Ten Yucatan miniature piglets were challenged with the human norovirus (NoV) GII.12/GII.3 CAU140599 strain and five piglets were used as negative controls. Stool, serum, and organs were collected and processed from two NoV-infected piglets and one negative piglet at 1, 2, 3, 5, and 7 days post-inoculation (dpi). NoV was detected in stool and serum samples by real-time RT-PCR. Mild diarrhea was observed at 1-3 dpi. Fecal shedding and viremia were detected intermittently at 1, 3, and 7 dpi. While interferon-α was significantly elevated at 2-3 dpi, interferon-γ was not changed. Immunohistochemistry demonstrated that the NoV capsid antigen was present in macrophages, lymphocytes, and dendritic cells of the stomach, intestines, lymph nodes, spleen, and tonsils. Intestinal epithelium did not exhibit a positive signal for NoV. In addition, negative-sense viral RNA was confirmed in immune cells by fluorescence in situ hybridization. Therefore, NoV might be associated with macrophages and lymphocytes in gastrointestinal tract and immune organs of experimentally infected miniature piglets.

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This study was conducted to develop a multiplex reverse transcription (RT) PCR for the detection of Anisakis allergens and to investigate the relationship between allergen profiles and anisakid larvae isolated from Scomber japonicus, Trichiurus lepturus, and Conger myriaster in Korea. The species of Anisakis was determined using Anisakis pegreffii-specific PCR and restriction fragment length polymorphism analysis. The prevalence and profiles of five Ani s allergens were examined by multiplex RTPCR. A. pegreffii and Anisakis typica accounted for 97.1 and 2.9%, respectively, of the 140 larvae examined. In A. pegreffii, allergen prevalence was 41.2% for Ani s 1, 72.1% for Ani s 2, 69.9% for Ani s 3, 86.7% for Ani s 4, and 93.4% for Ani s 5. Most A. pegreffii larvae had multiple allergen profiles, and 80.7% of A. pegreffii carried both Ani s 4 and Ani s 5, which are heat-resistant allergens. Fifty-two to 65% of A. pegreffii isolated from S. japonicus and C. myriaster carried all five Ani s allergens.