粪肠球菌生长曲线的测定及其对小鼠脑组织的影响

doi:10.16431/j.cnki.1671-7236.2018.04.026

Abstract

Abstract:

The objective of this trial was to determine the growth curve of Enterococcus faecalis that caused encephalitis in lambs and find a rapid and accurate method to determine the number of Enterococcus faecalis at different growth stages,and objectively evaluate its toxicity and effect on the brain tissue of mice.Plate counting method and D_λ value method were used to analyze the growth curve of Enterococcus faecalis. The relationship between the absorbance (D_{600 nm}) and the live bacteria (CFU) measured by plate colony counting method at the appropriate time period was explored.According to the data analysis,mice were infected with Enterococcus faecalis,and the death of mice was recorded.Finally,the Karber method was used to calculate the median lethal dose (LD₅₀) of the mice infected with Enterococcus faecalis.The mice were infected with the dose of LD₅₀,the brain tissues of the dead mice were collected in time and all the mice without death were sacrificed 72 h after infection,and the brain tissues were collected,too.Some of them were stained with smear and the pathological sections were made.Pathological changes were observed.PCR was used to identify bacteria.The results showed that the growth curves of Enterococcus faecalis were basically the same in both methods,the growth of Enterococcus faecalis was rapid at 2 to 8 h for logarithmic growth phase,the growth rate was slow for 8 to 14 h for stable period,the number of deaths increased after 14 h for decline phase.The relationship between D_{600 nm} and CFU of Enterococcus faecalis was studied,and the regression equation was established,y=20.769x-1.3422,R²=0.997.The LD₅₀ for the infection mice was 7.77×10¹¹ live bacteria.With this dose to infect mice,we could see gram-positive cocci in both brain tissue smear and staining.PCR result showed that there was a size of 112 bp band.Morphological observation of the brain tissue of mice found that the bacteria could lead to brain tissue congestion,bleeding,the formation of micro-thrombosis,meningeal congestion.The results of this study revealed that the establishment of the relationship between the growth curve and the D_{600 nm} and CFU could provide a Real-time monitoring of the number of Enterococcus faecalis,and provide an important theoretical basis for further study on the mechanism of Enterococcus faecalis crossing the blood-brain barrier.

Key words: Enterococcus faecalis; D_λ value method; flat colony counting method; LD₅₀

CLC Number:

S816.7

WANG Mengmeng, ZHANG Li, LI Hui, WANG Xiaoxuan, WEI Dianhua, GAO Jingwen, GAO Jianpeng, ZHANG Hualei, QI Yayin. Study on Growth Curve of Enterococcus faecalis and Its Effect on Brain Tissue of Mice[J]. , 2018, 45(4): 1041-1049.

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Study on Growth Curve of Enterococcus faecalis and Its Effect on Brain Tissue of Mice

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