Open Access
Volume 27, 2020
Article Number 65
Number of page(s) 8
Published online 24 November 2020

© X.-L. Zheng et al., published by EDP Sciences, 2020

Licence Creative CommonsThis is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Enterocytozoon bieneusi, a zoonotic intestinal pathogen, infects a wide range of species worldwide [20, 24]. Microsporidiosis occurs through the ingestion of infectious spores of E. bieneusi through contaminated soil, feces, surfaces, water, as well as by improper farming practices, such as using untreated animal manure as fertilizer directly on open crops or tillage land [20]. Enterocytozoon bieneusi has received considerable attention due to its known propensity to cause both water- and food-borne outbreaks of illness [44].

Sequence analysis of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene has revealed more than 500 genotypes (142 in humans, of which 49 were also identified in animals) [11, 20, 54]. Phylogenetic comparative analyses clustered all genotypes into eleven major genetic groups. Human cases have been reported to show infection with E. bieneusi genotypes from six groups, and more than 90% of human-pathogenic genotypes belonged to Group 1 or Group 2 [20, 54].

Thirty-eight studies from 14 countries have identified more than 80 genotypes in cattle, known carriers of E. bieneusi (Table 1). Among them, at least 17 genotypes (BEB4, BEB6, I, J, PtEb XI, EbpC, D, EbpA, M, Type IV, Peru 6, H, O, CS-4, CHN3, CHN4, and S7) have also been identified in humans [20]. Of the remaining 67 genotypes, 30 belonged to Group 1, and 27 belonged to Group 2, indicating the vital role of cattle in the epidemiology of E. bieneusi and their ability to transmit the pathogen to humans [20]. Therefore, cattle infected with E. bieneusi may pose a threat to public health.

Table 1

ITS genotypes of Enterocytozoon bieneusi of natural infection identified in cattle worldwide.

In China, cattle farming and dairy products are important economic industries. Previous studies on E. bieneusi in cattle in China focused on inland cities and did not include assessments in Hainan Province, the southernmost region of China, where, local yellow cattle breeding is very popular. Here, we evaluated the prevalence, genetic characteristics, and zoonotic potential of E. bieneusi in cattle from six cities of Hainan Province.

Materials and methods

Ethics statement

The study was initiated after obtaining written informed consent for animal use by farm owners. All animal experiments were reviewed and approved by the Ethics Committee of Hainan Medical University.

Fecal specimen collection

In all, 314 fecal samples were gathered from 10 cattle farms in six cities of Hainan Province between March and December 2019 (Fig. 1 and Table 2). The cattle farms were selected based only on the owners’ willingness to participate and the accessibility of animals for sampling. Samples were obtained from 30–50% of the total number of cattle on each farm. A sterile disposable latex glove was used to collect the fecal specimens immediately post defecation, and placed in individually labeled plastic bags. Cattle were divided into two groups: young aged ≤ 12 months (n = 18) and adults aged > 12 months (n = 296). Cattle were in good health at the time of sampling. Within 24 h of sampling, the labeled fecal bags were transported and stored in the laboratory at 4 °C and were processed within 48 h.

thumbnail Figure 1

Specific locations where samples were collected in this study. ▲: Sampling points.

Table 2

Prevalence and genotype distribution of E. bieneusi isolates in cattle in Hainan Province.

DNA extraction

All fecal specimens were filtered through sieve in distilled water, followed by centrifugation at 1500 ×g for 10 min. A QIAamp DNA stool mini kit (QIAgen, Germany) was used to isolate the genomic DNA of each processed specimen (approximately 200 mg), following the manufacturer’s instructions. A total of 200 mL AE elution buffer was used to elute the DNA, followed by storage at −20 °C before PCR analysis.

Polymerase chain reaction (PCR) amplification

Enterocytozoon bieneusi-specific nested primers and cycle parameters designed by Hamed Mirjalal were used to amplify a 410 bp sequence in the ITS region of the rRNA gene using TaKaRa Taq DNA Polymerase [25]. The PCR products were analyzed using 1.5% agarose gel electrophoresis, followed by GelRed (Biotium Inc., USA) staining.

