Skip to main content
Download PDF
  • Research article
  • Open access
  • Published:

Cryptosporidium andersoni as a novel predominant Cryptosporidium species in outpatients with diarrhea in Jiangsu Province, China

BMC Infectious Diseases volume 14, Article number: 555 (2014) Cite this article

Abstract

Background

Cryptosporidium hominis and C. parvum are usually considered to be the major pathogens responsible for human cryptosporidiosis. However, there have been few studies regarding the molecular epidemiology of Cryptosporidium in human infections in China. Here we investigated Cryptosporidium infection in patients with diarrhea, in Danyang Hospital of Jiangsu Province, China, at the genotype level.

Methods

A total of 232 stool specimens were collected from outpatients with diarrhea in Danyang Hospital of Jiangsu Province, China, from February 2012 to January 2013. Each specimen was stained from direct fecal smears and examined for Cryptosporidium using modified acid fast staining and microscopy. Moreover, genomic DNA of each fecal sample was screened for the presence of Cryptosporidium with nested PCR, which was genotyped by analyzing the DNA sequences of small subunit rRNA (SSU rRNA).

Results

The average infection rate of Cryptosporidium was 1.3% (3/232) by microscopy and subjected to PCR amplification of the SSU rRNA gene of Cryptosporidium, with 9.91% (23/232) being positive for Cryptosporidium with a significant peak in autumn. Based on the SSU rRNA gene, two Cryptosporidium spp. were identified, including C. andersoni (n =21) and C. hominis (n =2). Two types of C. andersoni, designated as A370 + and A370 - , were found in the SSU rRNA gene in our present study, which was 100% homologous to C. andersoni infections derived from dairy calves and goats, respectively. The clinical questionnaires showed no significant difference in age, gender and frequency of diarrhea, but duration of diarrhea was shorter for C. andersoni than that of C. hominis (mean, 2 vs. 4 days; p <0.01).

Conclusions

C. andersoni is the dominant species in Danyang City of Jiangsu Province. The fact that SSU rRNA sequences of C. andersoni obtained from human stools exhibited 100% homologous to those derived from dairy calves and goats supported that C. andersoni infection might be attributable to animal origin. The difference in the duration of diarrhea of C. andersoni and C. hominis indicated that different Cryptosporidium species might cause different clinical manifestations.

Peer Review reports

Background

Cryptosporidium spp. causes significant diarrheal disease in humans and animals worldwide [1]. Increasing numbers of molecular epidemiological studies on cryptosporidiosis have provided greater understanding of the diversity of the species infecting humans. To date, 30 species and more than 70 subtypes of Cryptosporidium have been identified in diverse vertebrate hosts [2]-[4]. Among these, C. hominis and C. parvum have been shown to be the two major human pathogens by numerous studies, responsible for approximate 90% of human cryptosporidiosis [5]-[8]. Moreover, there appear to be geographical differences in the distribution of Cryptosporidium spp. in humans from an epidemiological viewpoint. For example, C. parvum is the dominant species infecting humans in the Middle-East, while C. hominis is considered to be responsible for the majority of human infections in developing countries, and C. parvum and C. hominis are both observed in almost equal proportions in European countries [1],[9].

In China, since the first report of two human cases of cryptosporidiosis in 1987 [10], Cryptosporidium has been attracting increasing attention. Epidemiological data on human cryptosporidiosis have confirmed its common occurrence in at least 17 Chinese provinces or cities with a detection rate ranging from 1.4% to 10.4% based on morphological identification [11]. Identifying the parasite species infecting humans by molecular methods is important in determining the epidemiology of the disease and its likely transmission routes [12]. However there have been few studies conducted on the molecular epidemiology of Cryptosporidium in humans, with the exception of Shanghai [13], Henan Province [14], Hunan Province [15] and Tianjin [16]. Currently, there are only morphological data on human cryptosporidiosis in Jiangsu Province [17],[18]. The aim of the present study was to investigate Cryptosporidium infection in patients with diarrhea using molecular tools and sequencing, in Danyang Hospital of Jiangsu Province. We then further sought to investigate the possible relationship between epidemiological data and clinical manifestations of the predominant Cryptosporidium species detected.

