Objective:To determine the prevalence and species/genotype distribution of Cryptosporidium spp.in captive raccoon dogs in Hebei Province.Methods:A total of 389 fresh fecal samples were collected from captive raccoon dog farms in Hebei Province.Nested PCR based on the small subunit ribosomal RNA(SSU rRNA)gene of Cryptosporidium spp.was used to detect all samples.The 60 kDa glycoprotein(gp60)gene from the positive samples of Cryptosporidium SSU rRNA gene were amplified,and the obtained SSU rRNA and gp60 gene of Cryptosporidium spp.in this study were sequenced and analyzed.The constructed evolutionary trees based on these two genes separately in Mega(maximum-likelihood method)were used to analyze the molecular characteristics of the obtained Cryptosporidium isolates in this study.Results:A total of 6 positive samples of Cryptosporidium were detected by nested PCR in captive raccoon dogs in Hebei Province,with a Cryptosporidium infection rate of 1.5%(6/389).The infection rate of Cryptosporidium in>6-month-old raccoon dogs was 1.2%(1/85),while the infection rate of Cryptosporidium was 1.6%(5/304)in<6-month-old raccoon dogs,and no significant difference was seen(χ2=0.096,P=0.384).The infection rate of Cryptosporidium in raccoon dogs with abnormal feces(11.1%,4/36)was significantly higher than that in raccoon dogs with normal feces(0.6%,2/353)(χ2=23.18,P=0.001).Sequence and evolutionary tree analysis showed that all Cryptosporidium isolates in this study were identified as C.canis,and all isolates belonged to the XXa subtype family,including two known subtypes XXa2 and XXa4.Conclusion:The presence of two known zoonotic C.canis subtypes in captive raccoon dogs in Hebei Province implied that that raccoon dogs might be potential source of human Cryptosporidium infection.
关键词
隐孢子虫/核糖体小亚基rRNA/貉子/河北省
Key words
Cryptosporidium spp./SSU rRNA/Raccoon dog/Hebei Province
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