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目的观察棘阿米巴原虫对霍乱弧菌O139的环境生存影响。方法采用共培养方法, 通过染色和电镜超薄切片来观察霍乱弧菌O139在多噬棘阿米巴的滋养体和包囊内生存情况,经包囊传代及低温下生存耐受实验来观察阿米巴包囊能否提高霍乱弧菌O139的环境生存能力。结果与多噬棘阿米巴共培养8 h后,霍乱弧菌O139就能进入阿米巴滋养体内并在空泡内增殖。霍乱弧菌 O139可生存于胞浆空泡内和(或)包囊内壁间,经包囊传代培养能再分离培养出此菌。在30℃时,在棘阿米巴环境下120 d仍能检测出霍乱弧菌O139,而在阿米巴生理盐水液中45 d后就检测不出霍乱弧菌O139。在4℃时,实验组和对照组培养液30 d后均未能培养出霍乱弧菌O139;而被感染包囊分别在30、45、60和75 d后复苏培养能再检测出霍乱弧菌O139;被感染包囊90 d后发育成滋养体能力下降,未能再培养出活的霍乱弧菌O139,在电镜下可见包囊被裂解死亡。结论霍乱弧菌O139能在棘阿米巴的滋养体内繁殖和包囊内生存;阿米巴包囊能够提高霍乱弧菌O139的低温下生存能力;棘阿米巴包囊可能成为霍乱弧菌O139越冬的环境宿主。
Objective To observe the effect of Acanthamoeba protozoa on the environmental survival of Vibrio cholerae O139. Methods The co-culture method was used to observe the survival of Vibrio cholerae O139 in trophozoites and cysts of Amoeba polymorpha by staining and electron microscopy. The cytoplasm was passaged by cysts and survived at low temperature Miba cyst can improve the environmental viability of Vibrio cholerae O139. Results After co-culture with Acanthamoeba pharmcoccus for 8 h, Vibrio cholerae O139 could enter the amoebae trophozoites and proliferate in vacuoles. Vibrio cholerae O139 can survive within the cytoplasmic vacuoles and / or within the cyst wall, which can be isolated and cultured by cyst subculture. Vibrio cholerae O139 was still detectable at 120 days in Acanthamoeba at 30 ° C, whereas Vibrio cholera O139 was not detected in 45 days after amoebic saline. At 4 ℃, Vibrio cholerae O139 could not be cultured in the culture medium of experimental group and control group for 30 d, while Vibrio cholerae O139 was recovered after 30, 45, 60 and 75 days in infected cysts O139. After being infected for 90 days, the ability to develop into trophozoites decreased. Vibrio cholerae O139 could not be cultured any more. The cysts were lysed and died under the electron microscope. Conclusion Vibrio cholerae O139 can survive in the trophozoites of Acanthamoeba and survive within the cysts. The amoeba cyst can improve the viability of Vibrio cholerae O139 at low temperature. Overwintering environment host.