Methods

Ethics statement

Ethical approval for the collection and examination of human feces samples was obtained from the Ethics Committee of the Danyang Hospital of Jiangsu Province and the National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, China (reference no. 2012-12). The objectives, procedures and potential risks were verbally explained to all participants. Written informed consent was given to, and signed by all the study participants. Parents/guardians provided consent on behalf of all infant participants.

Fecal specimen collection and examination

Human stool specimens (n =232) were collected from Danyang Hospital of Jiangsu Province, China. All of the diarrhea patients investigated (1 month - 77 years old, medium 34.7 years old) in this study were recruited from outpatients departments during a 1-year period from February 2012 to January 2013. Patient details, including their age, gender, occurrence, duration and frequency of diarrhea, and consistency of stools, were recorded. The duration of an episode diarrhea was defined to be from the onset to attendance at the hospital. The frequency of diarrhea episodes was defined as the number of occurrences of diarrhea per day. Specimens were collected from patients with fecal excretion heavier than 200 mg and with no less than three events of diarrhea per day. Each specimen was stained from direct fecal smears and examined for Cryptosporidium using modified acid fast staining and microscopy [19],[20].

DNA extraction

Sufficient samples (200 mg of each fecal sample) were collected for DNA extraction and purification using a commercially available kit, QIAamp DNA Mini Stool Kit (Qiagen, Valencia, CA). The extracted DNA samples were stored at –30°C for PCR.

Genotyping of Cryptosporidium

All DNA preparations were screened for the presence of Cryptosporidium DNA by nested PCR amplification of a fragment (approximately 830 bp) of the SSU rRNA gene, and were identified to the species/genotype level as previously described [21],[22].

Each DNA preparation was performed three times by using 1 μl DNA per PCR. DNA from C. baileyi obtained from a chicken [23], provided by Prof. Longxian Zhang of Henan Agricultural University, was used as a positive control, and deionized water was used as a negative control. All nested PCR products were visualized by electrophoresis in 2% agarose gel stained with ethidium bromide before sequencing.

DNA sequence analysis

Positive secondary PCR products were subjected to two-directional sequencing with secondary primers by the Shanghai Sunny Biotechnology Co, Ltd. (Shanghai, China). The accuracy of the sequencing data was confirmed by sequencing in both directions. Amplified sequences were blasted against sequences in the NCBI database and then deposited in GenBank (accession numbers KF826294 to KF826316) and identified by direct comparisons using the MEGA5 software of the acquired nucleotide sequences with each other and with reference sequences downloaded from GenBank.

Statistical analysis

Significance testing was performed using χ2 or Fishers exact test for categorical variables. Differences were considered significant at p values of <0.05. All statistical analyses were performed using JMP software (SAS Institute, USA).

Results

Prevalence of cryptosporidiosis in patients with diarrhea in Jiangsu Province

Human stool specimens (n =232) were screened microscopically for the presence of Cryptosporidium oocysts. However, the average detection rate of Cryptosporidium was only 1.3% (3/232) by microscopy with average dimension of the Cryptosporidium oocysts of 7 μm × 5 μm. These specimens were also screened for the presence of Cryptosporidium by PCR (9.9%) collected from February 2012 to January 2013 in Danyang Hospital of Jiangsu Province (Table 1). The prevalence of Cryptosporidium in different seasons was also investigated using these specimens. The mean prevalence over the four seasons ranged from 2.2% to 29.1% and showed significant variation between seasons (χ2 = 9.753, p <0.05) (Table 1), with a peak in autumn. The prevalence of Cryptosporidium in all age categories (with the exception of children group) ranged from 7.7% to 12.5%, with no significant differences among different age groups (χ2 = 1.666, p >0.05) (Table 2).

