CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2022 , Vol. 40 >Issue 5: 594 - 602

DOI: https://doi.org/10.12140/j.issn.1000-7423.2022.05.005

ORIGINAL ARTICLES

Analysis of ribosomal and mitochondrial genes of Baylisascaris procyonis

  • WEI Kai-wen ,
  • ZENG Hong-xia ,
  • HE Mu ,
  • HU Jun-jie
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  • 1. School of Biological Sciences, Yunnan University, Kunming 650000, China
    2. Kunming Zoological Garden, Kunming 650000, China
    3. School of Ecology and Environmental Sciences and Yunnan Key Laboratory or Plateau Mountain Ecology and Restoration of Degraded Environments, Yunnan University, Kunming 650000, China

Received date: 2022-01-05

  Revised date: 2022-03-28

  Online published: 2022-10-17

Supported by

Scientific Fund of Education Department of Yunnan Province(2021Y044)

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Abstract

Objective To characterize the ribosomal and mitochondrial genes of Baylisascaris procyonis, and their phylogenetic relationship with other Baylisascaris spp. Methods Adult worms of B. procyonis were collected from the intestine of a deceased raccoon at the Kunming Zoo in Kunming, China. After morphological identification, 20 adult worms comprising 10 males and 10 females were selected for morphological measurement and related molecularbiological study. The DNA was extracted from the tissue sample of the parasite using a DNA extraction kit. For amplification of ribosome internal transcribed spacer (ITS), cytochrome c oxidase 1 (cox1), cox2, cox3, cytochrome b (cytb), nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1), nad5 and tRNA-nad2-tRNA by PCR, which were sequenced and aligned with the database deposited in GenBank using BLAST. Single nucleotide polymorphisms (SNP) were analyzed using MEGA X software, and phylogenetic trees based on ITS, cox1 and cox2 sequences were constructed respectively, using the maximum likelihood (ML) method. Results The adult worms were in ivory color, at a size of (9.15-14.50) cm × (0.20-0.32) cm for females and (6.45-8.70) cm × (0.15-0.20) cm for males. Adult male has copulatory spicule at the caudal ends. The eight genetic markers of B. procyonis were successfully sequenced, and all sequences were deposited in GenBank under the accession numbers MZ092850-MZ092855, MZ164969, MZ172987, MZ172988, MZ172982-MZ172986, MZ172989, MZ172981, and MZ172980 for ITS, cox1, cox2, cox3, cytb, nad1, nad5, and tRNA-nad2-tRNA, respectively. Of the eight loci, the newly obtained sequence shared identities of 98.6%-100%, 98.5%-99.5%, 99.4%-99.7%, 98.3%, 98.8%-99.0%, 98.2%, 97.8% and 99.2%, respectively, with those of B. procyonis previously deposited in GenBank. Additionally, the newly obtained ITS, cox1, cox2 and tRNA-nad2-tRNA sequences had identities of 98.6%-100%, 99.2%-100%, 99.4%-100% and 99.6%-99.9%, respectively, with those of B. columnaris previously deposited in GenBank. SNP analysis showed that the newly obtained ITS, cox1, cox2 and tRNA-nad2-tRNA sequences had 28, 7 and 12 SNP sites, respectively, compared to those of B. procyonis and B. columnaris previously deposited in GenBank. However, none of the SNP sites was suitable to distinguish B. procyonis from B. columnaris. Meanwhile, the newly obtained ITS sequences contained 7 to 9 GA tandem repeats, and the 7 and 8 repeats constituted the first records discovered in B. procyonis. Phylogenetic analysis inferred from ITS, cox1 or cox2 sequences indicated that B. procyonis and B. columnaris obtained from different geographical areas completely aggregated, and formed an individual clade, that clustered with B. devosi. Conclusion The markers sequences and SNP characteristics of B. raccoonus were highly similar to those of B. cylindrica, whereas phylogenetic analysis could not effectively differentiate B. procyonis from B. columnaris.

Key words： Raccoon; Baylisascaris procyonis; Nuclear and mitochondrial genes; Single nucleotide polymorphisms; Phylogeny

Cite this article

WEI Kai-wen , ZENG Hong-xia , HE Mu , HU Jun-jie . Analysis of ribosomal and mitochondrial genes of Baylisascaris procyonis[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022 , 40(5) : 594 -602 . DOI: 10.12140/j.issn.1000-7423.2022.05.005

