacin resistance development in Staphylococcus aureus. IFMBE Proceedings 95: 297-308. https://doi.org/10.1007/978-3-031-44630-6_24