Skip to main content Accessibility help

Phylogenetic analysis of Cryptosporidium isolates in Henan

  • Wang Jin-Chan (a1) (a2), Qi Wei-Wei (a3), Zhang Long-Xian (a1), Ning Chang-Shen (a1), Jian Fu-Chun (a1), Zhao Jin-Feng (a1) and Wang Ming (a3)...


The functional mitochondrial protein alternative oxidase (AOX) gene was used as a marker to analyse the phylogenetic relationship between Cryptosporidium isolates. This gene was characterized, and the phylogentic tree was established from Cryptosporidium isolates and compared to those generated from 18S rRNA and heat-shock protein 70 (HSP70) gene sequences. The present trial aimed at finding out whether the AOX gene is suitable for phylogenetic analysis of the Cryptosporidium genus. The results revealed that the genus Cryptosporidium contained the phylogenetically distinct species C. parvum, C. hominis, C. suis and C. baileyi, which were consistent with the biological characterization and host specificity reported earlier. Cryptosporidium species formed two clades: one included C. hominis, C. suis, C. parvum cattle genotypes and C. parvum mouse genotype; and the other comprised C. meleagridis and C. baileyi isolates. Within C. parvum, both the mouse genotype and the pig genotype I (also known as C. suis) isolates differed from cattle and human (also known as C. hominis) genotypes, based on the aligned nucleotide sequences. The sequence identity of the AOX gene was higher between C. meleagridis and C. baileyi than between C. meleagridis and C. parvum. The phylogenetic trees showed that C. meleagridis was closer to C. baileyi than to C. parvum. This result was inconsistent with the phylogenetic analysis deduced from 18S rRNA and HSP70 gene sequences, respectively. The present results suggest that the AOX gene is not only equally suitable for the phylogenetic analysis of Cryptosporidium, but also provides an outstanding and new approach in determining Cryptosporidium heredity.


Corresponding author

*Corresponding authors. E-mail: or


Hide All
Arrowood, M and Sterling, G (1987) Isolation of Cryptosporidium oocysts and sporozoites using discontinuous sucrose and isopycnic percoll gradients. Journal of Parasitology 73: 314319.
Bonnin, A, Fourmaux, M, Dubremetz, J, et al. (1996) Genotyping human and bovine isolates of Cryptosporidium parvum by polymerase chain reaction–restriction fragment length polymorphism analysis of a repetitive DNA sequence. FEMS Microbiology Letters 137: 207211.
Caccio, S, Homan, K, Van, D, et al. (1999) Genetic polymorphism at the beta-tubulin locus among human and animal isolates of Cryptosporidium parvum. FEMS Microbiology Letters 170: 173179.
Carraway, M, Tzipori, S and Widmer, G (1996) Identification of genetic heterogeneity in the Cryptosporidium parvum ribosomal repeat. Applied and Environmental Microbiology 62: 712716.
Fayer, R, Morgan, U and Upton, S (2000) Epidemiology of Cryptosporidium: transmission, detection and identification. International Journal of Parasitology 30: 13051322.
Fayer, R, Santin, M and Xiao, L (2005) Cryptosporidium bovis n. sp. (Apicomplexa: Cryptosporididae) in cattle (Bos taurus). Journal of Parasitology 91: 624629.
Liu, C, Widmer, G, Tzipori, S, et al. (2004) Complete genome sequence of the apicomplexan, Cryptosporidium parvum. Science 304: 441445.
Margarida, A, Xiao, L, Sulaiman, I, et al. (2003) Subgenotype analysis of Cryptosporidium isolates from humans, cattle, and zoo ruminants in Portugal. Journal of Clinical Microbiology 41: 27442747.
Morgan, U, Sargent, K, Deplazes, P, et al. (1998) Molecular characterization of Cryptosporidium from various hosts. Parasitology 117: 3137.
Morgan, U, Deplazes, P and Forbes, D (1999) Sequence and PCR-RFLP analysis of the internal transcribed spacers of the rDNA repeat unit in isolates of Cryptosporidium from different hosts. Parasitology 118: 4958.
Peng, M, Xiao, L, Freeman, A, et al. (1997) Genetic polymorphism among Cryptosporidium parvum isolates: evidence of two distinct human transmission cycles. Emerging Infectious Disease 3: 563573.
Ryan, U, Monis, P, Enemark, H, et al. (2004) Cryptosporidium suis n. sp. (Apicomplexa: Cryptosporididae) in pigs (Sus scrofa). Journal of Parasitology 90: 769773.
Schmidt, H, Strimmer, K, Vingron, M, et al. (2002) TREE-PUZZLE: maximum likelihood phylogenetic analysis using quarter and parallel computing. Bioinformatics 18: 502504.
Spano, F, Putignani, L and McLauchlin, J (1997) PCR-RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between Cryptosporidium wrairi and Cryptosporidium parvum, and between C. parvum isolates of human and animal origin. FEMS Microbiolology Letters 150: 209217.
Spano, F, Putignani, L, Guida, S, et al. (1998) Cryptosporidium parvum: PCR-RFLP analysis of the TRAP-C1 (thrombospondin-related adhesive protein of Cryptosporidium-1) gene discriminates between two alleles differentially associated with parasite isolates of animal and human origin. Experimental Parasitology 90: 195198.
Sreter, T, Kovacs, G and da Silva, AJ (2000) Morphologic culture and molecular characterization of a Hungarian Cryptosporidium meleagridis. Journal of Parasitology 86: 12441249.
Sulaiman, I, Morgan, U, Andrew-Thompson, R, et al. (2000) Phylogenetic relationships of Cryptosporidium parasites based on the 70-kilodalton heat shock protein (HSP70) gene. Applied and Environmental Microbiology 66: 23852391.
Swofford, D (2002) Phylogenetic analysis using parsimony (PAUP), version 4.0. Champaign: Illinois Natural History Survey, pp. 1144.
Thompson, J, Gibson, T, Plewniak, F, et al. (1997) The ClustalX windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis too. Nucleic Acids Research 24: 48764882.
Tzipori, S and Ward, H (2002) Cryptosporidiosis: biology, pathogenesis and disease. Microbiology Infection 4: 10471058.
Vasquez, J, Gooze, L, Kim, K, et al. (1996) Potential antifolate resistance determinants and genotypic variation in the bifunctional dihydrofolate reductase–thymidylate synthase gene from human and bovine isolates of Cryptospridium parvum. Molecular Biology of Parasitology 79: 153165.
Wang, Jin-chan, Jian, Fu-chun, Zhang, Long-xian, et al. (2007) Phylogenetic analysis of the Cryptosporidium based on 18S rRNA gene and HSP70 gene sequences. Acta Veterinaria et Zootechnica Sinica 38(9): 947953.
Xiao, L, Morgan, U, Limor, J, et al. (2000) Identification of species and sources of species and sources of Cryptosporidium oocysts in storm waters with a small-subunit rRNA-based diagnostic and genotyping tool. Applied and Environmental Microbiology 66: 54925498.
Xiao, L, Limor, J, Bern, C, et al. (2001) Tracking Cryptosporidium parvum by sequence analysis of small double-stranded RNA. Emerging Infectious Diseases 7: 141145.
Xiao, L, Fayer, R, Ryan, U, et al. (2004) Cryptosporidium taxonomy: recent advances and implications for public health. Clinical Microbiology 17: 7297.
Xu, Ping, Widmer, G, Wang, Y, et al. (2004) The genome of Cryptosporidium hominis. Nature 431: 11071112.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed