Indian Journal of Animal Research

  • Chief EditorM. R. Saseendranath

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.40

  • SJR 0.233, CiteScore: 0.606

  • Impact Factor 0.4 (2024)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Full Research Article

Occurrence and Zoonotic Evaluation of Cryptosporidium spp. and Enterocytozoon bieneusi in Dogs and Cats in Wenzhou, Southern China

Yongli Jian1, Suzhen Liu1, Qingsong Han1, Junke Song2, Peide Li1, Yongan Gao1, Yiqiang Tu1,*, Houqiang Luo1,*
  • 0000-0002-7030-7177, 0000-0002-2990-6012
1College of Animal Science, Wenzhou Vocational College of Science and Technology, Wenzhou 325006, People’s Republic of China.
2College of Veterinary Medicine, Northwest A and F University, Yangling 712100, People’s Republic of China.

ABSTRACT

Background: Cryptosporidium spp. and Enterocytozoon bieneusi are important enteric diarrheal pathogens commonly found in various animals and humans and have significant implications for public health. The purpose of present study was to investigate the prevalence, species and genotypes and zoonotic evaluation of the two parasitic pathogens in dogs and cats in Wenzhou, southern of China.

Methods: 89 fecal samples collected from 51 dogs and 38 cats were used to investigate the positive rates of Cryptosporidium spp. and Enterocytozoon bieneusi using the nested polymerase chain reaction amplification on methods, species and genotypes of them were conformed by sequencing and alignment.

Result: The positive rates of Cryptosporidium spp. and Enterocytozoon bieneusi were 7.8%(4/51), 17.6%(9/51) in dogs and 7.9%(3/38) ,10.5%(4/38) in cats, respectively. Sequence analysis revealed that Cryptosporidium canis, genotypes PtEb IX, CD9 and NED3 of E. bieneusi were confirmed in dogs, Cryptosporidium felis, Type IV in cats, respectively. C canis, C. felis and Type IV of E. bieneusi genotype belong to the zoonotic pathogens.The zoonotic species/genotype of these two pathogens identified in pets in this study indicated the potential zoonotic risk in pets-to- humans transmission. The effective interventions should be taken to prevent the spread of these enteric zoonotic pathogens.

INTRODUCTION

Cryptosporidium spp. and Enterocytozoon bieneusi are the most common enteric protists in humans, farms, companion animals and wild animals, mainly causing gastrointestinal issues, such as acute or chronic diarrhea, abdominal pain, malabsorption and irritable bowel syndrome, particularly in the hosts with compromised immune systems or getting sick (Feng et al., 2018; Prasertbun et al., 2019; Li et al., 2022; Jiang et al., 2023; Alarcón-Zapata  et al., 2023). The most important reasons for these pathogens infection are due to the fecal-oral transmission route, including the intimate contact with high-risk hosts infected with  zoonotic pathogens and ingestion of fecal-contaminated food or water (Ryan et al., 2018; Li et al., 2019; Gururajan et al., 2021; Swain et al., 2018).
       
Currently, at least 45 species and over 120 genotypes of Cryptosporidium have existed in animals and humans (Ryan et al., 2021; Yang et al., 2021), C. canis and C. felis are among the five zoonotic Cryptosporidium species,while the rest are host-specific pathogens (Yang et al., 2021). Similarly, there are over 500 E. bieneusi genotypes which are classified into 13 groups with divergent genetic diversity (Li and Xiao, 2020; Meng et al., 2023). Type IV, C and D genotypes in group 1 are commonly reported in animals and humans, suggesting the zoonotic importance (Santín and Fayer, 2011). BEB4, I and BEB6 genotypes in group 2 have been observed in broad range of hosts, indicating an increasing zoonotic risk (Li et al., 2019). However, the most genotypes in group 3-11 are host-specific, with low zoonotic potential (Li et al., 2020; Yang et al., 2023).
       
There are many records about Cryptosporidium spp. and E.bieneusi present in a variety of companion animals,including dogs and cats. While the most common identified species are Cryptosporidium canis in dogs and C. felis in cats (Barbosa et al., 2023), other species, like C. muris, C. parvum, C. ubiquitum and C. hominis were sporadically detected in these companion animals (Xu et al., 2016). Type IV, D, PtEb IX and EbpC were the predominant E. bieneusi genotypes in dogs (Cao et al., 2021), while genotypes I, Type IV and BEB6 were widely distributed in cats (Karim et al., 2014; Wang et al., 2021).

