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Parasites & Vectors

, 11:204 | Cite as

First report of Giardia duodenalis and Enterocytozoon bieneusi in forest musk deer (Moschus berezovskii) in China

  • Yuan Song
  • Wei Li
  • Haifeng Liu
  • Zhijun Zhong
  • Yan Luo
  • Yao Wei
  • Wenlong Fu
  • Zhihua Ren
  • Ziyao Zhou
  • Lei Deng
  • Jianguo Cheng
  • Guangneng Peng
Open Access
Short report
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Abstract

Background

Giardia duodenalis and Enterocytozoon bieneusi are widespread pathogens that can infect humans and various animal species. Thus far, there are only a few reports of G. duodenalis and E. bieneusi infections in ruminant wildlife. Thus, the objective of this study was to examine the prevalence of G. duodenalis and E. bieneusi in forest musk deer in Sichuan, China, as well as identifying their genotypes.

Results

In total, we collected 223 faecal samples from musk deer at the Sichuan Institute of Musk Deer Breeding in Dujiangyan (n = 80) and the Maerkang Breeding Institute (n = 143). Five (2.24%) faecal samples were positive for G. duodenalis; three belonged to assemblage E, and two belonged to assemblage A based on the sequence analysis of the β-giardin (bg) gene. One sample each was found to be positive based on the glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) gene, respectively. Thirty-eight (17.04%) faecal samples were found to be E. bieneusi-positive based on the internal transcribed spacer (ITS) sequence, and only SC03 genotype was identified, which belonged to the zoonotic group 1 according to the phylogenic analysis. The infection rates were significantly different among the different geographical areas and age groups but had no apparent association with gender or clinical symptoms.

Conclusions

To our knowledge, this was the first molecular characterisation of G. duodenalis and E. bieneusi in musk deer. Identification of the zoonotic genotypes indicated a potential public health threat, and our study suggested that the forest musk deer is an important carrier of these parasites.

Keywords

G. duodenalis E. bieneusi Zoonotic pathogens Musk deer China 

Abbreviations

ITS

Internal transcribed spacer

MLG

Multilocus genotype

MLGs

Multilocus genotypes

MLST

Multilocus sequence typing

SNPs

Single nucleotide polymorphisms

Background

Giardia spp. are parasites with a broad host range comprising economic, companion, and wildlife animals, ranging from mammals to amphibians and birds, and humans [1, 2]. These parasites can have various clinical manifestations such as diarrhoea and abnormalities in growth and development, particularly in young hosts. For example, giardiasis can develop into malabsorption syndromes and other chronic diseases, resulting in stunted growth or emaciation in children [3]. According to WHO, approximately 200 million people in Africa, Asia and Latin America have symptomatic Giardia infection [4]. Enterocytozoon bieneusi is another common intestinal parasite that infects the host’s enterocytes, causing gastrointestinal illness such as chronic diarrhoea in animals and humans, particularly in immunosuppressed groups, including organ-transplant recipients, children, the elderly, and patients with cancer, diabetes, or AIDS [5, 6]. Ingestion of water and food contaminated with oocyst-containing faeces is the principal route of transmission for these species [7].

Giardia duodenalis is the only species of Giardia infecting humans and is comprised of eight assemblages (A-H). Among them, assemblages A and B have a broad host range and zoonotic potential. In particular, subtypes A1, A2, A3, A4, B1, and B4 are closely associated with human infections. In contrast, assemblages C-H have strong host specificity and a narrow host range [1, 8, 9]. Approximately 90% of human microsporidiosis cases are caused by E. bieneusi [10, 11]. In addition to its detection in humans, E. bieneusi has been reported in various economic animals and wildlife, including snakes and birds [12, 13, 14]. Currently, over 240 genotypes of E. bieneusi have been identified and divided into eight groups (groups 1–8). Most genotypes in group 1 have zoonotic potential, whereas the other groups have narrow host range and higher host specificity [15].

