TW202218552A - 包含乳酸菌與脂肪酸的飼料及組成物 - Google Patents
包含乳酸菌與脂肪酸的飼料及組成物 Download PDFInfo
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- TW202218552A TW202218552A TW110135594A TW110135594A TW202218552A TW 202218552 A TW202218552 A TW 202218552A TW 110135594 A TW110135594 A TW 110135594A TW 110135594 A TW110135594 A TW 110135594A TW 202218552 A TW202218552 A TW 202218552A
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- acid
- fatty acid
- strain
- feed
- lactobacillus
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Abstract
本發明之課題係提供選自下列(1)至(4)之1者以上:(1)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之飼料、(2)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之IL-12產生增強用組成物、(3)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之免疫賦活用組成物、及(4)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之抑菌用組成物。
藉由上述飼料或組成物而解決上述課題。
Description
本發明係關於包含特定乳酸菌或其處理物與脂肪酸或其鹽之組合的飼料及組成物。
介白素12(IL-12:Interleukin-12)係1種藉由經活化之吞噬細胞(phagocytes)及樹突細胞所分泌之細胞激素,於多種免疫相關疾病之病因中扮演重要角色。已知某種乳酸菌具有免疫賦活作用及IL-12產生作用(專利文獻1)。
[先前技術文獻]
[專利文獻]
專利文獻1:日本特開2010-6801號
吾人尚未知曉藉由組合乳酸菌之胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽,可增強IL-12產生及
/或免疫賦活作用、及可得到抑菌作用。再者,迄今亦尚未知曉將此組合用於飼料。
本發明之目的係提供選自(1)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之飼料、(2)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之IL-12產生增強用組成物、(3)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之免疫賦活用組成物、及(4)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之抑菌用組成物之1者以上。
本發明者們發現,藉由組合乳酸菌之胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽,可增強IL-12產生及免疫賦活作用,更可得到抑菌作用,又此組合適用於飼料及組成物,再者,亦得知即便於脂肪酸或其鹽與乳酸菌之組合中,胚芽乳酸桿菌L-137株仍為優異。本發明者們經進一步反覆檢討而完成本發明。
亦即,本發明係如下所述者。
〔1〕一種飼料,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
〔2〕如〔1〕所述之飼料,其中,脂肪酸係選自由短鏈脂肪酸、中鏈脂肪酸及長鏈脂肪酸所組成之群組中之至少1種脂肪酸或此等之混合物。
〔3〕如〔2〕所述之飼料,其中,短鏈脂肪酸係選自由甲酸、乙酸、丙酸、丁酸、異丁酸、戊酸、異戊酸、乳酸、丙酮酸、己酸及琥珀酸所組成之群組中之至少1種。
〔4〕如〔1〕至〔3〕中任一項所述之飼料,其中,胚芽乳酸桿菌L-137株係死菌體。
〔5〕如〔1〕至〔4〕中任一項所述之飼料,其中,相對於全體飼料,胚芽乳酸桿菌L-137株或其處理物之含有率係0.0001質量%至0.1質量%。
〔6〕如〔1〕至〔5〕中任一項所述之飼料,其中,飼料中之胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之質量比係1:0.5至50000。
