Applicability Evaluation of Male-Specific Coliphage-Based Detection Methods for Microbial Contamination Tracking

Area of Science:

• Environmental microbiology
• Food safety science
• Virology

Background:

• Norovirus outbreaks linked to shellfish consumption pose a public health risk.
• Male-specific (F+) coliphage are potential indicators of viral contamination from waste sources.

Purpose of the Study:

• To compare the efficacy of double agar overlay and quantitative real-time PCR (RT-PCR) for F+ coliphage detection in shellfish.
• To evaluate F+ coliphage as a microbial contamination tracker in shellfish and seawater.

Main Methods:

• Shellfish samples were artificially contaminated with F+ coliphage.
• F+ coliphage detection was performed using both double agar overlay and RT-PCR methods.
• Virome analysis was conducted on contaminated oyster samples.

Main Results:

• RT-PCR yielded 1.6–39 times higher F+ coliphage plaque-forming unit (PFU) values compared to the double agar overlay method.
• RT-PCR detected both intact viruses and fragmented viral nucleic acids.
• Genomic diversity of F+ coliphages was observed, impacting detection specificity.

Conclusions:

• RT-PCR is a more sensitive method for detecting F+ coliphage in shellfish, potentially offering a more efficient contamination tracking tool.
• The genomic variability of F+ coliphages presents a limitation for precise microbial contamination tracking.
• Further development of specific primer sets is crucial for reliable F+ coliphage-based shellfish and seawater sanitation monitoring.