There have been much research on the prevalence and zoonotic potential of E. bieneusi and Cryptosporidium spp. broadly conducted in dogs and cats in various countries, including regions of China such as Xinjiang (Cao et al., 2021), Henan (Karim et al., 2014), Guangdong (Wang et al., 2020) and Shanghai (Xu et al., 2016). Wenzhou, a region known for its economic vitality in southern of China, has a population of over 9 million humans and 500 thousand of pets. The warm and humid climate in Wenzhou provides favorable conditions for the dissemination of these pathogens. However, there is scarce information available about the occurrence, species of Cryptosporidium spp. and E. bieneusi genotypes in these companion animals in Wenzhou. Therefore, the purpose of this survey was to ascertain the prevalence, species of Cryptosporidium spp. and E. bieneusi genotypes in dogs and cats and to evaluate potential risk for public health.

MATERIALS AND METHODS

Specimen collection, DNA extraction and nested amplification
 
From September in 2023 to August in 2024, there were 89 fresh fecal samples collected from dogs and cats in Wenzhou, China. Among these, 51 were from dogs and 38 from cats, containing 11 fecal samples from dogs and 9 from cats aged <12 months respectively. Each fecal sample was collected from the dogs or cats’ rectum with the sterile plastic gloves and immediately transferred to laboratory of Wenzhou Vocational College of Science and Technology under the cool condition and then placed into plastic tube containing 2.5% potassium dichromate, finally kept at 4°C for DNA extraction. To remove the large fecal particles and potassium dichromate, each fecal sample was sieved through wire screen and washed until the supernatant was clean. Genomic DNA was isolated from processed fecal materials of each sample in accordance with the manufacturer’s instructions (Omega, Norcross, GA, USA). The obtained DNA samples were kept at -20°C.
       
A nested PCR amplification and DNA sequencing for the SSU rRNA gene of Cryptosporidium spp. were used to detect the positive rate and species compositions in dogs and cats, as previously reported (Ghebremichael et al., 2023). Similarly, a nested PCR techniques and sequence analysis of Enterocytozoon bieneusi ITS region were used to examine the positive rate and genotypes in fecal samples (Yu et al., 2023). The nested PCRs were conducted in a 25 µL reaction systems: 2.5 µL of 10 x PCR Buffer, 0.15 µL rTaq enzyme (TaKaRa Co., Ltd., Beijing, China), 2 µL dNTP, 2.0 μL MgCl2,1 µL of primers (10 µM each), 1 µL of obtained DNA sample and 15.35 µL of nuclease-free water. The obtained secondary PCR products were performed by electrophoresis and visualized with the nucleic acid staining.

Sequence analysis
 
The positive obtained PCR products were sent to Sangon Biotech (Shanghai, China) for bi-directional sequencing. Each sequence obtained from the positive samples in present study was firstly assembled and edited using the tools online and then compared with the nucleotide sequence database in GenBank. To assess the relationships of Cryptosporidium spp. and E. bieneusi found in this study, phylogenetic trees were performed using MEGA V11.0 based on the maximum likelihood (ML) method and Bootstrap analysis calculated with 1000 replicates.

RESULTS AND DISCUSSION

In this investigation, out of the 51 fecal samples analyzed, 4 (7.8%, 4/51) tested positive for Cryptosporidium spp. and 9 (17.6%, 9/51) tested positive for E. bieneusi in dogs. While the positive rates of Cryptosporidium varied in different sites, age groups and deworming statuses, no significant differences were observed. Similarly, although the positive rates of E. bieneusi also varied, with notably high rates in the group without deworming (42.9%, 3/7), the differences for positive rates of E. bieneusi were not statistically significant among the different sites, age groups and deworming statuses. Notably, no positive samples for E. bieneusi were found in dogs from pet market (Table 1).

Table 1: Occurrence, distribution, species of Cryptosporidiumspp. and genotypes of E. bieneusi genotypesin dogs in Wenzhou, southern China.


       
3 out of 38 fecal samples (7.9%, 3/38) tested positive for Cryptosporidium spp. and 4 samples (10.5%, 4/38) tested positive for E. bieneusi in cats. While the positive rates for Cryptosporidium and E. bieneusi varied based on site, age and deworming status, the highest positive rate for Cryptosporidium spp. was observed in hospitals (11.8%, 2/17) and for E. bieneusi in no deworming group (22.2%, 2/9). However, no significant differences were found among different sites, age groups and deworming statuses. Surprisingly, cats in pet market and aged<12 months were neither infected with Cryptosporidium spp. nor E. bieneusi (Table 2).

