Food and Environmental Virology, ( ISI ), Volume (7), No (4), Year (2015-8) , Pages (381-389)

Title : ( Application of F+RNA Coliphages as Source Tracking Enteric Viruses on Parsley and Leek Using RT-PCR )

Authors: dina shahrampour , Masoud Yavarmanesh , Mohammad B Habibi Najafi , Mohebbat Mohebbi ,

Citation: BibTeX | EndNote

The objective of this study was to identify sources of fecal contamination in leek and parsley, by using four different FRNA coliphage genogroups (IV, I indicate animal fecal contamination and II, III indicate human fecal contamination). Three different concentrations (102, 104, 106 pfu/ml) of MS2 coliphage were inoculated on the surface of parsley and leek samples for detection of phage recovery efficiency among two methods of elution concentration (PEG-precipitation and Ultracentrifugation) by performing double agar layer (DAL) assay in three replications. Highest recovery of MS2 was observed in PEG method and in 106 inoculation concentration. Accordingly, the PEG method was used for washing and isolation of potentially contaminated phages of 30 collected samples (15 samples from the market and 15 samples from the farm). The final solutions of PEG method were tested for the enumeration of plaques by DAL assay. Total RNA was then extracted from recovered phages, and RT-PCR was performed by using four primer sets I, II, III, and IV. Incidence of FRNA coliphages was observed in 12/15 (80 %) and 10/15 (66/6 %) of samples were obtained from farm and market, respectively, using both DAL and RT-PCR test methods. Different genotypes (I, II, and IV) of FRNA coliphages were found in farm samples, while only genotype I was detected in market samples by using the primer sets. Due to the higher frequency of genotype I and IV, the absence of genotype III, and also the low frequency of genotype II, it is concluded that the contamination of vegetable (parsley and leek) in Neyshabour, Iran is most likely originated from animal sources.

Keywords

, Parsley and leek, Microbial source tracking, FRNA coliphage, PEG-precipitation, Ultracentrifugation,
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@article{paperid:1052738,
author = {Shahrampour, Dina and Yavarmanesh, Masoud and Habibi Najafi, Mohammad B and Mohebbi, Mohebbat},
title = {Application of F+RNA Coliphages as Source Tracking Enteric Viruses on Parsley and Leek Using RT-PCR},
journal = {Food and Environmental Virology},
year = {2015},
volume = {7},
number = {4},
month = {August},
issn = {1867-0334},
pages = {381--389},
numpages = {8},
keywords = {Parsley and leek; Microbial source tracking; FRNA coliphage; PEG-precipitation; Ultracentrifugation; RT-PCR},
}

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%0 Journal Article
%T Application of F+RNA Coliphages as Source Tracking Enteric Viruses on Parsley and Leek Using RT-PCR
%A Shahrampour, Dina
%A Yavarmanesh, Masoud
%A Habibi Najafi, Mohammad B
%A Mohebbi, Mohebbat
%J Food and Environmental Virology
%@ 1867-0334
%D 2015

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