As an endangered species, musk deer (Moschus spp.) is currently considered a class I-protected animal in China. Forest musk deer (Moschus berezovskii) is the largest species of musk deer and mainly found in the Sichuan and Guizhou provinces of China [16, 17]. Musk, which has a remarkably high economic and medicinal value, is secreted by the musk gland located in the groin of male forest musk deer [18]. Because of their pathological effects on forest musk deer, infection with G. duodenalis or E. bieneusi can result in a significant loss of musk yield. This study aimed to investigate the presence of these parasites in musk deer, which may pose a threat to the health of both forest musk deer and humans.

Methods

Fecal sample collection

In February 2017, 223 faecal samples were collected from forest musk deer at the Sichuan Institute of Musk Deer Breeding located in Dujiangyan and Maerkang, in the Sichuan Province of China. Immediately after defecation, fresh faecal samples were collected using sterile disposable latex gloves, numbered, and placed in individual plastic bags. During specimen collection, we only gathered the top layer of the faeces to ensure that there was no contamination. All samples were placed on ice in separate containers and immediately transported to the laboratory. Specimens were stored in 2.5% potassium dichromate at 4 °C in a refrigerator until analysis.

DNA extraction and nested PCR amplification

Faecal samples were washed with distilled water and centrifuged at 3000× g for 3 min. This process was repeated three times. Genomic DNA was then extracted from approximately 200 mg of each semi-purified product, using the E.Z.N.A. Stool DNA Kit (D4015–02; Omega Bio-Tek, Norcross, GA, USA). DNA samples were stored in 200 μl of the kit Solution Buffer at -20 °C until use.

G. duodenalis and E. bieneusi were identified using nested PCR amplification of the β-giardin (bg) gene and internal transcribed spacer (ITS) sequence, respectively. bg-positive specimens were subjected to further amplification of the glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes, whereas ITS-positive specimens were subjected to amplification of three microsatellites (MS1, MS3, and MS7) and one minisatellite (MS4). The primers and amplification conditions were as previously described [19, 20, 21] (Table 1). Each reaction was performed in a total volume of 25 μl that included 12.5 μl 2× Taq PCR Master Mix (KT201-02; Tiangen, Beijing, China), 8.5 μl deionized water (Tiangen), 2 μl DNA, and 1 μl each of upstream and downstream primers. Positive and negative controls were included in each test. Secondary PCR products were subjected to 1% agarose gel electrophoresis.
Table 1

Primers and annealing temperature for the identification of G. duodenalis and E. bieuensi

Gene locus

Primer sequences (5’–3’)

Annealing temperature (°C)

Expected product size (bp)

Reference

bg

F1: AAGCCCGACGACCTCACCCGCAGTGC

50

511

[19]

R1: GAGGCCGCCCTGGATCTTCGAGACGAC

   

F2: GAACGAACGAGATCGAGGTCCG

60

  

R2: CTCGACGAGCTTCGTGTT

   

gdh

F1: TTCCGTRTYCAGTACAACTC

50

530

[19]

R1: ACCTCGTTCTGRGTGGCGCA

   

F2: ATGACYGAGCTYCAGAGGCACGT

65

  

R2: GTGGCGCARGGCATGATGCA

   

tpi

F1: AAATIATGCCTGCTCGTCG

55

530

[19]

R1: CAAACCTTITCCGCAAACC

   

F2: CCCTTCATCGGIGGTAACTT

55

  

R2: GTGGCCACCACICCCGTGCC

   

ITS

F1: GATGGTCATAGGGATGAAGAGCTT

55

392

[20]

R1: AATACAGGATCACTTGGATCCGT

   

F2: AGGGATGAAGAGCTTCGGCTCTG

55

  

R2: AATATCCCTAATACAGGATCACT

   

MS1

F1: CAAGTTGCAAGTTCAGTGTTTGAA

58

676

[21]

R1: GATGAATATGCATCCATTGATGTT

   

F2: TTGTAAATCGACCAAATGTGCTAT

58

  

R2: GGACATAAACCACTAATTAATGTAAC

   

MS3

F1: CAAGCACTGTGGTTACTGTT

55

537

[21]

R1: AAGTTA GGGCATTTAATAAAATTA

   

F2: GTTCAAGTAATTGATACCAGTCT

55

  

R2: CTCATTGAATCTAAATGTGTATAA

   

MS4

F1: GCATATCGTCTCATAGGAACA

55

885

[21]

R1: GTTCATGGTTATTAATTCCAGAA

   

