Samples of feces and vomit collected from patients during 13 non-bacterial gastroenteritis outbreaks which occurred in Hokkaido between 1995 and 1998 were examined by electron microscopy (EM) and reverse-transcription polymerase chain reaction (RT-PCR) for evidence of infection with human caliciviruses (HuCVs). In 6 food-borne outbreaks, oysters were the probable source of infection, while the origin of HuCVs was not found out for the other 7 outbreaks. One-hundred-eleven of 214 stool, vomit and oyster specimens examined gave positive results by RT-PCR, while HuCVs were detected by EM in 36 of 121 stool specimens examined. We determined the nucleotide sequences of 470-bp or 373-bp PCR products amplified from the RNA polymerase region of the HuCV genomes with primer sets MR3/4 and Yuri22F/R , respectively. The sequences of different strains revealed great heterogenicity, with a range of 60 to 100% homology among strains. In a few cases, a mixed genotype was found in the same patient or same outbreak by means of nested PCR and cloning of PCR products into an appropriate vector. Of the 19 different strains found, 4 strains could be classified as Norwalk virus (genogroup 1) and the other 15 strains as Snow Mountain agent (genogroup 2) based on genotyping with homology analysis. Furthermore, the strains belonging to genogroup 2 could be classified into 4 subgroups with more than 93% homology in amino acids among strains in the subgroup .
A novel seafood paste was developed by the fermentation of scallop ovary using rice malt (koji) and yeast culture. Chemical analysis of the product showed the formation of high level of free amino acids and organic acids during the fermentation. The product color and flavor resembled to Japanese traditional soybean miso. The contents of total lipids (TLs) extracted from the fermented products were ranged from 9.18% to 11.59% or 11.38% to 13.57%/dry sample weight. Although the TL was rich in oxidatively unstable polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), little decrease was found in these PUFAs during the fermentation, showing the high oxidative stability of the TL from the fermented scallop ovary. Moreover, the oxidative stability of the TL extracted from the fermented products increased with increasing the fermentation time. This would be mainly due to the formation of lipid soluble antioxidants such as tocopherols, which might be derived from yeast used for fermentation.
To produce a novel Mozzarella cheese, the homofermentative lactic acid bacteria (LAB) Streptococcus salivarius ssp. thermophilus, isolated in Hokkaido, was chosen from the authors< originally constructed LAB library. Physiological analysis of this strain indicated relatively weak acid production. To use this LAB as starter bacteria to produce mozzarella-type cheese, citric acid was used as a supplement to decrease the pH. Thus, the manufacturing method using both lactic acid fermentation and pH adjustment was established for the mozzarella cheese. The mozzarella cheese made using this manufacturing method exhibited good flavor. Consequently, we demonstrated that the originally isolated strain could be applied to the cheese manufacturing process without the need to select variants that are tolerant to severe culture conditions. Therefore, this result indicated that wild strains might be applied to cheese manufacturing without the need for laborious pre-selection considerations.
In this study, we attempted to produce a probiotic alcoholic beverage from fermented sake cake with lactic acid bacteria. When a 10-fold dilute heat-sterilized sake cake was fermented using Lactobacillus plantarum HOKKAIDO at 30℃ for one day, lactic acid bacterial number and pH were 1.8 × 10 8 CFU/ml and pH 3.2, respectively. The fermented product also contained 0.21 mg/ml glucose and 7.03 mg/ml lactic acid, so the resulting product was slightly sweet with a strong acid taste. Moreover, we found that the fermented product contained higher ornithine and acquired taurine and citrulline, which are known as functional food materials. In a low-temperature storage test of the fermented product with alcohol for 35 days, it was observed that HOKKAIDO strain existed > 1 × 10 7 CFU/ml until the presence of 10% alcohol. The results suggest that a probiotic and any new functional low alcoholic beverage can be manufactured from fermented sake cake with lactic acid bacteria.
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