Skip to main content
Log in

Storage of gastrointestinal nematode infective larvae for species preservation and experimental infections

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Techniques to preserve the infective third-stage larvae (L3) of gastrointestinal nematodes are of considerable interest to preserve rare species and to maintain a stable source for routine experimental infections. This study compares the relative pros and cons of the two most common techniques, cryopreservation and refrigeration by comparing how they influence consequent infection outcome parameters in terms of life-history traits and fitness as a function of time using the gastrointestinal nematode of sheep Haemonchus contortus as a study species. Establishment capacity was found to be significantly reduced in cryopreserved stocks of L3 compared to refrigerated stocks, but this was followed by significant increases in their fecundity. Refrigeration did not affect L3 stocks consequent fitness by 16 months (the maximum examined) although they did incur a significant reduction in establishment, followed once again by an augmentation in fecundity. The study highlights potential areas for bias in comparing studies using L3 larvae maintained for different periods of time under different techniques.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baermann G (1917) Eine einfach methode zur auffindung von Ankylostomun—(Nematoda)—larven in erdproben. Welteureden Batavia Geneesk. Lab Feestbundel 41–47

  • Cabaret J, Ouhelli H (1984) Fertility of parasitic strongyles in the digestive system of sheep under natural conditions. Rev Med Vet 135:627–633

    Google Scholar 

  • Campbell WC, Blair LS, Egerton JR (1973) Unimpaired infectivity of the nematode Haemonchus contortus after freezing for 44 weeks in the presence of liquid nitrogen. J Parasitol 59:425–427

    Article  CAS  PubMed  Google Scholar 

  • Chehresa A, Beech RN, Scott ME (1997) Life-history variation among lines isolated from a population of Heligmosomoides polygyrus bakeri. Int J Parasitol 27:541–551

    Article  CAS  PubMed  Google Scholar 

  • Chylinski C, Lherminé E, Coquille M, Cabaret J (2014) Desiccation tolerance of gastrointestinal nematode third stage larvae: exploring the effects on fitness and the factors influencing survival. Parasitol Res. doi:10.1007/s00436-014-3938-1

    Google Scholar 

  • Chylinski C, Blanchard-Letort A, Neveu C, Cortet J, Cabaret J. Parasitic gastrointestinal nematodes coping with chemotherapy, resistant hosts and unfavourable climatic environments: an experimental evaluation. Submitted to Molecular Ecology. Under review

  • Gaba S, Chadoeuf J, Monestiez P, Sauve C, Cortet J, Cabaret J (2006) Estimation of abomasum strongyle nematode infections in sheep at necropsy: Tentative proposals for a simplified technique. Vet Parasitol 140:105–113

  • Gasnier N, Cabaret J, Moulia C (1992) Allozyme variation between laboratory reared and wild populations of Teladorsagia circumcincta. Int J Parasitol 22(5):581–587

    Article  CAS  PubMed  Google Scholar 

  • Kerboeuf D (1978a) The effects of time and temperature of storage on the infectivity of third-stage larvae of Heligmosomoides polygyrus (: Nematospiroides dubius). 1. Effects on the development to the adult stage in mice. Ann Rech Vet 9:153–159

    CAS  PubMed  Google Scholar 

  • Kerboeuf D (1978b) The effects of time and temperature of storage on the infectivity of third-stage larvae of Heligmosomoides polygyrus (: Nematospiroides dubius). 2. Studies on the fecundity of female worms as a function of the infectivity of third-stage larvae. Ann Rech Vet 9:161–168

    CAS  PubMed  Google Scholar 

  • MAFF. Ministry of Agriculture Fisheries and Food (1977) Technical Bulletin no 18 manual of veterinary parasitological laboratory techniques. Second Edition

  • MAFF: Ministry of Agriculture Fisheries and Food (1986) Reference book 418. Manual of veterinary parasitological laboratory techniques. Third edition

  • Mallet S, Kerboeuf D (1984) Relation entre l’activité enzymatique et le pouvoir infestant au cours du vieillissement des larves infestantes de Heligmosomoides polygyrus (Nematospiroides dubius). C R Acad Sci III 298:39–45

    CAS  Google Scholar 

  • Mallet S, Kerboeuf D (1985) Trichostrongylus colubriformis: relationship between ageing of infective larvae, infectivity and egg production by adult female worms. Ann Rech Vet 16(1):99–104