Table 1 The seasonal distribution of cryptosporidiosis in diarrhea outpatients in the Danyang hospital of Jiangsu province from February 2012 to January 2013
Full size table
Table 2 Feature distribution of Cryptosporidium spp. and subtype in Danyang hospital of Jiangsu province from February 2012 to January 2013
Full size table

Molecular analysis of Cryptosporidiumspecies

Nested PCR screening for Cryptosporidium identified 23 positive specimens. DNA sequencing of the SSU rRNA target fragments revealed the presence of two Cryptosporidium species: C. hominis (n =2) and C. andersoni (n =21) (Table 1). All three microscopy-positive specimens were further confirmed to be C. andersoni by PCR and sequencing of the SSU rRNA gene. In addition, two distinct C. andersoni sequences were classified at the SSU rRNA locus. Of the 21 SSU rRNA C. andersoni sequences obtained (771 bp in length), 12 sequences were assigned as A370 − type (KF826294, KF826295, KF826296, KF826298, KF826299, KF826303, KF826304, KF826305, KF826307, KF826310, KF826311, KF826313) and nine sequences as A370 + type (KF826297, KF826300, KF826301, KF826302, KF826306, KF826308, KF826309, KF826312, KF826314), based on the absence or presence, respectively, of an A at position 370 of the nucleotide sequence. A370 − type exhibited 100% homology with the C. andersoni isolated from dairy calves (JX515549), while A370 + type exhibited 100% homology with the C. andersoni isolated from a goat (EU926593).

Characteristics associated with C. andersoniinfection

According to Cryptosporidium infection among these age groups, C. hominis infection was found in two specimens from the adult group and elderly group, respectively, with an overall detection rate of 8.7% (Table 2). In contrast, C. andersoni infection was more commonly found in diarrheal patients, with the exception of children (Table 2). There were no significant differences in C. andersoni detection rates between the different age groups (χ2 = 1.82, p >0.05). However, according to our data, the detection rate of C. andersoni infection was 33.3% in infants and 38.1% in elderly patients. Further analysis of the results showed no significant difference in the overall number of Cryptosporidium-positive specimens between the genders (χ2 = 3.74, p >0.05); however the gender distribution of C. andersoni infection was four (19.05%) female and 17 (81.0%) male patients, compared with one female (KF826315) and one male patient (KF826316) for C. hominis (Table 2).

Analysis of associated information reported revealed that the outpatients with C. andersoni attended the hospital after a shorter duration of diarrhea compared to those with C. hominis infection (mean, 2 days and 4 days, respectively; p <0.01). Although there was no significant difference in the frequency of diarrhea episodes between these two species, we noticed a tendency for C. andersoni to result in more serious diarrhea, with a mean frequency of five times per day (Table 2).

Discussion

Knowledge of Cryptosporidium species is essential to the management of cryptosporidiosis and reduction in the risks of continued disease prevalence. In this study, the prevalence of Cryptosporidium spp. infection in our study was 1.3% according to microcopy versus 9.9% by PCR, and all three microscopy-positive specimens were confirmed as being C. andersoni by both PCR and sequencing of SSU rRNA gene. The result might be related to the fact that C. andersoni are larger than those of C. hominis. Meanwhile, no difference was observed morphologically between the dimensions of C. andersoni oocysts from humans in this study and those from animals reported previously [24]. Our molecular data was more sensitive and accurate than morphologic data, which was within the reported the range of 1.4%-10.4% of all patients with diarrhea for infections attributed to Cryptosporidium in China [10],[15]. However, our data for different age groups showed a detection rate of 12.5% in elderly patients, which was higher than that reported for elderly patients in Changchun (8.6%) [25]. This difference may be attributed to different detection methods in the same age group because PCR and microscopy detection were used in this study, while serological detection was used in the Changchun study. Our study indicates that molecular methods of detection have the advantage of increased sensitivity compared with conventional and immunological assays for detecting oocysts in feces. We also found an infection rate of 10.6% in infants in our study, which was higher than that of any pediatric hospital (0.4%-2.8%) in Shanghai [13]. These differences in prevalence may in part be related to differences in experimental design, specimen sources, and whether or not the patients were staying in hospital. In addition, our data also confirmed that seasonal variation can affect the prevalence of Cryptosporidium, with a peak in autumn [26],[27].