Nucleotide sequencing and analysis

The sequence accuracy of all E. bieneusi-positive PCR products (sequenced by Sangon Biotech Co., Ltd., China) was confirmed through bidirectional sequencing and the sequencing of additional PCR products. The Basic Local Alignment Search Tool (BLAST) and ClustalX 1.83 were used to compare the published GenBank sequences with the ones identified in this study to identify the genotypes of E. bieneusi. Genotypes that were identical to the genotypes deposited in the GenBank database were given the first published name, and those that generated ITS sequences with a single nucleotide substitution/deletion/insertion were identified as novel genotypes based on the DNA sequencing of minimum two PCR products [30]. The samples were labeled in the order of appearance by adding roman numerals after HNC (Hainan Cattle). A 243 bp part of the ITS region of the rRNA gene of E. bieneusi was used for naming reference, following the established nomenclature system [30].

Phylogenetic analysis

A neighbor-joining phylogenetic tree was built using Mega X software, and the Kimura-2-parameter model with 1000 replicates to evaluate the relationship between the novel ITS genotype and the known genotypes, and to confirm the gene group designation.

Statistical analysis

Fisher’s exact test and a Chi-square test were used to evaluate the difference in infection rates among different locations and ages, respectively, using SPSS v22.0 (IBM Corp., USA). A p-value < 0.05 was regarded as statistically significant.

Nucleotide sequence accession numbers

The GenBank database accession number of the identified nucleotide sequence was MT193626.

Results and discussion

Of the 314 fecal samples, 31 (9.9%) were E. bieneusi-positive, based on sequence analysis of the ITS region of the rRNA gene. A significant difference in the rate of occurrence of E. bieneusi was observed in cattle from the six cities (p < 0.05), with 20.5% (8/39) in Danzhou, 19.4% (6/31) in Wanning, 12.5% (12/96) in Lingshui, 4.9% (4/82) in Chengmai, 1.9% (1/54) in Haikou, and an absence of this parasite (0/12) in Ledong (Table 2).

Since the first report of E. bieneusi in calves in Germany, there have been 38 published epidemiological reports on E. bieneusi conducted in 14 countries, and the average infection rates in these countries range from 2.0% to 21.4% (Table 1). The infection rate of E. bieneusi, based on cattle from 16 provinces of China, falls in the range of 2.0–37.6% (Table 3). This study reports the occurrence of E. bieneusi in cattle from Hainan Province. The differences in prevalence might be related to the sensitivity and specificity of detection methods, the health status of hosts, the experimental design, the overall sample size, animal practices, and so on. Like in other animals and humans, age appears to be a significant factor affecting the occurrence of E. bieneusi in cattle [51]. In the present study, the prevalence of E. bieneusi was 22.2% (4/18) in young animals ≤ 12 months and 9.1% (27/296) in adult animals > 12 months. Although the infection rates in calves were higher than those in adults, the differences were not significant (χ 2 = 1.966, p > 0.05) (Table 2). A study by Ma et al. revealed E. bieneusi infection rates in juveniles, post-weaned calves, pre-weaned calves, and adults of 4.5% (6/134), 7.7% (8/104), 10% (1/10), and 3.9% (13/332), respectively [23]. Similarly, da Fiuza et al. reported that pre-weaned calves (27.6%, 21/76) and post-weaned calves (28.8%, 44/153) showed a higher rate of prevalence of E. bieneusi compared with heifers (14.1%, 12/85) and adults (1.4%, 2/138) [4]. Meanwhile, Li et al. showed that calves aged < 3 months (29.3%, 127/434) and 3–12 months (23.9%, 63/264) had higher infection rates than juveniles and adults (13.3%, 24/181) [16]. In accordance with these results, it was supposed that age was negatively correlated with the prevalence of E. bieneusi in cattle, probably due to the underdeveloped immune systems of the young animals.

Table 3

ITS genotypes of natural Enterocytozoon bieneusi infections identified in cattle in China.

