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All issues Volume 30 (2023) Parasite, 30 (2023) 45 Full HTML
Open Access
Issue
Parasite
Volume 30, 2023
Article Number 45
Number of page(s) 8
DOI https://doi.org/10.1051/parasite/2023048
Published online 02 November 2023

© W. Zhao et al., published by EDP Sciences, 2023

Licence Creative CommonsThis is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Introduction

Blastocystis sp. is a ubiquitous gut parasite that causes intestinal infections in humans and almost all animal species [12, 26]. The pathogenicity of Blastocystis sp. is controversial, and Blastocystis sp. infection is generally asymptomatic. Nonetheless, symptoms such as diarrhea, abdominal pain, weight loss, and bloating may occur in some patients [2]. Zoonotic, food and waterborne, and human-to-human transmission are the main routes through which Blastocystis sp. is transmitted [32]. In most instances, infection occurs via the fecal-oral route by drinking or eating the Blastocystis sp. cysts in contaminated food or water [21]. Identifying the source of Blastocystis sp. infections is key in preventing and controlling the spread of the parasite. However, there are limited data on the role of farmed animals in transmitting Blastocystis sp. to humans.

Molecular methods are widely used to detect Blastocystis sp. and are considered useful for identifying Blastocystis sp. reservoirs or hosts [29]. To date, approximately 40 Blastocystis sp. subtypes infecting humans and animals have been reported [12, 26, 30]. Rodents and carnivores may play important roles in the ecology and transmission of several Blastocystis sp. subtypes, including ST1–ST5, ST7, ST8, ST10, ST13, and ST17 (in rodents) and ST1–ST5, ST7, ST10, and ST14 (in carnivores) [12]. Except for ST13 and ST17, all the above Blastocystis sp. subtypes infecting rodents and carnivores have been detected in humans, suggesting that these animals are potential reservoirs of Blastocystis sp. [12].

Asiatic brush-tailed porcupines (Atherurus macrourus), bamboo rats (Rhizomys pruinosus), and masked palm civets (Paguma larvata) have recently been assessed for the IUCN Red List of Threatened Species, and in China, they are commonly farmed for food in Hainan Province. However, the role of these animals for Blastocystis sp. is not well understood. Therefore, this study used PCR to investigate the prevalence and genetic diversity of Blastocystis sp. in Asiatic brush-tailed porcupines, bamboo rats, and masked palm civets in Hainan Province to assess their zoonotic potential.

Material and methods

Ethics statement

This study was approved by the Research Ethics Committee and the Animal Ethical Committee of Wenzhou Medical University. Only excreted feces from the study animals were analyzed. No animal was injured during the research.

Sample collection

A total of 900 fecal samples were collected from 257 Asiatic brush-tailed porcupines, 360 bamboo rats, and 283 masked palm civets farmed in Hainan Province between December 1, 2017, and March 31, 2021 (Table 1). Asiatic brush-tailed porcupine samples were obtained from 6 farms in Baisha, Baoting, Lingshui, Wuzhishan, Dingan, and Qiongzhong; those of bamboo rats were sampled from 3 farms in Danzhou, Dingan, and Qiongzhong; and those of masked palm civets were sampled from 5 farms in Baisha, Dingan, Haikou, Lingshui, and Wuzhishan (Fig. 1). To minimize duplicate sampling, only one fecal sample (~15 g) was collected in each animal cage (each with 1 to 3 animals). Approximately 30–50% of animals on each farm were sampled. The farms were selected based on the willingness of the owners to participate in the study. All farms had a relatively good standard of environmental sanitation where animals are provided with clean tap water for drinking, 1 to 3 animals were kept in individual cages with minimal contact with other species. The management of animals on all fourteen farms was similar. Fresh fecal samples were collected in labeled sterile plastic bags and transported to the Key Laboratory of Tropical Translational Medicine of the Ministry of Education (Hainan Medical University) in containers packed with ice packs within 24 h and stored at 4 °C for processing within 12 h. None of the animals showed diarrhea at the time of sample collection.

thumbnail Figure 1

A map depicting the sampling sites for collecting fecal samples from Asiatic brush-tailed porcupines, bamboo rats and masked palm civets in Hainan Province, China.
Table 1

Prevalence and distribution of STs of Blastocystis in the three farmed animals in Hainan Province, China.

