Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Molecular Characterization of Nippostrongylus brasiliensis (Nematoda: Heligmosomatidae) from Mus musculus in India

  • Chaudhary, Anshu (Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University) ;
  • Goswami, Urvashi (Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University) ;
  • Singh, Hridaya Shanker (Molecular Taxonomy Laboratory, Department of Zoology, Chaudhary Charan Singh University)
  • Received : 2016.06.28
  • Accepted : 2016.09.26
  • Published : 2016.12.31
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Abstract

Mus musculus (Rodentia: Muridae) has generally been infected with a rodent hookworm Nippostrongylus brasiliensis. In this report, we present morphological and molecular identification of N. brasiliensis by light and scanning electron microscopy and PCR amplification of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and the protein sequences encoded by cox1 gene, respectively. Despite the use of N. brasiliensis in many biochemistry studies from India, their taxonomic identification was not fully understood, especially at the species level, and no molecular data is available in GenBank from India. Sequence analysis of cox1 gene in this study revealed that the present specimen showed close identity with the same species available in GenBank, confirming that the species is N. brasiliensis. This study represents the first record of molecular identification of N. brasiliensis from India and the protein structure to better understand the comparative phylogenetic characteristics.

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References

  1. Ogilvie BM, Jones VE. Nippostrongylus brasiliensis: A review of immunity and the host/parasite relationship in the rat. Exp Parasitol 1971; 29: 138-177. https://doi.org/10.1016/0014-4894(71)90021-X
  2. Croll NA, Wright KA. Observation on the movements and structure of the bursa of Nippostrongylus brasiliensis and Nematospiroides dubius. Can J Zool 1976; 54: 1466-1480. https://doi.org/10.1139/z76-170
  3. Camberis M, Le Gros G, Urban J Jr. Animal model of Nippostrongylus brasiliensis and Heligmosomoides polygyrus. Curr Protoc Immunol, Chapter 19: Unit 19.12. 2003; doi: 10.1002/0471142735.im1912s55.
  4. Ball G, Knox D. Vaccination of rats against the rodent hookworm Nippostrongylus brasiliensis with a recombinant superoxide dismutase fails to protect against infection. Acta Parasitol 2009; 54: 281-287.
  5. Milazzo C, Cagnin M, DI Bella C, Geraci F, Ribas A. Helminth Fauna of Commensal Rodents, Mus musculus (Linnaeus, 1758) and Rattus rattus (Linnaeus, 1758) (Rodentia, Muridae) in Sicily (Italy). Rev Ibero-Latinoam Parasitol 2010. 69: 194-198.
  6. Blouin MS. Molecular prospecting for cryptic species of nematodes: mitochondrial DNA versus internal transcribed spacer. Int J Parasitol 2002; 32: 527-531. https://doi.org/10.1016/S0020-7519(01)00357-5
  7. Singh N, Chaudhary A, Singh HS. Identification of two species of Binema Travassos, 1925 (Oxyurida: Travassosinematidae) based on morphological and sequence analysis of genomic (18S) and mitochondrial (Cox1) gene markers. J Nematode Morphol Syst 2013; 16: 173-180.
  8. Goswami U, Chaudhary A, Verma C, Singh HS. First Molecular Characterization of Aspiculuris tetraptera (Nematoda: Heteroxynematidae) from Mus musculus (Rodentia: Muridae) in India. Acta Parasitol 2015; 60: 553-556.
  9. Hebert PD, Ratnasingham S, deWaard JR. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc Biol Sci 2003; 270: S96-S99. https://doi.org/10.1098/rsbl.2003.0025
  10. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 1994; 3: 294-299.
  11. Thompson JD, Higgins DG, Gibson TJ. Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acid Res 1994; 22: 4673-4680. https://doi.org/10.1093/nar/22.22.4673
  12. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 2013; 30: 2725-2729. https://doi.org/10.1093/molbev/mst197
