Integrating time progression in ecoimmunology studies: beyond immune response intensity

Habitat alterations in agroecosystems may damage amphibian immune capacity. As agroecosystem extension is increasing worldwide, broader-context knowledge on the effects of agroecosystem stressful conditions on amphibian immune capacity is crucial for understanding and management of amphibian global declines. However, most studies on ecoimmunology assume synchronal among-group immune-response peaks, and focus on immune response after standardized time lapses, neglecting its progression. Herein, I compared inflamatory response short-term progression of agroecosystem and natural-habitat female and male natterjack toads Epidalea calamita, by measuring swelling response, once per hour, 6 h following an artificial immune challenge with innocuous antigen phytohemagglutinin. I also compared maximum magnitude of swelling response, irrespective of the moment when it was reached. Habitat differences arose only 3 h after challenge, when natural-habitat toads showed greater swelling response. Maximum magnitude of swelling response was similar in toads from both habitats. However, agroecosystem toads showed a delayed swelling response as compared with natural-habitat conspecifics, probably as a consequence of agroecosystem stressful conditions. Such a delay suggests a weaker immune capacity, and consequently impaired anti-pathogen performance. Regarding sex, swelling response magnitude did not differ between males and females. Female swelling response peaked earlier, but that of males was more sustained in time, which contradicts general across-taxa findings that males show impaired immune response. Interestingly, results suggest that measuring swelling response only after some standardized period following stimulation may be a simplistic approach and produce unrealistic results. Consequently, studies on ecoimmunology should implement swelling response progression in order to produce unbiased science.