Laser Treatment Increases the Antimicrobial Efficacy of Cyanobacterial Extracts against Staphylococcusaureus (SA) and Methicillin-resistantStaphylococcus aureus (MRSA)
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolation, Purification, and Morphological Identification of Cyanobacterial Samples
2.2. Pathogenic Bacterial Samples Used in Antimicrobial Bioassay
2.3. Initial Screening of Cyanobacterial Extracts and Antimicrobial Bioassays against Pathogenic Bacteria
2.4. Molecular Identification of Cyanobacterial Strains
2.4.1. DNA Extract of Cyanobacterial Strains
2.4.2. PCR Amplification of Cyanobacterial Genomic DNA
- Initial denaturation at 94 °C for 2 min.
- Amplification cycles (35×). Each cycle consists of:
- Denaturation at 94 °C for 20 s;
- Annealing at 55 °C for 30 s;
- Extension at 72 °C for 10 s;
- Final extension step at 72 °C for 10 min [47].
- The amplicons were purified and sequenced (Macrogen, South Korea), and the sequences were checked in Genbank for similarity check (Blastn). The sequences were deposited in GenBank to await accession number designation.
2.5. Laser Treatments Using Different Exposure Time Intervals and Distances
2.6. Measurement the Phycobiliprotein Pigments Quantity of Cyanobacterial Extracts Spectrophotometrically before and after Exposing to Laser
2.7. Statistical Analysis
3. Results
3.1. Molecular Identification of Cyanobacterial Isolate Strains
3.2. Phylogenetic Inference
- The filamentous unbranched nonheterocystous strain that was isolated from the thermal sulfur water spring belonged to Thermoleptolyngbya sp., which exhibited 92.92% sequence homology.
- The filamentous unbranched nonheterocystous strain that was isolated from the Hassawi rice field belonged to Leptolyngbya sp., which exhibited 94.58% sequence homology.
3.3. Molecular Identification of Bacteria
3.4. Antimicrobial Activity of Cyanobacterial Extracts against Pathogenic Bacteria
- Thermoleptolyngbya sp. extracts directly exposed to laser against Staphylococcus aureus, with an inhibition zone of 22 mm at 10 cm and 32 min.
- Thermoleptolyngbya sp. extracts directly exposed to the laser against Methicillin-resistant Staphylococcus aureus, with an inhibition zone of 19 mm at 5 cm and 16 min.
3.5. Measurement the Phycobiliprotein Pigments Quantification of Cyanobacteria by Spectrophotometric
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Cyanobacterial Strain | Distance | Exposure Time | Inhibition Zone of Staphylococcus aureus | |
---|---|---|---|---|
Extracts from Biomass Pre-Exposed to Laser | Extracts Directly Exposed to Laser | |||
Thermoleptolyngbya sp. | 5 cm | 4 min | 7.0 ± 0.0 | 15.5 ± 7.7 |
16 min | 14.0 ± 1.4 | 15.0 ± 0.0 | ||
32 min | 10.0 ± 1.4 | 15.0 ± 7.0 | ||
10 cm | 4 min | 12.0 ± 1.4 | 20.0 ± 4.2 | |
16 min | 8.5.0 ± 0.7 | 19.0 ± 4.2 | ||
32 min | 14.5 ± 3.5 | 22.0 ± 4.2 | ||
Control, no laser treatment | 11 ± 0.0 | 11.0 ± 0.0 | ||
Leptolyngbya sp. | 5 cm | 4 min | 8.0 ± 0.0 | 21.5 ± 3.5 |
16 min | 12.5 ± 3.5 | 11.5 ± 2.1 | ||
32 min | 8.5 ± 0.7 | 9.0 ± 0.0 | ||
10 cm | 4 min | 8.0 ± 1.4 | 11.0 ± 2.8 | |
16 min | 15.0 ± 2.8 | 10.0 ± 4.2 | ||
32 min | 9.5 ± 0.7 | 14.0 ± 1.4 | ||
Control, no laser treatment | 9.5 ± 0.0 | 9.5 ± 0.0 |
Cyanobacterial Strain | Distance | Exposure Time | Inhibition zone of MRSA-Staphylococcus aureus | |
---|---|---|---|---|
Extracts from Biomass Pre-Exposed to Laser | Extracts Directly Exposed to Laser | |||
Thermoleptolyngbya sp. | 5 cm | 4 min | 6.5 ± 0.7 | 13.5 ± 3.5 |
16 min | 10.0 ± 1.4 | 19.0 ± 1.4 | ||
32 min | 12.0 ± 4.9 | 13.5 ± 2.1 | ||
10 cm | 4 min | 8.5 ± 2.1 | 13.5 ± 3.5 | |
16 min | 9.0 ± 0.0 | 14.5 ± 4.9 | ||
32 min | 9.5 ± 0.7 | 16.0 ± 1.4 | ||
Control, no Laser treatment | - | - | 9.0 ± 0.0 | 9.0 ± 0.0 |
Leptolyngbya sp. | 5 cm | 4 min | 8.5 ± 0.7 | 13.0 ± 0.0 |
16 min | 8.5 ± 0.7 | 8.5 ± 0.7 | ||
32 min | 10.5 ± 2.1 | 11.0 ± 2.8 | ||
10 cm | 4 min | 11.0 ± 2.3 | 10.0 ± 2.1 | |
16 min | 8.5 ± 2.1 | 11.0 ± 2.8 | ||
32 min | 9.0 ± 0.0 | 9.0 ± 0.0 | ||
Control, no Laser treatment | 6.5 ± 0.0 | 6.5 ± 0.0 |
Species | Distance | Time | Phycocyanin (mg/mL) | Allophycocyanin (mg/mL) | Phycoerythrin (mg/mL) |
---|---|---|---|---|---|
Thermoleptolyngbya sp. | 0 (Control, no laser treatment) | 0 (Control, no laser treatment) | 0.0624704 | 0.0380794 | 0.205086 |
10 | 32 | 0.824993 | 0.118537 | 0.690715 | |
Leptolyngbya sp. | 0 (Control, no laser treatment) | 0 (Control, no laser treatment) | 0.0244599 | 0.0329147 | 0.206482 |
5 | 4 | 0.106869 | 0.0971033 | 0.527876 |
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Al Naim, H.M.; El Semary, N. Laser Treatment Increases the Antimicrobial Efficacy of Cyanobacterial Extracts against Staphylococcusaureus (SA) and Methicillin-resistantStaphylococcus aureus (MRSA). Int. J. Environ. Res. Public Health 2022, 19, 13305. https://doi.org/10.3390/ijerph192013305
Al Naim HM, El Semary N. Laser Treatment Increases the Antimicrobial Efficacy of Cyanobacterial Extracts against Staphylococcusaureus (SA) and Methicillin-resistantStaphylococcus aureus (MRSA). International Journal of Environmental Research and Public Health. 2022; 19(20):13305. https://doi.org/10.3390/ijerph192013305
Chicago/Turabian StyleAl Naim, Haifa M., and Nermin El Semary. 2022. "Laser Treatment Increases the Antimicrobial Efficacy of Cyanobacterial Extracts against Staphylococcusaureus (SA) and Methicillin-resistantStaphylococcus aureus (MRSA)" International Journal of Environmental Research and Public Health 19, no. 20: 13305. https://doi.org/10.3390/ijerph192013305