Although C. hominis and C. parvum account for more than 90% of human cases of cryptosporidiosis [2], other species that have also been associated with human disease globally include C. meleagridis, C. cuniculus, C. ubiquitum, C. felis, C. canis and C. andersoni as well as C. muris[5],[28]-[32]. There is considerable regional diversity in the distribution of Cryptosporidium species in China; C. hominis has been reported to be the predominant cause of infection in Eastern China [13],[16],[33], while C. parvum is the predominant species in Hunan Province [15]. However, the results of our study clearly showed that C. andersoni has become a novel predominant species (21 C. andersoni infections out of 23 cases) among outpatients with diarrhea in the geographical area investigated.

As a species with narrow host specificity [34], C. andersoni was usually detected primarily in domestic cattle [24]. Only a few cases have been published regarding human C. andersoni infections in France [35], Malawi [36], Iran [37], England [5] and Australia [38]. However, a recent study by our group [39] reported 34 C. andersoni in 252 human patients with diarrhea in Shanghai. And in this study, we also found 21 cases of C. andersoni infection. It is unclear why so many cases of human cryptosporidiosis are caused by C. andersoni in the investigated areas. The true source of C. andersoni infection needs to be elucidated by subtyping C. andersoni isolates from humans and animals in the future, as well as analyses of its transmission dynamics. Furthermore, these sequences of the 21 cases of C. andersoni were divided into two types, which 12 sequences belonging to A370 - type were identical to an isolate from a dairy calf (JX515549), while the others of 9 sequences belonging to A370 + type were identical to an isolate from a goat (EU926593). This suggests that C. andersoni has become the new major Cryptosporidium species to infect humans, and its zoonotic transmission may be from animals to humans either directly or indirectly. We have already assessed further genetic sources of human C. andersoni infections using multilocus sequence typing (MLST), which has also been used for exploring genetic sources of C. andersoni infection in zoonotic samples [34],[40] and confirmed at least two sites from cattle to be consistent with our current data (data not shown).

C. andersoni is also one of the main Cryptosporidium species found in contaminated water such as the Potomac River in the United States [41], the Huangpu River in Shanghai [42], and in the urban wastewater plants in Harbin [43]. It is reasonable to suggest that C. andersoni infected humans could shed C. andersoni oocysts into urban wastewater, thus further contaminating water supplies and resulting in anthroponotic transmission of C. andersoni. It is therefore essential to conduct further investigation into the transmission dynamics of Cryptosporidium between dairy calves, humans and water resources in different geographic locations.

Conclusion

In conclusion, the present findings, including distribution and genetic characterization of Cryptosporidium at the molecular level in patients with diarrhea, may be representative of the other molecular characteristics of human cryptosporidiosis. Whether or not the C. andersoni identified in patients with diarrhea in this study represented a natural infection needs to be confirmed with more systematic characterization of cryptosporidiosis and experimental infection studies.

Abbreviations

C. hominis :

Cryptosporidium hominis

C. parvum :

Cryptosporidium parvum

C. andersoni :

Cryptosporidium andersoni

SSU rRNA:

Small subunit rRNA

References

  1. Desai NT, Sarkar R, Kang G: Cryptosporidiosis: an under-recognized public health problem. Trop Parasitol. 2012, 2 (2): 91-98. 10.4103/2229-5070.105173.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Xiao L: Molecular epidemiology of cryptosporidiosis: an update. Exp Parasitol. 2010, 124 (1): 80-89. 10.1016/j.exppara.2009.03.018.

