Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 2. Intraeryrhrocytic culture of Babesia gibsoni by microaerophilous stationary phase(MASP)

Suh, Myung-deuk; Joo, Bo-hyun

Korean Journal of Veterinary Research 1998;38(2):359-365.

Myung-deuk Suh, Bo-hyun Joo

College of Veterinary Medicine, Gyeongsang National University

Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 2. Babesia gibsoni의 적혈구내 배양

서명득, 주보현

경상대학교 수의과대학

Abstract

This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

Key Words: Babesia gibsoni, IFA, intraerythrocytic culture, microaerophilous staionary phase, RPMI 1640 medium, protozoan poarasite