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Applied and Environmental Microbiology, December 1999, p. 5345-5349, Vol. 65, No. 12
Division of Comparative Pathology, University
of Miami School of Medicine, Miami, Florida
33136,1 and Avian and Exotics Animal
Medical Center, Miami, Florida 331562
Received 9 July 1999/Accepted 20 September 1999
An adult wild-caught corn snake (Elaphe guttata
guttata) was presented for humane euthanasia and necropsy because
of severe cryptosporidiosis. The animal was lethargic and >5%
dehydrated but in good flesh. Gastric lavage was performed prior to
euthanasia. Histopathologic findings included gastric mucosal
hypertrophy and a hemorrhagic erosive gastritis. Numerous 5- to
7-µm-diameter round extracellular organisms were associated with the
mucosal hypertrophy. A PCR, acid-fast stains, Giemsa stains, and an
enzyme immunoassay were all positive for Cryptosporidium
spp. PCR and restriction fragment length polymorphism (RFLP) analysis
on gastric lavage and gastric mucosal specimens, and subsequent
sequencing of the 18S rRNA gene, enabled a distinct molecular
characterization of the infecting organism as Cryptosporidium
serpentis. Until recently, studies on snake
Cryptosporidium have relied on host specificity and gross
and histopathologic observations to identify the infecting species. A
multiple alignment of our sequence against recently published sequences
of the 18S rRNA gene of C. serpentis (GenBank accession no.
AF093499, AF093500, and AF093501 [L. Xiao et al., unpublished data,
1998]) revealed 100% homology with the C. serpentis
(Snake) sequence (AF093499) previously described by
Xiao et al. An RFLP method to differentiate the five presently
sequenced strains of Cryptosporidium at this locus was developed. This assay, which uses SpeI and
SspI, complements a previously reported assay by
additionally distinguishing the bovine strain of
Cryptosporidium from Cryptosporidium wrairi.
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Molecular Analysis of the 18S rRNA Gene of Cryptosporidium
serpentis in a Wild-Caught Corn Snake (Elaphe guttata
guttata) and a Five-Species Restriction Fragment Length
Polymorphism- Based Assay That Can Additionally Discern C. parvum from C. wrairi
*
Corresponding author. Mailing address: Division of
Comparative Pathology, University of Miami School of Medicine, 1550 NW 10th Ave., Rm. 134, Miami, FL 33136. Phone: (305) 243-6640. Fax: (305)
243-5662. E-mail: debralee{at}hotmail.com.
Applied and Environmental Microbiology, December 1999, p. 5345-5349, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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