〔7〕一種IL-12產生增強用組成物,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
〔8〕一種免疫賦活用組成物,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
〔9〕一種抑菌用組成物,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
〔10〕一種組成物,係含有胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
〔11〕一種免疫賦活用組成物,係含有乳酸菌或其處理物與脂肪酸或其鹽。
〔12〕如〔11〕述之組成物,其中,組成物係醫藥、動物藥、準醫藥品(quasi drug)、飼料、飲食品或飲食品添加物。
於本記載中,較佳為可提供選自(1)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之飼料、(2)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之IL-12產生增強用組成物、(3)包含胚芽乳酸桿菌L-137株或其處
理物與脂肪酸或其鹽之免疫賦活用組成物、及(4)包含胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之抑菌用組成物中之1者以上。
圖1顯示於免疫細胞株添加本發明之乳酸菌與脂肪酸(短鏈脂肪酸)或其鹽之任一者、或者同時添加該等時產生IL-12。圖中之*,係顯示在比較單獨添加乳酸菌L-137以及同時添加乳酸菌L-137與脂肪酸的T檢定中,p值於「0.01≦p<0.05」之範圍內。
圖2顯示於免疫細胞株添加本發明之乳酸菌與脂肪酸(中鏈脂肪酸或長鏈脂肪酸)之任一者、或者同時添加該等時產生IL-12。圖中之*,係顯示在比較單獨添加乳酸菌L-137以及同時添加乳酸菌L-137與脂肪酸的T檢定中,p值於「0.01≦p<0.05」之範圍內。
圖3顯示對肉雞投予混合了本發明之乳酸菌與脂肪酸或其鹽之任一者、或者同時混合此等之飼料時的IL-12之基因表現強度。圖中之**,係顯示在比較T1區與T3區的T檢定中,p值為「p<0.01」。
〔乳酸菌之胚芽乳酸桿菌L-137株〕
本發明所使用之乳酸菌之胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)係以寄存編號FERM BP-08607號(移交自平成7年(1995年)11月30日寄存之FERM P-15317號)寄存於獨立行政法人產業技術綜合研究所特許生物寄存中心
(現:獨立行政法人製品評價技術基盤機構特許微生物寄存中心;住址:郵遞區號292-0818,日本國千葉縣木更津市上總鎌足2-5-8-120號室)。此外,即使是胚芽乳酸桿菌L-137之突變株,具備胚芽乳酸桿菌L-137之特徵者仍於胚芽乳酸桿菌L-137之範疇內。
此外,其他乳酸菌,特別是具有免疫賦活作用之乳酸菌亦可代替胚芽乳酸桿菌L-137株或與其一起併入本發明中。
上述乳酸菌之胚芽乳酸桿菌L-137株可為於天然培養基、合成培養基及半合成培養基等培養基所培養之任一者。於本發明中,乳酸菌之培養可依照習知方法、本身習知之方法或以該等為基準之方法進行。
前述培養基並無特別限制,例如,較佳為使用含有氮源及/或碳源者。前述氮源並無特別限制,可舉例如:肉汁、蛋白腖、麩質、酪蛋白、酵母萃取物、或胺基酸等。前述碳源並無特別限制,可舉例如:葡萄糖、木糖、果糖、肌醇、麥芽糖、水飴、麴汁、澱粉、甘蔗渣、小麥麩皮、糖蜜、或甘油等。此等亦可使用1種或組合2種以上使用。
前述培養基中,除了前述氮源及/或碳源,亦可進一步添加無機物質。前述無機物質並無特別限制,可舉例如:硫酸銨、磷酸鉀、氯化鎂、食鹽、鐵、錳、鉬或各種維生素類等,此等亦可使用1種或組合2種以上使用。
乳酸菌之胚芽乳酸桿菌L-137株之培養溫度及培養時間只要可有效率地實施培養則無特別限制,於本發明之一態樣中,培養溫度可為例如通常約25至40℃,較佳為約27至35℃,培養時間可為例如約12至48小時。再者,於本發明之一態樣中,乳酸菌之培養可藉由通氣震盪振盪而實施。再者,培養基之pH並無特別限制,於本發明之一態樣中,通常可為約pH3至6,較佳為約pH4至6。
〔乳酸菌之胚芽乳酸桿菌L-137株之處理物〕
所謂乳酸菌之胚芽乳酸桿菌L-137株之「處理物」,較佳為將該乳酸菌加工而成者或該乳酸菌之培養物,但不限於此等。再者,菌可為生菌,亦可為死菌體,但從安定性及操作容易性等觀點而言,較佳為使用死菌體。
於本發明中,可直接使用如此之處理物,亦可經冷凍乾燥、低溫乾燥、噴霧乾燥、或L-乾燥等、或是經此等之組合處理成粉末狀而使用。再者,關於此等之處理物(培養物)可以適合的溶劑(水、醇、有機溶劑等)稀釋使用,亦可添加適合的添加劑形成凝膠或固形製劑而使用。
