Research On The Detection Of Hematological Index To The Mice Infected By Eperythrozoon And Therapeutic Effects Of Drugs

[Objective] To detect the changes of hematological index of mice infected by Eperythrozoon to explore the etiopathogenesis of Eperythrozoon;to compare the therapeutic effects of different drugs treated the infection. [Methods]1. The detection of hematological index of mice infected by Eperythrozoon:â‘ Confirm the establishment of infected model by optical microscope examination, transmission electron microscope observation and PCR assay to amplify distinctive genes;â‘¡Evaluate the ATPase activity of erythrocyte membrane, routine hematological index, SOD activity, immune function and NO content of erythrocyte.2.The comparison of therapeutic effects of different drugs treated the infection:12 normal mice were grouped as negative control, and another 48 mice infected by Eperythrozoon were randomly divided to 4 groups(each 12): Group I was mice infected, Group II was treated with nothing as positive control, Group III was with Artesunate(60mg/kg), while Group IV with Artesunate(60mg/kg) associated with amikacin sulphate (400mg/kg).Drugs were injected through intravenous pass for 7 days. In order to evaluate rate of infection, blood samples of mice were collected to make blood slider during the treatment every day, which were tested under oil immersion lens. And the hematological index was detected at the same time.PCR assays were then undertaken to test the blood collected after the treatment. [Results] The morphosis of Eperythrozoon can be directly observed by microscopic examination of the blood slider and its Wright-Giemsa stain smear, while ultrastructural organization can be observed by transmission electron microscope. It was shown that Eperythrozoon attached to surface of erythrocyte. And 801 bp product was successfully achieved by PCR assay, which was guided by primers designed according to the distinct fragment of pig Eperythrozoon genome. The erythrocyte Number of Group I was significantly lower than Group negative(P<0.01); The erythrocyte number of Group I was significantly lower than Group negative(P<0.01); and the hemoglobin, ratio of RBC-C3b,ATPase activity of erythrocyte derived from Group I were also significantly lower than Group negative (P<0.05); However the hematocrite and SOD activity of Group I were less than Group negative, the data got no statistical significance(P>0.05),so was neutrophilic granulocyte number(P>0.05). In adverse, white blood cells ,lymphocytes and NO content of Group I were a little more than Group negative, but P>0.05. And the ratio of RBC-IC of Group I was significantly higher than Group negative(P<0.05).The erythrocyte can recover from infection after treating with artesunate associated with amikacin sulphate, whose infection rate was also decreased; With the artesunate alone erythrocyte can almost recover to normal status too(Group III); The infection rate of Group II(positive control) at the 7th day was 23.08%, which was significantly higher than Group III and IV(P<0.05); The erythrocytes and hemoglobin were significantly increased after infection being treated by drugs(P<0.05); Hematocrit index of five groups were statistically the same(P>0.05). The distinct fragment of PCR amplification can be achieved in Group I and Group II(positive control), while it got nothing in Group III and IV. [Conclusion] To verify organism is infected by Eperythrozoon, the way of microscopy examination of blood slider or Wright-Giemsa stain smear or transmission electron microscope examination can be carried out. But it is insufficient of optical microscope examination alone, which PCR assay and transmission electron microscope can be employed to detect the infection specially. 2.the purified Eperythrozoon can infect SPF Kunming mice. The infection model can be successfully established by the way which the 6-weeks Kunming mice being infected by purified human Eperythrozoon. 3. the erythrocyte membrane structure loses its normal shape if the Eperythrozoon is injected to experimental mice for infecting cells, and it gets dissolvable and fragile, which ultimately leads to changes of blood physiology and biochemistry.4.The therapeutic effect of artesunate associated with amikacin sulphate is better than artesunate alone for removing Eperythrozoon more quickly. And the number and function of erythrocyte can recover to normal status more fast if the infection is treated according to the associated-protocol.