DNA extraction

Each fecal sample was sieved, and the filtrates were washed three times with distilled water and then centrifuged for 10 min at 1500 ×g to concentrate the parasites. Genomic DNA was extracted from 180–200 mg of clean fecal samples using a QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany), according to the manufacturer’s instructions. The extracted DNA was eluted in 200 μL of AE buffer and stored at −20 °C prior to PCR analysis.

PCR amplification

The bamboo rats were identified to the species level by amplification of a 421-bp region of the cytb gene from fecal DNA using PCR [33]. To detect Blastocystis sp., an approximately 500 base pair fragment of the SSU rDNA gene was amplified by PCR as described by Santin et al. [29]. TaKaRa Taq DNA Polymerase (TaKaRa Bio Inc., Tokyo, Japan) was used for all PCR amplifications. Negative controls were included in all PCR tests for quality control. The PCR products were subjected to 1.5% agarose gel electrophoresis and were visualized on a Gel Doc EZ UV-gel imaging system (Bio-Rad Inc., USA). The gel was stained with GelRed (Biotium Inc., Hayward, CA) to aid in visualization.

Nucleotide sequencing and analysis

The PCR products of interest were sequenced by Sangon Biotech Co., Ltd (Shanghai, China). Sequence accuracy was confirmed by two-directional sequencing and further sequencing of PCR products for some DNA samples where necessary. Sequences of each strand were edited and aligned using DNASTAR Lasergene EditSeq v7.1.0 (http://www.dnastar.com/) and Clustal X v2.1 (http://www.clustal.org/) tools. Reference sequences were downloaded from the GenBank.

Phylogenetic analysis

Mega 7 was used to produce a phylogenetic tree for nucleotide sequences using the maximum likelihood method. One thousand bootstrap replicates were used to test the phylogenetic tree’s reliability and the statistical support for the topology. Evolutionary distances were calculated using the Tamura-3 parameter model [16].

Nucleotide sequence accession numbers

The novel nucleotide sequence of Blastocystis ST5 obtained in this study was deposited in the GenBank database under accession number: OP132525. Meanwhile, all the other sequences which similarity of 100% with sequences available in GenBank obtained here were submitted to GenBank under accession number: OR593325 for ST1, OR593326 to OR593328 for ST4, OR593329 for unST, and OR593330 to OR593339 for ST5.

Statistical analyses

Data were analyzed using SPSS version 22.0 (SPSS Inc., Chicago, IL, USA). The chi-square test at 95% confidence interval was used to compare the prevalence of Blastocystis sp. among the different animal species. P < 0.05 was considered statistically significant.

Results

Infection rates of Blastocystis sp.

Blastocystis sp. was detected in 47 (5.2%) of the 900 fecal samples from 27 (9.5%) masked palm civets, 12 (4.7%) Asiatic brush-tailed porcupines, and 8 (2.2%) bamboo rats (Table 1). Notably, the differences in the prevalence rates of Blastocystis sp. among the three species and across the farms sampled were statistically significant (χ2 = 17.4, df = 2, p < 0.001). Positive fecal samples of Asiatic brush-tailed porcupines were from four of the six farms surveyed, with the highest infection rate (20.0%) in Wuzhishan, followed by Baoting, Lingshui, and Baisha (Table 1). Positive fecal samples of bamboo rats and masked palm civets were from one out of all the farms surveyed. The prevalence of Blastocystis sp. in bamboo rats in Danzhou farm was 7.0%, and 48.2% in masked palm civets in Lingshui farm (Table 1).

Sequencing of PCR amplicons

PCR and sequencing analysis of cytb amplicons showed that all the bamboo rats were Rhizomys pruinosus and all cytb gene sequences had 100% identity with the reference sequence KY754146. Meanwhile all the 47 PCR amplicons were sequenced to determine Blastocystis sp. subtypes. Nucleotide sequence analysis revealed the presence of three distinct previously reported subtypes (ST1, ST4, and ST5) and one unnamed subtype (unST); there were no mixed infections detected. Among them, ST5 was the most common subtype detected at 55.3% (26/47), followed by ST4 at 40.4% (19/47). The prevalence of ST1 and the unST were each at 2.1% (1/47). While ST4 was detected in both Asiatic brush-tailed porcupines and bamboo rats, ST1 and ST5 were detected in masked palm civets only, and the unST was detected in one Asiatic brush-tailed porcupine (Table 1).

