Phylogenetic Relationships of Cryptosporidium Parasites Based on the 70-Kilodalton Heat Shock Protein (HSP70) Gene

Abstract

We have characterized the nucleotide sequences of the 70-kDa heat shock protein (HSP70) genes of Cryptosporidium baileyi, C. felis, C. meleagridis, C. muris, C. serpentis, C. wrairi, and C. parvum from various animals. Results of the phylogenetic analysis revealed the presence of several genetically distinct species in the genus Cryptosporidium and eight distinct genotypes within the species C. parvum. Some of the latter may represent cryptic species. The phylogenetic tree constructed from these sequences is in agreement with our previous results based on the small-subunit rRNA genes of Cryptosporidium parasites. The Cryptosporidium species formed two major clades: isolates of C. muris and C. serpentis formed the first major group, while isolates of C. felis, C. meleagridis, C. wrairi, and eight genotypes of C. parvum formed the second major group. Sequence variations were also observed between C. muris isolates from ruminants and rodents. The HSP70 gene provides another useful locus for phylogenetic analysis of the genus Cryptosporidium.

The prevalence and molecular characterisation of Cryptosporidium spp. in small ruminants in Zambia

“…One sample did not amplify on the 18S rRNA gene. In five lambs and in the goat kid sample C. parvum (genotype 2) was identified, based on a high homology (>96%) with previously published sequences, both for the 18SrRNA gene (Ryan et al, 2003) and for the HSP-70 gene (Sulaiman et al, 2000;Ryan et al, 2003). In one lamb the pig genotype of C. parvum or Cryptosporidium suis (Ryan et al, 2004) was identified (homology > 97%).…”

Identification of the Cryptosporidium Isolate from Chickens in Japan by Sequence Analyses

“…Furthermore, the same results were obtained when compared with three genes of C. baileyi Hungarian strain CBAI-1 (GenBank accession numbers: AJ276096 for 18S rRNA gene, AJ310765 for COWP gene and AJ310880 for HSP70 gene) (Data are not shown) [1]. In contrast, these three genes showed less than 91% homology with C. meleagridis genes (GenBank accession numbers: AF112574, AF221537, AF266266, respectively) [13,15,16] (Table 1). In addition, these genes were classified into the same cluster with C. baileyi other than C. meleagridis by phylogenetic analysis (Fig.…”

“…These partial sequences of 18S rRNA, COWP and HSP70 genes have been deposited in the GenBank database under accession numbers AB116663, AB116664 and AB116665, respectively. The 18S rRNA, COWP and HSP70 genes sequences from the chicken isolate showed nearly 100% homology with those of C. baileyi genes published by Xiao et al [15], Sulaiman et al [13] and Xiao et al [16], respectiv ely (G enB ank a ccessio n nu mb ers: AF 09 34 95 , AF221539, AF266276, respectively) ( Table 1). Furthermore, the same results were obtained when compared with three genes of C. baileyi Hungarian strain CBAI-1 (GenBank accession numbers: AJ276096 for 18S rRNA gene, AJ310765 for COWP gene and AJ310880 for HSP70 gene) (Data are not shown) [1].…”

“…Then DNA was extracted using the QIAamp DNA Mini Kit (Qiagen GmbH, Hilden, Germany) according to the manufacturers instructions. PCR and sequencing primers and amplification conditions for the 18S rRNA, COWP and HSP70 genes were determined using the methods reported by Johnson et al [6], Spano et al [12] and Sulaiman et al [13], respectively. Each PCR product was purified from agarose gel and sequencing reactions were carried out with the ABI PRISM BigDye Therminator Cycle Sequencing Ready Reaction kit (Applied Biosystems, Foster City, California).…”

A New Species of Cryptosporidium (Apicomplexa: Cryptosporidiidae) from Eastern Grey Kangaroos (Macropus giganteus)

“…sp., [Ryan et al, 2008]) was described after oocysts from a koala (Phascolarctos cincereus) were found to be genetically distinct from other described Cryptosporidium species. Cryptosporidium fayeri has since been characterized at 6 loci and described in a red kangaroo (M. rufous), eastern grey kangaroos (M. giganteus), a western barred bandicoot (Perameles bougainville), and a yellow footed rock wallaby (Petrogale xanthopus) (Morgan, Deplazes et al, 1999;Morgan, Monis et al, 1999;Xiao et al, 1999;Sulaiman et al, 2000;Power et al, 2003Power et al, , 2004. More recently, the Cryptosporidium marsupial ge-notype II was described from eastern grey kangaroos (Power et al, 2004).…”