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Biofilms of Pseudomonas and Lysinibacillus Marine Strains on High-Density Polyethylene

  • Microbiology of Aquatic Systems
  • Published:
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Abstract

Environmental pollution by plastic debris is estimated on a scale of 100 million metric tons, a portion of which is fragmented into micro- and nanoplastics. These fragments are often colonized by bacterial species in marine environments, possibly contributing to the biodegradation of such materials. However, further investigations are necessary to determine the impact of abiotic polymer weathering on biofilm adhesion, as well as the specific biofilm formation strategies employed by marine isolates. Here, we evaluate deep-sea sediment bacterial isolates for biofilm adhesion, extracellular matrix production, and polymer degradation ability. Our study focuses on high-density polyethylene (HDPE) fragments for their high durability and environmental persistence, subjecting fragments to abiotic weathering prior to bacterial colonization. Marine isolates identified as Pseudomonas sp. and Lysinibacillus sp. exhibited decreasing biofilm formation on weathered HDPE, especially over the first 24 h of incubation. This effect was countered by increased extracellular matrix production, likely improving cell adhesion to surfaces roughened by abiotic degradation. These adhesion strategies were contrasted with a reference Pseudomonas aeruginosa strain, which displayed high levels of biofilm formation on non-weathered HDPE and lower extracellular matrix production over the first 24 h of incubation. Furthermore, our results suggest that an increase in biofilm biomass correlated with changes to HDPE structure, indicating that these strains have a potential for biodegradation of plastic fragments. Therefore, this work provides a detailed account of biofilm formation strategies and bacteria-plastic interactions that represent crucial steps in the biodegradation of plastic fragments in marine environments.

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Acknowledgments

We thank Petróleo Brasileiro S.A. (PETROBRAS) for collecting the samples and financial support; Indústria Petroquímica Braskem S.A. for proving HDPE pellets; the Central Laboratory of Microscopy and Microanalysis (LabCEMM/PUCRS) for preparing and assisting the microscopic visualization; CNPq (Brazilian National Council for Scientific and Technological Development) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Brasil) for the scholarships of master’s students.

Funding

The financial support was provided by Conegás II Project (PETROBRAS TC N° 0050.0096017.15.9) and IPR/PUCRS Research Fund. Funding was also provided by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES - Finance Code 001) and Brazilian National Council for Scientific and Technological Development (CNPq - master’s scholarship).

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Authors and Affiliations

  1. Geobiology Laboratory, Institute of Petroleum and Natural Resources, Pontifical Catholic University of Rio Grande do Sul, PUCRS, Av. Ipiranga 6681, Building 96J, Porto Alegre, RS, 90619-900, Brazil

    Maiara Monteiro Oliveira, Audrey Menegaz Proenca, Francine Melise dos Santos, Letícia Marconatto & Renata Medina-Silva

  2. Immunology and Microbiology Laboratory, School of Health and Life Sciences, Pontifical Catholic University of Rio Grande do Sul, PUCRS, Av. Ipiranga 6681, Building 11, Porto Alegre, RS, 90619-900, Brazil

    Maiara Monteiro Oliveira, Audrey Menegaz Proenca, Eduardo Moreira-Silva & Renata Medina-Silva

  3. Biotechnology Division, Research and Development Center (CENPES), PETROBRAS, Av. Horácio Macedo 950, Rio de Janeiro, RJ, 21941-915, Brazil

    Aline Machado de Castro

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  1. Maiara Monteiro Oliveira
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  2. Audrey Menegaz Proenca
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  3. Eduardo Moreira-Silva
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Correspondence to Renata Medina-Silva.

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Supplementary Information

Fig S1

Biofilm formation pattern of each marine strain after 24 or 48 h of incubation with HDPE fragments. (a) Reference strain, (b) M3, (c) M4 and (d) M5 isolates showed an overall decrease in biofilm formation with HDPE weathering. The reference strain exhibited lower biofilm formation after 48 h, but the marine isolates showed similar biofilm levels after 24 or 48 h. Points represent mean ± SEM. (PNG 147 kb)

High resolution image (TIF 483 kb)

Fig S2

3D-AFM images of marine strains that showed more heterogeneous surfaces in different experimental conditions. Non-weathered HDPE: (a) M5 and (b) M3. 400-HDPE: (c) M5 and (d) M4. 600-HDPE: (e) reference strain and (f) M4. 800-HDPE: (g) M3 and (h) M5. Scale bar = 10 μm. (PNG 1656 kb)

High resolution image (TIF 6624 kb)

Fig S3

FTIR spectra of biofilm-colonized HDPE fragments obtained for all strains, surface weathering levels and incubation intervals. Absorbance values were vector-normalized, and spectra were shifted vertically for representation. RS = Reference strain (PNG 569 kb)

High resolution image (TIF 1830 kb)

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Oliveira, M.M., Proenca, A.M., Moreira-Silva, E. et al. Biofilms of Pseudomonas and Lysinibacillus Marine Strains on High-Density Polyethylene. Microb Ecol 81, 833–846 (2021). https://doi.org/10.1007/s00248-020-01666-8

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