Genetic diversity of Blastocystis subtypes

Except for one ST5 sequence (OP132525), which had a 99.8% identity with MK375243 isolated from a pig in Yunan Province, the rest of the 46 sequences have been previously described. The 25 ST5 displayed 100% genetic identity with previously reported subtypes: MN472792 (n = 7), MK244911 (n = 5), MK375238 (n = 3), MH634445 (n = 3), MK375235 (n = 2), MN493734 (n = 1), MK375243 (n = 1), MK375236 (n = 1), MK375241 (n = 1) and MK801407 (n = 1). A sequence alignment analysis of these ST5 sequences shows that there were 12 polymorphic sites among them (Fig. 2). A total of 19 nucleotide sequences were identified as ST4, with 13 of these from Asiatic brush-tailed porcupine (n = 5) and bamboo rats (n = 8) displaying 100% identity with Blastocystis sp. subtypes isolated from Rattus norvegicus in Japan (MH127500). The remaining six ST4 sequences were isolated from Asiatic brush-tailed porcupine and displayed 100% genetic identity with Blastocystis sp. subtype isolated from coypu in China (OK235459). ST1 and unST displayed an identity of 100% with a Blastocystis sp. subtype isolated in human from Mexico (KU147339) and a coypu from Henan Province (OK235452), respectively. Overall, nucleotide sequences of Blastocystis sp. subtypes obtained from this study clustered together with known subtypes in the phylogenetic tree except for the sequence of the unST, which fell into a distinct evolutionary branch (Fig. 3).

thumbnail Figure 2

Sequence variation in the small subunit (SSU) rRNA gene among of Blastocystis sp. ST5. SSU rRNA sequences of ST5 identified in this study were aligned. MN472792 as the reference sequence.
thumbnail Figure 3

Phylogenetic relationship of Blastocystis sp. STs based on SSU rRNA gene sequences. Clustal X and Mega 7 software were used to calculate genetic distance and build the phylogenetic tree. The phylogenetic tree was constructed using the maximum likelihood method, based on the evolutionary distances calculated by the Tamura-3 parameter model; all positions containing gaps and missing data were eliminated. Bootstrap values were obtained using 1,000 replicates. The blue and red fonts represent the known and novel sequences, respectively identified in this study.

Discussion

In the present study, Blastocystis sp. was detected in 5.2% (47/900) of animals, including 4.7% (12/257) Asiatic brush-tailed porcupine, 2.2% (8/360) bamboo rats, and 9.5% (27/283) masked palm civets sampled (Table 1). No study has reported on Blastocystis sp. in Asiatic brush-tailed porcupines and masked palm civets. Also, only one study has reported Blastocystis sp. infection in bamboo rats [31]. Unlike the low infection rate of 2.2% in bamboo rats sampled in this study, the single previous study on the same animal species from this area reported a higher prevalence rate of 4.58% [31]. Analysis of previous studies on the prevalence of Blastocystis sp. in wild and farmed animal species such as rodents and carnivores indicate geographical variations in infection rates, ranging from 3.1–100% [1, 3–10, 11, 13–15, 17–20, 22–25, 27, 28, 31, 34–36]. The differences in infection rates are possibly due to host variation in different regions. The prevalence of Blastocystis sp. in the two farmed rodents investigated in the present study was lower compared to other farmed rodents, such as in Myocastor coypus (14.3%) and in flying squirrels (30.4%) in China [9, 34], and in guinea pigs (35.4%) from Ecuador [11]. In addition to the animal species, the sample size, the health of the animals, the environment, geographical location, and the type of detection method may explain the variability in the prevalence rates. It is worth noting that only one study was done in 12 out of the 15 countries for rodents and seven out of eight countries for carnivores with extremely small sample sizes [1, 3–10, 11, 13–15, 17–20, 22–25, 27, 28, 31, 34–36]. The presence of Blastocystis sp. in rodents and carnivores is a public health concern, requiring large-scale investigations to assess their role in transmitting this parasite to humans.