具體說明調配乳酸菌之死菌體之方法。
於本發明中,前述死菌體之調配方法只要不喪失本發明之效果,則無特別限制,例如,可藉由(I)及(II)等之任一種方法調配:(I)於培養結束後,自培養液分離乳酸菌之生菌體後,將前述生菌體進行殺菌處理而成為死菌體狀態之方法;(II)於培養液中,將乳酸菌之生菌體進行殺菌處理而成為死菌體狀態,之後自培養液分離前述死菌體之方法。
自培養液分離菌體之方法可使用本領域中通常使用之各種方法,並無特別限制。於本發明之一態樣中,具體而言,可採用例如,將蒸餾水加入培養液後,經離心分離等手段去除上清液,藉此分離培養液與菌體之方法等。此外,於此態樣中,於培養液添加蒸餾水再進行離心並去除上清液後,因應所需,重複數次於去除上清液之殘留物中進一步添加蒸餾水再進行離心之操作。於本發明之一態樣中,分離操作可包含過濾步驟。
接著,具體說明乳酸菌之生菌體之殺菌處理方法。前述殺菌處理方法並無特別限制,可舉例如:加熱、紫外線照射、福馬林處理等。此外,前述殺菌處理可對所取得之生菌體進行,亦可對包含生菌體之培養液進行。
進行前述加熱處理時,加熱溫度並無特別限制,例如,通常可為約60℃至100℃,較佳為約70至90℃。加熱手段可使用習知方法,並無特別限制,例如可為加熱器等手段。加熱時間只要可充分完成殺菌處理,則無特別限制,例如,加熱時間可為達到期望之溫度後,通常約5至40分鐘,較佳為約10至30分鐘。
經上述處理所得之前述死菌體可進一步進行磨碎、破碎或冷凍乾燥處理等,形成死菌體處理物。於本發明中,前述死菌體處理物亦可作為死菌體而適當使用。
再者,於本發明中,亦可乳酸菌菌體之萃取物替代乳酸菌菌體或與菌體一起使用。前述萃取物之萃取方法並無特別限制,可依照習知方法、自體習知方法或以該等為基準之方法進行。具體而言,可舉例如將乳酸菌之生菌體或死菌體以(i)或(ii)之方法等萃取:(i)於常壓或加壓下,添加至室溫或經加溫之萃取溶劑中,一邊浸漬一邊攪拌而進行萃取之方法;(ii)於萃取溶劑中一邊回流一邊萃取之方法等。萃取溫度、萃取時間、使用之萃取溶劑種類可依萃取條件適當地選擇。
前述萃取溶劑並無特別限制,可使用例如:水、有機溶劑或此等之任意比例混合的溶劑。前述有機溶劑並無特別限制,可舉例如:低級醇(例如,甲醇、乙醇、正丙醇、異丙醇及正丁醇等)、多元醇(例如,1,3-丁二醇、丙二醇、甘油等)等室溫中為液體之醇類、醚類(例如,乙醚、丙醚等)、酯類(例如,乙酸乙酯、乙酸丁酯等)、酮類(例如,丙酮、甲基乙基酮等)、烴類(例如,己烷、二
甲苯、甲苯等)、氯仿等。此等亦可使用1種或組合2種以上使用。即使是於上述有機溶劑中,從操作性及對環境之影響等觀點而言,較佳為使用室溫中為液體之醇類,例如,碳原子數1至4之低級醇,從殘留溶劑之安全性之觀點而言,更佳為使用乙醇。
於本發明中,可將包含經前述萃取操作而得之萃取液及殘渣之混合物因應所需進行過濾或離心等,並將去除屬於固形成分之殘渣而得之萃取液直接用於本發明之前述載劑之調配中,亦可將前述萃取液藉由濃縮、冷凍乾燥或噴霧乾燥等方法進行乾燥、粉末化而使用。
本發明之飼料或組成物中之乳酸菌或其處理物的含量只要不喪失本發明之效果,則無特別限制,飼料或組成物100質量%中,例如可為約0.0001質量%至0.1質量%之範圍,較佳可為0.001至0.01質量%之範圍。本發明之飼料或組成物中之乳酸菌或其處理物與脂肪酸或其鹽的質量比,只要不喪失本發明之效果,則無特別限制,可為1:0.5至50000之範圍,較佳為1:0.5至2000之範圍,更佳為1:0.5至100,特佳為1:0.5至2.5之範圍。
〔脂肪酸〕
本發明之脂肪酸可使用例如飽和或不飽之直鏈或支鏈之脂肪酸。此等脂肪酸具體而言可列舉:甲酸、乙酸、丙酸、丁酸、戊酸、己酸、庚酸、辛酸、壬酸、癸酸、月桂酸、肉豆蔻酸、十五酸、棕櫚酸、棕櫚油酸、十七酸、硬脂酸、結核菌硬脂酸(tuberculostearic acid)、異硬脂酸、油酸、反油酸、牛油酸(vaccenic acid)、亞麻油酸、α-次亞麻油酸、γ-次亞麻油酸、桐酸(eleostearic acid)、十八碳四烯酸、蓖麻酸、花生酸、花生四烯酸、二高-γ-次亞麻油酸(dihomo-γ-linolenic acid)、二十碳五烯酸、二十二酸、芥子酸、二十二碳五烯酸、二十二碳六烯酸、二十四
酸、神經酸、蠟酸、二十八酸、蜂花酸、丙二酸、琥珀酸、蘋果酸、檸檬酸、戊二酸、己二酸、庚二酸、辛二酸、壬二酸、癸二酸等。
較佳之脂肪酸可舉例如選自由短鏈脂肪酸、中鏈脂肪酸及長鏈脂肪酸所組成之群組中之至少1種脂肪酸或此等之混合物。更佳為短鏈脂肪酸或中鏈脂肪酸,再更佳為短鏈脂肪酸。
再者,脂肪酸可具有取代基。取代基並無特別限制,可舉例如:烷基、環烷基、烷氧基、羧基、酮基、醛基、羰基、羥基、硝基、胺基、腈基、巰基、硝基、醚鍵、酯鍵、醯胺鍵、胺基甲酸乙酯鍵等通常用於醫藥、動物藥、食品、化妝品等領域中之習知基或鍵等。