Here, we identified one novel genotype (HNC-I) and five known genotypes (EbpC, BEB4, J, I, and CHG5). The novel genotype showed high similarity to genotype EbpC (AF076042), with one base variation at position 237 (C → T). Out of the six genotypes, the most prevalent genotype was EbpC (14 specimens), which was found in four of the six locations, followed by BEB4 (12 specimens), but this genotype was only found in Wanning. Genotype J was found in two cattle from Chengmai. The remaining three genotypes I, CHG5, and HNC-I were found in a single specimen, with the former from Danzhou and the latter two from Wanning. These results differed from those reported from the other regions of China. For example, in Gansu, Guangdong, Henan, Ningxia, Jiangsu, Shaanxi, and Xinjiang provinces, genotypes J and I were reported to be the dominant genotypes, and in Heilongjiang, genotype O was dominant (Table 3). Meanwhile, region-specific difference in genotype constitutions of E. bieneusi can also be observed in cattle in some studies, such as genotype D in Iran [16]. Therefore, the genotype distributions of E. bieneusi in cattle differed by region, but the reason behind this phenomenon is unclear.

In the present study, human-pathogenic genotypes EbpC, BEB4, J and I were observed with high occurrence (93.5%, 29/31). Genotype EbpC has been detected in humans, such as in cancer patients in Iran [25], in immunocompetent patients in the Czech Republic [29], in children in Peru and China [3, 45], and in HIV-positive patients in Peru, China, Iran, Thailand, and Vietnam [7, 18, 21, 25, 35, 41]. It was also found in more than 15 animal species and water samples [20]. Likewise, genotypes BEB4, J, and I were also found in humans [28, 47], non-human primates [15, 46], and other animals [20], and they have been documented in cattle (Table 1). This suggests that cattle infected with genotypes EbpC, BEB4, J, and I may facilitate transmission to other animals and humans.

The remaining genotype CHG5 and the novel genotype HNC-I were first identified in cattle here. Genotype CHG5 has been reported in goats with a wide distribution in China [34, 53]. We also observed this genotype in the Asiatic brush-tailed porcupines in Hainan Province [52]. Thus, the detection of the same genotype (CHG5) in multiple species (cattle, goats, and rodents) in the same region (Hainan, China) suggests a vast host range along with the possibility of cross-species transmission among cattle, goats and rodents.

The phylogenetic analysis revealed that EbpC and HNC-I, identified in this study, were divided into zoonotic Group 1, whereas genotypes BEB4, J, I, and CHG5 belong to Group 2 (Fig. 2). In total, 94.0% (79/84) of the genotypes identified in cattle clustered into Group 1 or 2 (except for genotypes CX1, CX2, TAR_fc3, CAM2, and S7) [20]. These findings suggest that E. bieneusi-infected cattle represent a potential threat to humans.

thumbnail Figure 2

Phylogenetic tree based on neighbor-joining (N-J) analysis of ITS sequences. Phylogenetic relationships between the E. bieneusi genotypes identified in cattle here and other known genotypes deposited in GenBank were inferred by an N-J analysis of ITS sequences based on genetic distance by the Kimura two-parameter model. The numbers on the branches are percent bootstrapping values from 1000 replicates. Each sequence is identified by its accession number, host origin, and genotype designation. Enterocytozoon bieneusi genotype CSK2 (KY706128) was used as the outgroup. The squares and triangles filled in black indicate novel and known genotypes identified in this study, respectively.


This study is the first evaluating the infection rates, genotype characteristics, and zoonotic potential of E. bieneusi in cattle from Hainan Province. Our results revealed a prevalence rate of 9.9% (31/314) for E. bieneusi within five of six cities in Hainan, China. We identified five known genotypes and a novel genotype. Genotype EbpC and novel genotype HNC-I were grouped into zoonotic Group 1, while genotypes BEB4, J, I and CHG5 were placed in Group 2. The observed high occurrence (93.5%, 29/31) of zoonotic genotypes (EbpC, BEB4, J, and I) emphasizes the possible role of cattle in the transmission of E. bieneusi to humans, which requires further investigations to reduce the threats posed by these animals to public health.