Table 2: Occurrence, distribution, species of Cryptosporidium spp. and genotypes of E. bieneusi genotypes in cats in Wenzhou, southern China.


       
All 4 Cryptosporidium-positive samples in dogs were successfully sequenced and identified as C. canis. Among the 4 C. canis isolates identified in present study, 3 were identical to the reference sequence (MN696800.1) from dogs in Guangzhou, southern China, with one nucleotide deletion.Meanwhile, there were two nucleotide deletions, a C to G substitution and two C to A nucleotide substitutions in the remaining isolate compared with this reference sequence (MN696800.1). Out of the 3 positive samples were sequenced for Cryptosporidium spp., only C. felis was identified. Among the 3 C. felis isolates, one was identical to a reference sequence from UK (OP935207.1) with 100% homology. One was 99% identical to an isolate from a cat in China (KJ194110.1), with one single nucleotide substitution, the last isolate had 99% similarity to another reference sequence (KX168415.1) (Fig 1).

Fig 1: Phylogenetic relationship of Cryptosporidium spp. identified (the filled bluecircle) in present study with the nucleotide sequence downloaded in database.


       
Of the 9 E. bieneusi-positive samples sequenced successfully in dogs, three known genotypes, namely PtEb IX, CD 9 and NED 3genotypes, were identified using the ITS gene locus sequencing method. PtEb IX (66.7%, 6/9)was the most predominant genotype found in dog isolates across all groups except the pet market. CD9 (22.2%, 2/9) was detected in samples from shelters and hospitals, while genotype NED3 (11.1%, 1/9) was only found in one specimen. Type IV was the only genotype identified in the 4 positive samples analyzed for E. bieneusi in cats (Fig 2).

​

Fig 2: Phylogenetic relationship of Enterocytozoon bieneusi obtained (the filled blue circle)with representative E. bieneusi genotypes deposited in GenBank conducted based above-mentioned method.


       
Analysis from the perspective of concurrent infections, there was one positive-sample co-infected with C. canis and NED3 genotype of E. bieneusi in dogs, two positive-samples were concurrently infected with Type IV genotype of E. bieneusi and C. felis in cats, the remaining positive-samples were mono-infections.
       
Cryptosporidium spp. and E. bieneusi are crucial zoonotic pathogens widely found in humans and various animals across different regions. Dogs and cats, commonly kept as pets in many households, have close contact with humans and could serve as important reservoirs of zoonotic pathogens, posing a potential risk for human health.
       
The results of this investigation revealed that the positive rate of Cryptosporidium spp. was 7.8% (4/51) in dogs and 7.9%  (3/38) in cats respectively, which was higher than that in Poland (3.4%, 2%) (Piekara-Stêpiñska  et al., 2021), Egypt (1.8%, 6.0%) (Elmahallawy et al., 2023), Yunnan in China (4.6%, 0.6%) (Wang et al., 2021), but was consistent with that in Brazil (7.8%, 5.4%) (de Oliveira et al., 2021), in Turkey (7.14%, 9.09%), (Ipek, 2023), in Saudi Arabia (8.5%) (Malk, 2021), Guangdong (6.9%, 6.2%) (Li et al., 2019), Shanghai (8.0%, 3.8% (Xu et al., 2016) and other regions in China.The discrepancy in Cryptosporidium infection in pets may be related to factors, like sample sizes,detection methods, geographic locations and living conditions.
       
Enterocytozoon bieneusi has been increasingly found in human and broad animals, including diverse intimate companions. In this survey, the positive rate of E. bieneusi was 17.6% in dogs and 10.5% in cats respectively, which was comparable to the positive rate of dogs (18.8%, 136/724) and cats (14.1%, 22/156) examined respectively in Sichuan province in China (Zhong et al., 2021), but was much higher than that in dogs (0.8%, 2/237) and cats( 3.0%, 3/99) in Northern Spain (Dashti et al., 2019) and was much lower than that in stray cats in Türkiye (50.15%, 170/339) (Erkunt et al., 2023). Variations in E. bieneusi prevalence among dogs and cats in different studies can be attributed to various risk factors such as living habits, age, sex,sample sizes and sources of the animals.
       
