Hepatitis A virus strains identified in jogaejeot associated with outbreaks in Seoul, South Korea.

Jogaejeot, seasoned Venerupis philippinarum, is a traditional Korean fermented food, and hepatitis A virus (HAV) can be transmitted through contaminated food, especially bivalve shellfish, causing acute gastroenteritis worldwide. Here, we carried out a phylogenetic analysis to identify and characterize HAV strains in jogaejeot samples associated with hepatitis A (HA) outbreaks in Seoul, South Korea, in 2019. The HAV strains were identified using blast and molecular analysis of the amplified HAV VP1-P2B genome region. The HAV strains identified in the five jogaejeot samples shared at least 99% sequence identity, were all classified as genotype IA and were most closely related to strains that are widespread in East Asia. These results support a link between the consumption of jogaejeot and the HA outbreaks observed in 2019 in Seoul. In addition, they indicate a need for more stringent enforcement of food safety regulations for the shellfish industry, especially against HAV, and the value of widespread vaccination.

Viruses in fermented foods: are they good or bad? Two sides of the same coin.

The emergence of Coronavirus disease 2019 as a global pandemic has increased popular concerns about diseases caused by viruses. Fermented foods containing high loads of viable fungi and bacteria are potential sources for virus contamination. The most common include viruses that infect bacteria (bacteriophage) and yeasts reported in fermented milks, sausages, vegetables, wine, sourdough, and cocoa beans. Recent molecular studies have also associated fermented foods as vehicles for pathogenic human viruses. Human noroviruses, rotavirus, and hepatitis virus have been identified in different fermented foods through multiple routes. No severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) virus or close members were found in fermented foods to date. However, the occurrence/persistence of other pathogenic viruses reveals a potential vulnerability of fermented foods to SARS-CoV-2 contamination. On the other side of the coin, some bacteriophages are being suggested for improving the fermentation process and food safety, as well as owing potential probiotic properties in modern fermented foods. This review will address the diversity and characteristics of viruses associated with fermented foods and what has been changed after a short introduction to the most common next-generation sequencing platforms. Also, the risk of SARS-CoV-2 transmission via fermented foods and preventive measures will be discussed.

Assessment of human norovirus inhibition in cabbage kimchi by electron beam irradiation using RT-qPCR combined with immunomagnetic separation.

Cabbage Kimchi, a traditional Korean fermented food, has occasionally been related to acute gastroenteritis caused by human norovirus (HuNoV). The present study examined the inhibitory effects of electron beam (e-beam) irradiation (1, 3, 5, 7, and 10 kGy) on HuNoV GII.4 in suspension or cabbage Kimchi using reverse transcription quantitative polymerase chain reaction combined with immunomagnetic separation (IMS/RT-qPCR). In addition, physicochemical and sensorial analyses were conducted to assess any change in the quality of cabbage Kimchi following e-beam irradiation. Following e-beam irradiation at 1 to 10 kGy, HuNoV significantly decreased to 0.28 to 2.08 log10 copy number/mL in suspension (P < 0.05). HuNoV levels in cabbage Kimchi were also significantly reduced to 0.26 to 1.57 log10 copy number/mL following irradiation with 1 to 10 kGy (P < 0.05) compared to positive control (6.0 log10 copy number/mL). The D-values for 1 log10 reduction (90% inhibition) of HuNoV in suspension and cabbage Kimchi were 4.94 and 6.96 kGy of e-beam, respectively. The pH and acidity in the irradiated cabbage Kimchi were 4.41 to 4.58 and 0.61% to 0.71%, respectively, indicating that e-beam did not affect the optimal pH or acidity. Although a slight increase of greenness was observed in the leaf portion of cabbage Kimchi irradiated with 7 to 10 kGy of e-beam, this color change was minimal and went undetected by panelists in the sensorial evaluation. The five properties of sensorial quality assessed were no different in the irradiated Kimchi sample compared with the control sample (nonirradiated cabbage Kimchi). Therefore, this study suggests that ≥6.96 kGy of e-beam could be applied in the cabbage Kimchi industry to obtain >90% of HuNoV without affecting the quality. PRACTICAL APPLICATION: As the most representative food in Korea, Kimchi needs the sanitation technology that can inhibit viral infection. Our findings suggest that e-beam irradiation can be used to reduce HuNoV effectively in Kimchi without changes in sensorial quality.

Viral community predicts the geographical origin of fermented vegetable foods more precisely than bacterial community.

Fermented foods are considered as an integral part of the global human diet. Fermented foods also have unique microbial communities such as bacteria, archaea, fungi, and viruses that are essential to the fermentation process and affect final product characteristics. Despite the ecological importance of virus, little is known about the diversity and ecological role of virus in the food ecosystem. In this study, the viral and host bacterial communities from 10 representative samples of Korean and Chinese kimchi were analyzed in triplicate using next-generation sequencing technology. The overall structures of bacterial and viral communities were dominated by lactic acid bacteria in phylum Firmicutes and bacteriophages in order Caudovirales, respectively. For the single-stranded DNA (ssDNA) viruses, bacteriophage in family Microviridae were dominant in Korean kimchi. After correction for multiple comparisons using false discovery rate (FDR, P < 0.05), the relative abundances of 6 bacterial taxa and 33 viral host taxa at the genus level were significantly different between Korean and Chinese kimchi. Notably, in beta-diversity analysis, viral communities were much more clearly separated according to their geographical origin (PERMANOVA pseudo-F = 11.57, P < 0.001 in Bray-Curtis PCoA) than bacterial communities (pseudo-F = 4.75, P < 0.001 in unweighted UniFrac PCoA). Thus, viral metagenomics represents a potentially useful in-depth analytical method for determining the geographical origins of fermented foods.

