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Cloning and sequence analysis of chitin synthase gene fragments of Demodex mites

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Abstract

To our knowledge, few reports on Demodex studied at the molecular level are available at present. In this study our group, for the first time, cloned, sequenced and analyzed the chitin synthase (CHS) gene fragments of Demodex folliculorum, Demodex brevis, and Demodex canis (three isolates from each species) from Xi’an China, by designing specific primers based on the only partial sequence of the CHS gene of D. canis from Japan, retrieved from GenBank. Results show that amplification was successful only in three D. canis isolates and one D. brevis isolate out of the nine Demodex isolates. The obtained fragments were sequenced to be 339 bp for D. canis and 338 bp for D. brevis. The CHS gene sequence similarities between the three Xi’an D. canis isolates and one Japanese D. canis isolate ranged from 99.7% to 100.0%, and those between four D. canis isolates and one D. brevis isolate were 99.1%–99.4%. Phylogenetic trees based on maximum parsimony (MP) and maximum likelihood (ML) methods shared the same clusters, according with the traditional classification. Two open reading frames (ORFs) were identified in each CHS gene sequenced, and their corresponding amino acid sequences were located at the catalytic domain. The relatively conserved sequences could be deduced to be a CHS class A gene, which is associated with chitin synthesis in the integument of Demodex mites.

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References

  • Ampasala, D.R., Zheng, S., Zhang, D., Ladd, T., Doucet, D., Krell, P.J., Retnakaran, A., Feng, Q., 2011. An epidermis- specific chitin synthase cDNA in Choristoneura fumiferana: cloning, characterization, developmental and hormonal-regulated expression. Arch. Insect Biochem. Physiol., 76(2):83–96. [doi:10.1002/arch.20404]

    Article  PubMed  CAS  Google Scholar 

  • Arakane, Y., Zhu, Y.C., Kramer, K.J., Charles, A., Michael, R., 2004. Characterization of two chitin synthase genes of the red flour beetle, Tribolium castaneum, and alternate exon usage in one of the genes during development. Insect Biochem. Mol. Biol., 34(3):291–304. [doi:10.1016/j.ibmb.2003.11.004]

    Article  PubMed  CAS  Google Scholar 

  • Ayres, S., Anderson, N.P., 1932. Demodex folliculorum—its role in the etiology of acne rosacea. Arch. Dermatol., 25(1):89–98. [doi:10.1001/archderm.1932.01450020097010]

    Article  Google Scholar 

  • Ayres, S.Jr., Ayres, S.3rd, 1961. Demodectic eruptions (demodicidosis) in the human. Arch. Dermatol., 83(5): 816–827. [doi:10.1001/archderm.1961.01580110104016]

    Article  PubMed  Google Scholar 

  • Chakrabarti, A., Pradhan, N.R., 1985. Demodicidosis in livestock in West Bengal (India). Int. J. Zoonoses, 12(4): 283–290.

    PubMed  CAS  Google Scholar 

  • Chen, X.F., Yang, X., Kumar, N.S., Tang, B., Sun, X.J., Qiu, X.M., Hu, J., Zhang, W.Q., 2007. The class A chitin synthase gene in Spodoptera exigua: molecular cloning and expressional patterns. Insect Biochem. Mol. Biol., 37(5):409–417. [doi:10.1016/j.ibmb.2007.01.006]

    Article  PubMed  CAS  Google Scholar 

  • Forton, F., Seys, B., 1993. Density of Demodex folliculorum in rosacea: a case-control study using standardized skin-surface biopsy. Br. J. Dermatol., 128(6):650–659. [doi:10.1111/j.1365-2133.1993.tb00261.x]

    Article  PubMed  CAS  Google Scholar 

  • Forton, F., Germaux, M.A., Brasseur, T., de Liever, A., Laporte, M., Mathys, C., Sass, U., Stene, J., Thibaut, S., Tytgat, M., et al., 2005. Demodicosis and rosacea: epidemiology and significance in daily dermatologic practice. J. Am. Acad. Dermatol., 52(1):74–87. [doi:10.1016/j.jaad.2004.05.034]

    Article  PubMed  Google Scholar 

  • Gagou, M.E., Kapsetaki, M., Turberg, A., Kafetzopoulos, D., 2002. Stage-specific expression of the chitin synthase DmeChSA and DmeChSB genes during the onset of Drosophila metamorphosis. Insect Biochem. Mol. Biol., 32(2):141–146. [doi:10.1016/S0965-1748(01)00101-1]

    Article  PubMed  CAS  Google Scholar 

  • Guindon, S., Dufayard, J.F., Lefort, V., Anisimova, M., Hordijk, W., Gascuel, O., 2010. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst. Biol., 59(3):307–321. [doi:10.1093/sysbio/syq010]

    Article  PubMed  CAS  Google Scholar 

  • Hogenkamp, D.G., Arakane, Y., Zimoch, L., Merzendorfer, H., Kramer, K.J., Beeman, R.W., Kanost, M.R., Specht, C.A., Muthukrishnan, S., 2005. Chitin synthase genes in Manduca sexta, characterization of a gut-specific transcript and differential tissue expression of alternately spliced mRNAs during development. Insect Biochem. Mol. Biol., 35(6):529–540. [doi:10.1016/j.ibmb.2005.01.016]