    Article  CAS  PubMed  Google Scholar 

  3. Elwin K, Hadfield SJ, Robinson G, Crouch ND, Chalmers RM: Cryptosporidium viatorum n. sp. (Apicomplexa: Cryptosporidiidae) among travellers returning to Great Britain from the Indian subcontinent, 2007-2011. Int J Parasitol. 2012, 42 (7): 675-682. 10.1016/j.ijpara.2012.04.016.

    Article  PubMed  Google Scholar 

  4. Slapeta J: Cryptosporidiosis and Cryptosporidium species in animals and humans: a thirty colour rainbow?. Int J Parasitol. 2013, 43 (12-13): 957-970. 10.1016/j.ijpara.2013.07.005.

    Article  PubMed  Google Scholar 

  5. Leoni F, Amar C, Nichols G, Pedraza-Diaz S, McLauchlin J: Genetic analysis of Cryptosporidium from 2414 humans with diarrhoea in England between 1985 and 2000. J Med Microbiol. 2006, 55 (Pt 6): 703-707. 10.1099/jmm.0.46251-0.

    Article  CAS  PubMed  Google Scholar 

  6. Elwin K, Hadfield SJ, Robinson G, Chalmers RM: The epidemiology of sporadic human infections with unusual cryptosporidia detected during routine typing in England and Wales, 2000-2008. Epidemiol Infect. 2012, 140 (4): 673-683. 10.1017/S0950268811000860.

    Article  CAS  PubMed  Google Scholar 

  7. Kvac M, Kvetonova D, Sak B, Ditrich O: Cryptosporidium pig genotype II in immunocompetent man. Emerg Infect Dis. 2009, 15 (6): 982-983. 10.3201/eid1506.071621.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Zhu H, Zhao J, Wang R, Zhang L: Molecular identification of a rare subtype of Cryptosporidium hominis in infants in China. PLoS One. 2012, 7 (8): e43682-10.1371/journal.pone.0043682.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ajjampur SS, Sarkar R, Sankaran P, Kannan A, Menon VK, Muliyil J, Ward H, Kang G: Symptomatic and asymptomatic Cryptosporidium infections in children in a semi-urban slum community in southern India. Am J Trop Med Hyg. 2010, 83 (5): 1110-1115. 10.4269/ajtmh.2010.09-0644.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Han F, Tan WX, Zhou XL: Two case reports of cryptosporidiosis in Nanjing. Jiangsu Med J. 1987, 13 (12): 692-

    Google Scholar 

  11. Lv S, Tian LG, Liu Q, Qian MB, Fu Q, Steinmann P, Chen JX, Yang GJ, Yang K, Zhou XN: Water-related parasitic diseases in China. Int J Environ Res Public health. 2013, 10 (5): 1977-2016. 10.3390/ijerph10051977.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Xiao L, Fayer R, Ryan U, Upton SJ: Response to the newly proposed species Cryptosporidium pestis. Trends Parasitol. 2007, 23 (2): 41-42. 10.1016/j.pt.2006.11.008.

    Article  PubMed  Google Scholar 

  13. Feng Y, Wang L, Duan L, Gomez-Puerta LA, Zhang L, Zhao X, Hu J, Zhang N, Xiao L: Extended outbreak of cryptosporidiosis in a pediatric hospital, China. Emerg Infect Dis. 2012, 18 (2): 312-314. 10.3201/eid1802.110666.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Wang L, Zhang H, Zhao X, Zhang L, Zhang G, Guo M, Liu L, Feng Y, Xiao L: Zoonotic Cryptosporidium species and Enterocytozoon bieneusi genotypes in HIV-positive patients on antiretroviral therapy. J Clin Microbiol. 2013, 51 (2): 557-563. 10.1128/JCM.02758-12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Yu Z, Li F, Zeng Z, Huang Z, Fan Z, Jin Y, Luo W, Xiang X, Deng Q: Prevalence and clinical significance of Cryptosporidium infection in patients with hepatitis B virus-associated acute-on-chronic liver failure. Int J Infect Dis. 2011, 15 (12): e845-e848. 10.1016/j.ijid.2011.08.006.