F2: CGAAGTGTACTACATGTCTCT

55

  

R2: GGACTTTAATAAGTTACCTATAGT

   

MS7

F1: GTTGATCGTCCAGATGGAATT

55

471

[21]

R1: GACTATCAGTATTACTGATTATAT

   

F2: CAATAGTAAAGGAAGATGGTCA

55

  

R2: CGTCGCTTTGTTTCATAATCTT

   

Nucleotide sequencing and analysis

Products of the expected size were sent for a two-directional sequencing analysis (Invitrogen, Shanghai, China). Assemblages and subtypes were determined by the alignment of the nucleotide sequences with known reference sequences for the bg, tpi, and gdh genes of G. duodenalis, and for the ITS, MS1, MS3, MS4, and MS7 sequences of E. bieneusi available in the GenBank database, using BLAST and Clustal X.

Neighbor-joining phylogenetic analysis of the aligned G. duodenalis and E. bieneusi sequences was utilised to assess genetic clustering of the genotypes. A total of 1000 replicates were used for the bootstrap analysis.

Nucleotide sequence GenBank accession numbers

All nucleotide sequences of the bg, gdh, and tpi genes of G. duodenalis, and ITS, MS1, MS3, MS4, and MS7 of E. bieneusi isolated from forest musk deer in this study were deposited in the GenBank database under accession numbers MF497406–MF497412 and MF942581–MF942596, respectively.

Results and discussion

G. duodenalis and E. bieneusi are emerging zoonotic pathogens. To our knowledge, this study is the first to report the presence of G. duodenalis and E. bieneusi in musk deer, with an infection rate of 2.24% (5/223) and 17.04% (38/223), respectively. G. duodenalis infection rate in the Dujiangyan breeding centre (3.75%) was slightly higher than that in the Maerkang breeding centre (1.40%), whereas E. bieneusi infection rate was much lower in Dujiangyan than in Maerkang (7.5% and 22.38%, respectively) (Table 2). This may be due to differences in the source of food and water used for feeding, or other environmental factors.
Table 2

Prevalence and distribution of G. duodenalis and E. bieneusi by location in Sichuan Province, China

Pathogen

Location (city)

No. of samples

No. positive (%)

Genotype (n)

G. duodenalis

Dujiangyan

80

3 (3.75)

assemblage E

Maerkang

143

2 (1.40)

assemblage A

Total

223

5 (2.24)

assemblage A (2); assemblage E (3)

E. bieneusi

Dujiangyan

80

6 (7.50)

SC03

Maerkang

143

32 (22.37)

SC03

Total

223

38 (17.04)

SC03

In this study, the infected forest musk deer ranged from less than one to eight years of age. Young individuals (≤ one-year old) accounted for more than half of the G. duodenalis- and E. bieneusi-positive samples (60% and 57.89%, respectively), which may be caused by incomplete development of the immune system of young animals compared with adult animals. The proportion of infected females and males was similar. Several infected animals had obvious diarrhoea (two and nine for G. duodenalis and E. bieneusi, respectively), which may be due to the individual’s low resistance to infection. There was no apparent age- or gender-associated difference for the infections in this study, in agreement with the findings of Zhang et al. [22]. Here, assemblage A of G. duodenalis was obtained only from young forest musk deer in the Maerkang breeding location, whereas assemblage E was obtained from adult forest musk deer in the Sichuan Institute of Musk Deer Breeding in Dujiangyan (Table 3).
Table 3

Genotypes of G. duodenalis isolates from musk deer in Dujiangyan and Maerkang in Sichuan Province, China at the bg, gdh and tpi loci