References

[1] Sapp SGH,, Gupta P,, Martin MK, et al. Beyond the raccoon roundworm: the natural history of non-raccoon Baylisascaris species in the New World[J]. Int J Parasitol Parasites Wildl, 2017, 6(2): 85-99.
[2] Camp LE,, Radke MR,, Shihabi DM, et al. Molecular phylogenetics and species-level systematics of Baylisascaris[J]. Int J Parasitol Parasites Wildl, 2018, 7(3): 450-462.
[3] Graeff-Teixeira C,, Morassutti AL,, Kazacos KR. Update on baylisascariasis, a highly pathogenic zoonotic infection[J]. Clin Microbiol Rev, 2016, 29(2): 375-399.
[4] Nadler SA. Phylogeny of some ascaridoid nematodes, inferred from comparison of 18S and 28S rRNA sequences[J]. Mol Biol Evol, 1992, 9(5): 932-944.
[5] Gatcombe RR,, Jothikumar N,, Dangoudoubiyam S, et al. Evaluation of a molecular beacon real-time PCR assay for detection of Baylisascaris procyonis in different soil types and water samples[J]. Parasitol Res, 2010, 106(2): 499.
[6] Blizzard EL,, Davis CD,, Henke S, et al. Distribution, prevalence, and genetic characterization of Baylisascaris procyonis in selected areas of Georgia[J]. J Parasitol, 2010, 96(6): 1128-1133.
[7] Testini G,, Papini R,, Lia RP, et al. New insights into the morphology, molecular characterization and identification of Baylisascaris transfuga (Ascaridida, Ascarididae)[J]. Vet Parasitol, 2011, 175(1/2): 97-102.
[8] Franssen F,, Xie KY,, Sprong H, et al. Molecular analysis of Baylisascaris columnaris revealed mitochondrial and nuclear polymorphisms[J]. Parasit Vectors, 2013, 6(1): 124.
[9] Choi Y,, Mason S,, Ahlborn M, et al. Partial molecular characterization of the mitochondrial genome of Baylisascaris columnaris and prevalence of infection in a wild population of striped skunks[J]. Int J Parasitol Parasites Wildl, 2017, 6(2): 70-75.
[10] Sharifdini M,, Heckmann RA,, Mikaeili F. The morphological and molecular characterization of Baylisascaris devosi Sprent, 1952 (Ascaridoidea, Nematoda), collected from pine marten (Martes martes) in Iran[J]. Parasit Vectors, 2021, 14(1): 33.
[11] Xie Y,, Zhang ZH,, Niu LL, et al. The mitochondrial genome of Baylisascaris procyonis[J]. PLoS One, 2011, 6(10): e27066.
[12] Untergasser A,, Cutcutache I,, Koressaar T, et al. Primer3: new capabilities and interfaces[J]. Nucleic Acids Res, 2012, 40(15): e115.
[13] Hu JJ,, Liu TT,, Liu Q, et al. Prevalence, morphology, and molecular characteristics of Sarcocystis spp. in domestic goats (Capra hircus) from Kunming, China[J]. Parasitol Res, 2016, 115(10): 3973-3981.
[14] Kumar S,, Stecher G,, Li M, et al. MEGA X: molecular evolutionary genetics analysis across computing platforms[J]. Mol Biol Evol, 2018, 35(6): 1547-1549.
[15] Sprent JF. Notes on Ascaris and Toxascaris, with a definition of Baylisascaris gen. nov[J]. Parasitology, 1968, 58(1): 185-198.
[16] Berry J. Phylogenetic relationship between Baylisascaris spp. Sprent, 1968(Nematoda ∶ Ascarididae) from skunks, raccoons and groundhogs in southern Ontario[D]. Guelph: University of Guelph, 1985: 86.
[17] Dangoudoubiyam S,, Vemulapalli R,, Kazacos KR. PCR assays for detection of Baylisascaris procyonis eggs and larvae[J]. J Parasitol, 2009, 95(3): 571-577.
[18] Sprent JFA. On the migratory behavior of the larvae of various Ascaris species in white mice: Ⅱ. longevity of encapsulated larvae and their resistance to freezing and putrefaction[J]. J Infect Dis, 1953, 92(2): 114-117.
[19] Sheng HL. Atlas of mammals of China[M]. Zhengzhou: Henan Science and Technology Press, 2005: 1-527. (in Chinese)
[19] ( 盛和林, 中国野生动物保护协会. 中国哺乳动物图鉴[M]. 郑州: 河南科学技术出版社, 2005: 1-527.)
[20] Zhang RZ. Zoogeography of China[M]. Beijing: Science Press, 1999: 1-502. (in Chinese)
[20] ( 张荣祖. 中国动物地理[M]. 北京: 科学出版社, 1999: 1-502.)
[21] Xie Y,, Zhou X,, Li M, et al. Zoonotic Baylisascaris procyonis roundworms in raccoons, China[J]. Emerg Infect Dis, 2014, 20(12): 2170-2172.
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