荧光光度法在瘤胃原虫对细菌吞噬作用研究中的应用

Application of Spectrofluorimetry in Studying Phagocytosis of Rumen Protozoa on Bacteria

本试验旨在探讨荧光光度法研究瘤胃原虫对细菌吞噬作用的可行性。试验选用4只装有永久性瘤胃瘘管的徐淮山羊采集瘤胃液以获取瘤胃原虫和细菌,以4,6-二脒基-2-苯基吲哚(DAPI)标记细菌的荧光强度为主要测定指标,研究山羊瘤胃中原虫对细菌的吞噬作用。结果表明,DAPI标记的瘤胃细菌浓度与其荧光强度之间存在线性相关(R2=0.9952)。在日粮粗精比为7∶3的条件下,徐淮山羊瘤胃原虫对细菌的吞噬速率为370.89cells/(cell.h),瘤胃内原虫对细菌的吞噬速率存在一定的波动性,吞噬量在60min达到饱和,其最大吞噬量为370.89cells/cell。结果提示,荧光光度法可应用于瘤胃原虫对细菌吞噬作用的研究。

The study was conducted to examine the feasibility of spectrofluorimetry used to study the phagocytosis of rumen protozoa on bacteria. Four healthy Xuhuai goats with permanent ruminal cannulas were selected as experimental animals to obtain rumen protozoa and bacteria. The fluorescence intensity of bacteria labeled by DAPA was tested to study the phagocytosis of rumen protozoa on bacteria. The results showed that there was linear correlation between the concentration of bacteria concentration labeled by DAPI and their fluorescence intensity （R2=0.995 2）. The grazing rate of protozoa was 370.89 cells/（cell·h） for Xuhuai goats fed the diet with forage∶concentration ratio of 7∶3. There was volatility on grazing rate of rumen protozoa on bacteria of Xuhuai goats. Grazing quantity reached saturation on 60 min and the largest grazing quantity was 370.89 cells/cell. It was concluded that spectrofluorimetry could be used in studying grazing rate of rumen protozoa on bacteria.