Complete Nucleotide Sequence Analysis of a Novel Bacillus subtilis-Infecting Bacteriophage BSP10 and Its Effect on Poly-Gamma-Glutamic Acid Degradation.

While the harmful effects of lactic acid bacterial bacteriophages in the dairy industry are well-established, the importance of Bacillus subtilis-infecting bacteriophages on soybean fermentation is poorly-studied. In this study, we isolated a B. subtilis-infecting bacteriophage BSP10 from Meju (a brick of dried fermented soybean) and further characterized it. This Myoviridae family bacteriophage exhibited a narrow host range against B. subtilis strains (17/52, 32.7%). The genome of bacteriophage BSP10 is 153,767 bp long with 236 open reading frames and 5 tRNAs. Comparative genomics (using dot plot, progressiveMauve alignment, heat-plot, and BLASTN) and phylogenetic analysis strongly suggest its incorporation as a new species in the Nit1virus genus. Furthermore, bacteriophage BSP10 was efficient in the growth inhibition of B. subtilis ATCC 15245 in liquid culture and in Cheonggukjang (a soybean fermented food) fermentation. Artificial contamination of as low as 10² PFU/g of bacteriophage BSP10 during Cheonggukjang fermentation significantly reduced bacterial numbers by up to 112 fold in comparison to the control (no bacteriophage). Moreover, for the first time, we experimentally proved that B. subtilis-infecting bacteriophage greatly enhanced poly-γ-glutamic acid degradation during soybean fermentation, which is likely to negatively affect the functionalities of Cheonggukjang.

Correlation between Changes in Microbial/Physicochemical Properties and Persistence of Human Norovirus during Cabbage Kimchi Fermentation.

Recently, cabbage kimchi has occasionally been associated with the foodborne diseases of enteric viruses such as human norovirus (HuNoV). This study aimed to evaluate the correlation between microbial/physicochemical properties and persistence of HuNoV in experimentally contaminated cabbage kimchi fermented and stored at 4°C or 10°C for 28 days. Changes in organic acid content, lactic acid bacteria (LAB), acidity, pH, and salinity were analyzed. The recovery of structurally intact HuNoV was examined for up to 28 days post-inoculation, using a NoV GII.4 monoclonal antibody-conjugated immuno-magnetic separation method combined with quantitative real-time reverse transcription polymerase chain reaction. On day 0, LAB loads were 4.70 log10 colony forming units/g and HuNoV GII.4 titers were 2.57 log10 genomic copies/µl, at both temperatures. After 28 days, intact HuNoV titers decreased to 1.58 (4°C) and 1.04 (10°C) log10 genomic copies/µl, whereas the LAB density increased. This correlated with a gradual increase in lactic acid and acetic acid at both temperatures. Our findings support a statistical correlation between changes in physicochemical properties and the recovery of structurally intact HuNoV GII.4. Moreover, we determined that the production of organic acid and low pH could affect HuNoV GII.4 titers in cabbage kimchi during fermentation. However, HuNoV GII.4 was not completely eliminated by microbial/physicochemical factors during fermentation, although HuNoV GII.4 was reduced. Based on this, we speculate that the persistence of HuNoV GII.4 may be affected by the continually changing conditions during kimchi fermentation.

Application of gamma radiation for the reduction of norovirus and the quality stability in optimally ripened cabbage kimchi.

Optimally ripened commercial cabbage kimchi is considered the main cause of enteric norovirus (NoV) outbreaks in Korea. This study investigated the effect of 1-10kGy gamma radiation on the inactivation of murine norovirus-1 (MNV-1; initial inoculum of 5-6log10PFU/ml), used as a human NoV surrogate, in kimchi. The effects of gamma radiation on the pH and acidity were also examined to address the index of quality and fermentation, respectively. Titers of MNV-1 significantly reduced (p<0.05) in kimchi subjected to increasing gamma radiation doses: MNV-1 titers in kimchi after 1, 3, 5, 7, and 10kGy were 4.82 (0.34-log10 reduction), 4.45 (0.71-log10 reduction), 4.18 (0.98-log10 reduction), 3.71 (1.45-log10 reduction), and 3.40 (1.76-log10 reduction) log10 PFU/ml, respectively. However, the values of pH (4.5-4.6) and acidity (0.6-0.7%) were not significantly different between non-irradiated and irradiated kimchi (p>0.05). The D-value (1-log reduction) for MNV-1 in kimchi, calculated using first-order kinetics, was 5.75kGy (R2=0.98, RMSE=0.10). Therefore, this study suggests that the use of ≥5.75kGy gamma radiation in the kimchi manufacturing industry could be very effective in reducing NoV contamination by >90% (1 log), without causing changes in quality and fermentation.