Santín Mónica, Trout James M, Fayer Ronald
Environmental Microbial Safety Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, United States Department of Agriculture, Beltsville, MD 20705, USA.
Vet Parasitol. 2008 Aug 1;155(1-2):15-23. doi: 10.1016/j.vetpar.2008.04.018. Epub 2008 May 10.
Fecal specimens were collected from 30 calves from birth to 24 months of age at a dairy farm in Maryland to determine the prevalence and age distribution of Cryptosporidium species/genotypes. After centrifugation to remove debris and concentrate oocysts, specimens were examined by immunofluorescence microscopy and polymerase chain reaction (PCR). Fragments of the SSU-rDNA gene amplified by PCR were purified and PCR products were sequenced. All 30 calves shed Cryptosporidium oocysts at some time during the 24 months of the study. Of 990 specimens, 190 were Cryptosporidium-positive (19.2%). The highest prevalence of infection was at 2 weeks of age when 29 of the 30 calves were excreting oocysts. Prevalence was higher in pre-weaned calves (1-8 weeks of age) (45.8%) than in post-weaned calves (3-12 months of age) (18.5%) and heifers (12-24 months of age) (2.2%). Sequence data for 190 PCR-positive specimens identified: C. parvum, C. bovis, the Cryptosporidium deer-like genotype and C. andersoni, with cumulative prevalences of 100, 80, 60, and 3.3%, respectively. C. parvum constituted 97% of infections in pre-weaned calves but only 4% and 0% of infections in post-weaned calves and heifers, respectively. All C. parvum GP60 nucleotide sequences were subtype IIaA15G2R1.
从马里兰州一个奶牛场的30头犊牛中收集粪便样本,这些犊牛从出生到24月龄,目的是确定隐孢子虫种类/基因型的流行率和年龄分布。离心去除碎片并浓缩卵囊后,通过免疫荧光显微镜检查和聚合酶链反应(PCR)对样本进行检测。对PCR扩增的小亚基核糖体DNA(SSU-rDNA)基因片段进行纯化,并对PCR产物进行测序。在研究的24个月期间,所有30头犊牛在某个时间都排出了隐孢子虫卵囊。在990份样本中,190份隐孢子虫呈阳性(19.2%)。感染率最高的是在2周龄时,30头犊牛中有29头排出卵囊。断奶前犊牛(1 - 8周龄)的感染率(45.8%)高于断奶后犊牛(3 - 12月龄)(18.5%)和小母牛(12 - 24月龄)(2.2%)。对190份PCR阳性样本的序列数据进行分析,鉴定出:微小隐孢子虫、牛隐孢子虫、隐孢子虫鹿样基因型和安氏隐孢子虫,其累积流行率分别为100%、80%、60%和3.3%。微小隐孢子虫在断奶前犊牛的感染中占97%,但在断奶后犊牛和小母牛的感染中分别仅占4%和0%。所有微小隐孢子虫GP60核苷酸序列均为IIaA15G2R1亚型。