Isolate

bg

gdh

tpi

DL12

E

E

Neg

DL26

E

Neg

Neg

DL29

E

Neg

Neg

ML55

A

Neg

A

ML117

A

Neg

Neg

Abbreviation: Neg negative

Although the distribution of G. duodenalis in musk deer has not been reported, there are few reports of these parasites infecting other species in the families Cervidae and Bovidae, in the same suborder as the forest musk deer. G. duodenalis identified in these animals was mainly assemblage A, and in several studies, the rate of infection in these species was higher than that in forest musk deer in our study. For example, Lalle et al. [23] reported that the prevalence of G. duodenalis was 11.5% in fallow deer (Dama dama) which was also higher in fawns than in older deer, and the genotype was assemblage A. García-Presedo et al. [24] reported that 8.9% of roe deer (Capreolus capreolus) samples were positive for G. duodenalis, and the genotype was AII. In Norway, 12.3% of moose (Alces alces), 1.7% of red deer (Cervus elaphus), 15.5% of roe deer, and 7.1% of reindeer (Rangifer tarandus) were found to be infected with G. duodenalis [25]. In the United States, one white-tailed deer (Odocoileus virginianus) was found positive for G. duodenalis, and assemblage A was identified [26]. Solarczyk et al. [27] reported that the sub-assemblage of G. duodenalis found in red deer and roe deer was AIII and zoonotic AI, respectively. Also, sheep faecal specimens from China were found to be positive for G. duodenalis assemblage A genotype [28]. Given that G. duodenalis assemblage A was previously identified in humans, forest musk deer can play a role in transmitting G. duodenalis to humans.

In the E. bieneusi analysis, ITS sequencing showed that all E. bieneusi isolates from Maerkang and Dujiangyan were characterised as SC03 (n = 38), which had been previously found in sika deer (Cervus nippon) at zoological gardens in China [29]. Other reference sequences in the same phylogenetic branch were from parasites isolated from racoons in eastern Maryland in the United States, goats in China, and patients with HIV/AIDS in the Henan Province of China [20, 30]. Based on the phylogenetic analysis of the ITS sequence, E. bieneusi isolated in this study belonged to group 1 (subgroup 1d) (Fig. 1), which suggested their zoonotic potential.
Fig. 1

Phylogenetic relationships of the ITS gene nucleotide sequences of the E. bieneusi genotypes identified in this study and other reported genotypes. The genotypes in this study are indicated by triangles for Maerkang and squares for Dujiangyan

From the 38 ITS-positive specimens, there were nine, five, one, and three isolates successfully sequenced at MS1, MS3, MS4, and MS7 loci, respectively. Analysis of sequence polymorphisms and single nucleotide polymorphisms (SNPs) at MS3 locus revealed two distinct types (type I and II) (Table 4). Zhang et al. [31] reported that 7.06% (23/326) of sika deer were positive for E. bieneusi with eight genotypes detected. Also, 34.0% (16/47) of faecal samples from Père David’s deer (Elaphurus davidianus) in China were E. bieneusi-positive [32]. Another report showed that 29 deer were infected with E. bieneusi, including 28 sika deer and one red deer [33]. Zhao et al. [34] reported seven genotypes of E. bieneusi in golden takins (Budorcas taxicolor bedfordi) in China, and Shi et al. [20] found E. bieneusi in 28.8% (176/611) of goats and 42.8% (177/414) of sheep, with 42 genotypes identified. Twenty-three (7.0%) yaks in China were E. bieneusi-positive; three genotypes (BEB4, I, and J) from group 2 that were previously reported in humans and two group 1 genotypes were identified [11]. Seventeen E. bieneusi genotypes were identified in 26 (32.5%) white-tailed deer in the United States [26]. Therefore, the E. bieneusi genotype we identified in forest musk deer, and most E. bieneusi genotypes reported in the Cervidae and Bovidae can infect both humans and animals. However, E. bieneusi isolated from forest musk deer appeared to be from a single genotype, in contrast to those found in other deer species, yaks, and goats.
Table 4

