The expansion of the in-house database is indispensable for improving the accuracy of microbial identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). To this end, it is important to ensure the high quality of the data to be registered in order to obtain accurate identification results. In this study, the effects of the formation of bacterial spores and yeast ascospores on mass spectra were investigated. A main spectrum (MSP, a spectrum that becomes a standard for registration) was prepared using spore-forming bacteria and ascospore-forming yeast cells cultured by different methods and for different incubation times. The score values indicating the similarities of the spectra were compared, and the culture conditions and sample preparation methods that can obtain accurate species identification results, even in the case of spore and ascospore formation, were determined. First, Bacillus subtilis and Bacillus cereus were investigated as spore-forming bacteria, and the spore formation was greatly affected by the oxygen supply amount and medium composition. When the rates of spore formation differed greatly, the mass spectra obtained also showed considerable differences. When cultured on a standard agar medium, by using the MSP obtained from bacterial cells on the third day of culture for both B. cereus and B. subtilis, a high score value was obtained, indicating accurate bacterial species identification results. This high score value was achieved even for the mass spectral data obtained from the bacterial cells on the first day without spore formation and on the fifth day when spore formation was observed to some extent. However, when ascospore-forming yeast Pichia manshurica was cultured on a potato dextrose agar (PDA) plate medium, the ascospore formation rate differed with the strains. In the ethanol-formic acid extraction method, which is the bacterial cell extraction method recommended by the manufacturer, the bacterial cell extraction rate decreased because of ascospore formation. Through cell crushing using beads conducted to increase the cell protein extraction efficiency, it was shown that species identification is possible even upon ascospore formation.

　食用カンナ葉部は，一部の地域で使用されているにも関わらず，成分や機能性に関する報告がない。そこで，本研究では，乳酸発酵前後の食用カンナの葉の栄養学的特性とその80％メタノール抽出物（N-MEECLおよびF-MEECL）の機能性について解析を行った。その結果，食用カンナの葉にはタンパク質とポリフェノールが豊富に含まれることが明らかになった。非発酵カンナ葉80％メタノール抽出物（N-MEECL）に含まれるポリフェノールとしてルチン，クロロゲン酸，ケルセチン，ピセアタンノール，ネオクロロゲン酸の5種類を同定した。発酵カンナ葉80％メタノール抽出物（F-MEECL）にはピセアタンノール以外の4種類のポリフェノールのほとんどが消失し，抗酸化能も低下した。また，N-MEECLおよびF-MEECLには遊離アミノ酸が含まれており，発酵後，量はわずかに増加した。機能性の評価として，ラット好塩基球白血病細胞（Rat basophilic leukemia（RBL-2H3）cells）を用い，脱顆粒抑制を指標とする抗アレルギー効果の評価を行った。N-MEECLおよびF-MEECLでは抗アレルギー効果が確認され，食用カンナ葉を乳酸発酵させたF-MEECLの脱顆粒抑制作用はN-MEECLよりも強かった。N-MEECLに含まれている5種類のポリフェノールは，N-MEECLよりもRBL-2H3に対して強い脱顆粒抑制作用を示したが，発酵によりピセアタンノールを除く4種類のポリフェノールが消失したため，F-MEECL中のRBL-2H3に対する強い脱顆粒抑制作用を示す化合物には，N-MEECLに含まれているピセアタンノール，及び4種類のポリフェノール（ルチン，クロロゲン酸，ケルセチン，ネオクロロゲン酸）以外の物質の関与が考えられた。また，21種類のアミノ酸およびイミノ酸標品によるRBL-2H3に対する脱顆粒抑制作用を検討したが，脱顆粒抑制作用はN-MEECL中のポリフェノールに及ばなかった。これらの結果から，食用カンナの葉を乳酸発酵させると，RBL-2H3の脱顆粒抑制作用を増強する新たな化合物が生成されることが示唆された。

　The total polyphenol content and H-ORAC values of 15 traditional Okinawan vegetables were determined and compared with those of spinach. Nishiyomogi, Hosobawadan, Botanboufu, and Suizenjina exhibited higher total polyphenol contents and H-ORAC values than those of spinach. The total polyphenol content and H-ORAC values exhibited a high correlation (r = 0.986, p < 0.01), suggesting that polyphenols are the major contributors to the antioxidant activity of Okinawan traditional vegetables in water systems. The effects of storage temperature on the H-ORAC values and total polyphenol content of six traditional Okinawan vegetables were also investigated. The behavior of the total polyphenol content and H-ORAC values during storage differed among the vegetables. In Nishiyomogi and Suizenjina, the total polyphenol content decreased on the fifth day of storage at 25 ℃. In contrast, for Botanboufu, significant increases in the total polyphenol content and H-ORAC values were observed after the second day of storage at 25 ℃ and on the fifth day of storage at 15 and 25 ℃ compared to the first day of storage. No increase or decrease in the total polyphenol content or H-ORAC values was observed in the other vegetables during storage.