This work was supported by the Young Talents Science and Technology Innovation Project of the Hainan Association for Science and Technology (QCXM201802); Hainan major science and technology project (ZDKJ2016017-01); the Innovation Research Team Project of the Hainan Natural Science Foundation (2018CXTD340); the National Natural Science Foundation of China (No. 81672072 and No. 81760378), and the Graduate Student Innovation Foundation of colleges and universities of Hainan Province, 2019 (Hys2019-287). The funders had no role in the study design, data collection, data interpretation, or the decision to submit the work for publication.


  1. Abu Samra N, Thompson PN, Jori F, Zhang H, Xiao L. 2012. Enterocytozoon bieneusi at the wildlife/livestock interface of the Kruger National Park, South Africa. Veterinary Parasitology, 190(3–4), 587–590. [CrossRef] [PubMed] [Google Scholar]
  2. Baroudi D, Khelef D, Hakem A, Abdelaziz A, Chen X, Lysen C, Roellig D, Xiao L. 2017. Molecular characterization of zoonotic pathogens Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in calves in Algeria. Veterinary Parasitology Regional Studies Reports, 8, 66–69. [CrossRef] [Google Scholar]
  3. Cama VA, Pearson J, Cabrera L, Pacheco L, Gilman R, Meyer S, Ortega Y, Xiao L. 2007. Transmission of Enterocytozoon bieneusi between a child and guinea pigs. Journal of Clinical Microbiology, 45(8), 2708–2710. [CrossRef] [PubMed] [Google Scholar]
  4. da Silva Fiuza VR, Lopes CW, de Oliveira FC, Fayer R, Santin M. 2016. New findings of Enterocytozoon bieneusi in beef and dairy cattle in Brazil. Veterinary Parasitology, 216, 46–51. [CrossRef] [PubMed] [Google Scholar]
  5. Del Coco VF, Córdoba MA, Bilbao G, de Almeida Castro P, Basualdo JA, Santín M. 2014. First report of Enterocytozoon bieneusi from dairy cattle in Argentina. Veterinary Parasitology, 199(1–2), 112–115. [CrossRef] [PubMed] [Google Scholar]
  6. Dengjel B, Zahler M, Hermanns W, Heinritzi K, Spillmann T, Thomschke A, Löscher T, Gothe R, Rinder H. 2001. Zoonotic potential of Enterocytozoon bieneusi. Journal of Clinical Microbiology, 39(12), 4495–4499. [CrossRef] [PubMed] [Google Scholar]
  7. Espern A, Morio F, Miegeville M, Illa H, Abdoulaye M, Meyssonnier V, Adehossi E, Lejeune A, Cam PD, Besse B, Gay-Andrieu F. 2007. Molecular study of microsporidiosis due to Enterocytozoon bieneusi and Encephalitozoon intestinalis among human immunodeficiency virus-infected patients from two geographical areas: Niamey, Niger, and Hanoi, Vietnam. Journal of Clinical Microbiology, 45(9), 2999–3002. [CrossRef] [PubMed] [Google Scholar]
  8. Fayer R, Santín M, Trout JM. 2007. Enterocytozoon bieneusi in mature dairy cattle on farms in the eastern United States. Parasitology Research, 102(1), 15–20. [CrossRef] [PubMed] [Google Scholar]
  9. Fayer R, Santin M, Macarisin D. 2012. Detection of concurrent infection of dairy cattle with Blastocystis, Cryptosporidium, Giardia, and Enterocytozoon by molecular and microscopic methods. Parasitology Research, 111(3), 1349–1355. [CrossRef] [PubMed] [Google Scholar]
  10. Feng Y, Gong X, Zhu K, Li N, Yu Z, Guo Y, Weng Y, Kváč M, Feng Y, Xiao L. 2019. Prevalence and genotypic identification of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in pre-weaned dairy calves in Guangdong, China. Parasites & Vectors, 12(1), 41. [CrossRef] [PubMed] [Google Scholar]
  11. Gong B, Yang Y, Liu X, Cao J, Xu M, Xu N, Yang F, Wu F, Li B, Liu A, Shen Y. 2019. First survey of Enterocytozoon bieneusi and dominant genotype Peru6 among ethnic minority groups in southwestern China’s Yunnan Province and assessment of risk factors. PLoS Neglected Tropical Diseases, 13(5), e0007356. [CrossRef] [PubMed] [Google Scholar]