    Article  PubMed  Google Scholar 

  16. Peng MM, Matos O, Gatei W, Das P, Stantic-Pavlinic M, Bern C, Sulaiman IM, Glaberman S, Lal AA, Xiao L: A comparison of Cryptosporidium subgenotypes from several geographic regions. J Eukaryot Microbiol. 2001, Suppl: 28S-31S. 10.1111/j.1550-7408.2001.tb00442.x.

    Article  CAS  PubMed  Google Scholar 

  17. Chen YG, Yao FB, Li HS, Shi WS, Dai MX, Lu M: Cryptosporidium infection and diarrhea in rural and urban areas of Jiangsu, People's Republic of China. J Clin Microbiol. 1992, 30 (2): 492-494.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Chen YG, Yao FB, Li HS, Shi WS, Dai MX, Lu M: [Epidemiological studies on human cryptosporidiosis in rural and urban areas of Jiangsu]. Chin J Parasitol Parasitic Dis. 1993, 11 (3): 207-210.

    CAS  Google Scholar 

  19. Zhang W, Yang F, Liu A, Wang R, Zhang L, Shen Y, Cao J, Ling H: Prevalence and genetic characterizations of Cryptosporidium spp. in pre-weaned and post-weaned piglets in Heilongjiang Province, China. PLoS One. 2013, 8 (7): e67564-10.1371/journal.pone.0067564.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ma P, Soave R: Three-step stool examination for cryptosporidiosis in 10 homosexual men with protracted watery diarrhea. J Infect Dis. 1983, 147 (5): 824-828. 10.1093/infdis/147.5.824.

    Article  CAS  PubMed  Google Scholar 

  21. Yin J, Shen Y, Yuan Z, Lu W, Xu Y, Cao J: Prevalence of the Cryptosporidium pig genotype II in pigs from the Yangtze River Delta, China. PLoS One. 2011, 6 (6): e20738-10.1371/journal.pone.0020738.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Xiao L, Bern C, Limor J, Sulaiman I, Roberts J, Checkley W, Cabrera L, Gilman RH, Lal AA: Identification of 5 types of Cryptosporidium parasites in children in Lima, Peru. J Infect Dis. 2001, 183 (3): 492-497. 10.1086/318090.

    Article  CAS  PubMed  Google Scholar 

  23. Wang R, Jian F, Sun Y, Hu Q, Zhu J, Wang F, Ning C, Zhang L, Xiao L: Large-scale survey of Cryptosporidium spp. in chickens and Pekin ducks (Anas platyrhynchos) in Henan, China: prevalence and molecular characterization. Avian Pathol. 2010, 39 (6): 447-451. 10.1080/03079457.2010.518314.

    Article  PubMed  Google Scholar 

  24. Lindsay DS, Upton SJ, Owens DS, Morgan UM, Mead JR, Blagburn BL: Cryptosporidium andersoni n. sp. (Apicomplexa: Cryptosporiidae) from cattle, Bos taurus. J Eukaryot Microbiol. 2000, 47 (1): 91-95. 10.1111/j.1550-7408.2000.tb00016.x.

    Article  CAS  PubMed  Google Scholar 

  25. Song JP, Zhao JX, Gao H, Liu Y, Yue HX, Zhang J, Su Y, Yin JG: [Serological detection of Cryptosporidium spp. infection in outpatients in Changchun]. Chin J Parasitol Parasitic Dis. 2011, 29 (3): 239-241.