又,本發明之脂肪酸之較佳來源可為例如藉由習知方法或自體習知方法從下列油獲得之動植物性游離脂肪酸等:大豆油、菜籽油、棉籽油、落花生油、棕櫚油、葵花籽油、小麥胚芽油、米糠油、玉米油、麻油、櫻桃籽油、紅花籽油、亞麻仁油、杏仁果油、印加果油(Sacha inchi oil)、荏胡麻油、橄欖油、橙子油、南瓜籽油、紫蘇油、茶油、椿油、花生油、葡萄籽油、夏威夷豆油、牛油、豬油、蛋黃油;自沙丁魚、鮭魚、鯖魚、秋刀魚、鯡魚、鮪魚等所得之魚油;魷魚、狹鱈(Theragra chalcogramma)等之魚肝油;鰹魚、鮪魚等之魚眼窩油;海豹油、磷蝦油(krill oil)等;亦可使用藉由化學合成等所得之市售品之脂肪酸等。
(脂肪酸之鹽)
於本說明書中,脂肪酸可依照習知方法轉換為期望之鹽。如此之鹽之較佳例可舉例如:與鈉、鉀、鋰、鈣、鎂等無機鹼之鹽;與氨、二甲胺、三甲胺、二環己胺等之有機鹽等,但不限於此等。
(短鏈脂肪酸)
於本說明書中,短鏈脂肪酸通常為碳數1至6之飽和或不飽和之直鏈或支鏈脂肪酸。較佳之短鏈脂肪酸之例可舉例如:甲酸、乙酸、丙酸、丁酸、異丁酸、異戊酸、乳酸、丙酮酸、戊酸、己酸、琥珀酸、反丁烯二酸、順丁烯二酸、蘋果酸、檸檬酸等,更佳為甲酸、乙酸、丙酸、丁酸、異丁酸、異戊酸、乳酸、丙酮酸、戊酸、己酸、琥珀酸,再更佳為乙酸、丙酸及丁酸,但不限於此等。
此外,就其他較佳例而言,短鏈脂肪酸係使用鹽之形態。如此之鹽可舉出上述與無機鹼之鹽,更佳為與鈉、鉀或鈣等之鹽。
(中鏈脂肪酸)
於本說明書中,中鏈脂肪酸通常為碳數7至11之飽和或不飽和之直鏈或支鏈脂肪酸。較佳之中鏈脂肪酸之例可舉例如:癸酸(羊脂酸)、庚酸、辛酸、壬酸等。較佳為中鏈脂肪酸以直鏈或飽和脂肪酸為佳,以直鏈之飽和脂肪酸為更佳。
(長鏈脂肪酸)
於本說明書中,長鏈脂肪酸通常為碳數12以上之飽和或不飽和之直鏈或支鏈脂肪酸。較佳之長鏈脂肪酸之例可舉例如:月桂酸、十三酸、肉豆蔻酸、棕櫚酸、硬脂酸、棕櫚油酸、牛油酸、二十烯酸、亞麻油酸、γ-次亞麻油酸、α-次亞麻油酸、花生四烯酸、油酸等。長鏈脂肪酸較佳為不飽和脂肪酸,更佳為ω-3、ω-6、ω-7或ω-9不飽和脂肪酸,再更佳為ω-7不飽和脂肪酸。此外,「ω-X不飽和脂肪酸」意指自脂肪酸之甲基末端算起之第X個鍵為不飽和鍵者。
〔飼料(飼料用組成物)〕
本發明之飼料(飼料用組成物)可舉例如:牛、馬、豬等之家畜用飼料;雞、鵪鶉、火雞、鴨、鵝等之家禽用飼料;幼鰤魚、鰤魚、鯛魚、蝦、牡蠣、蛤等魚貝等之養殖用飼料;狗、貓等之寵物用飼料等,但不限於此等。家禽用飼料較佳
為雞,更佳為肉雞、蛋雞、土雞之雞,並無特別限制。本發明之飼料除了於飼料中添加本發明之乳酸菌及脂肪酸之外,可藉由一般的飼料之製造方法適當地加工及製造。
〔IL-12產生增強用組成物及免疫賦活用組成物〕
如上所述,本發明之組成物之較佳係以具有IL-12產生增強作用及/或免疫賦活作用作為特長。本發明之組成物不是僅限於飼料者,其他較佳例係飲食品用及/或醫藥品用(亦包含動物藥)。就進一步之其他較佳例而言,本發明之組成物係使用為飲食品用添加劑。
飲食品用、飲食品用添加劑用或醫藥品用之組成物,係可將上述乳酸菌與脂肪酸,進一步適當地調配醫藥上可接受之載體、添加劑等而進行製劑化等。由於以往已充分地建立用於此之製劑化方法及製劑化技術,因此根據該等即可。例如,為醫藥品用時,具體可為:錠劑、包衣錠劑、丸劑、粉劑、顆粒劑、膠囊劑、液劑、懸浮劑、乳劑等口服劑;注射劑、輸液、栓劑、軟膏、貼劑等非口服劑。載體或添加劑之調配比例,只要是依據醫藥品或獸醫學領域中通常所採用之範圍而適當地設定即可。可調配之載體或添加劑並無特別限制,可舉例如:水、生理食鹽水、其他水性溶劑、水性或油性基劑等各種載體、酵素、pH調整劑、保存劑、殺菌劑、抗氧化劑、防霉劑、保鮮劑、漂白劑、光澤劑、香料、甜味劑、酸味劑、調味劑、苦味劑、乳化劑、增黏劑、穩定劑、膠凝化劑、糊劑、賦形劑、黏合劑、崩解劑、潤滑劑、著色劑、調味劑、香料等各種添加劑,但不限於此等。由於以往已充分地建立與此等相關之技術,因此於本發明中,依照此等技術即可。
此外,當投予本發明之IL-12產生增強用組成物或免疫賦活用組成物時,其對象較佳為除人以外,亦可為上述飼料之段落中所述之家畜、家禽或寵物等動物,但不限於此等。
再者,當本發明之組成物為飲食品用時,如此之飲食品中包含:健康食品、機能性顯示食品、特定保健用食品、患者用食品。飲食品之形態並無特別限制,於具體例可舉例如:作為所謂的營養補充品或補充劑之錠劑、顆粒劑、粉劑、保健飲料等。除此之外,可舉例如:茶、清涼飲料、碳酸飲料、能量飲料、果汁、乳酸飲料等飲料;蕎麥麵、烏龍麵、中華麵、泡麵等麵類;飴、糖果、口香糖、巧克力、小點心、餅乾、果凍、果醬、鮮奶油、烘焙點心、麵包等點心及麵包類;火腿、香腸、半片(魚漿)、竹輪等水產/畜產加工食品;加工乳、發酵乳等乳製品;沙拉油、天婦羅油、人造奶油、美乃滋、酥油、慣奶油(whipped cream)、沙拉醬等油脂及油脂加工食品;醬汁、沾醬(Tare sauce)等調味劑;咖哩、濃湯、丼飯、清粥、鹹粥等殺菌袋裝食品;冰淇淋、雪泥(sherbet)等冷凍點心等,但不限於此等。由於以往已充分建立與此等相關之技術,因此於本發明中,依照此等技術即可。
