Genotyping Of Cryptosporidium Spp. In Shanghai And Parts Of The Adjacent Areas

Cryptosporidiosis caused by Cryptosporidium spp. is transmitted through water and food, it is an emerging infectious disease, threats the health of infants and the immunodeficient individuals seriously. To date, there have been a number of outbreak cases worldwide. More than 20 Cryptosporidium spp. have been indentified, and 11 of them are zoonotic. However, about 60 genotypes remain undefined. Few studies about the distribution of Cryptosporidium spp. were performed in Shanghai and the adjacent areas. Therefore, it is important to carry out related researches.Objective To understand the Cryptosporidium species and genotypes infecting susceptible humans, animals and waters in Shanghai and parts of the adjacent areas. It supports to cryptosporidiosis risk assessment and source tracking, especially provides emergency technical reserves for public health emergency of cryptosporidiosis during large urban international activities (eg. Shanghai EXPO).Materials and Methods In the present study, (1) 1819 stool samples from susceptible population,208 samples from pigs,204 from cattle and 84 samples from other animals were collected in Shanghai, Shaoxing, Zhenjiang. (2) Besides,8 water samples from 3 tap water plants,21 samples from wastewater treatment plants,6 from rivers,10 from Shanghai zoo,23 from animal farms and 4 from farm nearby were collected in Shanghai and Shaoxing. USEPA1623 method was used to detect the Cryptosporidium oocyst in water samples except the inflows of wastewater treatment plants, wastewater from animal farms and nearby, and zoo swage. Then, QIAamp DNA Stool Mini Kit was applied to extract DNA from sediments of all samples, and amplified the Crypto.18S rRNA locus by nested PCR followed by 2% agarose gel electrophoresis. Finally, BLAST, ClustalX 1.83 and MEGA 4.1 were used to phylogenetic analysis with the positive sequences. Results In result, (1) 126 positive stool samples were identified. In Shanghai, 0.31% (3/947) of diarrheal patients were infected with Cryptosporidium,1.75% (2/114) of AIDS patients,29.10% (39/134) of pigs,17.65% (36/204) of cattles, and 27.37%(5/19) of dogs,1 sample from a takin; In Shaoxing,54.05% (40/74) of pigs were infected with Crypto sporidium. Analysis based on Crypto.18S rRNA gene sequences using the tree method of Neighbor Joining showed that C. meleagridis, C. hominis and C. felis infected in human, Cryptosporidium pig genotypeâ…¡and C. suis in pigs, C. andersoni and C. suis in cattle, C. canis in canies and. C. andersoni in a takin. (2) Among water samples,1 from a river in a district near Shanghai EXPO garden was positive by USEPA1623 method, and 8 samples were positive identified by nested PCR. And C. andersoni (3), C. canis (2) and Cryptosporidium pig genotypeâ…¡(3) were defined by phylogenetic analysis.Consclusions The present work suggested that the infection of many zoonotic Cryptosporidium spp. and genotype in human, several animals and waters poses a significant potential public health threat in Shanghai and the adjacent areas. So, much more attention must be paid to the management of animal feces, cryptosporidiosis surveillance, and the treatment of waters. It will also effectively support to prevent and control the potential cryptosporidiosis outbreak.