    CAS  PubMed  Google Scholar 

  • Maynard-Smith J (1989) Evolutionary genetics. Oxford University Press, Oxford

    Google Scholar 

  • O’Connor LJ, Walkden-Brown SW, Kahn LP (2006) Ecology of the free-living stages of major trichostrongylid parasites of sheep. Vet Parasitol 142:1–15

    Article  PubMed  Google Scholar 

  • Poulin R (1998) Evolutionary ecology of parasites; from individuals to communities. Champman and Hall, London

    Google Scholar 

  • Raynaud JP (1970) Etude de l’efficacité d’une technique de coproscopie quantitative pour le diagnostic le routine et le controle des infestations parasitaires des bovins, ovins, equins et porcins. Ann Parasitol 45:321–342

    CAS  Google Scholar 

  • Rew RS, Campbell WC (1983) Infectivity of Haemonchus controtus in sheep after freezing for 10 years over liquid nitrogen. J Parastiol 69:251–252

    Article  CAS  Google Scholar 

  • Roos MH, Otsen M, Hoekstra R, Veenstra JG, Lenstra JA (2004) Genetic analysis of inbreeding of two strains of the parasitic nematode Haemonchus contortus. Int J Parasitol 34:109–115

    Article  CAS  PubMed  Google Scholar 

  • Rossanigo CE (1992) Role de l’eau et de la température sur les taux de développement des nématodes parasites du tractus digestif des ruminants. PhD dissertation, Université Montpellier II

  • Siamba DN, Gatongi PM, Mulambalah CS, Ngeiywa MM, Wamae LW, Wambugu A (2011) Changes in lipids utilisation during moisture and temperature stress of infective (L3) and its implication on the epidemiology of Haemonchus contortus in arid and semi arid lands. Curr Res J Biol Sci 3(2):88–94

    Google Scholar 

  • Siamba DN, Mulambalah CS, Ngeiywa MM, Gatongi PM, Wamae LW (2012) Exsheathment characteristics and infectivity of revived anhydrobiotic L3 of Haemonchus contortus. Int J Anim Vet Adv 4(5):316–320

    Google Scholar 

  • Terefe G, Lacroux C, Andreoletti O, Grisez C, Prevot F, Bergeaud JP, Penicaud J, Rouillon V, Gruner L, Brunel JC, Francois D, Bouix J, Dorchies P, Jacquiet P (2007) Immune response to Haemonchus contortus infection in susceptible (INRA 401) and resistant (Barbados Black Belly) breeds of lambs. Parasite Immunol 29:415–424

    Article  CAS  PubMed  Google Scholar 

  • Van Wyk JA, Gerber HM, Van Aardt WP (1977) Cryopreservation of the infective larvae of the common nematodes of ruminants. Onderstepoort J Vet Res 44(3):173–194

  • Van Wyk JA, Gerber HM (1980) Survival and development of larvae of the common nematodes of ruminants after long-term cryopreservation and investigation of different routes of infestation. Onderstepoort J Vet Res 47:129–136

    PubMed  Google Scholar 

  • Van Wyk JA, Gerber HM (2000) Oral infectivity of cryopreserved Haemonchus contortus infective larvae that recovered relatively slowly when thawed after more than 15 years in liquid nitrogen. Onderstepoort J Vet Res 67:149–152

    PubMed  Google Scholar 

  • Van Wyk JA, Gerber HM, Villiers FJ (2000) Parenterally administered gastrointestinal nematode infective larvae viable after more than 15 years in liquid nitrogen. Vet Parasitol 88:239–247

    Article  PubMed  Google Scholar 

  • Vlassoff A, Leathwick DM, Health ACG (2001) The epidemiology of nematode infection in sheep. NZ Vet J 49(6):213–221

    Article  CAS  Google Scholar 

Download references

Acknowledgments

C. Chylinski is a grateful recipient of Marie Curie ITN funding “NematodeSystemHealth”. Thanks to T. Chaumeil and team for their care of the sheep.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to J. Cabaret.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chylinski, C., Cortet, J., Sallé, G. et al. Storage of gastrointestinal nematode infective larvae for species preservation and experimental infections. Parasitol Res 114, 715–720 (2015). https://doi.org/10.1007/s00436-014-4238-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-014-4238-5

Keywords

Navigation