〔抑菌用組成物〕
本發明之組成物較佳為進一步以具有抑制細菌增殖之抑菌作用作為特長。成為本發明之組成物所致之抑菌作用之對象的細菌較佳可舉例如:沙門氏菌、傷寒桿菌、副傷寒桿菌、包含O-157、O-111、O-121等之腸道出血性大腸桿菌、腸毒性大腸桿菌、腸道組織入侵性大腸桿菌等病原性大腸桿菌;霍亂弧菌、志賀桿菌、彎曲桿菌、梭菌、金黃色葡萄球菌、魏氏梭菌、腸炎弧菌等成為腸道感染症
之原因的菌,但不限於此等。沙門氏菌較佳可舉例如:傷寒桿菌、副傷寒桿菌、腸炎菌,但不限於此等。
本發明之組成物不是僅限於飼料者,就其他較佳例而言,可為飲食品用及/或醫藥品用(亦包含動物藥)。
飲食品用、飲食品用添加劑用或醫藥品用之組成物,係可將上述乳酸菌與脂肪酸,進一步適當地調配醫藥上可接受之載體、添加劑等而進行製劑化等。由於以往已充分建立用於此之製劑化方法及製劑化技術,因此依照該等即可。例如,為醫藥品用時,具體可為:錠劑、包衣錠劑、丸劑、粉劑、顆粒劑、膠囊劑、液劑、懸浮劑、乳劑等口服劑、注射劑、輸液、栓劑、軟膏、貼劑等非口服劑。載體或添加劑之調配比例,只要是基於醫藥品或獸醫學領域中通常所採用之範圍而適當地設定即可。可調配之載體或添加劑並無特別限制,可舉例如:水、生理食鹽水、其他水性溶劑、水性或油性基劑等各種載體;酵素、pH調整劑、保存劑、殺菌劑、抗氧化劑、防霉劑、保鮮劑、漂白劑、光澤劑、香料、甜味劑、酸味劑、調味劑、苦味劑、乳化劑、增黏劑、穩定劑、膠凝化劑、糊劑、賦形劑、黏合劑、崩解劑、潤滑劑、著色劑、調味劑、香料等各種添加劑,但不限於此等。由於以往已充分建立與此等相關之技術,因此於本發明中,依照此等技術即可。
再者,本發明之組成物為飲食品用時,如此之飲食品中包含:健康食品、機能性顯示食品、特定保健用食品、患者用食品。食品之形態並無特別限制,具體例可舉例如:作為所謂的營養補充品或補充劑之錠劑、顆粒劑、粉劑、保健飲料等。除此之外,可舉例如:茶飲料、清涼飲料、碳酸飲料、能量飲料、果汁、乳酸飲料等飲料;蕎麥麵、烏龍麵、中華麵、泡麵等麵類、飴、糖果、口香糖、巧克力、小點心、餅乾、果凍、果醬、鮮奶油、烘焙點心、麵包等點心及
麵包類;火腿、香腸、半片(魚漿)、竹輪等水產/畜產加工食品;加工乳、發酵乳等乳製品;沙拉油、天婦羅油、人造奶油、美乃滋、酥油、摜奶油、沙拉醬等油脂及油脂加工食品;醬汁、沾醬等調味劑;咖哩、濃湯、丼飯、清粥、鹹粥等殺菌袋裝食品;冰淇淋、雪泥等冷凍點心等,但不限於此等。由於以往已充分建立與此等相關之技術,因此於本發明中,依照此等技術即可。
雖詳細不明,但於本發明中,藉由向對象投予本發明之飼料或組成物,獲得IL-12產生被增強及/或免疫被賦活之結果,因而被認為是可抑制沙門氏菌等之增殖,產生抑菌作用者。
此外,當投予本發明之抑菌用組成物時,其對象較佳為除了人以外,亦可為上述飼料之段落中所述之家畜、家禽或寵物等動物,但不限於此等。
〔其他成分〕
於本發明之飼料或組成物中,只要不喪失本發明之效果,亦可含有例如醫學、藥學、獸醫學或食品等領域中已知之任意成分。
如此之成分可使用例如上述以外之已知具有免疫賦活活性之乳酸菌。例如,可使用上述胚芽乳酸桿菌L-137株以外之屬於乳酸桿菌屬、鏈球菌屬、腸球菌屬、乳球菌屬、或雙岐桿菌屬等之乳酸菌等。更詳細可列舉:胚芽乳酸桿菌(Lactobacillus plantarum)、嗜酸乳酸桿菌(Lactobacillus acidophilus)、短毛乳酸桿菌(Lactobacillus brevis)、酪蛋白乳酸桿菌(Lactobacillus casei)、發酵乳酸桿菌(Lactobacillus fermentum)、副乾酪乳酸桿菌(Lactobacillus paracasei)、布氏乳酸桿菌(Lactobacillus buchneri)、戴白氏乳酸桿菌(Lactobacillus delbrueckii)、鼠李糖乳酸桿菌(Lactobacillus rhamnosus)、嗜熱鏈球菌(Streptococcus thermophilus)、糞腸球菌(Enterococcus faecalis)、屎腸球菌(Enterococcus faecium)、乳酸乳球菌
(Lactococcus lactis)、胚芽乳球菌(Lactococcus plantarum)或嗜熱雙岐桿菌(Bifidobacterium thermophilum)、長雙歧桿菌(Bifidobacterium longum)或短雙歧桿菌(Bifidobacterium breve)等,但不限於此等。
