Characterization of the Bacterial Biofilm Communities Present in Reverse-Osmosis Water Systems for Haemodialysis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sampling
2.2. Scanning Electron Microscopy
2.3. Total and Heterotrophic Bacterial Counts
2.4. Polymerase Chain Reaction (PCR)-Denaturing Gradient Gel Electrophoresis (DGGE) and Analysis of DGGE Profiles
2.5. Genomic DNA Extraction and 16S rRNA Gene Massive Sequencing
2.6. Bacterial Isolation
2.7. Identification of Heterotrophic Aerobic Bacteria
2.8. Antimicrobial Susceptibility Assay
2.9. Statistical Analysis
3. Results
3.1. Scanning Electron Microscopy (SEM) Observation of RO Membranes
3.2. Total Cell and Viable Heterotrophic Bacterial Counts
3.3. Analysis of Similarity of Bacterial Communities by DGGE
3.4. Analysis of the Bacterial Communities’ Composition
3.5. Identification of Isolates
3.6. Antimicrobial Susceptibility Analysis
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Strain | Closest Relative Sequence | Phylum | Identity (%) | Gen Bank Access Number |
---|---|---|---|---|
DCN RO-3 | Pseudomonas sp. | Proteobacteria | 98 | MH114031.1 |
DCN RO-5 | Pseudomonas veronii | Proteobacteria | 95 | MH482963.1 |
DCN RO-6 | Stenotrophomonas maltophilia | Proteobacteria | 96 | FN395264.1 |
DCN RO-7 | Agrobacterium tumefaciens | Proteobacteria | 98 | MG575921.1 |
DCN RO-12 | Pigmentiphaga kullae | Proteobacteria | 98 | KR856352.1 |
DCN PW-4 | Ralstonia sp. | Proteobacteria | 97 | MH844635.1 |
DCN PW-10 | Arthrobacter sp. | Actinobacteria | 95 | JQ316230.1 |
DCN OW-7 | Bacteroides dorei | Bacteroidetes | 96 | CP008741.1 |
DCS RO-1 | Pseudomonas sp. | Proteobacteria | 98 | MK883238.1 |
DCS RO9 | Staphylococcus sp. | Firmicutes | 97 | LC435711.1 |
DCS RO-11 | Staphylococcus pasteuri | Firmicutes | 98 | MG996881.1 |
DCS RO-17 | Pseudomonas fluorescens | Proteobacteria | 98 | EU434450.1 |
DCS PW-7 | Pseudomonas poae | Proteobacteria | 97 | MG835964.1 |
DCS OW-3 | Stenotrophomonas sp. | Proteobacteria | 97 | KP663381.1 |
DCS OW-7 | Stenotrophomonas sp. | Proteobacteria | 98 | KY357351.1 |
Antibiotics | % Resistant Strains |
---|---|
Ampicillin | 69% |
Chloramphenicol | 55% |
Tetracycline | 24% |
Cefotaxime | 53% |
Ciprofloxacin | 15% |
Trimethoprim/sulfamethoxazole | 38% |
Gentamicin | 31% |
Bacterial Genus | AM 10 | CHL 30 | Te 30 | CTX 30 | CIP 5 | TSX 25 | GE 10 |
---|---|---|---|---|---|---|---|
Pseudomonas | 67% (7/11) | 55% (6/11) | 27% (3/11) | 67% (7/11) | 18% (2/11) | 36% (4/11) | 18% (2/11) |
Stenotrophomonas | 100% (4/4) | 50% (2/4) | 50% (2/4) | 50% (2/4) | 0% (0/4) | 25% (1/4) | 50% (2/4) |
Agrobacterium | 100% (0/2) | 0% (0/2) | 0% (0/2) | 100% (2/2) | 0% (0/2) | 0% (0/2) | 50% (1/2) |
Pigmentiphaga | 100% (2/2) | 0% (0/2) | 50% (1/2) | 100% (2/2) | 0% (0/2) | 0% (0/2) | 100% (2/2) |
Ralstonia | 100% (2/2) | 50% (1/2) | 0% (0/2) | 0% (0/2) | 0% (0/2) | 50% (1/2) | 0% (0/2) |
Archobacter | 0% (0/1) | 0% (0/1) | 0% (0/1) | 0% (0/1) | 0% (0/1) | 0% (0/1) | 0% (0/1) |
Bacteroides | 50% (1/2) | 0% (0/2) | 0% (0/2) | 0% (0/2) | 0% (0/2) | 0% (0/2) | 50% (1/2) |
Staphylococcus | 80% (4/5) | 40% (2/5) | 20% (1/5) | 40% (2/5) | 0% (0/5) | 100% (5/5) | 20% (1/5) |
* E. coli ATCC 25922 | R | S | S | S | S | S | S |
* P. aeruginosa ATCC 27853 | R | R | R | R | S | R | S |
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Cuevas, J.-P.; Moraga, R.; Sánchez-Alonzo, K.; Valenzuela, C.; Aguayo, P.; Smith, C.T.; García, A.; Fernandez, Í.; Campos, V.L. Characterization of the Bacterial Biofilm Communities Present in Reverse-Osmosis Water Systems for Haemodialysis. Microorganisms 2020, 8, 1418. https://doi.org/10.3390/microorganisms8091418
Cuevas J-P, Moraga R, Sánchez-Alonzo K, Valenzuela C, Aguayo P, Smith CT, García A, Fernandez Í, Campos VL. Characterization of the Bacterial Biofilm Communities Present in Reverse-Osmosis Water Systems for Haemodialysis. Microorganisms. 2020; 8(9):1418. https://doi.org/10.3390/microorganisms8091418
Chicago/Turabian StyleCuevas, Juan-Pablo, Ruben Moraga, Kimberly Sánchez-Alonzo, Cristian Valenzuela, Paulina Aguayo, Carlos T. Smith, Apolinaria García, Ítalo Fernandez, and Víctor L Campos. 2020. "Characterization of the Bacterial Biofilm Communities Present in Reverse-Osmosis Water Systems for Haemodialysis" Microorganisms 8, no. 9: 1418. https://doi.org/10.3390/microorganisms8091418