    Google Scholar 

  26. Leitch GJ, He Q: Cryptosporidiosis-an overview. J Biomed Res. 2012, 25 (1): 1-16. 10.1016/S1674-8301(11)60001-8.

    Article  PubMed  Google Scholar 

  27. Chalmers RM, Elwin K, Hadfield SJ, Robinson G: Sporadic human cryptosporidiosis caused by Cryptosporidium cuniculus, United Kingdom, 2007-2008. Emerg Infect Dis. 2011, 17 (3): 536-538. 10.3201/eid1703.100410.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Chalmers RM, Robinson G, Elwin K, Hadfield SJ, Xiao L, Ryan U, Modha D, Mallaghan C: Cryptosporidium sp. rabbit genotype, a newly identified human pathogen. Emerg Infect Dis. 2009, 15 (5): 829-830. 10.3201/eid1505.081419.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Gatei W, Ashford RW, Beeching NJ, Kamwati SK, Greensill J, Hart CA: Cryptosporidium muris infection in an HIV-infected adult, Kenya. Emerg Infect Dis. 2002, 8 (2): 204-206. 10.3201/eid0802.010256.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Gatei W, Suputtamongkol Y, Waywa D, Ashford RW, Bailey JW, Greensill J, Beeching NJ, Hart CA: Zoonotic species of Cryptosporidium are as prevalent as the anthroponotic in HIV-infected patients in Thailand. Ann Trop Med Parasitol. 2002, 96 (8): 797-802. 10.1179/000349802125002202.

    Article  CAS  PubMed  Google Scholar 

  31. Akiyoshi DE, Dilo J, Pearson C, Chapman S, Tumwine J, Tzipori S: Characterization of Cryptosporidium meleagridis of human origin passaged through different host species. Infect Immun. 2003, 71 (4): 1828-1832. 10.1128/IAI.71.4.1828-1832.2003.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Palmer CJ, Xiao L, Terashima A, Guerra H, Gotuzzo E, Saldias G, Bonilla JA, Zhou L, Lindquist A, Upton SJ: Cryptosporidium muris, a rodent pathogen, recovered from a human in Peru. Emerg Infect Dis. 2003, 9 (9): 1174-1176. 10.3201/eid0909.030047.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Wang R, Zhang X, Zhu H, Zhang L, Feng Y, Jian F, Ning C, Qi M, Zhou Y, Fu K, Wang Y, Sun Y, Wang Q, Xiao L: Genetic characterizations of Cryptosporidium spp. and Giardia duodenalis in humans in Henan, China. Exp Parasitol. 2011, 127 (1): 42-45. 10.1016/j.exppara.2010.06.034.

    Article  CAS  PubMed  Google Scholar 

  34. Feng Y, Yang W, Ryan U, Zhang L, Kvac M, Koudela B, Modry D, Li N, Fayer R, Xiao L: Development of a multilocus sequence tool for typing Cryptosporidium muris and Cryptosporidium andersoni. J Clin Microbiol. 2011, 49 (1): 34-41. 10.1128/JCM.01329-10.