Further, this study uncovered three known Blastocystis sp. subtypes (ST1, ST4, and ST5) and one unnamed subtype (unST). ST4 and ST5 were the predominant Blastocystis sp. subtypes (95.7%) detected in this study. Previous studies show that ST4 and ST5 are circulating in bamboo rats in the Hunan Province [31]. Moreover, Blastocystis sp. surveys in other studies have reported several other Blastocystis sp. subtypes, including potential zoonotic subtypes in rodents, coati, pandas, foxes, wolves, dogs, wild cats, and raccoons [1, 3–10, 11, 13–15, 17–20, 22–25, 27, 28, 31, 34–36]. The prior data show that ST5 is rare in carnivores, which is significantly different from the results of this study and reflects regional characteristics.

ST5 was the most prevalent Blastocystis sp. subtype in this study (55.3%). This subtype has previously been detected in multiple animal hosts, mostly pigs, with a 1.64% infection rate reported in human samples worldwide [26]. The ST5 prevalence rate in farmed masked palm civets suggests zoonotic transmission of this parasite from these animals to humans, pigs, and other animals, or, conversely, the ST5 harbored in the masked palm civets may also come from pigs or humans. Almost 95.0% of Blastocystis sp. detected in bamboo rats and porcupines were ST4. The global Blastocystis sp. ST4 prevalence in humans is 5.9%, although in Europe it is responsible for 19.8% of the cases reported [26]. This subtype has also been detected in rodents, birds, several deer species, and other wild mammals worldwide, indicating that it has a wide host range [27]. Given the prevalence of Blastocystis sp. ST4 in rodents (>17 rodent species), the subtype has possibly adapted to infect this type of host. Since most rodent species are peridomestic, they are considered potential sources of human Blastocystis sp. infections, needing extensive investigation. Although ST1 was only detected in one sampled animal (2.1%), the subtype is an important public health parasite because it is the second most common Blastocystis sp. subtype infecting humans and has been detected in animals worldwide [12, 26]. The unST detected in the present study has 92.4% identity with ST14. Given that this unST has been detected in a coypu in Henan [9], it potentially infects multiple hosts, including coypu and Asiatic brush-tailed porcupine.

Conclusion

This is the first report on the prevalence and genetic diversity of Blastocystis sp. in Asiatic brush-tailed porcupines, bamboo rats, and masked palm civets in Hainan Province. Our results indicate circulation of three known Blastocystis sp. subtypes (ST1, ST4, and ST5) and an unnamed Blastocystis sp. subtype in the animals mentioned above. These findings could provide a basis for preventing and controlling Blastocystis sp. infections in humans and the farmed animals in this region.

Conflict of interest

The authors do not have a commercial or other association that represents a conflict of interest.

Funding

This work was supported by the Department of Education Scientific Research Project of Zhejiang (Y202249687), National Natural Science Foundation of China (82060375), Research project of Hainan academician innovation platform (YSPTZX202004), Hainan talent development project (SRC200003), Major Science and Technology Program of Hainan Province (ZDKJ202003), Open Foundation of Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University (2020TTM004), High level talents project of Hainan Natural Science Foundation (822RC695) and, Science and Technology Plan of Hainan Province (Hainan Province Clinical Medical Center). The funding sponsors had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

#

These authors contributed equally to this work.