The findings, only C. canis identified in dogs and C. felis in cats respectively in present survey, were consistent with previous research. However, in some other recent records, there were presence of several other Cryptosporidium species occasionnally infecting dogs and cats, like Cryptosporidium parvum, Cryptosporidium ubiquitum, Cryptosporidium muris and the Cryptosporidium rat genotype IV (Gil et al., 2017; Li et al., 2019). C. canis and C. felis, generally regarded as host-specific pathogens, were commonly detected in some human cases worldwide and mainly found in immunocompromised individuals and low-income countries (Yang et al., 2021). The result suggests that people should take protective measures to minimize zoonotic pathogen transmission from their pets.
       
There were 4 genotypes of E. bieneusi, PtEb IX, CD9, NED3 in dogs and Type IV in cats identified in this study. PtEb IX, which was the main E. bieneusi genotype and regarded as the dog-specific with widespread globally in dogs (Xu et al., 2016; Wang et al., 2020), was also the dominant genotype (6/9) in present survey. However, CD9 was the second dominant genotype in this survey (2/9), which was also the predominant E. bieneusi genotype in Sichuan province (Zhong et al., 2021). NED3, identified in this study,is recognized as a dog-adapted genotype.
       
Type IV was the sole genotype identified in adult cats in this study, this consequence was also consistent with its prevalence in cats from other sources (Sürgeç  et al., 2023).This genotype, known for its pathogenicity in humans and global presence, has also been found in a wide variety of animals, such as Lynx pardinus (Ávalos  et al., 2024), foxes and raccoon dogs (Zhang et al., 2021; Chen et al., 2023), zoo animals (Zhang et al., 2021), pigeons (Holubová  et al., 2024) and domestic animals. Moreover, there were some reports about Type IV of E. bieneusi infections in humans in Shanghai (Jiang et al., 2023), Ningbo (Liu et al., 2023), southern China and northeast Egypt respectively (Naguib et al., 2022). These findings suggest that animals carrying zoonotic Type IV genotype may serve as significant sources of human microsporidiosis. In contrast, the PtEb IX genotype was predominantly found in cats in other research (Wang et al., 2020).

CONCLUSION

The molecular epidemiological survey of Cryptosporidium spp. and E. bieneusi in dogs and cats in Wenzhou, southern China, revealed positive rates of 7.8% (4/51) and 17.6% (9/51) for Cryptosporidium spp. and E. bieneusi in dogs and 7.9% (3/38), 10.5% (4/38) in cats, respectively. Cryptosporidium canis, genotypes PtEb IX, CD 9 and  NED3 of E. bieneusi were confirmed in dogs, Cryptosporidium felis, Type IV in cats, respectively, C. canis, C. felis and Type IV of E. bieneusi genotype belong to the zoonotic pathogens. These findings suggest the need for effective interventions to minimize cross-infection of these two pathogens between pets and humans. Further research should explore the prevalence and species/genotypes of these two pathogens in pet owners, veterinarians and other animals in the same habitat and assess the potential public health risks.

ACKNOWLEDGEMENT

The present study was supported byWenzhou Engineering Research Center of Pet (WP01), the Service technology innovation projects of Wenzhou Association for Science and Technology (kjfw08) and Wenzhou Science and Technology Commissioner Projects (X2023074).
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
   
Informed consent
 
All animal procedures for experiments were approved by the Committee of Experimental Animal care and handling techniques were approved by the Wenzhou Vocational College of Science and Technology of Animal Care Committee.
 

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

REFERENCES


  1. Alarcón-Zapata, Antonio, M., Romero, S.D., Chaparro, G., Jovana, J., González, H., Milagros, O.C., Melina, M. and Serrano, S.A. (2023). Frequency of Giardia spp. and Cryptosporidium spp. in domestic and captive wild animals in the north of Veracruz, Mexico. Pakistan Veterinary Journal. 43: 814-818. 

  2. Ávalos, G., Caballero, G.J., Matas, M.P., Castro, S.S., Jiménez, M.D., Köster, P.C., Santín, M., Bailo, B., Cano, T.D., González, B.D., Mateo, M., García, B.I., Dashti, A., Nájera,  F. and Carmena, D. (2024). Detection and genotyping of  zoonotic microsporidia in the endangered Iberian lynx (Lynx pardinus). Medical Mycology. 62: myae027.

