Standardization of prey immunomarking: does a positive test always indicate predation?

A prey immunomarking procedure (PIP) in combination with generic anti-rabbit and anti-chicken immunoglobulin G (IgG) enzyme-linked immunosorbent assays (ELISAs) are used frequently to study arthropod predation. This study was conducted to: (1) further standardize the PIP as a tool for predator gut analysis research, (2) investigate the most effective means for administering IgG marks to prey items, and (3) assess the possibility of the PIP yielding false positive reactions as a consequence of a predator obtaining a mark by incidental contact with, or by a failed predation attempt on, a protein-marked prey item. The pest Lygus hesperus Knight (Hemiptera: Miridae) was tagged with either an external rabbit IgG mark, an internal chicken IgG mark, or a double (external rabbit IgG and internal chicken IgG) mark treatment. Then, the variously marked prey items were fed to chewing and piercing-sucking type predators and their gut contents were examined for the presence of IgG remains. Data revealed that all three marking treatments were highly effective at tagging targeted prey. However, ELISA results showed that the prey items should only be marked internally to maximize the likelihood of detecting prey remains while minimizing the risk of obtaining false positive errors. The merits and limitations of using the generic PIP for predator gut analysis research are discussed.