  12. Hu S, Liu Z, Yan F, Zhang Z, Zhang G, Zhang L, Jian F, Zhang S, Ning C, Wang R. 2017. Zoonotic and host-adapted genotypes of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in dairy cattle in Hebei and Tianjin, China. Veterinary Parasitology, 248, 68–73. [CrossRef] [PubMed] [Google Scholar]
  13. Jiang Y, Tao W, Wan Q, Li Q, Yang Y, Lin Y, Zhang S, Li W. 2015. Zoonotic and potentially host-adapted Enterocytozoon Bieneusi genotypes in sheep and cattle in northeast china and an increasing concern about the zoonotic importance of previously considered ruminant-adapted genotypes. Applied & Environmental Microbiology, 81(10), 3326–3335. [CrossRef] [PubMed] [Google Scholar]
  14. Juránková J, Kamler M, Kovařčík K, Koudela B. 2013. Enterocytozoon bieneusi in Bovine Viral Diarrhea Virus (BVDV) infected and noninfected cattle herds. Research in Veterinary Science, 94(1), 100–104. [CrossRef] [PubMed] [Google Scholar]
  15. Karim MR, Dong H, Li T, Yu F, Li D, Zhang L, Li J, Wang R, Li S, Li X, Rume FI, Ning C. 2015. Predomination and new genotypes of Enterocytozoon bieneusi in captive nonhuman primates in zoos in China: high genetic diversity and zoonotic significance. PLoS One, 10(2), e0117991. [CrossRef] [PubMed] [Google Scholar]
  16. Kord-Sarkachi E, Tavalla M, Beiromvand M. 2018. Molecular diagnosis of microsporidia strains in slaughtered cows of southwest of Iran. Journal of Parasitic Diseases, 42(1), 81–86. [CrossRef] [Google Scholar]
  17. Lee JH. 2007. Prevalence and molecular characteristics of Enterocytozoon bieneusi in cattle in Korea. Parasitology Research, 101(2), 391–396. [CrossRef] [PubMed] [Google Scholar]
  18. Leelayoova S, Subrungruang I, Suputtamongkol Y, Worapong J, Petmitr PC, Mungthin M. 2006. Identification of genotypes of Enterocytozoon bieneusi from stool samples from human immunodeficiency virus-infected patients in Thailand. Journal of Clinical Microbiology, 44(8), 3001–3004. [CrossRef] [PubMed] [Google Scholar]
  19. Li J, Luo N, Wang C, Qi M, Cao J, Cui Z, Huang J, Wang R, Zhang L. 2016. Occurrence, molecular characterization and predominant genotypes of Enterocytozoon bieneusi in dairy cattle in Henan and Ningxia, China. Parasites & Vectors, 9, 142. [CrossRef] [PubMed] [Google Scholar]
  20. Li W, Feng Y, Santin M. 2019. Host specificity of Enterocytozoon bieneusi and public health implications. Trends in Parasitology, 35(6), 436–451. [CrossRef] [PubMed] [Google Scholar]
  21. Liu H, Jiang Z, Yuan Z, Yin J, Wang Z, Yu B, Zhou D, Shen Y, Cao J. 2017. Infection by and genotype characteristics of Enterocytozoon bieneusi in HIV/AIDS patients from Guangxi Zhuang autonomous region,China. BMC Infectious Diseases, 17(1), 684. [CrossRef] [PubMed] [Google Scholar]
  22. Lobo ML, Xiao L, Cama V, Stevens T, Antunes F, Matos O. 2006. Genotypes of Enterocytozoon bieneusi in mammals in Portugal. Journal of Eukaryotic Microbiology, 53(Suppl 1), S61–S64. [CrossRef] [Google Scholar]
  23. Ma J, Li P, Zhao X, Xu H, Wu W, Wang Y, Guo Y, Wang L, Feng Y, Xiao L. 2015. Occurrence and molecular characterization of Cryptosporidium spp. and Enterocytozoon bieneusi in dairy cattle, beef cattle and water buffaloes in China. Veterinary Parasitology, 207(3–4), 220–227. [CrossRef] [PubMed] [Google Scholar]
  24. Matos O, Lobo ML, Xiao L. 2012. Epidemiology of Enterocytozoon bieneusi infection in humans. Journal of Parasitology Research, 2012, 981424. [CrossRef] [PubMed] [Google Scholar]
  25. Mirjalali H, Mirhendi H, Meamar AR, Mohebali M, Askari Z, Mirsamadi ES, Rezaeian M. 2015. Genotyping and molecular analysis of Enterocytozoon bieneusi isolated from immunocompromised patients in Iran. Infection, Genetics and Evolution, 36, 244–249. [CrossRef] [Google Scholar]
