Experimental Study On Physical And Chemical Analysis Of Echinococcus Multilocularis And Echinococcus Granulosus

ObjectiveTo explore the physical and chemical properties of Echinococcus multilocularis protoscoleces and Echinococcus granulosus protoscoleces in vitro.Methods1.The collection of Echinococcus granulosus protoscoleces: Fresh Echinococcus granulosus protoscoleces were extracted from liver lesions of sheep infected with cystic echinococcosis.After treatment with eosin staining,the activity of protoscoleces was observed.Then the number was counted under microscope.Only those with more than 98% of the activity can be further cultured and measured.2.The Grouping of Echinococcus granulosus protoscoleces: The protoscoleces were divided into 4groups according to the density after counting and then added to the culture bottle with 15 ml medium.The medium used by Echinococcus granulosus protoscoleces and the medium used by Echinococcus multilocularis protoscoleces were both from the same batch.The density was 0/15 ml in the blank group,10,000/15 ml in the control group,20,000/15 ml in the control group,and 30,000/15 ml in the control group.The Echinococcus granulosus protoscoleces and the Echinococcus multilocularis protoscoleces shared the same blank group.3.The collection of Echinococcus multilocularis protoscoleces: The Echinococcus multilocularis protoscoleces were obtained from the abdominal cavity of the breeding mice for more than half a year.After treatment with eosin staining,the activity of protoscoleces was observed.Then the number was counted under microscope.Only those with more than 98% of the activity can be further cultured and measured.4.The Grouping of Echinococcus multilocularis protoscoleces: The protoscoleces were divided into 4groups according to the density after counting and then added to the culture bottle with 15 ml medium.The density was 0/15 ml in the blank group,10,000/15 ml in the control group,20,000/15 ml in the control group,and 30,000/15 ml in the control group.5.The method of measurement and the choice of time node: In order to ensure that physicochemical indexes of the culture medium were not changed,medium change was not allowed in the whole process of culturing and measurement.The Measurement was on the premise that the original culture density was not changed.A pipettor was used to blow the culture medium quickly and uniformly.And the medium containing the protoscoleces was absorbed for the next measurement.The first measurement was made after the protoscoleces was packed,and the follow-up measurement was made after 1 day,2 days,3 days,4days and 5 days.6.The measurement of PH,lactic acid,calcium ion and glucose: The supernatant of the culturemedium was absorbed and transferred to a dry sterile EP tube.Medical blood gas analyzer was used to measure the values of the above indexes.7.The measurement of glycogen: After the supernatant of the culture medium was discarded,the glycogen content in procephalus was measured by using glycogen reagent detection box.Results1.The supernatant PH of two kinds of protoscolex both decreased in vitro.And with the higher density,the decreasing trend was more obvious.2.The supernatant lactic acid and calcium of two kinds of protoscolex both increased in vitro.And with the higher density,the increasing trend was more obvious.3.The supernatant glucose of two kinds of protoscolex both decreased in vitro.And with the higher density,the decreasing trend was more obvious.4.The glycogen content in the two kinds of protoscolex gradually accumulated with the growth,but they both decreased at the later stage.And with the higher density,the decreasing trend was more obvious.5.By statistical analysis,the above indicators showed statistically significant differences in the two groups(P<0.05),but there were no statistically significant differences between the two groups(P>0.05).Conclusion1.The physical and chemical properties of the two kinds of echinococcus in vitro culture were similar.2.In the metabolic process of the two kinds of echinococcus,acid represented by lactic acid was produced and then the PH of the external environment decreased.3.Calcium accumulation occurred during the growth of the two kinds of echinococcus.4.The surrounding glucose was the main nutrient during the growth of the two kinds of echinococcus.And the glucose taken by the echinococcus was synthesized into glycogen for growth and development.