Multi-locus sequence typing of E. bieneusi in musk deer

Code

Multi-locus sequence genotype

ITS

MS1

MS3

MS4

MS7

MLGs

LD1

SC03

ns

ns

ns

Type I

MLG1

LD48

SC03

Type I

ns

ns

ns

MLG2

LD49

SC03

ns

ns

Type I

ns

MLG3

LM50

SC03

Type I

ns

ns

Type I

MLG4

LM61

SC03

Type I

Type I

ns

ns

MLG5

LM81

SC03

ns

Type I

ns

ns

MLG6

LM83

SC03

ns

Type I

ns

ns

MLG6

LM97

SC03

ns

Type I

ns

ns

MLG6

LM102

SC03

Type I

ns

ns

ns

MLG7

LM103

SC03

Type I

ns

ns

ns

MLG7

LM104

SC03

Type I

ns

ns

ns

MLG7

LM105

SC03

Type I

ns

ns

ns

MLG7

LM123

SC03

Type I

Type II

ns

Type I

MLG8

LM131

SC03

Type I

ns

ns

ns

MLG7

Others

SC03

ns

ns

ns

ns

MLG9

Abbreviation: ns not successfully sequenced or unsuccessful, PCR amplification

Although the genetic heterogeneity of G. duodenalis and E. bieneusi is well described, their method of transmission is still not clear. Investigations on their epidemiology, detection methods, and diagnosis are required to provide experimental bases for ensuring the health and safety of both animals and humans.

Conclusions

This study demonstrated the prevalence of G. duodenalis and E. bieneusi in forest musk deer in China. Furthermore, to our knowledge, this is the first report of G. duodenalis and E. bieneusi infections in musk deer and thus demonstrating that the host range of these parasites is wider than previously reported. Zoonotic genotypes identified in this study showed the transmission potential of G. duodenalis and E. bieneusi from forest musk deer to humans or other animals. Currently, there is no known effective vaccine or drug to treat infection with these parasites. Hence, measures should be taken to prevent humans and animals from being infected by these parasites.

Notes

Acknowledgments

We thank Jie Xiao, Jing-xin Yao, Yu-ying Cao, Lian-yu Huang, Meng Wang, Lei-qiong Xiang, Xue Luo and Wen-ping Xia for DNA extraction and nested PCR amplification.

Funding

This study was supported by Chengdu Giant Panda Breeding Research Foundation (CPF2015-07, CPF2015-09, CPF2014-12).

Availability of data and materials

The datasets supporting the conclusions of this article are included within the article. Representative sequences are submitted to the GenBank database under accession numbers: MF497406–MF497412 and MF942581–MF942596.

Authors’ contributions

GP, WL and YS conceived and designed the research. JC, WF, YW and LD collected samples. YS, WL, HL and ZZ performed the experiments. YS, WL, YL and ZR analysed the data. YS wrote the paper. All authors read and approved the final manuscript.

Ethics approval and consent to participate

This study was performed in accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the Ministry of Health, China. Before experiments, the protocol of the current study was reviewed and approved by the Institutional Animal Care and Use Committee of the Sichuan Agricultural University under permit number DYY-S20174605. No animals were harmed during the sampling process. Permission was obtained from the relative institutions of the forest musk deer before the collection of faecal specimens.