〔投予量〕
胚芽乳酸桿菌L-137之攝取量,在經口或注射投予時,可依據攝取者之年齡及體重、症狀、投予時間、劑形、投予方法、藥劑之組合等而決定。例如,將胚芽乳酸桿菌L-137作為乾燥死菌體,成人每人(約60kg)每日攝取較佳為約0.5至200mg,更佳為約1至100mg,再更佳為約2至50mg。或者,將胚芽乳酸桿菌L-137以生菌數換算時,較佳為以使成人每人(約60kg)每日攝取較佳為約5×108至2×1011cfu(Colony forming unit;菌落形成單位),更佳為約1×109至1×1011cfu之方式進行設定。攝取次數可為1日1次或分成複數次進行。
當外用塗佈時,可因應施用之皮膚面積而適當地選擇胚芽乳酸桿菌L-137之塗佈量,通常,相對於施用部位之面積約10cm2,該塗佈量係每日較佳為約0.01至2.5mg,更佳為約0.02至1mg。可將前述投予量以1日1次或分成數次投予或施用。
〔製造方法〕
本發明係包含飼料等之本發明組成物之製造方法,該製造方法例如包含:將胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物及脂肪酸,因應所需,與其他成分混合之步驟。於混合步驟中,為了使胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物及脂肪酸或其鹽與其他成分於飼料或組成物全體中均勻,較佳為藉由習知方法或自體習知方法混合或攪拌。由於本領域中已充分建立混合或攪拌之方法,因此依照該方法即可。
〔IL-12產生增強效果、免疫賦活效果及抑菌效果之確認〕
確認本發明之飼料或組成物之效果之方法可舉例如:相較於不含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物及脂肪酸或其鹽皆之飼料或組成物,確認含有胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物及脂肪酸或其鹽之本發明之飼料或其他組成物具有較優異的L-12產生、免疫賦活活性或抑菌效果之方法。IL-12產生及免疫賦活活性之確認方法,由於本領域中已充分建立ELISA等,因此依照該方法即可。抑菌效果之確認方法,由於本領域中已充分建立以目視或顯微鏡等確認細菌增殖之有無,以及利用適合的培養基培養而測定增殖性之細菌類所產生之酵素量之測定、PCR法等,因此依照該方法即可。
此外,免疫賦活活性具體可舉例如:B細胞、T細胞、巨噬細胞、自然殺手(NK)細胞、樹突細胞等細胞之IL-12等介白素、或者IFN-β或IFN--γ等干擾素等細胞激素的產生促進活性等,但不限於此等。
此等之上述方法之一例可參照後述之實施例。
將本發明之組成物調配成飲食品、動物用飼料、醫藥品(亦包含動物用醫藥)、或準醫藥品之形態時,於該飲食品、飼料、醫藥品、或準醫藥品、其附加仿單或其包裝盒等,有鑑於本發明之組成物之作用,可標示係具有IL-12產生增強作用或免疫賦活活性之乳酸菌或其處理物的要旨。
本發明之飼料中之乳酸菌或其處理物及脂肪酸或其鹽之質量比(乳酸菌或其處理物:脂肪酸或其鹽),例如,較佳為1:0.5至50000,更佳為1:0.5至2000,再更佳為1:0.5至100,特佳為1:0.5至2.5。
本發明之醫藥品(亦包含動物用醫藥)、飲食品、準醫藥品等組成物中之乳酸菌或其處理物及脂肪酸或其鹽之質量比(乳酸菌或其處理物:脂肪酸或其鹽),例如,較佳為1:0.5至50000,更佳為1:0.5至2000,再更佳為1:0.5至100,特佳為1:0.5至2.5。
[實施例]
以下舉出實施例及試驗例以更具體地說明本發明,但本發明不限於此等。
〔實施例1〕於免疫細胞株添加有本發明之乳酸菌與脂肪酸或其鹽時之IL-12產生增強效果之確認
實驗方法:以使胚芽乳酸桿菌L-137乾燥死菌體以2mg/mL懸浮於PBS中之方式,添加含有10%FBS之RPMI1640培養基稀釋成40μg/mL而作為試驗液1。以無Dnase水(短鏈脂肪酸)或EtOH(中鏈、長鏈脂肪酸)使脂肪酸(中鏈、長鏈)或脂肪酸之鈉鹽(短鏈)溶解成0.5M後,添加含有10%FBS之RPMI1640培養基稀釋成400μM而作為試驗液2。使類巨噬細胞之細胞株之J774.1細胞(細胞編號JCRB9108;日本國立研究開發法人醫藥基盤健康營養研究所JCRB細胞銀行)各自以成為1.0×106細胞/mL之方式懸浮於含有10%FBS之RPMI1640培養基,作為細胞株懸浮液。將各50μL試驗液1、2與各100μL細胞株懸浮液添加至96孔培養盤(菌體最終濃度:10μg/mL,脂肪酸最終濃度:100μM,細胞最終濃度:5.0×105/mL),於5%CO2培養箱內以37℃培養48小時。培養結束後之上清液中之IL-12p40濃度係經ELISA法測定。此外,檢測極限濃度為0.07ng/mL。結果如下述表1及圖1、圖2所示。
