兩種綠膿桿菌噬菌體的分辨及特性

Abstract

中文摘要 由污水中分離得兩種綠膿桿菌噬菌體，將其命名為 L1及S2。 L1噬菌體產生圓形、中心透明周圍有模糊暈輪的大溶菌斑，直徑約 3－4 毫米； S2噬菌體產生區形、小而模糊的溶菌斑，直徑約 0 . 5－ 1.0毫米。於電予顯微鏡觀察顯示L1 及S2 ，噬菌體均為蝌蚪狀的構造。L1 噬菌體有長形的頭部，大小為 89 ×57 nm, 尾部為 167 ×5nm； S2噬菌體頭部則為多角形，大小為 159 × 152nm，尾部為 167 ×25nm ，且頸項、尾鞘及尾端之基部平板亦可見。 兩種噬繭體的寄主特異性都很窄，對於其他不同屬的菌種均不產生溶菌斑。測定 L1噬菌體所採用之細菌懸浮液以振盪培養 7 小時之細菌濃度為最適宜，而S2 噬菌體則以 6 小時為佳。二者最適合培養的溫度均為 37 ℃。 對 L1和S2噬菌體處理十分鐘之不活性溫度分別為 65℃和 70 ℃ 。經紫外?照射一分鐘， L 1噬菌體約有75％不活化：而 S2噬菌體約 80％不活化。二種噬菌體均於弱酸性溶液中較穩定， L1噬菌體的範圍則為 pH7至 pH9 ，S2噬菌體則為 pH7 至 p H 11 。二者對氯仿均不敏感，氯仿處理後 L1 噬菌體力價增加約43.1％，而S2噬菌體力價增加22.5％左右。 兩種噬菌體的感染均會使寄主細菌發生溶菌的現象。以測遊離噬菌體的方法求得 L 1及 S2噬菌體之平均吸附速率常數分別為 4.11 × 10－10和 5.54 × 10 －9me/min 。

Abstract Two bacceriophaes of Pseudomonas aeruginosa Holloway strain were isolated from sewage, named L1 and S2 respectively. Phage L1 produced large plaques of 3—4 mm in diameter, with clear centers surrounded. by broad turbid. halos; while S2 formed small turbid plaques of 0.5—1.0mm in diameter. Electron microscopic study revealed that both L and S2 were tadpolclike structure. Phage L1 possessed an elongated head of 89 x67 nm and tail of 167 x 5nm.S2had a polygonal head of 159 x 152nm, with a tail of 167 x25nm in dimension; neck, tail sheath and baseplate also could be seen. Both L1 and S2 showed limited host range, they could not form plaque on bacteria other than the genus of Pseudomonas. The best bacteria1 culture age for L1 and S2 were seven—hour and six—hour respectively. The optimum incubation temperature for both were 37°C The thermal inactivation temperature of ten—minute treatment for L1 and 2 were 65° and 70°C respectively. About 75% of L1 phages and 80% of S2 phages were inactivated by treatment with ultraviolet light for 60 seconds. Both phages were stable on week .alkaline solution, the range for L1 was at pH7 to pH9, while S2 at pH7 to pH11.Both phages were insensitive to chloroform. The chloroform treatment increased the titers of L1 and S2 stocks by approximately 43.1% and 22.5% respectively. Infection by either phage L1 or S2, lysed the host cells. Using the method of assaying free phages, the average adsorption rate constants for L1 and S2 were 4.11x10－10 and 5.54x10－9 ml/min respectively.