  3. Barbosa, A.D.,Egan, S.,Feng, Y., Xiao, L. and Ryan, U.(2023). Cryptosporidium and Giardia in cats and dogs: What is the real zoonotic risk?Current Research in Parasitology and Vector-Borne Diseases. 4: 100158. 

  4. Cao,Y., Tong, Q., Zhao, C., Maimaiti, A., Chuai, L., Wang, J., Ma, D.and Qi, M. (2021). Molecular detection and genotyping of Enterocytozoon bieneusi in pet dogs in Xinjiang, Northwestern China. Parasite. 28: 57. 

  5. Chen, M.H., Wang, H.D., Li, X., Guo, Y., Lu,Y., Zheng, L., Liang, G., Sui,Y., Wang, B., Dai, H., Dong, H. and Zhang, L. (2024). Molecular epidemiology of Enterocytozoon bieneusi from

  6. foxes and raccoon dogs in the Henan and Hebei provinces in China. BMC Veterinary Research. 20(1): 53.

  7. Dashti, A., Santín, M., Cano, L., de Lucio, A., Bailo, B., de Mingo, M.H., Köster, P.C., Fernández-Basterra, J.A., Aramburu-Aguirre, J., López-Molina, N., Fernández-Crespo, J.C., Calero- Bernal,R. and Carmena, D. (2019). Occurrence and genetic diversity of Enterocytozoon bieneusi (Microsporidia) in owned and sheltered dogs and cats  in Northern Spain. Parasitology Research. 118: 2979- 2987. 

  8. De Oliveira, A.G., Sudré, A.P., Bergamo, T.C.and Carneiro, H.L. (2021). Molecular characterization of Cryptosporidium spp. in dogs and cats in the city of Rio de Janeiro, Brazil, reveals potentially zoonotic species and genotype. PLoS One.16: e0255087. 

  9. Elmahallawy, E.K., Gareh, A., Abu-Okail, A., Köster, P.C., Dashti, A., Asseri, J., Gouda, A.A., Mubaraki, M.A., Mohamed, S.A., Mohamed, Y.M., Hassan, E.A., Elgendy, M., Hernández- Castro, C., Bailo, B., González-Barrio, D., Xiao, L.H. and Carmena, D. (2023). Molecular characteristics and zoonotic potential of enteric protists in domestic dogs and cats in Egypt. Frontiers in Veterinary Science. 10: 1229151.

  10. Erkunt, A.S.,Can, H.Y.,Deðirmenci, D.A.,Sürgeç, E., Güvendi, M., Ün, C., Döºkaya, M., Gürüz, A.Y.and Karakavuk, M. (2023). Molecular prevalence of Enterocytozoon bieneusi in stray cats of Ýzmir, Türkiye. Comparative Immunology, Microbiology and Infectious Diseases. 100: 102037. 

  11. Feng, Y., Ryan, U.M. and Xiao, L. (2018).Genetic diversity and population structure of Cryptosporidium. Trends in Parasitology. 34(11): 997-1011.  

  12. Gil, H., Cano, L., de Lucio, A., Bailo, B., de Mingo, M.H., Cardona, G.A., Fernández-Basterra, J.A, Aramburu-Aguirre, J., López- Molina, N.and Carmena, D. (2017). Detection  and molecular diversity of Giardia duodenalis and Cryptosporidium spp. in sheltered dogs and cats in Northern  Spain. Infection, Genetics and Evolution. 50: 62-69. 

  13. Ghebremichael, S.T., Meng, X., Yang, Y. andegiorgish, A.K., Wu, Z., Chen, J., Wei, J., Li, T., Bao, J., Zhou, Z. and Pan, G. (2023). First identification and coinfection detection

  14. of  Enterocytozoon bieneusi, Encephalitozoon spp., Cryptosporidium spp. and Giardia duodenalis in diarrheic pigs in Southwest China. BMC Microbiology. 23(1): 334. Gururajan, A., Rajkumari, N., Devi, U. and Borah, P. (2021). Cryptosporidium and waterborne outbreaks - A mini review. Tropical Parasitology. 11(1): 11-15.

  15. Holubová, N., Zikmundová,V., Kicia, M., Zaj¹czkowska, ¯., Rajský, M., Koneèný, R., Rost, M., Mravcová, K., Sak, B.and Kváè, M. (2024). Genetic diversity of Cryptosporidium spp., Encephalitozoon spp. and Enterocytozoon bieneusi in feral and captive pigeons in Central Europe. Parasitology Research. 123(3):158. 

