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研究生:張詩芷
研究生(外文):Shi-zhi Chang
論文名稱:鮪魚香腸中組織胺相關衛生品質、組織胺生產菌之分離及禽畜肉摻假之探討
論文名稱(外文):Histamine-Related Hygienic Quality, Histamine-Forming Bacteria and Adulteration of Pork and Poultry in Tuna Sausage Products
指導教授:龔賢鳳
指導教授(外文):Hsien-feng Kung
學位類別:碩士
校院名稱:大仁科技大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:146
中文關鍵詞:保色劑摻入組織胺生產菌鮪魚香腸生物胺組織胺
外文關鍵詞:Color fasting agentAdulterationHistamine-forming bacteriaTuna sausageBiogenic amineHistamine
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本研究自台灣傳統市場、超級市場及網路宅配各販售點購買25件鮪魚香腸製品,進行生物胺含量分析、組織胺生產菌分離及禽畜肉摻入之鑑定。研究結果顯示所有樣品生物胺含量之平均値皆低於2.0 mg/100 g。在組織胺生產菌之分離,計篩出13株組織胺生產菌,以Trypticase soy broth含1.0% L-histidine(TSBH)中進行增殖培養,得知其組織胺生成量在12.1-1261.4 ppm之間,其中Raoultella ornithinolytica(1株)、Enterobacter aerogenes(1株)與Staphylococcus pasteuri(2株)屬於高組織胺生產菌。
以多套式PCR進行鮪魚香腸製品掺入肉品物種鑑定分析,顯示 25 件樣品均含魚肉,80%(20/25)的樣品含豬肉,4%(1/25)含禽肉。進一步對鮪魚香腸製品之魚種進行分析,得知有22 件樣品(88%)魚種分析結果為黃鰭鮪(Thunnus albacares),而長鰭鮪(T. alalunga)及黑鮪(T. thynnus)各有1 件(4%),然有1件樣品則未檢出鮪魚成分,卻檢出黑皮旗魚(Makaira nigrican)之魚肉存在。
在保色劑硝酸鹽及亞硝酸鹽對高組織胺生產菌之生長及組織胺生成影響之探討,將高組織胺生產菌E. aerogenes、R. ornithinolytica與S. pasteuri以2.3 log CFU/mL菌量接種於不同濃度硝酸鈉 (0 ppm、70 ppm、200 ppm、300 ppm)及亞硝酸鈉(0 ppm、40 ppm、70 ppm、100 ppm、200 ppm)之TSBH培養液中,於4℃及35℃的貯存試驗,得知亞硝酸鈉對高組織胺生產菌的生長及組織胺生成之抑制效果優於硝酸鈉,在35℃貯存條件下硝酸鈉及亞硝酸鈉對革蘭氏陽性組織胺生產菌S. pasteuri之生長與組織胺蓄積之抑制效果較革蘭氏陰性菌E. aerogenes、R. ornithinolytica佳。此外,於4℃的冷藏溫度下,可有效的抑制延緩組織胺生產菌之生長及組織胺生成,故以70 ppm亞硝酸鈉並配合4℃的冷藏可分別延緩高組織胺生產菌E. aerogenes及R. ornithinolytica之生長及組織胺之生成。
Twenty-five tuna sausage products were purchased from retail markets in Taiwan. Occurrence of biogenic amines, histamine-forming bacteria, and adulteration of pork and poultry were determined. Average content of various biogenic amines in all tested samples was less than 2.0 mg/100g. Thirteen histamine-producing bacterial strains isolated from tested samples produced 12.1 ppm to 1261 ppm of histamine in trypticase soy broth supplemented with 1.0% L-histidine (TSBH). Among them, the Raoultella ornithinolytica (one strain), Enterobacter aerogenes(one strain)and Staphylococcus pasteuri (two strains) isolates were prolific histamine-formers. Assay of multiplex polymerase chain reaction(PCR)revealed adulteration rates were 80% (20/25) and 4% (1/25) for pork and poultry, respectively, in tuna sausage. Tuna species in tuna sausage samples were identified as Thunnus albacares for 22 samples (88%), Thunnus alalunga for one sample (4%) and Thunnus thynnus for one sample(4%), whereas the other one was identified as Makaira nigrican(blue marlin).