    Article  CAS  PubMed  Google Scholar 

  35. Guyot K, Follet-Dumoulin A, Lelievre E, Sarfati C, Rabodonirina M, Nevez G, Cailliez JC, Camus D, Dei-Cas E: Molecular characterization of Cryptosporidium isolates obtained from humans in France. J Clin Microbiol. 2001, 39 (10): 3472-3480. 10.1128/JCM.39.10.3472-3480.2001.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Morse TD, Nichols RA, Grimason AM, Campbell BM, Tembo KC, Smith HV: Incidence of cryptosporidiosis species in paediatric patients in Malawi. Epidemiol Infect. 2007, 135 (8): 1307-1315. 10.1017/S0950268806007758.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Agholi M, Hatam GR, Motazedian MH: HIV/AIDS-associated opportunistic protozoal diarrhea. AIDS Res Hum Retroviruses. 2013, 29 (1): 35-41. 10.1089/aid.2012.0119.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Waldron LS, Dimeski B, Beggs PJ, Ferrari BC, Power ML: Molecular epidemiology, spatiotemporal analysis, and ecology of sporadic human cryptosporidiosis in Australia. Appl Environ Microbiol. 2011, 77 (21): 7757-7765. 10.1128/AEM.00615-11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Liu H, Shen Y, Yin J, Yuan Z, Jiang Y, Xu Y, Pan W, Hu Y, Cao J: Prevalence and genetic characterization of Cryptosporidium, Enterocytozoon. Giardia and Cyclospora in diarrheal outpatients in China. BMC Infect Dis. 2014, 14: 25-10.1186/1471-2334-14-25.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Wang R, Jian F, Zhang L, Ning C, Liu A, Zhao J, Feng Y, Qi M, Wang H, Lv C, Zhao G, Xiao L: Multilocus sequence subtyping and genetic structure of Cryptosporidium muris and Cryptosporidium andersoni. PLoS One. 2012, 7 (8): e43782-10.1371/journal.pone.0043782.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Yang W, Chen P, Villegas EN, Landy RB, Kanetsky C, Cama V, Dearen T, Schultz CL, Orndorff KG, Prelewicz GJ, Brown MH, Young KR, Xiao L: Cryptosporidium source tracking in the Potomac River watershed. Appl Environ Microbiol. 2008, 74 (21): 6495-6504. 10.1128/AEM.01345-08.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Feng Y, Zhao X, Chen J, Jin W, Zhou X, Li N, Wang L, Xiao L: Occurrence, source, and human infection potential of Cryptosporidium and Giardia spp. in source and tap water in shanghai, china. Appl Environ Microbiol. 2011, 77 (11): 3609-3616. 10.1128/AEM.00146-11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Liu A, Ji H, Wang E, Liu J, Xiao L, Shen Y, Li Y, Zhang W, Ling H: Molecular identification and distribution of Cryptosporidium and Giardia duodenalis in raw urban wastewater in Harbin, China. Parasitol Res. 2011, 109 (3): 913-918. 10.1007/s00436-011-2333-4.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the Chinese Special Program for Scientific Research of Public Health (No. 201302004, to Y.S.), the Shanghai Public Health Outstanding Academic Leader (No. GWDTR201214, to Y.S.) and the National S & T Major Program (No. 2012ZX10004-201, to J.C.).

Author information

Authors and Affiliations

  1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Key Laboratory of Parasite and Vector Biology, Ministry of Health, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, 200025, China

    Yanyan Jiang, Zhongying Yuan, Hua Liu, Wei Pan, Jianping Cao & Yujuan Shen

  2. Department of Infectious Diseases, Danyang Hospital of Jiangsu Province, Danyang, 212300, China

    Jinhua Ren, Hong Zhao, Lei Chu & Yijin Lin

  3. Department of Parasitology, Harbin Medical University, Harbin, 150081, China

    Aiqin Liu

Authors
  1. Yanyan Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jinhua Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhongying Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Aiqin Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hong Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hua Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lei Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Wei Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jianping Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yijin Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yujuan Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yijin Lin or Yujuan Shen.

Additional information

Competing interests

The authors declare that they and their institutions have no financial or personal relationships with other people or organizations that could inappropriately influence (bias) their actions.

Authors' contributions

YS, JC and YL conceived and designed the experiments; YJ, ZY, HZ, HL, LC and WP performed the experiments; YJ, JR and YS analyzed the data, statistical analysis and writing up the manuscript; JC, AL and YS critically revised the manuscript. All authors read and approved the final version of the manuscript.

Rights and permissions

Open Access  This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.

The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.

The Creative Commons Public Domain Dedication waiver (https://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jiang, Y., Ren, J., Yuan, Z. et al. Cryptosporidium andersoni as a novel predominant Cryptosporidium species in outpatients with diarrhea in Jiangsu Province, China. BMC Infect Dis 14, 555 (2014). https://0-doi-org.brum.beds.ac.uk/10.1186/s12879-014-0555-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://0-doi-org.brum.beds.ac.uk/10.1186/s12879-014-0555-7

Keywords

BMC Infectious Diseases

ISSN: 1471-2334

Contact us