    Article  PubMed  CAS  Google Scholar 

  • Ibrahim, G.H., Smartt, C.T., Kiley, L.M., Christensen, B.M., 2000. Cloning and characterization of a chitin synthase cDNA from the mosquito Aedes aegypti. Insect Biochem. Mol. Biol., 30(12):1213–1222. [doi:10.1016/S0965-1748(00)00100-4]

    Article  PubMed  CAS  Google Scholar 

  • Izdebska, J.N., 2010. Demodex sp. (Acari, Demodecidae) and Demodecosis in dogs: characteristics, symptoms, occurrence. Bull. Vet. Inst. Pulawy, 54(3):335–338.

    Google Scholar 

  • Kumar, N.S., Tang, B., Chen, X.F., Tian, H.G., Zhang, W.Q., 2008. Molecular cloning, expression and comparative analysis of chitin synthase gene B in Spodoptera exigua. Comp. Biochem. Physiol. B: Biochem. Mol. Biol., 149(3): 447–453. [doi:10.1016/j.cbpb.2007.11.005]

    Article  Google Scholar 

  • Mederle, N., Darabu, G., Oprescu, I., Morariu, S., Ilie, M., Indre, D., Mederle, O., 2010. Diagnosis of canine demodicosis. Sci. Parasitol., 11(1):20–23.

    Google Scholar 

  • Merzendorfer, H., Zimoch, L., 2003. Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases. J. Exp. Biol., 206(24):4393–4412. [doi:10.1242/jeb.00709]

    Article  PubMed  CAS  Google Scholar 

  • Piao, D.H., Fan, D., 2008. Cloning and sequence analysis of chitin synthase partial cDNA from Mamestra brassicae (L.). J. Northeast Agric. Univ., 39(9):53–57 (in Chinese).

    CAS  Google Scholar 

  • Qu, M., Liu, T., Yang, J., Yang, Q., 2011. The gene, expression pattern and subcellular localization of chitin synthase B from the insect Ostrinia furnacalis. Biochem. Biophys. Res. Commun., 404(1):302–307. [doi:10.1016/j.bbrc.2010.11.111]

    Article  PubMed  CAS  Google Scholar 

  • Ravera, I., Altet, L., Francino, O., Bardagí, M., Sánchez, A., Ferrer, L., 2011. Development of a real-time PCR to detect Demodex canis DNA in different tissue samples. Parasitol. Res., 108(2):305–308. [doi:10.1007/s00436-010-2062-0]

    Article  PubMed  Google Scholar 

  • Rojas, M.D., Riazzo, C., Callejón, R., Guevara, D., Cutillas, C., 2012. Morphobiometrical and molecular study of two populations of Demodex folliculorum from humans. Parasitol. Res., 110(1):227–233. [doi:10.1007/s00436-011-2476-3]

    Article  PubMed  Google Scholar 

  • Smith, H.J., 1961. Demodicidosis in a flock of goats. Can. Vet. J., 2(6):231–233.

    PubMed  CAS  Google Scholar 

  • Tamura, K., Dudley, J., Nei, M., Kumar, S., 2007. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol., 24(8):1596–1599. [doi:10.1093/molbev/msm092]

    Article  PubMed  CAS  Google Scholar 

  • Tellam, R., Vuocolo, T., Johnson, S.E., Jarmey, J., Pearson, R.D., 2000. Insect chitin synthase cDNA sequence, gene organization and expression. Eur. J. Biochem., 267(19): 6025–6043. [doi:10.1046/j.1432-1327.2000.01679.x]

    Article  PubMed  CAS  Google Scholar 

  • Thompson, J.D., Gibson, T.J., Plewni, F., Jeanmougin, F., Higgins, D.G., 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res., 25(24): 4876–4882. [doi:10.1093/nar/25.24.4876]

    Article  PubMed  CAS  Google Scholar 

  • Toops, E., Blagburn, B., Lenaghan, S., Kennis, R., MacDonald, J., Dykstra, C., 2010. Extraction and characterization of DNA from Demodex canis. Intern. J. Appl. Res. Vet. Med., 8(1):31–43.

    CAS  Google Scholar 

  • Xu, H.Z., Xiao, F.P., Qin, Z.Y., Ye, X.F., Ai, Y.P., 2009. Early diagnosis, prevention and cure of goat Demodicidosis. Guizhou Agric. Sci., 37(11):125–126 (in Chinese).

    CAS  Google Scholar 

  • Zhang, J., Zhu, K.Y., 2006. Characterization of a chitin synthase cDNA and its increased mRNA level associated with decreased chitin synthesis in Anopheles quadrimaculatus exposed to diflubenzuron. Insect Biochem. Mol. Biol., 36(9):712–725. [doi:10.1016/j.ibmb.2006.06.002]

    Article  PubMed  CAS  Google Scholar 

  • Zhao, Y.E., Cheng, H., 2009. RAPD analysis and sequence alignment of genomic DNA of hair follicle mites Demodex folliculorum and D. brevis (Acari: Demodicidae). Acta Entomol. Sin., 52(11):1273–1279 (in Chinese).

