Detection ofCryptosporidiumspecies in feces or gastric contents from snakes and lizards as determined by polymerase chain reaction analysis and partial sequencing of the 18S ribosomal RNA gene

Abstract

Abstract. Cryptosporidiosis is a well-known gastrointestinal disease of snakes and lizards. In the current study, 672 samples (feces and/or gastric contents or regurgitated food items) of various snakes and lizards were examined for the presence of cryptosporidia by polymerase chain reaction (PCR) assay targeting a part of the 18S ribosomal RNA gene. A consecutive sequencing reaction was used to identify the cryptosporidian species present in PCR-positive samples. Cryptosporidium varanii (saurophilum) was detected in 17 out of 106 (16%) samples from corn snakes (Pantherophis guttatus) and in 32 out of 462 (7%) samples from leopard geckos (Eublepharis macularius). Cryptosporidium serpentis was found in 8 out of 462 (2%) leopard gecko samples, but in no other reptile. The Cryptosporidium sp. "lizard genotype" was present in 1 leopard gecko sample, and 1 sample from a corn snake showed a single nucleotide mismatch to this genotype. Pseudoparasitic cryptosporidian species were identified in 5 out of 174 (3%) ophidian samples, but not in lizards. Other sequences did not show complete similarity to previously published Cryptosporidium sequences. The results stress the importance for diagnostic methods to be specific for Cryptosporidium species especially in snakes and show a relatively high prevalence of C. varanii in leopard geckos and corn snakes.

<i>Cryptosporidium varanii</i> infection in leopard geckos (<i>Eublepharis macularius</i>) in Argentina

“…oocysts, identifying C. varanii (n = 3), C. serpentis (n = 2) and C. parvum (n = 2). A similar study performed in Austria, detected C. varanii and C. serpentis (32/74 and 8/74 leopard geckos, respectively) (Richter et al, 2011). In the present study, molecular identification of Cryptosporidium spp.…”

“…were detected by Ziehl-Neelsen staining. Previous studies highlight the importance of molecular diagnostic methods to identify species level within the genus Cryptosporidium (Richter et al, 2011). In Spain, Pedraza-Diaz et al (2009) performed PCR and sequencing from seven leopard geckos stool samples with Cryptosporidium spp.…”

Cryptosporidiosis outbreak in captive chelonians (Testudo hermanni) with identification of two Cryptosporidium genotypes

“…The pooled sample and additional feces from each animal collected 5 days later were examined for Cryptosporidium oocysts with a CST for oocysts, 12 a commercial coproantigen ELISA f (enzyme immunoassay [EIA], previously described 5 and used as a commercially provided kit), and a PCR assay targeting a part of the 18S ribosomal RNA (rRNA) gene. 18 Four samples were positive with the CST, 5 samples were positive and 1 had a threshold value with the EIA, and 6 samples were positive and 2 had threshold values with the PCR (Table 1). The 2 initially presented tortoises with symptoms consistent with cryptosporidiosis (505110, 510210) tested positive with the EIA and PCR or only with the PCR.…”

“…Cryptosporidium varanii has also been described in ophidian species, and a recent study found a 16% prevalence of C. varanii in fecal samples from corn snakes (Pantherophis guttatus). 18 Clinical signs have mainly been described for lizards and include emaciation, anorexia, and regurgitation. 4 14 An Italian study found 7 of 21 captive Testudo spp.…”

“…Two tortoises (504910, 510510) were examined for nematode eggs by fecal flotation, and oxyurid eggs (Pharyngodonidae family) were detected. The PCR amplicons of the Cryptosporidium-positive samples were further analyzed by nucleotide sequencing, 18 and sequence spans correlating in the respective forward and reverse sequences were submitted to a basic local alignment search tool (BLAST; http://www.ncbi.nlm.nih.gov/blast/Blast. cgi) for comparison with published GenBank sequences.…”