Consent for publication

Not applicable

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. 1.
    Ryan U, Caccio SM. Zoonotic potential of Giardia. Int J Parasitol. 2013;43(12-13):943–56.CrossRefPubMedGoogle Scholar
  2. 2.
    Cunha MJRD, Cury MC, Santín M. Molecular identification of Enterocytozoon bieneusi, Cryptosporidium, and Giardia in Brazilian captive birds. Parasitol Res. 2017;116(2):487.CrossRefPubMedGoogle Scholar
  3. 3.
    Reynosorobles R, Poncemacotela M, Rosaslópez LE, Ramosmorales A, Martínezgordillo MN, Gonzálezmaciel A. The invasive potential of Giardia intestinalis in an in vivo model. Sci Rep. 2015;5:15168.CrossRefGoogle Scholar
  4. 4.
    Escobedo AA, Hanevik K, Almirall P, Cimerman S, Alfonso M. Management of chronic Giardia infection. Expert Rev Anti Infect Ther. 2014;12(9):1143–57.CrossRefPubMedGoogle Scholar
  5. 5.
    Matos O, Lobo ML, Xiao L. Epidemiology of Enterocytozoon bieneusi infection in humans. J Parasitol Res. 2012;2012(4):981424.Google Scholar
  6. 6.
    Didier ES, Weiss LM. Microsporidiosis: not just in AIDS patients. Curr Opin Infect Dis. 2011;24(5):490–5.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Ye J, Xiao L, Li J, Huang W, Amer SE, Guo Y, et al. Occurrence of human-pathogenic Enterocytozoon bieneusi, Giardia duodenalis and Cryptosporidium genotypes in laboratory macaques in Guangxi, China. Parasitol Int. 2014;63(1):132–7.Google Scholar
  8. 8.
    Minetti C, Taweenan W, Hogg R, Featherstone C, Randle N, Latham SM, et al. Occurrence and diversity of Giardia duodenalis assemblages in livestock in the UK. Transbound Emerg Dis. 2015;61(6):e60–7.CrossRefGoogle Scholar
  9. 9.
    Feng Y, Xiao L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev. 2011;24(1):110–40.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Zhao W, Zhang W, Yang D, Zhang L, Wang R, Liu A. Prevalence of Enterocytozoon bieneusi and genetic diversity of ITS genotypes in sheep and goats in China. Infect Genet Evol. 2015;32:265–70.CrossRefPubMedGoogle Scholar
  11. 11.
    Ma J, Cai J, Ma J, Feng Y, Xiao L. Enterocytozoon bieneusi genotypes in yaks (Bos grunniens) and their public health potential. J Eukaryot Microbiol. 2015;62(1):21–5.CrossRefPubMedGoogle Scholar
  12. 12.
    Karim MR, Yu F, Li J, Li J, Zhang L, Wang R, et al. First molecular characterisation of enteric protozoa and the human pathogenic microsporidian, Enterocytozoon bieneusi, in captive snakes in China. Parasitol Res. 2014;113(8):3041–8.CrossRefPubMedGoogle Scholar
  13. 13.
    Li W, Li Y, Li W, Yang J, Song M, Diao R, et al. Genotypes of Enterocytozoon bieneusi in livestock in China: high prevalence and zoonotic potential. PloS One. 2014;9(5):e97623.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Zhao W, Zhang W, Yang F, Cao J, Liu H, Yang D, et al. High prevalence of Enterocytozoon bieneusi in asymptomatic pigs and assessment of zoonotic risk at the genotype level. Appl Environ Microbiol. 2014;80(12):3699–707.Google Scholar
  15. 15.
    Wang L, Zhang H, Zhao X, Zhang L, Zhang G, Guo M, et al. Zoonotic Cryptosporidium species and Enterocytozoon bieneusi genotypes in HIV-positive patients on antiretroviral therapy. J Clin Microbiol. 2013;51(2):557–63.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Yang Q, Meng X, Xia L, Feng Z. Conservation status and causes of decline of musk deer (Moschus spp.) in China. Biol Conserv. 2003;109(3):333–42.CrossRefGoogle Scholar
  17. 17.
    Green MJB. Aspects of the ecology of the Himalayan Musk Deer. Cambridge: The University of Cambridge; 1985.Google Scholar
  18. 18.
    Homes V. On the scent: conserving musk deer - the uses of musk and Europe's role in its trade. Brussels: Traffic Europe; 1999.Google Scholar
  19. 19.
    Zhang XX, Zheng WB, Ma JG, Yao QX, Zou Y, Bubu CJ, et al. Occurrence and multilocus genotyping of Giardia intestinalis assemblage C and D in farmed raccoon dogs, Nyctereutes procyonoides, in China. Parasit Vectors. 2016;9(1):471.Google Scholar
  20. 20.
    Shi K, Li M, Wang X, Li J, Karim MR, Wang R, et al. Molecular survey of Enterocytozoon bieneusi in sheep and goats in China. Parasit Vectors. 2016;9(1):23.Google Scholar