使用之試劑等之廠商及產品名如下述。
乙酸鈉(S5636,Sigma-Aldrich)、丙酸鈉(P5436,Sigma-Aldrich)、N-丁酸鈉(B5887,Sigma-Aldrich)、癸酸(21409,Sigma-Aldrich)、棕櫚油酸(P9417、Sigma-Aldrich)、FBS(SH30071.03,Hyclone)、RPMI1640培養基(23400-021,Thermo Fisher Scientific)、ELISA用一次抗體(505202,Biolegend)、ELISA用二次抗體(BAF419,R & D Systems)、ELISA用標準品(577009,Biolegend)
結果及考察:
如上述表1及圖1、圖2所示,單獨添加脂肪酸時,任一脂肪酸(短鏈脂肪酸、中鏈脂肪酸、長鏈脂肪酸)或其鹽均無法使免疫細胞產生IL-12。而且,雖乳酸菌之胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137株)對於免疫細胞具有一定的IL-12產生效果,但同時添加乳酸菌與脂肪酸時,所增強之IL-12之產生具有單獨添加脂肪酸時完全看不到的顯著差異。從此等之結果可知,藉由組合乳酸菌與脂肪酸,可增進IL-12之產生。此外,增進IL-12之產生被認為亦對免疫賦活作用有良好影響。
本發明係首次發現藉由將單獨時不具有IL-12產生活性之脂肪酸與乳酸菌之胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137株)組合,所增強之IL-12之產生具有顯著差異的初例。
〔實施例2〕對動物投予本發明之乳酸菌與脂肪酸或其鹽時之IL-12產生增強效果及抑菌效果之確認
又,於實施例2中,對動物(肉雞)投予含有胚芽乳酸桿菌L-137與脂肪酸之飼料(試驗餐),並進行其評估。
實驗方法:
(1)關於試驗準備及飼養方法
於實施例2中,將64隻肉雞(Ross308,雄性,購自泰國南奔府正大食品)分成下列試驗區(T1、T2及T3),並進行評估。
區(T1):給予不含胚芽乳酸桿菌L-137乾燥死菌體粉末及丙酸鈉(100% Sodium Propionate,CALTECH CORPORATION LIMITED)之試驗餐(粉末飼料),
區(T2):給予混有10ppm濃度之胚芽乳酸桿菌L-137乾燥死菌體之試驗餐,
區(T3):給予混有5ppm濃度之胚芽乳酸桿菌L-137乾燥死菌體與2000ppm濃度之丙酸鈉之試驗餐。
(以下試驗中,若T3之有效性與T2同等以上,則視為得到協同效果)
對肉雞自1日齡至21日齡給予各區試驗餐,自22日齡至42日齡以通常飼料飼養。關於試驗餐及通常飼料之細節係如下述表2。
(2)IL-12產生增強效果之確認
42日齡時從各區之8隻肉雞採取盲腸扁桃腺,並進行RNA萃取。自該RNA檢體合成cDNA,以cDNA為模板實施qPCR。所使用之引子之序列如下述表3。
對所採取之盲腸扁桃腺之基因表現進行解析,並測量及統計T1區、T2區及T3區之各自肉雞中之IL-12之表現強度。結果如圖3及下述表4。
(3)對沙門氏菌(Salmonella spp.)之抑菌效果之確認
42日齡時自各區之6隻肉雞以無菌方式採取盲腸內容物,並測量沙門氏菌之菌數。結果如表5所示。表中之值為平均值。
結果及考察:
從盲腸扁桃腺基因表現解析的結果(圖3及表4)可知,相較於T2區,T3區中,IL-12的表現量顯著較高。再者,採取並分析盲腸內容物時,關於沙門氏菌(Salmonella spp.)之數量,相較於T2區,T3區中,菌數較少。
亦即,在投予了包含胚芽乳酸桿菌L-137乾燥死菌體及丙酸兩者飼料之肉雞中,確認了協同的IL-12表現作用及抑菌。詳細理由雖不明,但咸信是因為胚芽乳酸桿菌L-137乾燥死菌體及丙酸鈉的併用,使得作為宿主之肉雞的IL-12產生被增強,結果,抑制沙門氏菌的增殖,而產生抑菌作用者。
再者,由沙門氏菌的菌數(表5)可知,含有乳酸菌之胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137株)或其處理物與脂肪酸或其鹽之組成物,不僅於生體外(in vitro),於動物(in vivo)中也顯示有效的IL-12產生增強作用及免疫賦活作用。
[產業上之可利用性]
本發明之飼料及組成物係具有IL-12產生增強作用、免疫賦活作用或抑菌作用,因此可用為飲食品、飼料、醫藥品或準醫藥品、及化妝品等。
【生物材料寄存】
【寄存國家】JP日本
【寄存機構】獨立行政法人製品評價技術基盤機構特許微生物寄託中心
【寄存日期】1995/11/30
【寄存號碼】FERM BP-08607
<110> 日商好侍健康食品股份有限公司(HOUSE WELLNESS FOODS CORPORATION)
