Establishment Of PCR Detection Method And The Study Of Clinical Pathological Changes Of Dogs Infected With B.Gibsoni

Canine babesia is a kind of dogs severe protozoan diseases,which caused by genus Babesia.Canine babesiosis is characterized by anemia,fever,jaundice,splenomegaly,hemoglobinuria,visual mucosa pale and other symptoms,following with serious with acute renal failure,acute respiratory distress syndrome,DIC,hepatopathy and neurotic symptoms.Clinical and pathological examination showed mainly hemolytic anemia,parasitemia induced erythrocyte osmotic fragility,hemolysis and anemia.Additionally soluble parasite antigens may adhere to the surface of some noninfected erythrocytes and platelets,ehich can contribute to the immune-mediated anemia.In this tudy,we established the detection method of canine Babesia containing amplified internal standard.On the basis of accurate diagnosis,clinical and pathological parameters were detected.The effect of splenectomy on canine babesiosis was also studied.These results are very useful for the clinical diagnosis and treatment of canine babesiosis.1 Establishment of PCR method included an internal amplification control of canine babesiosisTo effictively avoid the false negative PCR detection results of Babesia the experiment by adding the internal standard of Babesia PCR detection system to build the PCR detection method with internal standard.In the experiment,the recombinant plasmid was constructed by using the gene fragment of chicken,and the amplified internal had the same primer as the PCR detection method of Babesia.The results suggested that the detection sensitivity was 106copies/25 Ri,and the reaction system was 105 copies which added to the 25?L reaction system.In conclusion,the experiment showed the PCR detection method based on the amplified internal standard can be quickly accuately and easily detect the infection of canine babeisa and it also avoided the false negative phenomenon efficiently.2 Clinical pathological changes of babesiosis infected with B.gibsoniThe experiment through infected B.gibsoni by intravenously,then monitored body temperature,the blood smear,blood parameters and blood biochemical indexes and clinical pathological change.We taked blood samples for PCR detection whether infected succeeds.The results showed that on day 9 detected the gene fragment by PCR.All dogs after infected with B.gibsoni showed severe anemia,decline in red blood cell count,hemoglobin decrease,reduction of neutropenil,lymphopenia,thrombocyopenia.At the same time,triglyceride,total cholesterol,LDH and ALP increased,creatine kinase increased,and serum potassium increased,blood glucose,serum sodium and chlorine decreased in the exoeriment.Two of the experimental dogs died 27 days,30 days respectively.The results showed that babesia gibsoni caused the function of serious heart,liver and kidney injured.3 The effects of splenectomy on babesiosisTo study the effect the role of spleen in canine babesia gibsoni,the experiment divided into three groups.Group A was splenectomy when healed after infected.Group B was splennotomy when appeared severe jaundice and the infection rate of parasites less than 1%.Group C was without splenectomy,and then detected the changes of blood temperature,the hematological,and blood biochemical.The results showed that the rate of prasites in dogs of splenectomy healed after the infection was not high,and the rate of prasites in dogs of splenectomy when infected with severe jaundice and the paraties level less than 1%increased.Removing the spleen aggravated the situation of anemia,and the group when infection of jaundice and the prasites level less than 1%of spenectomy red blood cells decreased more;platelets have a transient rise,after the decline of more powerful.The rate of parasites of when healed was lower than that of the non spleen group.After the removal of the spleen,the rate of prasites was increased.Splenectomy group of infection when appeared severe jaundice and parasites rate less than 1%,the total bilirubin,direct bilirubin,alanine aminotransferase and aspartate aminotransferase in infection on the 21st day increased.In the splenectomy groups,the triglyceride,lactate dehydrogenase and creatine kinase increased,and the cholesterol decreased more than the non splenectomy group.Blood glucose in the splenectomy group was lower than that in the group without splenectomy.In the experiment,spkenectomy had little effect on serum sodium and chloride,calcium.The results of this experiment showed that removal of the spleen can lead to more serious damage to the kidney and heart.In the experiment both of splenectomy when appeared severe jaundice and anemia were dead.