Molecular Characterization of Chitin Synthase Gene of Demodex canis from Mizoram, India

Abstract

Aims

Canine demodicosis is a parasitic condition affecting the skin of dogs. The present study was designed to characterize chitin synthase gene of Demodex canis. The molecular technique was used for better understanding of this gene.

Methods

A total of 75 dogs which are reared as pets with or without showing any skin lesions were examined during the study period. Skin scrapings were examined by indirect method using 10% potassium hydroxide solution under 10 × microscope. DNA samples were extracted from positive skin samples and were subjected to PCR for molecular identification.

Results

A total of 25 dogs irrespective of age, sex, breed or coat showed positive result for D. canis. The PCR revealed a single amplified product of 339 bp length which exactly matched with D. canis. The chitin synthase gene was amplified by PCR, subsequently cloned, sequenced, and compared with available data in GenBank for the particular gene of D. canis. Only one single nucleotide polymorphism (SNP) was noticed at 231 position of the chitin synthase gene sequence when compared to other isolates.

Conclusion

The molecular technique confirms with the morphological identity of D. canis. This report signifies the value of peculiar tool to identify ‘follicular mite’ even from apparently healthy skin.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  1. 1.

    Day M.J. 1997. An immune-histochemical study of the lesions of demodecosis in the dog. Journal of Comparative Pathology, 116, 203–216.

  2. 2.

    Esch K.J., Juelsgaard R., Martinez P. A. et al. 2013. Programmed death 1-mediated T cell exhaustion during visceral leishmaniasis impairs phagocyte function. Journal of Immunology, 191, 5542–5550.

    CAS  Article  Google Scholar 

  3. 3.

    Forton F.M. 2012. Papulopustular rosacea, skin immunity and Demodex: pityriasis folliculorum as a missing link. Journal of European Academy of Dermatology and Venereology, 26, 19–28.

    CAS  Article  Google Scholar 

  4. 4.

    Gortel K. 2006. Update on canine demodecosis. Veterinary Clinics of North America: Small Animal Practice, 36 (1), 229–241.

  5. 5.

    Izdebska J.N. 2010. Demodex sp. (Acari:Demodicidae) and Demodicosis in dogs: characteristics, symptoms, occurrence. Bulletin of the Veterinary Institute Pulawy, 54(3), 335–338.

  6. 6.

    Jarmuda S., O’Reilly N., Żaba R., Jakubowicz O., Szkaradkiewicz A., Kavanagh K. 2012. Potential role of Demodex mites and bacteria in the induction of rosacea. Journal of Medical Microbiology, 61, 1504–1510.

    Article  Google Scholar 

  7. 7.

    Kuznetsova E., Bettenay S., Nikolaeva L. et al. 2012. Influence of systemic antibiotics on the treatment of dogs with generalized demodicosis. Veterinary Parasitology, 188, 148–155.

  8. 8.

    Mederle N., Darabu G., Oprescu I., Morariu S., Ilie M., Indre D., et al. 2010. Diagnosis of canine demodicosis. Scientia Parasitologica, 11(1), 20–23.

    Google Scholar 

  9. 9.

    Merzendorfer H., Zimoch L. 2003. Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases. Journal of Experimental Biology, 206(24), 4393–4412. https://doi.org/10.1242/jeb.00709.

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Miller W.H., Griffin C.E., Campbell K.L. 2013. Parasitic diseases. In: Muller and Kirk’s small animal dermatology, 7th edition. Philadelphia, PA: W.B. Saunders Co. 284–342.

    Google Scholar 

  11. 11.

    Muller G., Kirk R. 2013. Parasitic skin disease. In: Miller, W.H., Griffin, C.E., Campbell, K.L. (Eds.), Small Animal Dermatology., 7th ed. Saunders Elsevier, St. Louis, pp. 304–313.

    Google Scholar 

  12. 12.

    Mumprecht S., Sch€ urch C., Schwaller J.,et al. 2009. Programmed death 1 signaling on chronic myeloid leukemia-specific T cells results in T-cell exhaustion and disease progression. Blood, 114, 1528–1536.

    CAS  Article  Google Scholar 

  13. 13.

    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. Parasitology Research, 108(2), 305–308. https://doi.org/10.1007/s00436-010-2062-0.

    Article  PubMed  Google Scholar 

  14. 14.

    Ravera I., Altet L., Francino O., Sanchez A., Roldan W., Villanueva S., et al. 2013. Small Demodex populations colonize most parts of the skin of healthy dogs. Advances in Veterinary Dermatology. https://doi.org/10.1111/j.1365-3164.2012.01099.x.

    Article  Google Scholar 

  15. 15.

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

    Article  PubMed  Google Scholar 

  16. 16.

    Tater K. C., Patterson A.C. 2008. Canine and Feline demodicosis. Veterinary Medicine. 444–461.

  17. 17.

    Toops E, Blagburn B, Lenaghan S, Kennis R, MacDonald J, Dykstra C. 2010. Extraction and characterization of DNA from Demodex canis. International Journal of Applied Research in Veterinary Medicine, 8(1), 31–43.

    CAS  Google Scholar 

  18. 18.

    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 Agricultural Sciences, 37(11), 125–126.

    CAS  Google Scholar 

  19. 19.

    Zhao Y.E., Ma J.X., Hu L., Wa L.P., de Rojas M. 2013. Discrimination between Demodex folliculoru m (Acari: Demodicidae)isolates from China and Spain based on mitochondrial cox1 sequences. Journal of Zhejiang University Science B (Biomed and Biotechnol), 14(9), 829–836.

    Article  Google Scholar 

  20. 20.

    Zhao Y.E., Wu L.P. 2012. Phylogenetic relationships in Demodex mites (Acari: Demodicidae) based on mitochondrial 16S rDNA partial sequences. Parasitology Research, https://doi.org/10.1007/s00436-012-2941-7.

  21. 21.

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

  22. 22.

    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). Experimental Parasitology, 131(1), 45-51. https://doi.org/10.1016/j.exppara.2012.02.025.

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are also grateful to the Veterinary Officers posted in different hospitals throughout the state for collection of samples. The authors are also grateful to the Dean, College of Veterinary Sciences and A.H., CAU, Selesih, Aizawl, India for his necessary support to conduct the experiment.

Funding

The study was funded by Central Agricultural University, Imphal, Manipur, India under laboratory research to the first author.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Gautam Patra.

Ethics declarations

Conflict of interest

No competing interests have been declared.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Patra, G., Behera, P., Ghosh, S. et al. Molecular Characterization of Chitin Synthase Gene of Demodex canis from Mizoram, India. Acta Parasit. 64, 57–62 (2019). https://doi.org/10.2478/s11686-018-00008-6

Download citation

Keywords

  • Demodex canis
  • Chitin synthase gene
  • Cloning and sequencing
  • Phylogenetic analysis