Characteristics of Quaternary Ammonium Resistant Bacteria from a Fine Papermachine System

紙機系統抗四級銨鹽殺菌劑好氧菌株之分離與鑑定

柯淳涵(Chun-Han Ko) ； 黃啟裕(Chi-Yu Huang) ； 汪淮(Hweig Wang) ； 陳柏昇(Bo-Sheng Chen)

《國立臺灣大學生物資源暨農學院實驗林研究報告》 17卷2期 (2003/06) Pp. 87-95

https://doi.org/10.6542/EFNTU.200306_17(2).0003

摘要

本研究於生產文化用紙之紙機進行採樣，共分離出兼氣性厭氧菌株（facultative anaerobe）與好養菌株（aerobes）51株。初步以四級銨鹽殺菌劑進行液態抗藥性試驗，濃度範圍設定為30-120ppm，以分光光度計於波長600nm下測量菌液吸光度，作為菌株生長監測。試驗結果，51株分離菌株中具四級銨鹽殺菌劑抗藥性菌株共43株，其中抗藥性濃度可達30ppm共計22株，60ppm為13株，90ppm為6株，120ppm為2株。從中選取抗藥性最強之菌株作為研究菌株，菌株編號分別為HB22與HB45。再經由API 20E與16S rRNA gene sequencing進行菌種鑑定，並進行形態特性與生化特性之研究，期望能對於紙機中具有抵抗四級銨鹽殺菌劑之菌株特性有更深入之瞭解，並進一步對抗藥性菌株進行最佳抗藥性生長條件評估之研究。菌株HB22與Morganella morganii由資料比對分析上應可判斷屬於同一菌種。菌株HB45與Pseudomonas cf. monteilii, Pseudomonas mosselii及Pseudomonas putida於資料比對分析上有相似之處。菌株HB22為革蘭氏陰性桿菌，具有鞭毛，有移動性，催化酶反應陽性，氧化酶反應陰性，Indole反應陽性，不可產生硫化氫，為兼氣性厭氧菌株，生長之最佳溫度為35℃，最佳pH為7.0。菌株HB45為革蘭氏陰性桿菌，具有鞭毛，有移動性，可產生催化酶反應陽性，氧化酶反應陰性，Indole反應陰性，不可產生硫化氫，為好氧性菌株，最佳生長溫度為30℃，最佳pH為7.4。菌株HB22在35℃-40℃有較佳之抗藥性，最大抗藥濃度為150ppm，其抑菌時間可達5-10小時。菌株HB45在25℃-35℃有較佳的抗藥性，最大抗藥濃度可達200ppm，其抑菌時間為6-10小時。菌株HB22只可利用D(+)-Mannose與glucose兩種基質進行生長。菌株HB45可利用的基質包含L-Arabinose, L(+)-Rhamnose, D(+)-Mannose, Glucose, Glycerol, Lactose, Maltose, Raffinose, Xy-lose, Cellulose 及Xylan。其中菌株HB45與Pseudomonas putida於基因親緣上甚為接近，而Pseudomonas putida已被証明能形成生物膜，並對紙業廢水有良好分解能力，因此HB45將有相當大的應用潛力。

關鍵字

兼氣性厭氧菌 ； 好氧菌 ； 殺菌劑 ； 四級銨鹽 ； 菌泥

並列摘要

To evaluate the biocide effect of quaternary ammonium chloride (QAC), a whitewater sample was taken from a fine papermachine headbox. By plate spreading method, 51 strains of facultative anaerobes were isolated morphologically. Forty-three strains among the isolates were found resistant to the biocide, quaternary ammonium chloride. Then the strains were separately transferred to basal medium and were incubated before the beginning of log phase. To identify strains with different QAC resistance, 30-120 ppm of N-Alkyl-benzyl-dimethyl ammonium chloride were added to basal medium. Biocide effect was evaluated by comparing bacterial growth, which can be monitored by 600 nm light absorbance of basal medium suspension. Among 51 strains, only 2 strains can survive QAC concentration up to 120 ppm. API 20E and 16S rRNA gene sequencing technique were applied to identify two strains with highest (120 ppm) QAC resistance. One strain (HB22) was identified as Morganella morganii. HB22 can resist QAC concentration up to 150 ppm. HB22 is Gram-negative rod, motile with flagella, catalase positive, oxidase negative, Indole positive, H2S production negative, facultative anaerobe. HB22 has optimum growth in condition of 35℃ and pH 7.0. HB22 can catabolize only glucose and D(+)-Mannose. The other (HB45) was identified with high similarity among Pseudomonas cf. monteili or Pseudomonas mosselii or Pseudomonas putida. HB45 can resist QAC concentration up to 200 ppm. HB45 is Gram-negative rod, motile with flagella, catalase positive, oxidase negative, Indole negative, H2S production negative, aerobe. HB45 has optimum growth in condition of 30℃ and pH 7.4. HB45 can catabolize L-Arabinose, L(+)-Rhamnose, D(+)-Mannose, Glucose, Glycerol, Lactose, Maltose, Raffinose, Xylose, Cellulose and Xylan. The applicability of this work to paper industry is also discussed.

並列關鍵字

Facultative anaerobes ； aerobes ； biocides ； quaternary ammonium chlorides ； slimes