  26. Qi M, Jing B, Jian F, Wang R, Zhang S, Wang H, Ning C, Zhang L. 2017. Dominance of Enterocytozoon bieneusi genotype J in dairy calves in Xinjiang, Northwest China. Parasitology International, 66(1), 960–963. [CrossRef] [PubMed] [Google Scholar]
  27. Rinder H, Thomschke A, Dengjel B, Gothe R, Löscher T, Zahler M. 2000. Close genotypic relationship between Enterocytozoon bieneusi from humans and pigs and first detection in cattle. Journal of Parasitology, 86(1), 185–188. [CrossRef] [Google Scholar]
  28. Sak B, Brady D, Pelikánová M, Květoňová D, Rost M, Kostka M, Tolarová V, Hůzová Z, Kváč M. 2011. Unapparent microsporidial infection among immunocompetent humans in the Czech Republic. Journal of Clinical Microbiology, 49(3), 1064–1070. [CrossRef] [PubMed] [Google Scholar]
  29. Sak B, Kváč M, Kučerová Z, Květoňová D, Saková K. 2011. Latent microsporidial infection in immunocompetent individuals – a longitudinal study. PLoS Neglected Tropical Diseases, 5(5), e1162. [CrossRef] [PubMed] [Google Scholar]
  30. Santín M, Fayer R. 2009. Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence: a consensus. Journal of Eukaryotic Microbiology, 56(1), 34–38. [CrossRef] [Google Scholar]
  31. Santín M, Fayer R. 2009. A longitudinal study of Enterocytozoon bieneusi in dairy cattle. Parasitology Research, 105(1), 141–144. [CrossRef] [PubMed] [Google Scholar]
  32. Santín M, Trout JM, Fayer R. 2005. Enterocytozoon bieneusi genotypes in dairy cattle in the eastern United States. Parasitology Research, 97(6), 535–538. [CrossRef] [PubMed] [Google Scholar]
  33. Santín M, Dargatz D, Fayer R. 2012. Prevalence and genotypes of Enterocytozoon bieneusi in weaned beef calves on cow-calf operations in the USA. Parasitology Research, 110(5), 2033–2041. [CrossRef] [PubMed] [Google Scholar]
  34. Shi K, Li M, Wang X, Li J, Karim MR, Wang R, Zhang L, Jian F, Ning C. 2016. Molecular survey of Enterocytozoon bieneusi in sheep and goats in China. Parasites & Vectors, 9, 23. [CrossRef] [PubMed] [Google Scholar]
  35. Sulaiman IM, Bern C, Gilman R, Cama V, Kawai V, Vargas D, Ticona E, Vivar A, Xiao L. 2003. A molecular biologic study of Enterocytozoon bieneusi in HIV-infected patients in Lima, Peru. Journal of Eukaryotic Microbiology, 50(Suppl), 591–596. [CrossRef] [PubMed] [Google Scholar]
  36. Sulaiman IM, Fayer R, Yang C, Santin M, Matos O, Xiao L. 2004. Molecular characterization of Enterocytozoon bieneusi in cattle indicates that only some isolates have zoonotic potential. Parasitology Research, 92(4), 328–334. [CrossRef] [PubMed] [Google Scholar]
  37. Tang C, Cai M, Wang L, Guo Y, Li N, Feng Y, Xiao L. 2018. Genetic diversity within dominant Enterocytozoon bieneusi genotypes in pre-weaned calves. Parasites & Vectors, 11(1), 170. [CrossRef] [PubMed] [Google Scholar]
  38. Tao WF, Ni HB, Du HF, Jiang J, Li J, Qiu HY, Ye L, Zhang XX. 2020. Molecular detection of Cryptosporidium and Enterocytozoon bieneusi in dairy calves and sika deer in four provinces in Northern China. Parasitology Research, 119(1), 105–114. [CrossRef] [PubMed] [Google Scholar]
  39. Udonsom R, Prasertbun R, Mahittikorn A, Chiabchalard R, Sutthikornchai C, Palasuwan A, Popruk S. 2019. Identification of Enterocytozoon bieneusi in goats and cattle in Thailand. BMC Veterinary Research, 15(1), 308. [CrossRef] [PubMed] [Google Scholar]
  40. Valenčáková A, Danišová O. 2019. Molecular characterization of new genotypes Enterocytozoon bieneusi in Slovakia. Acta Tropica, 191, 217–220. [CrossRef] [PubMed] [Google Scholar]
  41. Wang L, Zhang H, Zhao X, Zhang L, Zhang G, Guo M, Liu L, Feng Y, Xiao L. 2013. Zoonotic Cryptosporidium species and Enterocytozoon bieneusi genotypes in HIV-positive patients on antiretroviral therapy. Journal of Clinical Microbiology, 51(2), 557–563. [CrossRef] [PubMed] [Google Scholar]