References

  1. AbuOdeh R, Ezzedine S, Madkour M, Stensvold CR, Samie A, Nasrallah G, AlAbsi E, ElBakri A. 2019. Molecular subtyping of Blastocystis from diverse animals in the United Arab Emirates. Protist, 170(5), 125679. [CrossRef] [PubMed] [Google Scholar]
  2. Asghari A, Sadeghipour Z, Hassanipour S, Abbasali Z, Ebrahimzadeh-Parikhani H, Hashemzaei M, Alimardani V, Hatam G. 2021. Association between Blastocystis sp. infection and immunocompromised patients: a systematic review and meta-analysis. Environmental Science and Pollution Research International, 28(43), 60308–60328. [CrossRef] [PubMed] [Google Scholar]
  3. Betts EL, Gentekaki E, Tsaousis AD. 2020. Exploring micro-eukaryotic diversity in the gut: Co-occurrence of Blastocystis subtypes and other protists in zoo animals. Frontiers in Microbiology, 11, 288. [CrossRef] [PubMed] [Google Scholar]
  4. Betts EL, Gentekaki E, Thomasz A, Breakell V, Carpenter AI, Tsaousis AD. 2018. Genetic diversity of Blastocystis in non-primate animals. Parasitology, 145(9), 1228–1234. [CrossRef] [PubMed] [Google Scholar]
  5. Calero-Bernal R, Santín M, Maloney JG, Martín-Pérez M, Habela MA, Fernández-García JL, Figueiredo A, Nájera F, Palacios MJ, Mateo M, Balseiro A, Barral M, Lima-Barberoi JF, Köster PC, Carmena D. 2020. Blastocystis sp. subtype diversity in wild carnivore species from Spain. Journal of Eukaryotic Microbiology, 67(2), 273–278. [CrossRef] [PubMed] [Google Scholar]
  6. Chai Y, Deng L, Liu H, Yao J, Zhong Z, Fu H, Shen L, Zhou Z, Deng J, Hu Y, Peng G. 2020. First subtyping of Blastocystis sp. from pet rodents in southwestern China. International Journal for Parasitology, Parasites and Wildlife, 11, 143–148. [CrossRef] [Google Scholar]
  7. Cian A, El Safadi D, Osman M, Moriniere R, Gantois N, Benamrouz-Vanneste S, Delgado-Viscogliosi P, Guyot K, Li LL, Monchy S, Noël C, Poirier P, Nourrisson C, Wawrzyniak I, Delbac F, Bosc S, Chabé M, Petit T, Certad G, Viscogliosi E. 2017. Molecular epidemiology of Blastocystis sp. in various animal groups from two French zoos and evaluation of potential zoonotic risk. PloS One, 12(1), e0169659. [CrossRef] [PubMed] [Google Scholar]
  8. Deng L, Yao JX, Liu HF, Zhou ZY, Chai YJ, Wang WY, Zhong ZJ, Deng JL, Ren ZH, Fu HL, Yan X, Yue CJ, Peng GN. 2019. First report of Blastocystis in giant pandas, red pandas, and various bird species in Sichuan province, southwestern China. International Journal for Parasitology, Parasites and Wildlife, 9, 298–304. [CrossRef] [Google Scholar]
  9. Deng L, Yao J, Chen S, He T, Chai Y, Zhou Z, Shi X, Liu H, Zhong Z, Fu H, Peng G. 2021. First identification and molecular subtyping of Blastocystis sp. in zoo animals in southwestern China. Parasites & Vectors, 14(1), 11. [CrossRef] [PubMed] [Google Scholar]
  10. Farah Haziqah MT, Mohd Zain SN, Chandrawathani P, Premaalatha B, Mohd Khairul Nizam M.K., Arutchelvan R., Suresh K.. 2018. Genetic diversity of rodent Blastocystis sp. from Peninsular Malaysia. Tropical Biomedicine, 35(2), 586–592. [PubMed] [Google Scholar]
  11. González-Ramírez LC, Vázquez CJ, Chimbaina MB, Djabayan-Djibeyan P, Prato-Moreno JG, Trelis M, Fuentes MV. 2021. Ocurrence of enteroparasites with zoonotic potential in animals of the rural area of San Andres, Chimborazo, Ecuador. Veterinary Parasitology, Regional Studies and Reports, 26, 100630. [CrossRef] [PubMed] [Google Scholar]
  12. Hublin JSY, Maloney JG, Santin M. 2021. Blastocystis in domesticated and wild mammals and birds. Research in Veterinary Science, 135, 260–282. [CrossRef] [PubMed] [Google Scholar]
  13. Kaczmarek A, Sobociński W, Wesołowska M, Gołąb E, Kołodziej-Sobocińska M, Sałamatin R. 2021. Blastocystis occurrence and subtype diversity in wild European terrestrial mammals – The case of Białowieża Primeval Forest (NE Poland). International Journal for Parasitology, Parasites and Wildlife, 16, 120–125. [CrossRef] [Google Scholar]
  14. Katsumata M, Yoshikawa H, Tokoro M, Mizuno T, Nagamoto T, Hendarto J, Asih PBS, Rozi IE, Kimata I, Takami K, Syafruddin D. 2018. Molecular phylogeny of Blastocystis isolates from wild rodents captured in Indonesia and Japan. Parasitology Research, 117(9), 2841–2846. [CrossRef] [PubMed] [Google Scholar]
  15. Köster PC, Dashti A, Bailo B, Muadica AS, Maloney JG, Santín M, Chicharro C, Migueláñez S, Nieto FJ, Cano-Terriza D, García-Bocanegra I, Guerra R, Ponce-Gordo F, Calero-Bernal R, González-Barrio D, Carmena D. 2021. Occurrence and genetic diversity of protist parasites in captive non-human primates, zookeepers, and free-living sympatric rats in the Córdoba zoo conservation centre, Southern Spain. Animals, 11(3), 700. [CrossRef] [PubMed] [Google Scholar]
  16. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. 2018. MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Molecular Biology and Evolution, 35(6), 1547–1549. [CrossRef] [PubMed] [Google Scholar]
  17. Li J, Yuan Y, Jiang Y, Wang W, Chao L, Sun R, Li J, Karim MR, Qi M. 2020. Molecular identification and subtyping of Blastocystis sp. in laboratory rats in China. Parasite, 27, 35. [CrossRef] [EDP Sciences] [PubMed] [Google Scholar]