<120> 包含乳酸菌與脂肪酸的飼料及組成物
<130> T09F7134
<140> TW 110135594
<141> 2021-09-24
<150> JP 2020-161335
<151> 2020-09-25
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> 引子
<400> 1
<210> 2
<211> 29
<212> DNA
<213> 人工序列
<220>
<223> 引子
<400> 2
<210> 3
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引子
<400> 3
<210> 4
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 引子
<400> 4
Claims (12)
- 一種飼料,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
- 如請求項1所述之飼料,其中,脂肪酸係選自由短鏈脂肪酸、中鏈脂肪酸及長鏈脂肪酸所組成之群組中之至少1種脂肪酸或此等之混合物。
- 如請求項2所述之飼料,其中,短鏈脂肪酸係選自由甲酸、乙酸、丙酸、丁酸、異丁酸、戊酸、異戊酸、乳酸、丙酮酸、己酸及琥珀酸所組成之群組中之至少1種。
- 如請求項1至3中任一項所述之飼料,其中,胚芽乳酸桿菌L-137株係死菌體。
- 如請求項1至4中任一項所述之飼料,其中,相對於全體飼料,胚芽乳酸桿菌L-137株或其處理物之含有率係0.0001質量%至0.1質量%。
- 如請求項1至5中任一項所述之飼料,其中,飼料中之胚芽乳酸桿菌L-137株或其處理物與脂肪酸或其鹽之質量比係1:0.5至50000。
- 一種IL-12產生增強用組成物,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
- 一種免疫賦活用組成物,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
- 一種抑菌用組成物,係包含胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
- 一種組成物,係含有胚芽乳酸桿菌L-137株(Lactobacillus plantarum L-137)或其處理物與脂肪酸或其鹽。
- 一種免疫賦活用組成物,係含有乳酸菌或其處理物與脂肪酸或其鹽。
- 如請求項11所述之免疫賦活用組成物,其中,免疫賦活用組成物為醫藥、動物藥、準醫藥品、飼料、飲食品或飲食品添加物。
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EP (1) | EP4218936A1 (zh) |
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CN (1) | CN116322350A (zh) |
BR (1) | BR112023003812A2 (zh) |
CA (1) | CA3193303A1 (zh) |
CL (1) | CL2023000858A1 (zh) |
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JP4813902B2 (ja) | 2003-03-26 | 2011-11-09 | ハウスウェルネスフーズ株式会社 | 抗ストレス剤 |
JP2008195713A (ja) | 2007-01-19 | 2008-08-28 | Shunkosha:Kk | 免疫調節剤 |
JP5207313B2 (ja) | 2008-05-30 | 2013-06-12 | 有限会社バイオメディカルリサーチグループ | 乳酸菌配合物及びその製造方法 |
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CN116322350A (zh) | 2023-06-23 |
CA3193303A1 (en) | 2022-03-31 |
CL2023000858A1 (es) | 2023-09-08 |
JPWO2022065268A1 (zh) | 2022-03-31 |
WO2022065268A1 (ja) | 2022-03-31 |
JP7342338B2 (ja) | 2023-09-12 |
EP4218936A1 (en) | 2023-08-02 |
BR112023003812A2 (pt) | 2023-04-04 |
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