  16. Ipek, D.N.(2023). Molecular characterization and zoonotic significance of Cryptosporidium spp. and Giardia  duodenalis in asymptomatic adult stray cats and dogs in Turkey. Indian Journal of Animal Research. doi:10.18805/ IJAR.BF-1707.

  17. Jiang,Y., Liu,L., Yuan, Z., Liu, A., Cao, J.and Shen, Y. (2023). 

  18. Molecular identification and genetic characteristics of Cryptosporidium spp., Giardia duodenalis and Entero- cytozoon bieneusi in human immunodeficiency virus/ acquired immunodeficiency syndrome patients in Shanghai, China. Parasites and Vectors. 16(1): 53. 

  19. Karim, M.R., Dong, H., Yu, F., Jian, F., Zhang, L., Wang, R., Zhang, S., Rume, F.I., Ning, C.and Xiao, L. (2014). Genetic diversity in Enterocytozoon bieneusi isolates from dogs and cats in China: host specificity and public health implications. Journal of Clinical Microbiology. 52(9): 3297-3302. 

  20. Li, J., Dan, X., Zhu, K., Li, N., Guo, Y., Zheng, Z., Feng, Y.and Xiao, L. (2019). Genetic characterization of Cryptosporidium spp. and Giardia duodenalis in dogs and cats in Guangdong, China. Parasites and Vectors. 12(1): 571.

  21. Li, J., Shi, K., Sun, F., Li, T., Wang, R., Zhang, S., Jian, F., Ning, C. and Zhang, L. (2019). Identification of human pathogenic Enterocytozoon bieneusi, Cyclospora cayetanensis and Cryptosporidium parvum on the  surfaces of vegetables and fruits in Henan, China. International Journal of Food Microbiology. 307: 108292. 

  22. Li, W., Feng, Y.and Santin, M. (2019). Host Specificity of Entero- cytozoon bieneusi and public health implications.Trends in Parasitology. 35(6): 436-451. 

  23. Li, W., Feng, Y. and Xiao, L. (2022). Enterocytozoon bieneusi. Trends in Parasitology. 38(1): 95-96.

  24. Li, W. and Xiao, L. (2020). Ecological and public health significance of Enterocytozoon bieneusi. One Health. 12: 100209. 

  25. Li, W., Feng, Y. and Xiao, L. (2020). Diagnosis and molecular typing of Enterocytozoon bieneusi: The significant role of domestic animals in transmission of human microsporidiosis. Research in Veterinary Science. 133: 251-261.

  26. Liu, H., Ni, H., Liu, S., Shen,Y., Wang, R., Cao, J. and Yin, J. (2023). First report on occurrence and genotypes of Entero- cytozoon bieneusi, Cryptosporidium spp. and Cyclospora cayetanensis from diarrheal outpatients in Ningbo, Southeast China. Microbial Pathogenesis.174: 105952. 

  27. Malk, Al.S. (2021). Prevalence of Intestinal Parasite Infections in Stray Dogs in Taif Region, KSA. Indian Journal of Animal Research. 55(12): 1491-1497. doi: 10.18805/IJAR.BF-1421.

  28. Meng, X., Chu,W., Tang,Y., Wang,W., Chen,Y., Li,N., Feng,Y., Xiao,L. and Guo ,Y. (2023). High zoonotic potential and heavy environmental burden of Cryptosporidium spp. and Enterocytozoonbieneusi in farmed and pet African pygmy hedgehogs (Atelerixal biventris). One Health. 16:100532.

  29. Naguib, D., Roellig, D.M., Arafat, N. and Xiao, L. (2022). Prevalence and genetic characterization of Enterocytozoon bieneusi in children in Northeast Egypt. Parasitology Research. 121(7): 2087-2092.  

  30. Piekara-Stêpiñska, A., Piekarska, J. and Gorczy kowski, M.  (2021) Cryptosporidium spp. in dogs and cats in Poland. Annal of Agriculturaland Environmental Medicine. 28(2): 345-347. 

  31. Prasertbun, R., Mori, H., Sukthana,Y., Popruk, S., Kusolsuk, T., Hagiwara, K. and Mahittikorn, A.(2019). Enterocytozoon bieneusi and Cryptosporidium: A cross-sectional study conducted throughout Thailand. BMC Infectious Diseases. 19(1): 808. 

