Species and genotype identification of Cryptosporidium in dairy cattle in Kaifeng, China
ZHAO Jin-feng1, 2, QI Meng2, WANG Hai-yan3, HUA Jun3, ZHANG Na2, ZHANG Kun-po2, LI Jun-qiang2, LIU Qun1
1.College of Veterinary Medicine, China Agriculture University, Beijing 100193, China; 2.College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; 3.Department of Animal Science, Henan Vocational College of Agriculture, Zhongmu 451450, China
Abstract:To identify Cryptosporidium species and subtype in dairy cattle in Kaifeng, China, fresh fecal samples were detected by using the Sheather’s sugar flotation method, and the Cryptosporidium-positive samples were identified with a restriction fragment length polymorphism analysis of the small subunit ribosomal RNA. C. parvum was subtyped by using 60 kDa glycoprotein (gp60) gene. The infection rates of Cryptosporidium spp. was 8.4% (52/622). Significant differences were observed between the infection rate of sampling sites, age and feeding condition (P<0.01). In the 18S rRNA gene, 50 isolates were successfully amplified, and the PCR product was digested with restriction enzymes Ssp I and Mbo II, respectively. The result showed that C. andersoni was the dominant species (n=23), as well as other species of Cryptosporidium were C. parvum (n=5), C. bovis (n=12) and C. ryanae (n=6), and the occurrence of infections with mixed C. bovis and C. ryanae (n=4). Sequence analyses of gp60 gene revealed that five isolates were zoonotic subtype IIdA19G1. Dairy cattle infected zoonotic C. parvum subtype in Kaifeng, which is of capability of potential transmission between human and animal.
赵金凤, 齐萌, 王海燕, 化军, 张娜, 张昆坡, 李俊强, 刘群. 开封地区奶牛隐孢子虫种类及基因型鉴定[J]. 中国人兽共患病学报, 2015, 31(8): 733-736.
ZHAO Jin-feng, QI Meng, WANG Hai-yan, HUA Jun, ZHANG Na, ZHANG Kun-po, LI Jun-qiang, LIU Qun. Species and genotype identification of Cryptosporidium in dairy cattle in Kaifeng, China. Chinese Journal of Zoonoses, 2015, 31(8): 733-736.
[1] Xiao L. Molecular epidemiology of cryptosporidiosis: An update[J]. Exp Parasitol, 2010, 124(1): 80-89. doi:10.1016/j.exppara.2009.03.018 [2] Kvac M, Hofmannova L, Hlaskova L, et al. Cryptosporidium erinacei n. sp. (Apicomplexa: Cryptospo- ridiidae) in hedgehogs[J]. Vet Parasitol, 2014, 201(1-2): 9-17. doi:10.1016/j.vetpar.2014.01.014 [3] Widmer G, Lee Y, Hunt P, et al. Comparative genome analysis of two Cryptosporidium parvum isolates with different host range[J]. Infect Genet Evol, 2012, 12(6): 1213-1221. doi:10.1016/j.meegid.2012.03.027 [4] Xiao L, Feng Y. Zoonotic cryptosporidiosis[J]. FEMS Immunol Med Microbiol, 2008, 52(3): 309-323. doi:10.1111/j.1574-695X.2008.00377.x [5] Ralston B, Thompson RC, Pethick D, et al. Cryptosporidium andersoni in Western Australian feedlot cattle[J]. Aust Vet J, 2010, 88: 458-460. [6] Feng Y, Ortega Y, He G, et al. Wide geographic distribution of Cryptosporidium bovis and the deer-like genotype in bovines[J]. Vet Parasitol, 2007, 144(1-2): 1-9. [7] Widmer G, Sullivan S. Genomics and population biology of Cryptosporidium species[J]. Parasite Immunol, 2012, 34(2-3): 