[Isolation and identification of an isolate of cow-origin Cryptosporidium sp].
OBJECTIVE
To isolate and identify Cryptosporidium oocysts from feces of naturally infected cow.


METHODS
Fecal samples were collected from Cryptosporidium infected cows confirmed by modified acid-fast staining method. Oocysts were isolated and purified with Sheathed sucrose density gradient centrifugation technique. Genomic DNA was isolated with Chelex-100. Both primers were designed to amplify Cryptosporidium small subunit ribosome RNA gene (SSU rRNA) and Cryptosporidium oocyst wall protein gene (COWP), respectively. The PCR products were cloned into pGEM-T and pGEM-T Easy vector and sequenced subsequently. Homology and phylogeny were analyzed with BLASTn and MEGA software.


RESULTS
The results suggested that the size of oocysts was (7.4+/-0.32) microm by (5.4+/-0.21) microm and the ratio of length and width was 1.37+/-0.07 (n=20). BLASTn revealed that the identity of SSU rRNA and COWP gene of Cryptosporidium isolated from cow to the counterparts of andersoni was 100% and 99% respectively. Phylogenetic reconstruction placed the isolated Cryptosporidium within the C. andersoni clade based on the sequence of SSU rRNA and COWP gene.


CONCLUSIONS
What isolated from naturally infected cow feces has been identified as C. andersoni.
Liu
Hai-Peng
HP
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, MOH, Shanghai 200025, China.
Cao
Jian-Ping
JP
Shen
Yu-Juan
YJ
Chen
You-Gui
YG
Li
Xiao-Hong
XH
Lu
Wei-Yuan
WY
Xu
Yu-Xin
YX
Liu
Yi-Sheng
YS
Liu
Shu-Xian
SX
Zhou
Xiao-Nong
XN
Tang
Lin-Hua
LH
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology*; D003457:Cryptosporidiosis / Q000469:parasitology*; D003458:Cryptosporidium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology; D008969:Molecular Sequence Data; D033661:Oocysts / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D015800:Protozoan Proteins; D012335:RNA, Ribosomal; C106305:oocyst wall protein, Cryptosporidium