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Cellular alterations in Pectobacterium carotovorum treated with nanostructured formulations during the incubation time

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Abstract

Pectobacterium carotovorum was incubated in formulations of chitosan nanoparticles or thyme essential oil-loaded chitosan nanoparticles for a maximum period of 48 h time. The cellular changes and viability were evaluated by transmission electron microscopy (TEM), and two colorimetric assays 3-(4,5 dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide (MTT) and alamar blue (AMB), respectively. The incubation time and the addition of the secondary metabolite to the formulation were key factors to the cell damage and cell inhibitory effects on P. carotovorum, TEM observations overall demonstrated on the treated bacterium, cell surface alterations such as deforming and disappearance of the cell wall and the plasmatic membrane, with agglomeration of nanoparticles outside and inside of the cells, loss of cell content and lysis. Cell viability was reduced about 80% (MTT) and 100% (AMB) in the applied treatment of chitosan-loaded thyme essential oil nanoparticles after 48 h incubation, in addition, the total cell inhibition was shown from 6 h incubation onwards with the AMB assay. The TEM micrographs and the cell viability assays provided enough evidence of the antimicrobial activity of the nanostructured formulations compared with the control where no damage was observed.

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Acknowledgements

The authors acknowledge the financial support of the Secretary of Research and Postgraduates of the National Polytechnic Institute, through the project SIP-20160468 and Dr. Eva Ramón-Gallegos and Rocío Cruz-Muñoz of the National Polytechnic Institute for the technical guidance of the cell viability assays.

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Authors and Affiliations

  1. Instituto Politécnico Nacional, Centro de Desarrollo de Productos Bióticos (CEPROBI), Carretera Yautepec-Jojutla, Km 6, San Isidro, CP 62730, Yautepec, Morelos, Mexico

    Maria Elena Sotelo-Boyás, Maria Luisa Corona-Rangel & Silvia Bautista-Baños

  2. Consejo Nacional de Ciencia y Tecnología and CEPROBI, Carretera Yautepec-Jojutla, Km 6, San Isidro, CP 62730, Yautepec, Morelos, Mexico

    Zormy Correa-Pacheco

  3. Instituto Politécnico Nacional, Unidad Profesional Interdisciplinaria de Biotecnología, Avenida Acueducto s/n, Barrio La Laguna Ticomán, CP 07340, Mexico, Mexico

    Ramón Villanueva-Arce

Authors
  1. Maria Elena Sotelo-Boyás
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  2. Zormy Correa-Pacheco
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  3. Maria Luisa Corona-Rangel
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  4. Ramón Villanueva-Arce
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  5. Silvia Bautista-Baños
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Correspondence to Silvia Bautista-Baños.

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The authors declare that they have no competing interest.

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This study did not involve humans, human data or animals; no ethics approval or consent is required to publish the results.

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Communicated by Erko Stackebrandt.

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Sotelo-Boyás, M.E., Correa-Pacheco, Z., Corona-Rangel, M.L. et al. Cellular alterations in Pectobacterium carotovorum treated with nanostructured formulations during the incubation time. Arch Microbiol 201, 615–622 (2019). https://doi.org/10.1007/s00203-019-01628-w

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  • DOI: https://doi.org/10.1007/s00203-019-01628-w

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