Abstract
Murine immunological responses to experimental infection with Trichuris muris and the effects of the resident microbiota on these responses are of increasing interest. For these studies, accurate dose delivery and improved sterilization of inocula are essential to prevent co-infection with unknown contaminants. We found that washing T. muris eggs with antibiotics may not be sufficient for sterilization of inocula. However, washing eggs in 6.25% hypochlorite/bleach eliminated bacteria and fungi, as determined by culture and PCR, did not harm viable T. muris eggs and reduced the number of non-viable eggs in the inocula. A hatching assay and propidium iodide staining method were developed and found to increase the accuracy for assessing T. muris egg viability prior to infection for rapid dose evaluation. In addition, metal gavage feeding needles increased the accuracy and precision of the dose delivered to the mice compared to flexible rubber tubes. These methods will improve experimental Trichuris studies by decreasing the variability in outcome due to unintended carryover of adherent microorganisms and unrecognized variation in inocula.
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Acknowledgements
We thank Dr. Julia Bell for the help with the statistical methods and for the review of the manuscript. The project was funded in whole with federal funds from the NCRR, NIH, and the Department of Health and Human Services under grant no. 5K26RR023080. Some of the mice were produced with federal funds from the NIAID, NIH, and the Department of Health and Human Services under contract no. N01-AI-30058.
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Kopper, J.J., Mansfield, L.S. Development of improved methods for delivery of Trichuris muris to the laboratory mouse. Parasitol Res 107, 1103–1113 (2010). https://doi.org/10.1007/s00436-010-1978-8
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DOI: https://doi.org/10.1007/s00436-010-1978-8