中文摘要....................................................................................................... I
英文摘要....................................................................................................... II
誌謝............................................................................................................... III
目錄............................................................................................................... IV
圖目錄........................................................................................................... VII
表目錄........................................................................................................... IX
第壹章、前言................................................................................................ 1
第貳章、文獻回顧 3
一、台灣常見之鮪屬魚種介紹.................................................... 3
(一) 鮪屬魚種之分類................................................................... 3
(二) 台灣常見之鮪魚................................................................... 3
(三) 我國鮪魚之漁獲量與主要利用情形................................... 5
二、香腸之種類介紹..................................................................... 6
(一) 香腸製程之種類................................................................... 6
(二) 台灣中式香腸之起源........................................................... 7
(三) 中式香腸之特性與製作原理............................................... 8
(四) 香腸中保色劑之硝酸鹽及亞硝酸鹽................................... 8
(五) 香腸中防腐劑之己二烯酸鈉............................................... 12
三、魚介類之鮮度指標................................................................. 13
(一) 魚介類死後生化學變化....................................................... 14
(二) 總揮發性鹽基態氮 (Total volatile base nitrogen, TVBN)....... 15
(三) pH值....................................................................................... 16
(四) 總生菌數 (Aerobic plate count, APC)..................................... 17
四、食品中之生物胺.................................................................... 17
(一) 生物胺之生成與生理功能................................................... 17
(二) 食品中生物胺之代謝............................................................ 19
五、組織胺與食品中毒之關係..................................................... 20
(一) 食品中組織胺來源............................................................... 20
(二) 組織胺中毒症狀與劑量…................................................... 20
(三) 食品中組織胺限量標準....................................................... 21
(四) 食品中組織胺生產菌之分離................................................ 22
(五) 影響魚類組織胺形成之因素................................................ 23
(六) 貯藏條件對魚肉中組織胺生成之影響............................... 25
(七) 食品中組織胺中毒之預防.................................................... 25
(八) 食品中組織胺之分析方法.................................................... 26
六、一般魚種鑑定之方法............................................................... 29
(一) 外觀特徵鑑種法................................................................... 29
(二) 以蛋白質分析法來鑑定魚種............................................... 29
七、利用 PCR 技術鑑定物種......................................................... 31
(一) 直接定序法........................................................................... 31
(二) PCR- RAPD分析.................................................................. 32
(三) PCR- SSCP分析................................................................... 32
(四) Species- specific PCR分析.................................................. 33
(五) Multiplex- PCR分析技術.................................................... 34
(六) AFLP分析............................................................................ 35
(七) PCR- RFLP鑑種技術之應用............................................... 35
第參章、材料與方法
一、實驗材料與藥品........................................................................... 38
二、鮪魚香腸製品之微生物品質分析............................................... 41
三、鮪魚香腸製品中一般成份及生物胺含量分析........................... 43
四、食品中組織胺生產菌之分離及保存........................................... 48
五、食品中組織胺生產菌之鑑定....................................................... 49
六、組織胺生產菌16S rDNA序列鑑定菌種...................................... 49
七、鮪魚香腸製品中摻入禽畜肉品之基因鑑定............................... 52
八、鮪魚香腸製品中鮪魚魚種之基因鑑定....................................... 53
九、鮪魚香腸製品中鰹魚魚種之基因鑑定....................................... 54
十、鮪魚香腸製品中旗魚魚種之基因鑑定....................................... 54
十一、保色劑對組織胺生產菌之生長及組織胺生成之影響........... 55
第肆章、結果與討論
一、鮪魚香腸製品之衛生品質與組織胺生產菌之分離................... 56
(一) 市售鮪魚香腸製品之微生物品質分析..................................... 56
(二) 市售鮪魚香腸製品之化學品質分析......................................... 57
二、鮪魚香腸製品之生物胺分析與組織胺生產菌之分離............... 58
(一) 鮪魚香腸製品之生物胺含量..................................................... 58
(二) 鮪魚香腸製品之組織胺生產菌分離及鑑定............................. 58
三、鮪魚香腸製品之物種鑑定及鮪魚魚種之分析........................... 59
(一) 鮪魚香腸製品之禽畜肉摻入鑑定............................................. 59
(二) 鮪魚香腸製品之鮪魚魚種基因鑑定......................................... 60
(三) 鮪魚香腸製品之鰹魚基因鑑定................................................. 61
(四) 鮪魚香腸製品之旗魚基因鑑定與旗魚魚種基因鑑定…... 61
四、高組織胺生產菌在不同濃度硝酸鹽及亞硝酸鹽之TSBH條件
下總生菌數及組織胺生成量之探討........................................... 61
(一) 不同濃度亞硝酸鈉對E. aerogenes之生長及組織胺生成 之影響................................................................................... 62
(二) 不同濃度亞硝酸鈉對R. ornithinolytica之生長及組織胺生成之影響........................................................................... 63
(三) 不同濃度亞硝酸鈉對S. pasteuri之生長及組織胺生成之影響....................................................................................... 64
(四) 不同濃度硝酸鈉對E. aerogenes之生長及組織胺生成之影響....................................................................................... 64
(五) 不同濃度硝酸鈉對R. ornithinolytica之生長及組織胺生成之影響............................................................................... 65
(六) 不同濃度硝酸鈉對S. pasteuri之菌數及組織胺生成之影響........................................................................................... 66
第伍章、結論................................................................................................ 67
第陸章、參考文獻........................................................................................ 116
第柒章、附錄................................................................................................. 145
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