  42. Wang R, Li N, Jiang W, Guo Y, Wang X, Jin Y, Feng Y, Xiao L. 2019. Infection patterns, clinical significance, and genetic characteristics of Enterocytozoon bieneusi and Giardia duodenalis in dairy cattle in Jiangsu, China. Parasitology Research, 118(10), 3053–3060. [CrossRef] [PubMed] [Google Scholar]
  43. Wang HY, Qi M, Sun MF, Li DF, Wang RJ, Zhang SM, Zhao JF, Li JQ, Cui ZH, Chen YC, Jian FC, Xiang RP, Ning CS, Zhang LX. 2019. Prevalence and population genetics analysis of Enterocytozoon bieneusi in dairy cattle in China. Frontiers in Microbiology, 10, 1399. [CrossRef] [PubMed] [Google Scholar]
  44. Weiss LM, Becnel JJ. 2014. Epidemiology of microsporidian human infections, in Microsporidia: Pathogens of Opportunity, Weiss LM, Becnel JJ, Editors, 1st ed. John Wiley & Sons Inc: Chichester. [Google Scholar]
  45. Yang J, Song M, Wan Q, Li Y, Lu Y, Jiang Y, Tao W, Li W. 2014. Enterocytozoon bieneusi genotypes in children in Northeast China and assessment of risk of zoonotic transmission. Journal of Clinical Microbiology, 52(12), 4363–4367. [CrossRef] [PubMed] [Google Scholar]
  46. Yu F, Qi M, Zhao Z, Lv C, Wang Y, Wang R, Zhang L. 2019. The potential role of synanthropic rodents and flies in the transmission of Enterocytozoon bieneusi on a dairy cattle farm in China. Journal of Eukaryotic Microbiology, 66(3), 435–441. [CrossRef] [Google Scholar]
  47. Zhang X, Wang Z, Su Y, Liang X, Sun X, Peng S, Lu H, Jiang N, Yin J, Xiang M, Chen Q. 2011. Identification and genotyping of Enterocytozoon bieneusi in China. Journal of Clinical Microbiology, 49(5), 2006–2008. [CrossRef] [PubMed] [Google Scholar]
  48. Zhang Y, Koehler AV, Wang T, Haydon SR, Gasser RB. 2019. Enterocytozoon bieneusi genotypes in cattle on farms located within a water catchment area. Journal of Eukaryotic Microbiology, 66(4), 553–559. [CrossRef] [Google Scholar]
  49. Zhang Q, Zhang Z, Ai S, Wang X, Zhang R, Duan Z. 2019. Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis from animal sources in the Qinghai-Tibetan Plateau Area (QTPA) in China. Comparative Immunology, Microbiology and Infectious Diseases, 67, 101346. [CrossRef] [PubMed] [Google Scholar]
  50. Zhao W, Zhang W, Yang F, Zhang L, Wang R, Cao J, Shen Y, Liu A. 2015. Enterocytozoon bieneusi in dairy cattle in the Northeast of China: genetic diversity of its gene and evaluation of zoonotic transmission potential. Journal of Eukaryotic Microbiology, 62(4), 553–560. [CrossRef] [Google Scholar]
  51. Zhao W, Zhou H, Jin H, Sun L, Li P, Liu M, Qiu M, Xu L, Li F, Ma T, Wang S, Yin F, Li L, Cui X, Chan JF, Lu G. 2020. Genotyping of Enterocytozoon bieneusi among captive long-tailed macaques (Macaca fascicularis) in Hainan Province: high genetic diversity and zoonotic potential. Acta Tropica, 201, 105211. [CrossRef] [PubMed] [Google Scholar]
  52. Zhao W, Zhou H, Yang L, Ma T, Zhou J, Liu H, Lu G, Huang H. 2020. Prevalence, genetic diversity and implications for public health of Enterocytozoon bieneusi in various rodents from Hainan Province, China. Parasites & Vectors, 13(1), 438. [CrossRef] [PubMed] [Google Scholar]
  53. Zhou HH, Zheng XL, Ma TM, Qi M, Cao ZX, Chao Z, Wei LM, Liu QW, Sun RP, Wang F, Zhang Y, Lu G, Zhao W. 2019. Genotype identification and phylogenetic analysis of Enterocytozoon bieneusi in farmed black goats (Capra hircus) from China’s Hainan Province. Parasite, 26, 62. [CrossRef] [EDP Sciences] [PubMed] [Google Scholar]
  54. Zhou HH, Zheng XL, Ma TM, Qi M, Zhou JG, Liu HJ, Lu G, Zhao W. 2020. Molecular detection of Enterocytozoon bieneusi in farm-raised pigs in Hainan Province, China: infection rates, genotype distributions, and zoonotic potential. Parasite, 27, 12. [CrossRef] [EDP Sciences] [PubMed] [Google Scholar]

