title: [Isolation and identification of an isolate of cow-origin Cryptosporidium sp].
abstract: 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.
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Key Laboratory of Parasite and Vector Biology, MOH, Shanghai 200025, China.
authors.forename: Hai-Peng
authors.initials: HP
authors.lastname: Liu
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Cao
authors.forename: Yu-Juan
authors.initials: YJ
authors.lastname: Shen
authors.forename: You-Gui
authors.initials: YG
authors.lastname: Chen
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Li
authors.forename: Wei-Yuan
authors.initials: WY
authors.lastname: Lu
authors.forename: Yu-Xin
authors.initials: YX
authors.lastname: Xu
authors.forename: Yi-Sheng
authors.initials: YS
authors.lastname: Liu
authors.forename: Shu-Xian
authors.initials: SX
authors.lastname: Liu
authors.forename: Xiao-Nong
authors.initials: XN
authors.lastname: Zhou
authors.forename: Lin-Hua
authors.initials: LH
authors.lastname: Tang
mesh_terms: 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
chemical_list: D015800:Protozoan Proteins; D012335:RNA, Ribosomal; C106305:oocyst wall protein, Cryptosporidium