  18. Li XD, Zou Y, Pan J, Liang QL, Zeng Z, Meng YM, Wang XL, Wang HN, Zhu XQ. 2020. Prevalence and subtypes of Blastocystis sp. infection in zoo animals in three cities in China. Parasitology Research, 119(2), 465–471. [CrossRef] [PubMed] [Google Scholar]
  19. Liu X, Ge Y, Wang R, Dong H, Yang X, Zhang L. 2021. First report of Blastocystis infection in Pallas’s squirrels (Callosciurus erythraeus) in China. Veterinary Research Communications, 45(4), 441–445. [CrossRef] [PubMed] [Google Scholar]
  20. Liu X, Ni F, Wang R, Li J, Ge Y, Yang X, Qi M, Zhang L. 2022. Occurrence and subtyping of Blastocystis in coypus (Myocastor coypus) in China. Parasites & Vectors, 15(1), 14. [CrossRef] [PubMed] [Google Scholar]
  21. Ma JY, Li MY, Qi ZZ, Fu M, Sun TF, Elsheikha HM, Cong W. 2022. Waterborne protozoan outbreaks: An update on the global, regional, and national prevalence from 2017 to 2020 and sources of contamination. Science of the Total Environment, 806(Pt 2), 150562. [CrossRef] [Google Scholar]
  22. Martinez-Hernandez F, Martinez-Ibarra JA, Lopez-Escamilla E, Villanueva-Garcia C, Muñoz-Garcia CI, Rendon-Franco E, Maravilla P, Villalobos G. 2020. Molecular genotyping of Blastocystis spp. in wild mammals from Mexico. Parasitology Research, 119(1), 97–104. [CrossRef] [PubMed] [Google Scholar]
  23. Masuda A, Wada M, Saho H, Tokunaga K, Kikuchi Y, Yamasaki F, Matsumoto J. 2021. Prevalence and molecular characterization of the zoonotic enteric protozoans Cryptosporidium spp., Enterocytozoon bieneusi, and Blastocystis from Pallas’s Squirrels (Callosciurus erythraeus) in Kanagawa Prefecture, Japan. Microbiology. Spectrum, 9(3), e0099021. [Google Scholar]
  24. Mohammadpour I, Bozorg-Ghalati F, Gazzonis AL, Manfredi MT, Motazedian MH, Mohammadpour N. 2020. First molecular subtyping and phylogeny of Blastocystis sp. isolated from domestic and synanthropic animals (dogs, cats and brown rats) in southern Iran. Parasites & Vectors, 13(1), 365. [CrossRef] [PubMed] [Google Scholar]
  25. Oliveira-Arbex AP, David ÉB, Tenório MDS, Cicchi PJP, Patti M, Coradi ST, Lucheis SB, Jim J, Guimarães S. 2020. Diversity of Blastocystis subtypes in wild mammals from a zoo and two conservation units in southeastern Brazil. Infection, Genetics and Evolution, 78, 104053. [CrossRef] [PubMed] [Google Scholar]
  26. Popruk S, Adao DEV, Rivera WL. 2021. Epidemiology and subtype distribution of Blastocystis in humans: A review. Infection, Genetics and Evolution, 95, 105085. [CrossRef] [PubMed] [Google Scholar]
  27. Ramírez JD, Sánchez LV, Bautista DC, Corredor AF, Flórez AC, Stensvold CR. 2014. Blastocystis subtypes detected in humans and animals from Colombia. Infection, Genetics and Evolution, 22, 223–228. [CrossRef] [PubMed] [Google Scholar]
  28. Roberts T, Stark D, Harkness J, Ellis J. 2013. Subtype distribution of Blastocystis isolates from a variety of animals from New South Wales, Australia. Veterinary Parasitology, 196(1–2), 85–89. [CrossRef] [PubMed] [Google Scholar]
  29. Santín M, Gómez-Muñoz MT, Solano-Aguilar G, Fayer R. 2011. Development of a new PCR protocol to detect and subtype Blastocystis spp. from humans and animals. Parasitology Research, 109(1), 205–212. [CrossRef] [PubMed] [Google Scholar]
  30. Santin M, Figueiredo A, Molokin A, George NS, Köster PC, Dashti A, González-Barrio D, Carmena D, Maloney JG. 2023. Division of Blastocystis ST10 into three new subtypes: ST42-ST44. Journal of Eukaryotic Microbiology, e12998. [CrossRef] [PubMed] [Google Scholar]
  31. Song J, Yang X, Ma X, Wu X, Wang Y, Li Z, Liu G, Zhao G. 2021. Molecular characterization of Blastocystis sp. in Chinese bamboo rats (Rhizomys sinensis). Parasite, 28, 81. [CrossRef] [EDP Sciences] [PubMed] [Google Scholar]
  32. Stensvold CR, Clark CG. 2016. Current status of Blastocystis: A personal view. Parasitology International, 65(6 Pt B), 763–771. [CrossRef] [PubMed] [Google Scholar]
  33. Verma SK, Singh L. 2003. Novel universal primers establish identity of an enormous number of animal species for forensic application. Molecular Ecology Notes, 3, 28–31. [CrossRef] [Google Scholar]
  34. Wang J, Gong B, Liu X, Zhao W, Bu T, Zhang W, Liu A, Yang F. 2018. Distribution and genetic diversity of Blastocystis subtypes in various mammal and bird species in northeastern China. Parasites & Vectors, 11(1), 522. [CrossRef] [PubMed] [Google Scholar]
  35. Xiao X, Zhou SH, Jiang N, Tian DZ, Zhou ZM, Zhang M, Ke H, Jiang XC, Lv WL, Gao QH. 2019. First record of Leptospira and Blastocystis infections in captive flying squirrels (Trogopterus xanthipes) from Enshi County, China. Acta Tropica, 197, 105065. [CrossRef] [PubMed] [Google Scholar]
  36. Yoshikawa H, Tokoro M, Nagamoto T, Arayama S, Asih PB, Rozi IE, Syafruddin D. 2016. Molecular survey of Blastocystis sp. from humans and associated animals in an Indonesian community with poor hygiene. Parasitology International, 65(6 Pt B), 780–784. [CrossRef] [PubMed] [Google Scholar]

