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Rapid quantitative detection of Enteric viruses in River Nile and drainage water, Egypt

Rapid quantitative detection of Enteric viruses in River Nile and drainage water, Egypt

Mohamed I. Azzam1, Safaa M. Ezzat1, K. A. El-Dougdoug2, Badawi A. Othman2

1 Central Lab. for Environmental Quality Monitoring, National Water Research Center.
2 Microbiology Department, Faculty of Agriculture, Ain Shams University

ABSTRACT

Contamination of surface water with enteric viruses is a major public health concern that
warrants the need to develop reliable indicators for enteric viruses contamination. This
work assesses the bio-diversity of coliphages to use as bio-indicators for viral water
pollution. The study elucidates the occurrence of enteric viruses at various locations in
Rosetta Branch of the Nile River in addition to five main drains located on its sides which
selected for confirmation the eco-diversity of aquatic viral isolates. The evaluation was
carried out using real time-quantitative reverse transcriptase - polymerase chain reaction
(rt-qRT-PCR). Eight coliphage isolates were detected in both Rosetta Branch and
drainage water samples. Transmission Electron microscopy revealed that, the isolated
coliphages have an isometric head and long-contractile tail; some particles revealed a
short tail with full heads, resembling those belong to the myoviridae and siphoviridae
family. Restriction enzymes by EcoRI, HindIII and BamHI showed the presence of
double stranded (ds) DNA as well as heterogeneity among these phages. The results
showed that EcoRI produced 7, 6, 2, 9, 10, 8, 10, 7 unique fragments and HindIII
produced 2, 3, 0, 1, 2, 5, 4, 3 unique fragments while BamHI produced only 4, 0, 1, 1, 0,
4, 2, 1 unique fragments, for the eight phage isolates, respectively. Out of fifteen tested
sites, two only (El-Rahawy drain outlet and Sabal drain outlet) were found to be polluted
with enteroviruses with rate of 3.6X104 and 3.4X104 gene copies per microliters (GC μl-
1), respectively using rt-qRT-PCR.

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Journal of Virological Sciences

July

Vol. 3, Iss. 1

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