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Survival of Nonculturable Human Noroviruses during High Pressure Processing

Lou, Fangfei
http://rave.ohiolink.edu/etdc/view?acc_num=osu1408727004

Abstract Details

2014, Doctor of Philosophy, Ohio State University, Food Science and Technology.
Human norovirus (NoV) is responsible for more than 90% of non-bacterial acute gastroenteritis worldwide and over 60% of foodborne illnesses in the US. Currently, there is no effective measure to eliminate human NoV in high risk foods. High pressure processing (HPP) has been considered a promising non-thermal processing technology to inactivate foodborne viral pathogens while maintaining food quality. However, the effectiveness of HPP on inactivating human NoV remains poorly understood because human NoV cannot be grown in cell culture and lacks a small animal model. The objective of this project was to determine the survival of human NoV during HPP using three novel approaches. First, the stability of human NoV to HPP was estimated by targeting the major capsid protein of the virus using human NoV virus-like particles (VLPs) as a model since VLPs are structurally and antigenically similar to native virions and possess authentic receptor binding activity. Inactivation was evaluated based on the structural integrity of VLPs and binding ability to histo-blood group antigens (HBGAs), the functional receptors for human NoV. Second, the survival of human NoV was estimated by the ability to bind to porcine gastric mucin, which contains HBGA-like molecules, conjugated to magnetic beads, followed by quantification of bound virions using real-time reverse transcription polymerase chain reaction (RT-PCR). Third, the infectivity of human NoV after HPP was determined in a gnotobiotic pig model by evaluation of viral shedding in feces, the presence of intestinal lesions, and viral replication in the small intestines. In summary, we effectively developed multiple approaches to determine the survival of uncultivable human NoV during HPP treatment. We concluded that (i) the mechanism of human NoV inactivation by HPP is the disruption of the integrity of viral capsid structure and viral receptor binding activity; (ii) different human NoV strains have variable susceptibilities to HPP; (iii) processing parameters (such as initial temperature, pH, salt, and pressure) significantly impacted the effectiveness of HPP on the inactivation of human NoV; (iv) human NoV VLPs were more stable than viable human NoV; (v) the combination of detection of receptor binding activity and viral genomic RNA is an improved method to estimate viral survival; and (vi) gnotobiotic piglets are a useful model to evaluate the infectivity of human NoV in vivo. Our results demonstrate that HPP is capable of effectively inactivating human NoV in aqueous medium and oysters under commercially acceptable pressures with a short holding time at 4°C. Thus, HPP has the potential to be an effective intervention strategy for eliminating human NoVs in high risk foods and ensuring food safety.
Jianrong Li (Advisor)
191 p.
Food Science

Recommended Citations

Citations

  • Lou, F. (2014). Survival of Nonculturable Human Noroviruses during High Pressure Processing [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1408727004

    APA Style (7th edition)

  • Lou, Fangfei. Survival of Nonculturable Human Noroviruses during High Pressure Processing . 2014. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1408727004.

    MLA Style (8th edition)

  • Lou, Fangfei. "Survival of Nonculturable Human Noroviruses during High Pressure Processing ." Doctoral dissertation, Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1408727004

    Chicago Manual of Style (17th edition)

osu1408727004
292
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This open access ETD is published by The Ohio State University and OhioLINK.