Synergistic effects of nisin and CLF36 antimicrobial peptides in vitro

Document Type : Full Research Paper

Authors

1 Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

2 Department of Food Science & Technology, Ferdowsi University of Mashhad

3 Department of Animal Science, Faculty of Agriculture, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

   Globally, clinical mastitis (CM) is a significant disease in the dairy industry and has a tremendous economic impact because of the significantly reduced amount of milk production. Antibiotics are medicines used to treat infections, particularly those of bacterial origin. Misusing these common drugs leads to emerging resistance bacteria and public health concerns in human and animal medicine worldwide. Therefore, the use of natural alternatives with low effects is recommended. This article aims to evaluate the antibacterial activity of peptide nisin on mastitis bacteria and its synergistic effect with CLF36 peptide in vitro. Recombinant peptide nisin and CLF36 were obtained from previous research. The minimum inhibitory concentration (MIC) test was performed using the microdilution method on bacteria extracted from animal mastitis, such as Clostridium perfringens, Proteus mirabilis, Enterococcus faecalis, Salmonella typhimurium, Pseudomonas aeruginosa, and Listeria monocytogenes. This study showed that nisin and CLF36 alone had an inhibitory effect on all the studied bacteria in the 64-32 and 256-32 μg/ml concentration range, respectively. Also, the combination of nisin and CLF36 showed a general synergistic interaction for all used strains (FIC I < 0.5). This study demonstrated that combining these two peptides could reduce the risk of developing drug resistance by creating synergistic effects with a minimum concentration of each of the two peptides.

