Abstract
Purpose
Acanthamoeba spp. are free-living amoebas with worldwide distribution and play an important role as disease-causing agents in humans. Drug inability to completely eradicate these parasites along with their toxic effects suggest urgent need for new antimicrobials. Nisin is a natural antimicrobial peptide produced by Lactococcus lactis. Nisin is also the only bacteriocin approved for use in food preservation. In this work, we analyzed the effect of nisin on the growth of Acanthamoeba castellanii trophozoites.
Methods
A total of 8 × 104 trophozoites were exposed to increasing concentrations of nisin to determine its activity. Changes in cell membrane and cellular cycle of trophozoites were investigated by flow cytometry, and nisin cytotoxicity in mammalian cells was evaluated in L929 cells by MTT method.
Results
After 24 h exposure to increasing nisin concentrations, an IC50 of 4493.2 IU mL−1 was obtained for A. castellanii trophozoites. However, after 72 h a recovery in amoebic growth was observed, and it was no longer possible to determine IC50. Flow cytometry analysis showed that nisin has no effect on the membrane integrity. Treatment with nisin induced cell-cycle arrest during G1 and S phases in A. castellanii trophozoites, which recovered their growth after 72 h.
Conclusion
This is one of the first studies showing the effect of internationally approved nisin against A. castellanii trophozoites. Nisin caused cell-cycle arrest in trophozoites, momentarily interfering with the DNA replication process. The data highlight the amoebostatic activity of nisin, and suggest its use as an adjuvant for the treatment of infections caused by Acanthamoeba spp.
References
Mahmoudi MR, Rahmati B, Seyedpour SH, Karanis P (2015) Occurrence and molecular characterization of free-living amoeba species (Acanthamoeba, Hartmannella, and Saccamoeba limax) in various surface water resources of Iran. Parasitol Res 114:4669–4674. https://doi.org/10.1007/s00436-015-4712-8
Duggal SD, Rongpharpi SR, Duggal AK et al (2017) Role of Acanthamoeba in granulomatous encephalitis: a review. J Infect Dis Immune Ther 1:1
Harrison WT, Lecky B, Hulette CM (2018) Fatal granulomatous amebic encephalitis in a heart transplant patient: clinical, radiographic, and autopsy findings. J Neuropathol Exp Neurol 77:1001–1004. https://doi.org/10.1093/jnen/nly089
Matsui T, Maeda T, Kusakabe S et al (2018) A case report of granulomatous amoebic encephalitis by group 1 Acanthamoeba genotype T18 diagnosed by the combination of morphological examination and genetic analysis. Diagn Pathol 13:1–6. https://doi.org/10.1186/s13000-018-0706-z
Robaei D, Carnt N, Minassian DC, Dart JKG (2014) The impact of topical corticosteroid use before diagnosis on the outcome of Acanthamoeba keratitis. Ophthalmology 121:1383–1388. https://doi.org/10.1016/j.ophtha.2014.01.031
Maycock NJR, Jayaswal R (2016) Update on Acanthamoeba keratitis: diagnosis, treatment, and outcomes. Cornea 35:713–720. https://doi.org/10.1097/ICO.0000000000000804
CDC (2017) Acanthamoeba keratitis fact sheet for healthcare professionals. https://www.cdc.gov/parasites/acanthamoeba/health_professionals/acanthamoeba_keratitis_hcp.html. Accessed 20 October 2020
Papa V, Rama P, Radford C et al (2020) Acanthamoeba keratitis therapy: time to cure and visual outcome analysis for different antiamoebic therapies in 227 cases. Br J Ophthalmol 104:575–581. https://doi.org/10.1136/bjophthalmol-2019-314485
Ehlers N, Hjortdal J (2004) Are cataract and iris atrophy toxic complications of medical treatment of Acanthamoeba keratitis? Acta Ophthalmol Scand 82:228–231. https://doi.org/10.1111/j.1600-0420.2004.00237.x
Herz NL, Matoba AY, Wilhelmus KR (2008) Rapidly progressive cataract and iris atrophy during treatment of Acanthamoeba keratitis. Ophthalmology 115:866–869. https://doi.org/10.1016/j.ophtha.2007.05.054
Carrijo-Carvalho LC, Sant’ana VP, Foronda AS et al (2017) Therapeutic agents and biocides for ocular infections by free-living amoebae of Acanthamoeba genus. Surv Ophthalmol 62:203–218. https://doi.org/10.1016/j.survophthal.2016.10.009
Chikindas ML, Weeks R, Drider D et al (2018) Functions and emerging applications of bacteriocins. Curr Opin Biotechnol 49:23–28. https://doi.org/10.1016/j.copbio.2017.07.011