Cite this article as: Zhao W, Zhang Y, Li J, Ren G, Qiang Y, Wang Y, Lai X, Lei S, Liu R, Chen Y, Huang H, Li W, Lu G & Tan F. 2023. Prevalence and distribution of subtypes of Blastocystis in Asiatic brush-tailed porcupines (Atherurus macrourus), bamboo rats (Rhizomys pruinosus), and masked palm civets (Paguma larvata) farmed in Hainan, China. Parasite 30, 45.

All Tables

Table 1

Prevalence and distribution of STs of Blastocystis in the three farmed animals in Hainan Province, China.

In the text

All Figures

thumbnail Figure 1

A map depicting the sampling sites for collecting fecal samples from Asiatic brush-tailed porcupines, bamboo rats and masked palm civets in Hainan Province, China.
In the text
thumbnail Figure 2

Sequence variation in the small subunit (SSU) rRNA gene among of Blastocystis sp. ST5. SSU rRNA sequences of ST5 identified in this study were aligned. MN472792 as the reference sequence.
In the text
thumbnail Figure 3

Phylogenetic relationship of Blastocystis sp. STs based on SSU rRNA gene sequences. Clustal X and Mega 7 software were used to calculate genetic distance and build the phylogenetic tree. The phylogenetic tree was constructed using the maximum likelihood method, based on the evolutionary distances calculated by the Tamura-3 parameter model; all positions containing gaps and missing data were eliminated. Bootstrap values were obtained using 1,000 replicates. The blue and red fonts represent the known and novel sequences, respectively identified in this study.
In the text

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