  32. Ryan, U. M., Feng, Y., Fayer, R. and Xiao, L. (2021). Taxonomy and molecular epidemiology of Cryptosporidium and Giardia- a 50 year perspective (1971-2021). International Journal for Parasitology. 51(13-14): 1099-119. 

  33. Ryan, U., Hijjawi, N. and Xiao, L. (2018). Foodborne cryptosporidiosis. International Journal for Parasitology. 48(1): 1-12.  

  34. Santín, M. and Fayer, R. (2011). Microsporidiosis: Enterocytozoon bieneusi in domesticated and wild animals. Research in Veterinary Science. 90(3): 363-371. 

  35. Sürgeç, E., Güvendi, M., Karakavuk, M., Erkunt, A.S., Deirmenci, D.A., Ün, C., Döskaya, M., Gürüz, A.Y. and Can, H. (2023). Genotyping of Enterocytozoon bieneusi isolates detected in stray cats of izmir, Türkiye. Parasitology Research. 122(11): 2729-2735.  

  36. Swain, K., Routray, A., Sahoo, S., Ganguly, S.(2018). Cryptosporidium oocyst shedding in buffalo calves in Haryana: A case study. Indian Journal of Animal Research. 53(2): 275- 277. doi: 10.18805/ijar.B-3487.

  37. Wang, H., Lin, X., Sun,Y., Qi, N., Lv, M., Xiao,W., Chen, Y., Xiang, R., Sun, M. and Zhang, L. (2020). Occurrence, risk factors and genotypes of Enterocytozoon bieneusi in dogs and cats in Guangzhou, southern China: High genotype diversity and zoonotic concern. BMC Veterinary Research. 16(1): 201. 

  38. Wang, Y., Zou,Y., Yu, Z., Chen, D., Gui, B., Yang, J., Zhu, X., Liu, G. and Zou, F. (2021). Molecular investigation of zoonotic intestinal protozoa in pet dogs and cats in yunnan province, Southwestern China. Pathogens. 10(9): 1107.

  39. Xu, H., Jin,Y., Wu, W., Li, P., Wang, L., Li, N., Feng, Y. and Xiao, L. (2016). Genotypes of Cryptosporidium spp., Entero- cytozoonbieneusi and Giardia duodenalis in dogs and cats in Shanghai, China. Parasites and Vectors. 9: 121.

  40. Yang, X., Guo, Y., Xiao, L. and Feng, Y. (2021). Molecular epidemiology of human cryptosporidiosis in low- and middle-income Countries. Clinical Microbiology Reviews. 34(2): e00087-19. 

  41. Yang, X., Wang, J., Huang, S., Song, J., Fan,Y. and Zhao, G. (2023).Molecular characterization of Cryptosporidium

  42. spp., Giardiaduodenalis, Enterocytozoon bieneusi 

  43. and Escherichia coli in dairy goat kids with diarrhea in partial regions of shaanxi province, China. Animals.13(18): 2922. 

  44. Yu, X., Wang, H., Li, Y., Mu, X., Yuan, K., Wu, A., Guo, J., Hong,Y. and Zhang, H. (2023). Occurrence and Genotypic Identification of Blastocystis spp., Enterocytozoon bieneusi 

  45. and Giardia duodenalis in Leizhou Black Goats in Zhanjiang City, Guangdong Province, China. Animals.13(17): 2777. 

  46. Zhang, K., Zheng, S., Wang, Y., Wang, K., Wang, Y., Gazizova, A., Han, K., Yu, F., Chen, Y. and Zhang, L. (2021). Occurrence and molecular characterization of Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi and Blasto- cystissp. in captive wild animals in zoos in Henan,China. BMC Veterinary Research. 17(1): 332. 

  47. Zhang, Y., Xin, L., Zhao, A., Xu, C., Wang, T., Jing, B. and Qi, M. (2021). Molecular detection and genotypes of Entero- cytozoonbieneusi in farmed mink (Neovisonvison), blue foxes (Alopex lagopus) and raccoon dogs (Nyctereute- sprocyonoides) in Xinjiang, China. International Journal for Parasitology: Parasites and Wildlife. 14: 211-215. 

  48. Zhong, Y., Zhou, Z., Deng, L., Liu, H., Zhong, Z., Ma, X., Zhang, K., Wang, Y., Fu, H. and Peng, G. (2021). Prevalence and new genotypes of Enterocytozoon bieneusi in sheltered dogs and cats in Sichuan province, southwestern China. Parasite. 28: 31.
Reviewed By

Download Article
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)