Modugno C, Loupiac C, Bernard A et al (2018) Effect of high pressure on the antimicrobial activity and secondary structure of the bacteriocin nisin. Innov Food Sci Emerg Technol 47:9–15. https://doi.org/10.1016/j.ifset.2018.01.006
Ogaki MB, Furlaneto MC, Maia LF (2015) Review: General aspects of bacteriocins. Braz J Food Technol 18:267–276. https://doi.org/10.1590/1981-6723.2215
Bali V, Panesar PS, Bera MB, Kennedy JF (2016) Bacteriocins: recent trends and potential applications. Crit Rev Food Sci Nutr 56:817–834. https://doi.org/10.1080/10408398.2012.729231
Hussein AR, Khalaf ZZ, Kadhim MJ (2017) The antibiofilm activity of bacteriocin produced by Proteus mirabilis against some bacterial species. Current Res Microbiol Biotechnol 5(3):1071–1077
Atanaskovic I, Kleanthous C (2019) Tools and approaches for dissecting protein bacteriocin import in gram-negative bacteria. Front Microbiol 10:1–12. https://doi.org/10.3389/fmicb.2019.00646
Drider D, Bendali F, Naghmouchi K, Chikindas ML (2016) Bacteriocins: not only antibacterial agents. Probiotics Antimicrob Proteins 8:177–182. https://doi.org/10.1007/s12602-016-9223-0
Lewies A, Du Plessis LH, Wentzel JF (2018) Antimicrobial peptides: the Achilles’ heel of antibiotic resistance? Probiotics Antimicrob Proteins 11:370–381. https://doi.org/10.1007/s12602-018-9465-0
Gharsallaoui A, Oulahal N, Joly C, Degraeve P (2016) Nisin as a food preservative: part 1: physicochemical properties, antimicrobial activity, and main uses. Crit Rev Food Sci Nutr 56:1262–1274. https://doi.org/10.1080/10408398.2013.763765
Shin JM, Gwak JW, Kamarajan P et al (2016) Biomedical applications of nisin. J Appl Microbiol 120:1449–1465. https://doi.org/10.1111/jam.13033
Benitez LB, Caumo K, Brandelli A, Rott MB (2011) Bacteriocin-like substance from Bacillus amyloliquefaciens shows remarkable inhibition of Acanthamoeba polyphaga. Parasitol Res 108:687–691. https://doi.org/10.1007/s00436-010-2114-5
Anacarso I, Bondi M, Condo C (2014) Amoebicidal effects of three bacteriocin like substances from lactic acid bacteria against Acanthamoeba polyphaga. J Bacteriol Parasitol 6:8–11. https://doi.org/10.4172/2155-9597.1000201
de Santos IGA, Scher R, Rott MB et al (2016) Amebicidal activity of the essential oils of Lippia spp. (Verbenaceae) against Acanthamoeba polyphaga trophozoites. Parasitol Res 115:535–540. https://doi.org/10.1007/s00436-015-4769-4
Britta EA, Barbosa Silva AP, Ueda-Nakamura T et al (2012) Benzaldehyde thiosemicarbazone derived from limonene complexed with copper induced mitochondrial dysfunction in Leishmania amazonensis. PLoS ONE 7:1–12. https://doi.org/10.1371/journal.pone.0041440
Mukherjee C, Clark CG, Lohia A (2008) Entamoeba shows reversible variation in ploidy under different growth conditions and between life cycle phases. PLoS Negl Trop Dis 2:1–9. https://doi.org/10.1371/journal.pntd.0000281
Uzlikova M, Nohynkova E (2014) The effect of metronidazole on the cell cycle and DNA in metronidazole-susceptible and—resistant giardia cell lines. Mol Biochem Parasitol 198:75–81. https://doi.org/10.1016/j.molbiopara.2015.01.005
ISO (2009) Biological evaluation of medical devices. Part 5: tests for in vitro cytotoxicity. 10993-5
Amer EI, Mossallam SF, Mahrous H (2014) Therapeutic enhancement of newly derived bacteriocins against giardia lamblia. Exp Parasitol 146:52–63. https://doi.org/10.1016/j.exppara.2014.09.005
Oyeyemi O, Adegbeyeni O, Oyeyemi I et al (2018) In vitro ovicidal activity of poly lactic acid curcumin-nisin co-entrapped nanoparticle against Fasciola spp. eggs and its reproductive toxicity. J Basic Clin Physiol Pharmacol 29:73–79. https://doi.org/10.1515/jbcpp-2017-0045
Martínez-García M, Bart JM, Campos-Salinas J et al (2018) Autophagic-related cell death of Trypanosoma brucei induced by bacteriocin AS-48. Int J Parasitol Drugs Drug Resist 8:203–212. https://doi.org/10.1016/j.ijpddr.2018.03.002
Martín-Escolano R, Cebrián R, Martín-Escolano J et al (2019) Insights into Chagas treatment based on the potential of bacteriocin AS-48. Int J Parasitol Drugs Drug Resist 10:1–8. https://doi.org/10.1016/j.ijpddr.2019.03.003
