Development of a standardized protein immunomarking protocol for insect mark–capture dispersal research

A field study was conducted to test the marking efficiency of broadcast spray applications of protein marks on stationary (represented by cadavers) and free‐roaming lady beetles Hippodamia convergens Guérin‐Méneville that were strategically placed in blooming alfalfa plots. The marks tested included three different concentrations of egg albumin from chicken egg white, casein from bovine milk and trypsin inhibitor from soy milk. The cadaver and free‐roaming beetle treatments served to measure the acquisition and retention of each protein treatment regime by direct contact with the spray solution and by residual contact with protein‐marked residue on alfalfa, respectively. In addition, the vertical distribution of marking efficacy was determined by sampling alfalfa plant tissue and beetle cadavers that were located on the upper and lower portion of the plant canopy. The data indicated that the backpack spray apparatus was very effective at uniformly administering the various protein marks, regardless of the concentration, throughout the entire plant canopy. Also, the free‐roaming beetles readily self‐marked by contact exposure to protein‐treated plants. We also identified concentrations of each protein type that will mark about 90% of the resident beetle population. Moreover, if a mark–capture‐type study only requires two unique protein marks, we determined that concentrations of 25% for egg white and 100% for bovine milk could be used to mark 98% of the population. Our results provide a significant step towards standardizing protein immunomarking protocols for insect mark–capture dispersal research. In addition, we identify several areas of research that are needed to further standardize the protein mark–capture procedure.     

Evaluating Drosophila suzukii immunomarking for mark‐capture research

Drosophila suzukii Matsumura (Diptera: Drosophilidae) utilizes ‘Himalaya’ blackberry, Rubus armeniacus Focke (Rosaceae), as a host and may invade berry and stone fruit crops from field margins containing this invasive weed. Laboratory and semi‐field studies were conducted to determine (1) the persistence of protein marks including 10% chicken egg whites (egg albumin protein), 20% bovine milk (milk casein protein), and 20% soy milk (soy trypsin inhibitor protein) on topically sprayed D. suzukii, (2) protein retention on blackberry leaves, and (3) D. suzukii acquisition of protein after exposure to marked blackberry leaves for up to 14 days after application. All flies and leaves were assayed for the presence of the protein marks using protein‐specific enzyme‐linked immunosorbent assays. Egg albumin, milk casein, and soy trypsin proteins persisted on 94, 49, and 25% of the topically marked D. suzukii, respectively, throughout the 14‐day study period. Egg albumin was retained on 100% of treated leaves for 14 days, regardless of environmental conditions. At least 50% of flies exposed residually to egg albumin‐treated leaves were marked for 3 days, regardless of exposure time and environmental conditions. However, increasing fly exposure time to treated leaves in April and June appeared to improve protein mark acquisition. Acquisition of protein by flies from treated leaves for milk casein was inconsistent, and poor for soy trypsin, despite detectable levels on treated leaves. Egg albumin had the longest and most consistent persistence on flies, leaves, and flies exposed to leaves in laboratory and semi‐field studies, under a variety of environmental conditions and exposure times.