  13. Milne I, Lindner D, Bayer M, Husmeier D, McGuire G, Marshall DF, Wright F. TOPALi v2: a rich graphical interface for evolutionary analyses of multiple alignments on HPC clusters and multi-core desktops. Bioinformatics 2009; 25: 126-127. https://doi.org/10.1093/bioinformatics/btn575
  14. Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A. Protein Identification and Analysis Tools on the ExPASy Server. In Walker JM ed, The Proteomics Protocols Handbook. Humana Press. 2005, pp 571-607.
  15. Geourjon C, Deleage G. SOPMA: significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments. Comput Appl Biosci 1995; 11: 681-684.
  16. Robert X, Gouet P. Deciphering key features in protein structures with the new ENDscript server. Nucleic Acids Res 2014; 42: 320-324.
  17. Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T, Kiefer F, Gallo Cassarino T, Bertoni M, Bordoli L, Schwede T. SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res 2014; 42: 252-258. https://doi.org/10.1093/nar/gku340
  18. Kelley LA, Mezulis S, Yates CM, Wass MN, Sternberg MJ. The Phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc 2015; 10: 845-858. https://doi.org/10.1038/nprot.2015.053
  19. Yang J, Yan R, Roy A, Xu D, Poisson J, Zhang Y. The I-TASSER Suite: Protein structure and function prediction. Nat Methods 2015; 12: 7-8. https://doi.org/10.1038/nmeth.3213
  20. Phillips JC, Braun R, Wang W, Gumbart J, Tajkhorshid E, Villa E, Chipot C, Skeel RD, Kale L, Schulten K. Scalable molecular dynamics with NAMD. J Comput Chem 2005; 26: 1781-1802. https://doi.org/10.1002/jcc.20289
  21. Buchan DW, Minneci F, Nugent TC, Bryson K, Jones DT. Scalable web services for the PSIPRED Protein Analysis Workbench. Nucleic Acids Res 2013; 41: 349-357. https://doi.org/10.1093/nar/gkt381
  22. Lovell SC, Davis IW, Arendall WB 3rd, de Bakker PI, Word JM, Prisant MG, Richardson JS, Richardson DC. Structure validation by Calpha geometry: phi,psi and C beta deviation. Proteins 2003; 50: 437-450. https://doi.org/10.1002/prot.10286
  23. Wiederstein M, Sippl MJ. ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res 2007; 35: 407-410.
  24. Zhang Y, Skolnick J. TM-align: a protein structure alignment algorithm based on the TM-score. Nucleic Acids Res 2005; 33: 2302-2309. https://doi.org/10.1093/nar/gki524
  25. Blouin MS, Yowell CA, Courtney CH, Dame JB. Substitution bias, rapid saturation, and the use of mtDNA for nematode systematics. Mol Biol Evol 1998; 15: 1719-1727. https://doi.org/10.1093/oxfordjournals.molbev.a025898
  26. Gissi C, Iannelli F, Pesole G. Evolution of the mitochondrial genome of Metazoa as exemplified by comparison of congeneric species. Heredity 2008; 101: 301-320. https://doi.org/10.1038/hdy.2008.62
  27. Blaxter ML, De Ley P, Garey JR, Liu LX, Scheldeman P, Vierstraete A, Vanfleteren JR, Mackey LY, Dorris M, Frisse LM, Vida JT, Thomas WK. A molecular evolutionary framework for the phylum Nematoda. Nature 1998; 392: 71-75. https://doi.org/10.1038/32160
  28. Zalesny G, Hildebrand J, Paziewska-Harris A, Behnke JM, Harris PD. Heligmosomoides neopolygyrus Asakawa & Ohbayashi, 1986, a cryptic Asian nematode infecting the striped field mouse Apodemus agrarius in Central Europe. Parasit Vectors 2014; 7: 457. https://doi.org/10.1186/s13071-014-0457-y
  29. Sotillo J, Sanchez-Flores A, Cantacessi C, Harcus Y, Pickering D, Bouchery T, Camberis M, Tang SC, Giacomin P, Mulvenna J, Mitreva M, Berriman M, LeGros G, Maizels RM, Loukas A. Secreted proteomes of different developmental stages of the gastrointestinal nematode Nippostrongylus brasiliensis. Mol Cell Proteomics 2014; 13: 2736-2751. https://doi.org/10.1074/mcp.M114.038950
  30. Wang ZO, Pollock DD. Coevolutionary patterns in cytochrome c oxidase subunit I depend on structural and functional context. J Mol Evol 2007; 65: 485-495. https://doi.org/10.1007/s00239-007-9018-8

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