The globally re-emerging norovirus GII.2 manifests higher heat resistance than norovirus GII.4 and Tulane virus.

AIMS: To compare the heat stability of two globally prevalent human norovirus (HuNoV) strains (GII.2[P16] and GII.4[P16]) and a commonly used HuNoV surrogate, Tulane virus (TV). METHODS AND RESULTS: With the use of a newly developed zebrafish larvae platform, we measured the change of infectivity of HuNoV GII.2[P16] and GII.4[P16] toward mild heat treatment at 55°C for 5 min. TV was tested with the same experimental design. As a result, the virus infectivity measurement clearly indicated the higher heat resistance of HuNoV GII.2[P16] (no reduction) than GII.4[P16] (>0.8-log TCID50  ml-1 reduction) and TV (2.5-log TCID50  ml-1 reduction). Further exploration revealed higher virus structural stability of HuNoV GII.2 than GII.4 strains by the use of different clinical samples with different evaluation methods. CONCLUSION: The inactivation data generated from the surrogate virus TV cannot be used directly to describe the inactivation of HuNoV. The phylogenetic classification of HuNoVs may correlate with the virus stability and/or circulation dynamics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is expected to serve as an important reference when revisiting the numerous previous data evaluating HuNoV inactivation conditions in foods with the use of TV as the cultivable surrogate or with genuine HuNoV but using molecular methods. The higher resistance of NoV GII.2 strains than GII.4 strains toward inactivation treatment supplies a possible explanation for the global re-emerging of NoV GII.2 epidemic in recent years.

No Clinical Symptom Experienced after Consumption of Berry Fruits with Positive RT-qPCR Signals of Human Norovirus.

Human noroviruses (hNoVs) are the most important foodborne viruses, and soft berries are one of the most common food sources of hNoV outbreaks and contamination. This paper presents a human volunteer study in order to investigate the correlation between molecular detection results of hNoV in berries with the public health risks. The participants with diverse histo-blood group antigens (HBGAs) phenotypes were required to consume self-purchased berries and meanwhile submit aliquots of the products for reverse transcription-quantitative polymerase chain reaction (RT-qPCR) detection. As a result, none of the 20 participants reported any hNoV infection-like symptoms after six independent consumptions (120 consumptions in total). In contrast, within the 68 berry samples with >1% virus recoveries, 28 samples were detected to be positive for hNoV GI and/or GII (the positive rate at 41%). All of the positive signals were below the limit of quantification (<120 genome copies/g) except one fresh strawberry sample at 252 genome copies/g. It is expected that this study would contribute to the definition of quantitative standards for risk assessment purposes in the future.

Fucoidan But Not 2'-Fucosyllactose Inhibits Human Norovirus Replication in Zebrafish Larvae.

Human noroviruses (hNoVs) cause heavy disease burden worldwide and there is no clinically approved vaccination or antiviral hitherto. In this study, with the use of a zebrafish larva in vivo platform, we investigated the anti-hNoV potentials of fucoidan (from brown algae Fucus vesiculosus) and 2'-Fucosyllactose (2'-FL). As a result, although both fucoidan and 2'-FL were able to block hNoV GII.4 virus-like particle (VLPs) from binding to type A saliva as expected, only fucoidan, but not 2'-FL, was able to inhibit the replication of hNoV GII.P16-GII.4 in zebrafish larvae, indicating the possible needs of higher molecular weights for fucosylated carbohydrates to exert anti-hNoV effect.

Influence of fucosidase-producing bifidobacteria on the HBGA antigenicity of oyster digestive tissue and the associated norovirus binding.

Bivalve molluscan shellfish such as oysters are filter feeders and are able to accumulate human noroviruses (NoVs) largely due to the presence of human histo-blood group antigens (HBGAs)-like carbohydrates in their intestine. Since the fucose contents play a key role in the binding of NoVs to HBGAs, this study intended to investigate the influence of fucosidase-producing bifidobacteria on the HBGA antigenicity of oyster digestive tissue and the associated NoV binding. On the contrary to the expected, after a treatment of the oyster digestive tissue extracts with Bifidobacterium bifidum strain JCM 1254, the binding of human NoV GII.4 virus like particles (VLPs) to the oyster digestive tissue extracts enhanced significantly (OD450 from 1.15 ± 0.05 to 1.51 ± 0.02, P < 0.001) in an in vitro direct binding assay. The accumulation of human NoV GII·P16-GII.4 also enhanced significantly in the intestine of B. bifidum JCM 1254 treated oysters from 4.27 ± 0.25 log genomic copies/g oyster digestive tissue to 5.25 ± 0.29 log genomic copies/g oyster digestive tissue (P < 0.005) as observed in an in vivo test. Correspondingly, the type A antigenicity of the oyster digestive tissue extracts enhanced (OD450 from 0.77 ± 0.04 to 1.06 ± 0.05, P < 0.01) after the treatment with B. bifidum JCM 1254. These results could be explained by the substrate specificity of the B. bifidum JCM 1254 associated fucosidases. This study identified an indirect interaction possibly happening between the bacterial microbiota with human NoVs during their transmission in the food systems. We also supplied a potential strategy to mitigate the NoV contamination from shellfish, suppose bacterial strains with specified fucosidase production could be obtained in the future.

Biomonitoring and Subsequent Risk Assessment of Combined Exposure to Phthalates in Iranian Children and Adolescents.

This study aimed to estimate the exposure and related health risks of phthalates, and to assess the health risks from combined exposure to three of the phthalates sharing the same mode of action (anti-androgenicity) in children. We determined the internal exposure of 56 Iranian children and adolescents aged 6 to 18 years by analyzing seven urinary metabolites of five phthalates. The estimated daily intake values derived from the biomonitoring data ranged from 0.01 µg/kg bw/day for butyl benzyl phthalate (BBP), to 17.85 µg/kg bw/day for di(2-ethylhexyl) phthalate (DEHP). The risk assessment revealed that not only the exposure to the individual phthalates, but also the combined exposure to the three anti-androgenic phthalates (DEHP, DBP, BBP) did not raise a safety concern (hazard index values averaged 0.2). The range of maximum cumulative ratio values varied from around 1 for most individuals to around 2 in some individuals, indicating that the combined exposures were dominated by one and in some cases by two of the three anti-androgenic phthalates, especially dibutyl phthalate (DBP) and/or DEHP. Based on biomonitoring data, the overall combined exposure of Iranian children to phthalates does not raise a concern, while reduction of exposure is best focused on DEHP and DBP that showed the highest hazard quotient.