Techniques to Conduct Morphological and Molecular Investigations on Nematodes

  • Aasha Rana
  • Anshika Yadav
  • Aashaq Hussain Bhat
  • Ashok Kumar Chaubey
  • Sandeep K. Malhotra
Part of the Springer Protocols Handbooks book series (SPH)


The intriguing variety in the form and size of roundworms leading free-living as well as parasitic life has mesmerized scientists world over. One most common happening during developmental cycle of roundworms is moulting, and this characteristic is remarkably retained irrespective of the presence or absence of intermediate hosts (direct life cycle) in the life cycle of round worms in certain specified cases. The application of fixatives, in particular, therefore becomes a challenging exercise to suit the body form at different stages of development. However, looking at the severity of pathogenic influence of animal-parasitic and plant-parasitic nematodes necessitated appropriate fixation procedures of these worms infesting separate variety of hosts. Such procedures are enumerated in the text of this chapter with meticulous details on morphology based on scanning electron microscopic examination of certain worms, wherever required. Prominent techniques with molecular applications in taxonomy of roundworms to segregate arthropod-parasitic, freshwater, marine, and soil taxa have been outlined. The methods for effective barcode analyses of these round worms from terrestrial or aquatic vertebrate hosts have been discussed briefly.


Nematodes Liver habitat Molecular taxonomy Plant-parasitic nematodes Entomopathogenic Zoonotic DNA barcoding Genomic DNA extraction 


  1. 1.
    Seinhorst JW (1962) Extraction methods for nematodes inhabiting soil. In: Murphy PW (Ed.), Progress in Soil Zoology, Butterworths, London 243–256Google Scholar
  2. 2.
    De Grisse AT (1969) Redescription ou modifications de quelques technique utilisées dans 1’étude des nématodes phytoparasitaires. Meded. Fac. Landb Wettens 34: 351–369Google Scholar
  3. 3.
    Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98Google Scholar
  4. 4.
    Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874. Scholar
  5. 5.
    Floyd RM, Rogers AD, Lambshead PJD, Smith CR (2005) Nematode-specific PCR primers for the 18S small subunit rRNA gene. Mol Ecol Notes 5:611–612CrossRefGoogle Scholar
  6. 6.
    Sandeep MK (1986) Bioecology of the parasites of high altitude homeothermic host–parasite systems. I. Influence of season and temperature on infection by strobilocerci of three species of Hydatigera in Indian rat host. J Helminthol 60:15–20CrossRefGoogle Scholar
  7. 7.
    Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3:294–299Google Scholar
  8. 8.
    Thompson JD, Gibson TJ, Plewniak F et al (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25(24):4876–4882CrossRefGoogle Scholar
  9. 9.
    Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425Google Scholar
  10. 10.
    Tamura K, Peterson D, Peterson N et al (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28(10):2731–2739CrossRefGoogle Scholar
  11. 11.
    Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Aasha Rana
    • 1
  • Anshika Yadav
    • 2
    • 3
  • Aashaq Hussain Bhat
    • 1
  • Ashok Kumar Chaubey
    • 1
  • Sandeep K. Malhotra
    • 2
  1. 1.Nematology Laboratory, Department of ZoologyChaudhary Charan Singh UniversityMeerutIndia
  2. 2.Parasitology Laboratory, Department of ZoologyUniversity of AllahabadAllahabadIndia
  3. 3.Wild Life Section, Zoological Survey of India, M-Block, New AliporeKolkataIndia

Personalised recommendations