Detection of Viruses in Coastal Seawater Using Mytilus Galloprovincialis as an Accumulation Matrix

Abstract

The purpose of this study was to demonstrate that shellfish can be used to detect enteric viruses in marine waters where they are present at very low concentrations. Aqua-cultured mussels were placed in the sea just off the mouth of a drainage channel affected by human and animal faecal contamination. Samples were taken from the channel, the sea and the mussels at intervals over two 4-week periods. The samples were tested to verify the presence of both rotaviruses and E. coli. Rotaviruses were detected by Real Time-PCR, typed by multiplex PCR and subsequently sequenced. E. coli was enumerated in water matrices by a filtering method and in mussels by the MPN method. The presence of E. coli in the examined matrices demonstrates contamination of faecal origin throughout the studied environments. Rotaviruses were recorded in channel waters, but not in sea water. In both experiments, rotaviruses were detected in mussels 21 and 28 days after being placed in the sea water off the channel mouth. The use of mussels thus enabled the detection of rotaviruses in waters where the high dilution rendered direct investigation impossible. This study indicates that mussels can be used in marine virological surveillance programs.

Detection of Human Bocavirus Species 2 and 3 in Bivalve Shellfish in Italy

“…HBoV can be discharged in untreated wastewaters through virus shedding and, consequently, disperse throughout water environments, as shown recently in Italy (30,31) and elsewhere (26,28,32). Bivalve mollusks concentrate several human pathogens transmitted by the fecal-oral route, including a wide range of enteric viruses (35)(36)(37), whose presence in these aquatic organisms may be evaluated to assess the safety of the product for human consumption (38) or for biomonitoring purposes (39,40).…”

Identification of HEV in symptom‐free migrants and environmental samples in Italy

“…Viruses were concentrated from water samples by the tangential flow ultrafiltration technique using polypropylene membranes with a 10‐KDa molecular weight cut‐off and a pore size of 0.001–0.01 μm. These filters have been used in previous studies to concentrate viruses present in water, particularly enteroviruses and bacteriophages, with a sensitivity of 1 viral particle/ mL of water sample analysed . To eliminate particulate matter, the samples were prefiltered with 12‐μm polypropylene membranes.…”

Rotavirus Occurrence in Shellfish with Low Levels of E. coli

“…1). Here, previous studies had revealed the widespread presence of bacterial indicators of fecal contamination and RV (Grassi et al 2010;De Donno et al 2012).…”

“…The results of the investigations carried out in a 5-week period showed that the waters of the site in which the study was performed are affected by a continuous, although low, contamination of fecal origin deducible from concentration of E. coli. As previously described (Grassi et al 2010;De Donno et al 2012), such contamination could be caused by Castellana-Trafili channel that could carry into the bay several microorganism of fecal origin including Rotavirus that however was never directly detected in seawater. The salinity values, appreciably lower than the control point, confirm the presence of significant input of fresh water in marine environment.…”

“…In this field study, the accumulation of RV in transplanted individuals of M. galloprovincialis was evaluated and compared with the concentration of E. coli. RV determination in mussels was performed using a previously discussed method of RNA-extraction which guaranteed a good recovery of RNA from the homogenate hepatopancreas of shellfish (Ribao et al 2004), combined with a Real-Time PCR method already used (De Donno et al 2012) for the amplification of rotaviral RNA extracted from both aqueous matrices and tissues of mussels.…”