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Abstract
In order to investigate the occurrence of norovirus in rivers and beaches, a total of 81 samples were tested at seven sites of Oncheon stream, Suyeong river and Gwanganri beach in Busan from January to November, 2017. To improve the detection of norovirus from sea water, we applied the inorganic cation-coated filter method which showed 48.8% ± 12.2% (n=3) and 27.4% ± 6.0% (n=3) recovery yields from river water and sea water inoculated with Norovirus, respectively. Norovirus was detected in a total of four samples (4.9%), which all were GII genotype. Norovirus GII was detected in three samples at two waste water treatment plants (WWTP) outlet and one sample at about 500 meter downstream from WWTP in both the winter and spring seasons. We also monitored fecal indicator organisms, Escherichia coli (E. coli), Enterococcus and coliphages [somatic coliphages (SC), male-specific coliphages (MSC)] to analyze the potential transmission of enteritis causative agent in dry and wet days. Bacterial influences were found at the site of the WWTP effluents in the dry days and spread further to the costal beach in the wet days. But no viral influences were found in the river downstream in both dry and wet days.
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Figure 1.
Location of the sample collection sites. Location of the sites in which water samples were collected was indicated as S1~S7. S, site.
Figure 2.
Seasonal variation in the distribution of microorganism indicators at all study sites. E. coli, Escherichia coli; SC, Somatic coliphage; MSC, Male-specific coliphage. Error bars indicate standard deviation and E. coli (a) and SC (b) are showed significant difference between summer and winter according to t-test (p<0.05).
Figure 3.
Comparison of fecal bacteria and coliphages in dry days and wet days by sample location. It showed significant difference between Dry days and Wet days according to t-test (p<0.05).
Figure 4.
Variability of microorganism indicator in dry days and wet days at study area. The color shows the changes of concentration level of the indicator microorganism. Concentration units are shown above each color bars.
Table 1.
Characteristics of host cells of coliphages
Table 2.
The sequences of oligonucleotides used for the detection of norovirus
Table 3.
Recovery of Norovirus from natural seawater and stream water spiked with Norovirus GII
| Sample | Initial concentration (total 300 mL water) viral copies/ml | Recovered concentration (total 1 mL concentrate) viral copies/ml | Recovery ratea (%±SD) |
|---|---|---|---|
| Seawaterb | 6.9 × 104 | 1.9 ± 0.4 × 104 | 27.4 ± 6.0 |
| Stream waterb | 6.9 × 104 | 3.4 ± 0.8 × 104 | 48.8 ± 11.2 |
Table 4.
Parameter characteristics of norovirus (NoV) GII positive samples
Table 5.
Seasonal characteristics in physiochemical factors and waterborne microbial indicators
| Season | N | Temp. (℃) | pH | DO (mg/l) | Salinity (PSU) | Turbidity (NTU) | E. coli (MPN/ 100 ml) | Enterococcus (MPN/ 100 ml) | Coliphage (PFU/100 ml) | |
|---|---|---|---|---|---|---|---|---|---|---|
| SC | MSC | |||||||||
| Spring | 31 | 16.5 | 7.2 | 8.0 | 11.63 | 3.17 | 2.0×103 | 4.2×103 | 1.2×103 | 4.0×101 |
| (Dry days)a | (19) | 16.9 | 7.2 | 7.8 | 11.68 | 2.40 | 5.8×103 | 1.0×103 | 4.8×102 | 1.0×101 |
| (Wet days)b | (12) | 15.8 | 7.3 | 8.4 | 11.56 | 4.40 | 4.2×104 | 9.3×103 | 2.3×103 | 7.5×101 |
| Summer (Wet days)c | 14 | 23.7 | 7.2 | 6.6 | 13.88 | 2.75 | 3.0×104 | 4.2×103 | 1.4×103 | 2.5×101 |
| Autumn (Dry days) | 21 | 18.1 | 7.3 | 7.6 | 14.12 | 2.30 | 8.2×103 | 3.3×103 | 2.1×103 | 9.0×100 |
| Winter (Dry days) | 15 | 9.5 | 7.4 | 10.9 | 14.07 | 2.07 | 2.8×103 | 2.1×103 | 1.2×102 | 9.0×100 |
| Total | 81 | 16.9 | 7.3 | 8.2 | 13.10 | 2.7 | 1.6×104 | 3.6×103 | 1.3×103 | 2.2×101 |
Temp, Temperature; DO, Dissolved oxygen; PSU, Practical salinity unit; NTU, Nephelometric turbidity unit; E. coli, Escherichia coli; SC, Somatic coliphage; MSC, Male-specifc (F+) coliphage; MPN, most probable number; PFU, plaque forming unit; N, number of samples; Spring months: March, April, and May; Summer months: June, July, and August; Autumn months: September, October, and November; Winter months: December, January, and February.
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