Detection of Enteric Viruses and Bacterial Indicators in a Sewage Treatment Center and Shallow Water Bay
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Collection
2.2. Virus Concentration
2.3. Viral RNA Extraction and Synthesis of Complementary DNA
2.4. Detection of Enteric Viruses by PCR
2.5. Microbial Indicators
2.6. Statistical Analysis
3. Results
3.1. Enteric Viruses Detection
3.2. Microbiological Indicators
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Virus | Primer Sequence (5′→3′) | Amplicon Size (bp) | Map Position | Reference |
---|---|---|---|---|
EV | FW: 5′-CAAGCACTTCTGTTTCCCCGG-3′ RV: 5′-ATTGTCACCATAAGCAGCCA-3′ | 434 | 162–182 577–596 | [28] |
FW: 5′-TCCGGCCCCTGAATGCGG-3′ RV: 5′-CACCGGATGGCCAATCCAAT-3′ | 196 | 446–463 623–642 | [28] | |
AV | FW: 5′-CGTCATTATTTGTTGTCATACT-3′ RV: 5′-ACATGTGCTGCTGTTACTATG-3′ | 289 | 1182–1203 1450–1470 | [29] |
HAV | FW: 5′-CAGCACATCAGAAAGGTGAG-3′ RV: 5′-CTCCAGAATCATCTCCAAC-3′ | 192 | 2035–2054 2208–2226 | [27] |
RV | FW:5′-GGCTTTAAAAGAGAGAATTTCCGTCTGG-3′ RV: 5′-GATCCTGTTGGCCATCC-3′ | 392 | 1–28 376–392 | [28] |
Virus | PCR Conditions (Temperature and Time) | |||||
---|---|---|---|---|---|---|
Cycles | Initial Denaturation | Template Denaturation | Primer Annealing | Primer Extension | Final Extension | |
EV | 35 | 94 °C, 3 min | 94 °C, 45 s | 55 °C, 30 s | 72 °C, 60 s | 72 °C, 10 min |
AV | 30 | 94 °C, 3 min | 94 °C, 30 s | 50 °C, 20 s | 72 °C, 30 s | 72 °C, 5 min |
HAV | 35 | 94 °C, 3 min | 94 °C, 1 min | 51 °C, 1:30 min | 72 °C, 1 min | 72 °C, 7 min |
RV | 40 | 94 °C, 3 min | 94 °C, 30 s | 50 °C, 20 s | 72 °C, 30 s | 72 °C, 5 min |
Detected Bacteria | Mean Total Count of Bacteria (cfu/mL) 1 | Mean Titer of Bacteria-Specific Phages (pfu/mL) 2 | |
---|---|---|---|
Influent | Effluent | ||
E. coli | 3.20 × 103 | 8.2 × 1010 | 2.15 × 108 |
Salmonella spp. | 1.32 × 103 | 5.6 × 105 | 2.77 × 103 |
Shigella spp. | 1.92 × 103 | ND * | ND * |
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Janahi, E.M.; Mustafa, S.; Parkar, S.F.D.; Naser, H.A.; Eisa, Z.M. Detection of Enteric Viruses and Bacterial Indicators in a Sewage Treatment Center and Shallow Water Bay. Int. J. Environ. Res. Public Health 2020, 17, 6483. https://doi.org/10.3390/ijerph17186483
Janahi EM, Mustafa S, Parkar SFD, Naser HA, Eisa ZM. Detection of Enteric Viruses and Bacterial Indicators in a Sewage Treatment Center and Shallow Water Bay. International Journal of Environmental Research and Public Health. 2020; 17(18):6483. https://doi.org/10.3390/ijerph17186483
Chicago/Turabian StyleJanahi, Essam M., Sakina Mustafa, Saba F. D. Parkar, Humood A. Naser, and Zaki M. Eisa. 2020. "Detection of Enteric Viruses and Bacterial Indicators in a Sewage Treatment Center and Shallow Water Bay" International Journal of Environmental Research and Public Health 17, no. 18: 6483. https://doi.org/10.3390/ijerph17186483