What are the relative costs and benefits of mounting immune responses? Practitioners of ecoimmunology have grappled with this central question since the field's inception with the main tension being how to make tractable methodological choices that maintain the ecological relevance of induced and measured immune costs. Here, we point out two methodological approaches that we feel are underrepresented in the field, describe risks associated with neglecting these methods, and suggest modern techniques that maximize both the diversity and ecological relevance of collected data. First, it is commonly assumed that frequently used and experimentally convenient immune stimulants will induce ecologically relevant immune responses in study organisms. This can be a dangerous assumption. Even if a stimulant's general immune response properties are well characterized, it is critical to also measure the type and scale of immune responses induced by live pathogens. Second, patterns of immune defenses evolve like other traits, thus a comparative approach is essential to understand what forces shape immune variation. Finally, we describe modern genetic and immunological approaches that will soon become essential tools for ecoimmunologists, and present case studies that exemplify the utility of our recommendations.
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