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Validated flow cytometry allows rapid quantitative assessment of immune responses in amphibians

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Assessments of amphibian immune state have been commonly made through indirect methods like phytohemagglutinin (PHA) injections, or by direct methods like cell counts from blood smears. Here we validate a simple method to assess immune responses in amphibians by means of flow cytometry with a fluorescent lipophilic dye (3,3′ Dipentyloxacarbocyanine), which removes the need for specific antibodies. We experimentally altered the immunological state of Pelobates cultripes tadpoles by exposing some to exogenous corticosterone. We then determined the immune state of each tadpole through both blood smears and flow cytometry. We found that both techniques showed similar patterns of the proportion of white blood cells. Once validated, flow cytometry also allowed quantitation of changes in absolute number of white cells. We discuss the suitability of both techniques attending to the accuracy of each technique, body size requirements, or the tractability in field studies.

Affiliations: 1: Doñana Biological Station, CSIC, Avda. Américo Vespucio s/n, 41092 Seville, Spain

*Corresponding author; e-mail:

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