[Continuous axenic cultivation of Pneumocystis carinii isolated from the bronchoalveolar lavage fluid of infected rat].
OBJECTIVE
To establish axenic cultivation of Pneumocystis carinii (P.c).


METHODS
The organisms of P.c were isolated from the bronchoalveolar lavage fluid (BALF) of the rats with Pneumocystis carinii pneumonia (PCP) and cultured in a medium which was based on IMDM(GIBCO) supplemented with S-adenosyl-L-methionine, putrescine, N-acetyl glucosamine, putrescine, L-cysteine and L-glutamine, and newborn calf serum. The organisms cultured in the system were identified by observing the morphology of cysts in smears stained with Gomori's methenamine silver nitrate stain (GMS). Ultrastructure of the cysts/trophozoites was examined by transmission electron microscopy. The sequences of mitochondrial large ribosomal DNA subunit of the cultured organisms were compared with the Pneumocystis carinii f.sp. ratti variant isolate (GenBank No U20173) and Pneumocystis carinii f.sp.hominis (GenBank No M58605).


RESULTS
Five isolates of P. carinii received from BALF of 8 rats with PCP were cultured axenically and continuously in the system. The cultured organisms could be stored in frozen condition and used to reinitiate culture, and were amplified by 19-22 times within 72 h. The morphology, ultrastructure and gene sequencing of the cultured organisms confirmed that the isolated organisms were P. carinii.


CONCLUSIONS
Five continuously and axenicly cultured isolates of P. carinii have been received.
Huang
Min-Jun
MJ
Beijing Tropical Medicine Research Institute, Beijing 100050, China.
An
Yi-Jun
YJ
Li
Shu-Zhen
SZ
Lu
Si-Qi
SQ
Guo
Zeng-Zhu
ZZ
D000818:Animals; D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D004195:Disease Models, Animal; D005260:Female; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D045363:Pneumocystis carinii / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D011020:Pneumonia, Pneumocystis / Q000382:microbiology*; D012335:RNA, Ribosomal / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D012335:RNA, Ribosomal