Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii

doi:10.1007/s11686-021-00436-x

. 2022 Mar;67(1):511-517.

doi: 10.1007/s11686-021-00436-x. Epub 2021 Jun 22.

Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii

Affiliations

¹ Laboratório de Entomologia e Parasitologia Tropical, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, 49100-000, Brazil.
² Laboratório de Biologia e Imunologia de Câncer e Leishmania, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, Brazil.
³ Programa de Pós-Graduação em Biotecnologia Industrial, Universidade Tiradentes, Aracaju, Sergipe, Brazil.
⁴ Departamento de Microbiologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.
⁵ Laboratório de Bacteriologia, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, 49100-000, Brazil. ana.barbosaufs@yahoo.com.br.
⁶ Laboratório de Entomologia e Parasitologia Tropical, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, 49100-000, Brazil. dolabellaufs@gmail.com.

PMID: 34156633
DOI: 10.1007/s11686-021-00436-x

Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii

Marianna de Carvalho Clímaco et al. Acta Parasitol. 2022 Mar.

. 2022 Mar;67(1):511-517.

doi: 10.1007/s11686-021-00436-x. Epub 2021 Jun 22.

Affiliations

¹ Laboratório de Entomologia e Parasitologia Tropical, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, 49100-000, Brazil.
² Laboratório de Biologia e Imunologia de Câncer e Leishmania, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, Brazil.
³ Programa de Pós-Graduação em Biotecnologia Industrial, Universidade Tiradentes, Aracaju, Sergipe, Brazil.
⁴ Departamento de Microbiologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.
⁵ Laboratório de Bacteriologia, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, 49100-000, Brazil. ana.barbosaufs@yahoo.com.br.
⁶ Laboratório de Entomologia e Parasitologia Tropical, Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, Sergipe, 49100-000, Brazil. dolabellaufs@gmail.com.

PMID: 34156633
DOI: 10.1007/s11686-021-00436-x

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.

Keywords: Acanthamoeba castellanii; Bacteriocins; Free-living amoeba; Mode of action.

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References

1. 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 - DOI - PubMed
1. Duggal SD, Rongpharpi SR, Duggal AK et al (2017) Role of Acanthamoeba in granulomatous encephalitis: a review. J Infect Dis Immune Ther 1:1
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 - DOI - PubMed
1. 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 - DOI
1. 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 - DOI - PubMed

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[1] 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 - DOI - PubMed

[2] 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 - DOI - PubMed

[3] Duggal SD, Rongpharpi SR, Duggal AK et al (2017) Role of Acanthamoeba in granulomatous encephalitis: a review. J Infect Dis Immune Ther 1:1

[4] Duggal SD, Rongpharpi SR, Duggal AK et al (2017) Role of Acanthamoeba in granulomatous encephalitis: a review. J Infect Dis Immune Ther 1:1

[5] 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 - DOI - PubMed

[6] 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 - DOI - PubMed

[7] 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 - DOI

[8] 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 - DOI

[9] 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 - DOI - PubMed

[10] 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 - DOI - PubMed

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Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii

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Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii

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