61-71. doi:10.1111/j.1365-3024.2011.01301.x. [8] Waldron LS, Ferrari BC, Power ML. Glycoprotein 60 diversity in C. hominis and C. parvum causing human cryptosporidiosis in NSW Australia[J]. Exp Parasitol, 2009, 122(2): 124-127. doi:10.1016/j.exppara.2009.02.006. [9] Xiao L, Escalante H, Yang CF, et al. Phylogenetic analysis of Cryptosporidium parasites based on the small-subunit rRNA gene locus[J]. Appl Environ Microbiol, 1999, 65(4): 1578-1583. [10] Alves M,Xiao L, Sulaiman I, et al. Subgenotype analysis of Cryptosporidium isolates from humans,cattle and zoo ruminants in Portugal[J]. J Clin Microbiol, 2003, 41(6): 2744-2747. [11] Xiao L, Bern C, Limor J, et al. Identification of 5 types of Cryptosporidium parasites in children in Lima, Peru[J]. J Infect Dis, 2001, 183(3): 492-497. [12] Lu QB, Qiu SX, Ru BR, et al. Epidemiological investigation of cryptosporidiosis in dairy calves in some prefectures of Henan Province[J]. Chin Vet Sci, 2008, 38(3): 261-267. (in Chinese) 卢庆斌, 仇书兴, 茹宝瑞,等.河南省部分地区乳牛隐孢子虫病的流行病学调查[J].中国兽医科学, 2008, 38(3):261-267. [13] Xu WL, Li PY, Gu YF, et al. Epidemiological survey of cow cryptosporidiosis in Anhui Province[J]. J Anhui Sci Technol Univ, 2007, 21(6): 9-11. (in Chinese) 徐文龙, 李培英, 顾有方, 等.安徽省奶牛隐孢子虫病流行病学调查[J]. 安徽科技学院学报, 2007, 21(6):9-11. [14] Liu A, Wang R, Li Y, et al. Prevalence and distribution of Cryptosporidium spp. in dairy cattle in Heilongjiang Province, China[J]. Parasitol Res, 2009, 105(3): 797-802. doi:10.1007/s00436-009-1457-2 [15] Liang N, Jian FC, Li JC, et al. Species identification of cryptosporidia isolated from dairy cattle by PCR-RFLP and phylogenetic analysis[J]. Chin Vet Sci, 2009, 39(1): 22-28. (in Chinese) 梁楠, 菅复春, 李家诚, 等.乳牛源隐孢子虫种类的PCR-RFLP和分子种系发育分析鉴定[J]. 中国兽医科学, 2009, 39(1):22-28. [16]Lin J, Zhou RQ, Tan YC, et al. Isolation and genotype identification of Cryptosporidium spp.in dairy calves in Western Chongqing[J]. Acta Veterinaria Et Zootechnica Sinica, 2013, 44(3): 495-500. (in Chinese) 林洁, 周荣琼, 谭燕财, 等. 渝西地区奶牛隐孢子虫的分离和基因型鉴定[J].畜牧兽医学报, 2013, 44(3): 495-500. [17] Wang R, Wang H, Sun Y, et al. Characteristics of Cryptosporidium transmission in preweaned dairy cattle in henan, China[J]. J Clin Microbiol, 2011, 49(3): 1077-1082. doi:10.1128/JCM.02194-10 [18] Langkjaer RB, Vigre H, Enemark HL, et al. Molecular and phylogenetic characterization of Cryptosporidium and Giardia from pigs and cattle in Denmark[J]. Parasitology, 2007, 134(Pt 3): 339-350. [19] Wang R, Ma G, Zhao J, et al. Cryptosporidium andersoni is the predominant species in post-weaned and adult dairy cattle in China[J]. Parasitol Int, 2011, 60(1): 1-4. doi:10.1016/j.parint.2010.09.002 [20] Muhid A, Robertson I, Ng J, et al. Prevalence of and management factors contributing to Cryptosporidium sp. infection in pre-weaned and postweaned calves in Johor, Malaysia[J]. Exp Parasitol, 2011, 127(2): 534-538. doi:10.1016/j.exppara.2010.10.015 [21] Amer S, Zidan S, Adamu H, et al. Prevalence and characterization of Cryptosporidium spp. in dairy cattle in Nile River delta provinces, Egypt[J]. Exp Parasitol, 2013, 135(3): 518-523. doi:10.1016/j.exppara.2013.09.002