Keywords


1. Ashari, D.A., Nissa, A., Nursiwi, A., Sari, A.M. and Utami, R., 2019. Antimicrobial effect of Zingiber officinale var. officinale essential oil and nisin against pathogenic and spoilage microorganisms. In IOP Conference Series: Materials Science and Engineering (Vol. 633, No. 1, p. 012005). IOP Publishing.
2. Atef Yekta, M., Verdonck, F., Van Den Broeck, W., Goddeeris, B., Cox, E. and Vanrompay, D., 2010. Lactoferrin inhibits E. coli O157: H7 growth and attachment to intestinal epithelial cells. Veterinarni Medicina, 55(8), pp.359-368.
3. Churklam, W., Chaturongakul, S., Ngamwongsatit, B. and Aunpad, R., 2020. The mechanisms of action of carvacrol and its synergism with nisin against Listeria monocytogenes on sliced bologna sausage. Food Control, 108, p.106864.
4. Cirioni, O., Silvestri, C., Ghiselli, R., Orlando, F., Riva, A., Mocchegiani, F., Chiodi, L., Castelletti, S., Gabrielli, E., Saba, V. and Scalise, G., 2008. Protective effects of the combination of α-helical antimicrobial peptides and rifampicin in three rat models of Pseudomonas aeruginosa infection. Journal of Antimicrobial Chemotherapy, 62(6), pp.1332-1338.
5. Dielbandhoesing, S.K., Zhang, H., Caro, L.H.P., Van Der Vaart, J.M., Klis, F.M., Verrips, C.T. and Brul, S., 1998. Specific cell wall proteins confer resistance to nisin upon yeast cells. Applied and Environmental Microbiology, 64(10), pp.4047-4052.
6. EFSA Panel on Animal Health and Welfare (AHAW), Saxmose Nielsen, S., Alvarez, J., Bicout, D.J., Calistri, P., Depner, K., Drewe, J.A., Garin‐Bastuji, B., Gonzales Rojas, J.L., Gortázar Schmidt, C. and Michel, V., 2020. Health and welfare of rabbits farmed in different production systems. EFSA Journal, 18(1), p.e05944.
7. Embleton, N.D., Berrington, J.E., McGuire, W., Stewart, C.J. and Cummings, S.P., 2013, June. Lactoferrin: Antimicrobial activity and therapeutic potential. In Seminars in Fetal and Neonatal Medicine (Vol. 18, No. 3, pp. 143-149). WB Saunders.
8. Garde, S., Ávila, M., Medina, M. and Nuñez, M., 2004. Fast induction of nisin resistance in Streptococcus thermophilus INIA 463 during growth in milk. International Journal of Food Microbiology, 96(2), pp.165-172.
9. Holtenius, K., Waller, K.P., Essén-Gustavsson, B., Holtenius, P. and Sandgren, C.H., 2004. Metabolic parameters and blood leukocyte profiles in cows from herds with high or low mastitis incidence. The Veterinary Journal, 168(1), pp.65-73.
10. Jahangiri, A., Neshani, A., Mirhosseini, S.A., Ghazvini, K., Zare, H. and Sedighian, H., 2021. Synergistic effect of two antimicrobial peptides, Nisin and P10 with conventional antibiotics against extensively drug-resistant Acinetobacter baumannii and colistin-resistant Pseudomonas aeruginosa isolates. Microbial Pathogenesis, 150, p.104700.
11. Javadmanesh, A., Mohammadi, E., Mousavi, Z., Azghandi, M. and Tanhaiean, A., 2021. Antibacterial effects assessment on some livestock pathogens, thermal stability and proposing a probable reason for different levels of activity of thanatin. Scientific Reports, 11(1), pp.1-10.
12. Liu, H., Pei, H., Han, Z., Feng, G. and Li, D., 2015. The antimicrobial effects and synergistic antibacterial mechanism of the combination of ε-Polylysine and nisin against Bacillus subtilis. Food Control, 47, pp.444-450.
13. Magalhães, L. and Nitschke, M., 2013. Antimicrobial activity of rhamnolipids against Listeria monocytogenes and their synergistic interaction with nisin. Food Control, 29(1), pp.138-142.
14. Mousavi, Z., Rashidian, Z., Zeraatpisheh, Y. and Javadmanesh, A., 2022. Molecular docking of bacteriocin enterocin P peptide with mastitis-causing E. coli antigen in cattle. Veterinary Researches & Biological Products.
15. Mousavi, Z., Tahmoorepur, M., Sekhavati, M.H., and Javadmanesh, A., 2018. Evaluation of antibacterial properties of camel lactoferrampin-lactoferricin recombinant peptide on the growth rate of Staphylococcus aureus bacteriacauses of mastitis in Holstein dairy cows. Journal of Veterinary Microbiology, 14(2), pp.37-47.
16. Mousavi, Z., Tanhaeian, A. and Javadmanesh, A., 2022. Evaluation of in vitro antifungal properties of Thanatin peptide and Chinaberry extract on fungal pathogens of bovine mastitis. Veterinary Researches & Biological Products, 35(1), pp.215-221.
17. Murdock, C.A., Cleveland, J., Matthews, K.R. and Chikindas, M.L., 2007. The synergistic effect of nisin and lactoferrin on the inhibition of Listeria monocytogenes and Escherichia coli O157: H7. Letters in Applied Microbiology, 44(3), pp.255-261.
18. Ndlovu, B., Schoeman, H., Franz, C.M.A.P. and Du Toit, M., 2015. Screening, identification and characterization of bacteriocins produced by wine‐isolated LAB strains. Journal of Applied Microbiology, 118(4), pp.1007-1022.
19. Oliver, S.P. and Murinda, S.E., 2012. Antimicrobial resistance of mastitis pathogens. Veterinary Clinics: Food Animal Practice, 28(2), pp.165-185.
20. Roshanak, S., Shahidi, F., Yazdi, F.T., Javadmanesh, A. and Movaffagh, J., 2020. Evaluation of antimicrobial activity of Buforin I and Nisin and the synergistic effect of their combination as a novel antimicrobial preservative. Journal of Food Protection, 83(11), pp.2018-2025.
21. Roshanak, S., Shahidi, F., Yazdi, F.T., Javadmanesh, A. and Movaffagh, J., 2021. Buforin I an alternative to conventional antibiotics: Evaluation of the antimicrobial properties, stability, and safety. Microbial Pathogenesis, 161, p.105301.
22. Shahidi, F., Roshanak, S., Javadmanesh, A., Tabatabaei Yazdi, F., Pirkhezranian, Z. and Azghandi, M., 2020. Evaluation of antimicrobial properties of bovine lactoferrin against foodborne pathogenic microorganisms in planktonic and biofilm forms (in vitro). Journal of Consumer Protection and Food Safety, 15(3), pp.277-283.
23. Tafreshi, S.Y.H. and Mirdamadi, S., 2015. Survey study of lipid effect on nisin nanoliposome formation and 
application in pasteurized milk as a food model. Applied Food Biotechnology, 2(2), pp.7-14..
24. Tanhaian, A., Azghandi, M., Razmyar, J., Mohammadi, E. and Sekhavati, M.H., 2018. Recombinant production of a chimeric antimicrobial peptide in E. coli and assessment of its activity against some avian clinically isolated pathogens. Microbial Pathogenesis, 122, pp.73-78.
25. Yedery, R.D. and Reddy, K.V.R., 2005. Antimicrobial peptides as microbicidal contraceptives: prophecies for prophylactics–a mini review. The European Journal of Contraception & Reproductive Health Care, 10(1), pp.32-42.