Abengózar MÁ, Cebrián R, Saugar JM et al (2017) Enterocin AS-48 as evidence for the use of bacteriocins as new leishmanicidal agents. Antimicrob Agents Chemother 61:1–13. https://doi.org/10.1128/AAC.02288-16
Rose NL, Sporns P, Stiles ME, McMullen LM (1999) Inactivation of nisin by glutathione in fresh meat. J Food Sci 64:759–762. https://doi.org/10.1111/j.1365-2621.1999.tb15906.x
Bhatti M, Veeramachaneni A, Shelef LA (2004) Factors affecting the antilisterial effects of nisin in milk. Int J Food Microbiol 97:215–219. https://doi.org/10.1016/j.ijfoodmicro.2004.06.010
Grisi TCSDL, Gorlach-Lira K (2005) Action of nisin and high pH on growth of Staphylococcus aureus and Salmonella sp. in pure culture and in the meat of land crab (Ucides cordatus). Braz J Microbiol 36:151–156. https://doi.org/10.1590/S1517-83822005000200010
Khunkitti W, Avery SV, Lloyd D et al (1997) Effects of biocides on Acanthamoeba castellanii as measured by flow cytometry and plaque assay. J Antimicrob Chemother 40:227–233. https://doi.org/10.1093/jac/40.2.227
Borazjani RN, May LL, Noble JA et al (2000) Flow cytometry for determination of the efficacy of contact lens disinfecting solutions against Acanthamoeba spp. Appl Environ Microbiol 66:1057–1061. https://doi.org/10.1128/AEM.66.3.1057-1061.2000
Mogoa E, Bodet C, Legube B, Héchard Y (2010) Acanthamoeba castellanii: cellular changes induced by chlorination. Exp Parasitol 126:97–102. https://doi.org/10.1016/j.exppara.2009.12.005
Heredero-Bermejo I, Copa-Patiño JL, Soliveri J et al (2013) In vitro evaluation of the effectiveness of new water-stable cationic carbosilane dendrimers against Acanthamoeba castellanii UAH-T17c3 trophozoites. Parasitol Res 112:961–969. https://doi.org/10.1007/s00436-012-3216-z
Imayasu M, Tchedre KT, Dwight Cavanagh H (2013) Effects of multipurpose solutions on the viability and encystment of Acanthamoeba determined by flow cytometry. Eye Contact Lens 39:228–233. https://doi.org/10.1097/ICL.0b013e31828af147
Heredero-Bermejo I, Copa-Patiño JL, Soliveri J et al (2015) Evaluation of the activity of new cationic carbosilane dendrimers on trophozoites and cysts of Acanthamoeba polyphaga. Parasitol Res 114:473–486. https://doi.org/10.1007/s00436-014-4205-1
Sang Y, Blecha F (2008) Antimicrobial peptides and bacteriocins: alternatives to traditional antibiotics. Anim Health Res Rev 9:227–235. https://doi.org/10.1017/S1466252308001497
Stohr M, Bommert K, Schulze I, Jantzen H (1987) The cell cycle and its relationship to development in Acanthamoeba castellanii. J Cell Sci 88:579–590
Jantzen H, Schulze I, Stöhr M (1988) Relationship between the timing of DNA replication and the developmental competence in Acanthamoeba castellanii. J Cell Sci 91(Pt 3):389–399
Byers TJ, Kim BG, King LE, Hugo ER (1991) Molecular aspects of the cell cycle and encystment of Acanthamoeba. Rev Infect Dis 13:S373–S384
Yang SC, Lin CH, Sung CT, Fang JY (2014) Antibacterial activities of bacteriocins: application in foods and pharmaceuticals. Front Microbiol 5:1–10. https://doi.org/10.3389/fmicb.2014.00241
Cotter PD, Ross RP, Hill C (2013) Bacteriocins—a viable alternative to antibiotics? Nat Rev Microbiol 11:95–105. https://doi.org/10.1038/nrmicro2937
Rodrigues FAR, Bomfim IS, Cavalcanti BC et al (2014) Mefloquine-oxazolidine derivatives: a new class of anticancer agents. Chem Biol Drug Des 83:126–131. https://doi.org/10.1111/cbdd.12210
Acknowledgements
This study was partly financed by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES-Finance Code 001).
Author information
Authors and Affiliations
Contributions
SSD and AATB conceived the study and designed the experiments. MCC, YLMO, JRS and ARSTS performed the experiments. RS and CBC participated in the flow cytometry experiments and guided data interpretation. SSD, AATB, MCC, ACSR, SJ and MBR analyzed data and participated in the writing and revision of the manuscript. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
de Carvalho Clímaco , M., de Oliveira, Y.L.M., Ramos, A.C.S. et al. Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii. Acta Parasit. 67, 511–517 (2022). https://doi.org/10.1007/s11686-021-00436-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11686-021-00436-x