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In vitro hatching of Trichuris suis eggs

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Abstract

Eggs of the pig whipworm, Trichuris suis ova (TSO), are currently tested in human clinical trials for their potential immunomodulatory capacity. The biological potency of TSO (egg viability and infectivity) is traditionally assessed in Göttingen minipigs as the establishment of intestinal larvae after inoculation with a known number of eggs. To minimize testing in animal models, development of an in vitro egg hatching assay is proposed as a reliable, cost-effective, and a faster alternative to test the egg viability. The present study aimed to investigate the influence of different chemical, physical, and biological factors on egg hatching. Thus, in a series of experiments and in different combinations, the eggs were stimulated with glass beads, artificial gastric juice, bile salt and trypsin solution, fermentation gut medium, or stimulated with mucosal scrapings from the ileum and the large intestine of the infected and uninfected Göttingen minipig. Mechanical stimulation with glass beads presented a simple and reproducible method for egg hatching. However, incubation of eggs with mucosal scrapings from the ileum, caecum, and colon for 24 h at 38 °C significantly increased hatching.

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Acknowledgments

Laboratory technicians Charlotte Fischer and Niels Peter Hansen are thanked for their excellent technical assistance. We would like to express our thanks to Ellegaard Göttingen Minipigs A/S (Dalmose, Denmark) for supplying minipigs for the study. Our gratitude goes to Dr. Christophe Chassard from the Institute of Food, Nutrition and Health (Swiss Federal Institute of Technology Zürich, Switzerland) for supplying fermented gut medium used in the experiment and for fruitful discussion. Dr. Branko Kokotovic at the Section for Bacteriology, Pathology and Parasitology (National Veterinary Institute, DTU, Denmark) is acknowledged for assisting with the use of an anaerobic chamber.

Ethical approval

The study with minipigs was conducted under approval from The Animal Experiments Inspectorate, Danish Ministry of Food, Agriculture and Fisheries (license number: 2012-15-2934-00641).

Compliance with Ethical Standards

S. M. Thamsborg, H. Kringel, J. M. Bruun, and C. M. O. Kapel are employed by Parasite Technologies A/S, a company that is GMP-licensed by the Danish Medicines Agency for production of T. suis eggs.

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Authors and Affiliations

  1. Section for Organismal Biology, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

    Nermina Vejzagić & Christian M. O. Kapel

  2. Parasite Technologies A/S, Dr. Neergaards Vej 3, 2970, Hørsholm, Denmark

    Stig Milan Thamsborg, Helene Kringel, Allan Roepstorff, Johan Musaeus Bruun & Christian M. O. Kapel

  3. Parasitology and Aquatic Diseases, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Dyrlægevej 100, 1870, Frederiksberg C, Denmark

    Stig Milan Thamsborg

Authors
  1. Nermina Vejzagić
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  2. Stig Milan Thamsborg
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  3. Helene Kringel
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  4. Allan Roepstorff
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  5. Johan Musaeus Bruun
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  6. Christian M. O. Kapel
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Correspondence to Nermina Vejzagić.

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Vejzagić, N., Thamsborg, S.M., Kringel, H. et al. In vitro hatching of Trichuris suis eggs. Parasitol Res 114, 2705–2714 (2015). https://doi.org/10.1007/s00436-015-4476-1

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  • DOI: https://doi.org/10.1007/s00436-015-4476-1

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