  21. 21.
    Feng Y, Li N, Dearen T, Lobo ML, Matos O, Cama V, et al. Development of a multilocus sequence typing tool for high-resolution genotyping of Enterocytozoon bieneusi. Appl Environ Microbiol. 2011;77(14):4822–8.Google Scholar
  22. 22.
    Zhang W, Ren G, Zhao W, Yang Z, Shen Y, Sun Y, et al. Genotyping of Enterocytozoon bieneusi and subtyping of Blastocystis in cancer patients: relationship to diarrhoea and assessment of zoonotic transmission. Front Microbiol. 2017;8:1835.Google Scholar
  23. 23.
    Lalle M, DRA F, Poppi L, Nobili G, Tonanzi D, Pozio E, et al. A novel Giardia duodenalis assemblage A subtype in fallow deer. J Parasitol. 2007;93(2):426–8.Google Scholar
  24. 24.
    Ignacio GP, Susana PD, Marta GW, Mercedes M, Mercedes GB, Luis Miguel OM, et al. The first report of Cryptosporidium bovis, C. ryanae and Giardia duodenalis sub-assemblage A-II in roe deer (Capreolus capreolus) in Spain. Vet Parasitol. 2013;197(3-4):658–64.Google Scholar
  25. 25.
    Hamnes IS, Gjerde B, Robertson L, Vikoren T, Handeland K. Prevalence of Cryptosporidium and Giardia in free-ranging wild cervids in Norway. Vet Parasitol. 2006;141(1-2):30–41.Google Scholar
  26. 26.
    Santin M, Fayer R. Enterocytozoon bieneusi, Giardia, and Cryptosporidium infecting white-tailed deer. J Eukaryot Microbiol. 2015;62(1):34–43.Google Scholar
  27. 27.
    Solarczyk P, Majewska AC, Moskwa B, Cabaj W, Dabert M, Nowosad P. Multilocus genotyping of Giardia duodenalis isolates from red deer (Cervus elaphus) and roe deer (Capreolus capreolus) from Poland. Folia Parasitol. 2012;59(3):237–40.Google Scholar
  28. 28.
    Ye J, Xiao L, Wang Y, Guo Y, Roellig DM, Feng Y. Dominance of Giardia duodenalis assemblage A and Enterocytozoon bieneusi genotype BEB6 in sheep in Inner Mongolia, China. Vet Parasitol. 2015;210(3-4):235–9.Google Scholar
  29. 29.
    Li W, Deng L, Yu X, Zhong Z, Wang Q, Liu X, et al. Multilocus genotypes and broad host-range of Enterocytozoon bieneusi in captive wildlife at zoological gardens in China. Parasit Vectors. 2016;9(1):395.Google Scholar
  30. 30.
    Sulaiman IM, Fayer R, Lal AA, Trout JM, Iii FWS, Xiao L. Molecular characterization of microsporidia indicates that wild mammals harbor host-adapted Enterocytozoon spp. as well as human-pathogenic Enterocytozoon bieneusi. Appl Environ Microbiol. 2003;69(8):4495–501.Google Scholar
  31. 31.
    Zhang XX, Cong W, Liu GH, Ni XT, Ma JG, Zheng WB, et al. Prevalence and genotypes of Enterocytozoon bieneusi in sika deer in Jilin Province, northeastern China. Acta Parasitol. 2016;61(2):382–8.Google Scholar
  32. 32.
    Zhang Z, Huang J, Karim MR, Zhao J, Dong H, Ai W, et al. Zoonotic Enterocytozoon bieneusi genotypes in Père David's deer (Elaphurus davidianus) in Henan, China. Exp Parasitol. 2015;155:46–8.Google Scholar
  33. 33.
    Zhao W, Zhang W, Wang R, Liu W, Liu A, Yang D, et al. Enterocytozoon bieneusi in sika deer (Cervus nippon) and red deer (Cervus elaphus): deer specificity and zoonotic potential of ITS genotypes. Parasitol Res. 2014;113(11):4243–50. Google Scholar
  34. 34.
    Zhao GH, Du SZ, Wang HB, Hu XF, Deng MJ, Yu SK, et al. First report of zoonotic Cryptosporidium spp., Giardia intestinalis and Enterocytozoon bieneusi in golden takins (Budorcas taxicolor bedfordi). Infect Genet Evol. 2015;34:394–401.Google Scholar

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© The Author(s). 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors and Affiliations

  • Yuan Song
    • 1
  • Wei Li
    • 1
  • Haifeng Liu
    • 1
  • Zhijun Zhong
    • 1
  • Yan Luo
    • 1
  • Yao Wei
    • 1
  • Wenlong Fu
    • 2
  • Zhihua Ren
    • 1
  • Ziyao Zhou
    • 1
  • Lei Deng
    • 1
  • Jianguo Cheng
    • 2
  • Guangneng Peng
    • 1
  1. 1.The Key Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary MedicineSichuan Agricultural UniversityChengduChina
  2. 2.Sichuan Institute of Musk Deer BreedingDujiangyanChina

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