Cite this article as: Zheng X-L, Zhou H-H, Ren G, Ma T-M, Cao Z-X, Wei L-M, Liu Q-W, Wang F, Zhang Y, Liu H-L, Xing M-P, Huang L-I, Chao Z & Lu G. 2020. Genotyping and zoonotic potential of Enterocytozoon bieneusi in cattle farmed in Hainan Province, the southernmost region of China. Parasite 27, 65.

All Tables

Table 1

ITS genotypes of Enterocytozoon bieneusi of natural infection identified in cattle worldwide.

Table 2

Prevalence and genotype distribution of E. bieneusi isolates in cattle in Hainan Province.

Table 3

ITS genotypes of natural Enterocytozoon bieneusi infections identified in cattle in China.

All Figures

thumbnail Figure 1

Specific locations where samples were collected in this study. ▲: Sampling points.

In the text
thumbnail Figure 2

Phylogenetic tree based on neighbor-joining (N-J) analysis of ITS sequences. Phylogenetic relationships between the E. bieneusi genotypes identified in cattle here and other known genotypes deposited in GenBank were inferred by an N-J analysis of ITS sequences based on genetic distance by the Kimura two-parameter model. The numbers on the branches are percent bootstrapping values from 1000 replicates. Each sequence is identified by its accession number, host origin, and genotype designation. Enterocytozoon bieneusi genotype CSK2 (KY706128) was used as the outgroup. The squares and triangles filled in black indicate novel and known genotypes identified in this study, respectively.

In the text

Current usage metrics show cumulative count of Article Views (full-text article views including HTML views, PDF and ePub downloads, according to the available data) and Abstracts Views on Vision4Press platform.

Data correspond to usage on the plateform after 2015. The current usage metrics is available 48-96 hours after online publication and is updated daily on week days.

Initial download of the metrics may take a while.