    CAS  Google Scholar 

  • Zhao, Y.E., Wu, L.P., 2012a. Phylogenetic relationships in Demodex mites (Acari: Demodicidae) based on mitochondrial 16S rDNA partial sequences. Parasitol. Res., 111(3):1113–1121. [doi:10.1007/s00436-012-2941-7]

    Article  PubMed  Google Scholar 

  • Zhao, Y.E., Wu, L.P., 2012b. RAPD-SCAR marker and genetic relationship analysis of three Demodex species (Acari: Demodicidae). Parasitol. Res., 110(6):2395–2402. [doi:10.1007/s00436-011-2778-5]

    Article  PubMed  Google Scholar 

  • Zhao, Y.E., Guo, N., Wu, L.P., 2009a. The effect of temperature on the viability of Demodex folliculorum and Demodex brevis. Parasitol. Res., 105(6):1623–1628. [doi:10.1007/s00436-009-1603-x]

    Article  PubMed  Google Scholar 

  • Zhao, Y.E., Cheng, H., Xun, M., Wu, L.P., 2009b. Extraction and random primer PCR detection of genomic DNA of parasitic mites Demodex folliculorum and Demodex brevis (Acari: Demodicidae). Acta Entomol. Sin., 52(8): 929–933 (in Chinese).

    CAS  Google Scholar 

  • Zhao, Y.E., Wu, L.P., Peng, Y., Cheng, H., 2010. Retrospective analysis of the association between Demodex infestation and rosacea. Arch. Dermatol., 146(8):896–902. [doi:10.1001/archdermatol.2010.196]

    Article  PubMed  Google Scholar 

  • Zhao, Y.E., Peng, Y., Wang, X.L., Wu, L.P., Wang, M., Yan, H.L., Xiao, S.X., 2011a. Facial dermatosis associated with Demodex: a case-control study. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 12(12):1008–1015. [doi:10.1631/jzus.B1100179]

    Article  Google Scholar 

  • Zhao, Y.E., Guo, N., Wu, L.P., 2011b. The influence of temperature and medium on viability of Demodex folliculorum and Demodex brevis (Acari: Demodicidae). Exp. Appl. Acarol., 54(4):421–425. [doi:10.1007/s10493-011-9445-5]

    Article  PubMed  Google Scholar 

  • Zhao, Y.E., Guo, N., Xun, M., Xu, J.R., Wang, M., Wang, D.L., 2011c. Sociodemographic characteristics and risk factor analysis of Demodex infestation (Acari: Demodicidae). J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 12(12): 998–1007. [doi:10.1631/jzus.B1100079]

    Article  Google Scholar 

  • Zhao, Y.E., Wu, L.P., Hu, L., Xu, J.R., 2012a. Association of blepharitis with Demodex: a meta-analysis. Ophthalmic Epidemiol., 19(2):95–102. [doi:10.3109/09286586.2011.642052]

    Article  PubMed  Google Scholar 

  • Zhao, Y.E., Xu, J.R., Hu, L., Wu, L.P., Wang, Z.H., 2012b. Complete sequence analysis of 18S rDNA based on genomic DNA extraction from individual Demodex mites (Acari: Demodicidae). Exp. Parasitol., 131(1):45–51. [doi:10.1016/j.exppara.2012.02.025]

    Article  PubMed  CAS  Google Scholar 

  • Zhao, Y.E., Hu, L., Wu, L.P., Ma, J.X., 2012c. A meta-analysis of association between acne vulgaris and Demodex infestation. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 13(3):192–202. [doi:10.1631/jzus.B1100285]

    Article  Google Scholar 

  • Zhao, Y.E., Wu, L.P., Hu, L., Xu, Y., Wang, Z.H., Liu, W.Y., 2012d. Sequencing for complete rDNA sequences (18S, ITS1, 5.8S, ITS2, and 28S rDNA) of Demodex and phylogenetic analysis of Acari based on 18S and 28S rDNA. Parasitol. Res., in press. [doi:10.1007/s00436-012-3058-8]

  • Zhu, Y.C., Specht, C.A., Dittmer, N.T., Muthukrishnan, S., Kanost, M.R., Kramer, K.J., 2002. Sequence of a cDNA and expression of the gene encoding a putative epidermal chitin synthase of Manduca sexta. Insect Biochem. Mol. Biol., 32(11):1497–1506. [doi:10.1016/S0965-1748(02)00070-X]

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Ya-e Zhao or Ji-ru Xu.

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Project supported by the National Natural Science Foundation of China (No. 81271856) and the National College of Innovative Experimental Projects of China (No. 101069819)

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Zhao, Ye., Wang, Zh., Xu, Y. et al. Cloning and sequence analysis of chitin synthase gene fragments of Demodex mites. J. Zhejiang Univ. Sci. B 13, 763–768 (2012). https://doi.org/10.1631/jzus.B1200155

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  • DOI: https://doi.org/10.1631/jzus.B1200155

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