绿色荧光蛋白标记的病原性河流弧菌在青石斑肠道的黏附定植研究
Adhesion and colonization of fluorescence labeled pathogenic Vibrio fluvialis to intestine of Einephelus awoara
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摘要: 通过灌胃将绿色荧光蛋白标记的病原性河流弧菌导入青石斑鱼消化道内,追踪标记菌在青石斑肠道中的黏附和定植及对肠道消化酶活性的影响。结果表明,青石斑前肠和中肠在河流弧菌灌胃12 h即可检出,前肠在24 h达到峰值,大约为7.1×105 cfu/g,中肠则在36 h达到峰值,约为5.39×105 cfu/g,而后肠则是在24 h才开始检出,且检出时即达到峰值,但仅为2.44×104 cfu/g,说明青石斑鱼的前肠和中肠都是河流弧菌的主要黏附部位。灌胃96 h后前肠和中肠仍有标记菌存在,但在后肠已检测不到标记菌,反映出标记菌已在前肠和中肠定植下来,但没有在后肠定植。此外,标记河流弧菌的定植能够显著降低前肠蛋白酶活性和淀粉酶活性以及后肠脂肪酶活性。研究结果说明,病原性河流弧菌可通过黏附定植于鱼类肠道影响鱼类肠道消化酶活力,这可能是病原性河流弧菌引发鱼类病害的原因之一。Abstract: The fluorescence labeled bacterium EGFP-Vibrio fluvialis were introduced into the digestive tract of Einephelus awoara by intragastic injection in order to trace this bacterium colonization and its effect on digestive enzyme in fish intestine. The results showed that labeled bacterium could be detected in forgut and midgut 12 h after intragastic injection,and the number of the labeled bacteria in forgut reached a peak value 7.1×105cfu/g 24 h after intragastic injection,in midgut the peak value 5.39×105 cfu/g was observed 36 h after intragastic injection,in hindgut the labeled bacterium was not detected until 24 h after intragastic injection and the peak value was only 2.44×104 cfu/g. This result indicated V. fluvialis tended to adhere in forgut and midgut. Bacterium was still detected in forgut and midgut 96 h after intragastic injection,but not detected in hindgut. This indicated that forgut and midgut were the main location for bacteria colonization. The results also showed that the activity of alkaline protease and amylase in forgut and midgut and lipase in hindgut were significantly decreased after V. fluvialis colonization. This result implied that V. fluvialis could affect activity of digestive enzyme in the fish gut after colonization,which might be one of the pathogenesis of V. fluvialis.
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Key words:
- Vibrio fluvialis /
- Epinephelus awoara,adhesion /
- colonization /
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