A Monoclonal Antibody to Pink Bollworm (Lepidoptera: Gelechiidae) Egg Antigen: A Tool for Predator Gut Analysis
ABSTRACT We describe the development, selection, and application of a monoclonal antibody (MAb) to eggs of pink bollworm, Pectinophora gossypiella (Saunders). We tested this MAb against all pink bollworm life stages and the egg stage of 10 other insect species using an enzyme-linked immunosorbent assay (ELISA). In all cases, the MAb was highly specific to pink bollworm egg and adult female antigens. A Western blot analysis showed that the MAb reacted with two egg polypeptides with molecular weights between 46 and 60 kDa. Predation studies were conducted in the laboratory to test the usefulness of this MAb for studying predator-prey interactions. Most predators fed either one or two pink bollworm eggs responded positively to the MAb in a serological analysis of gut contents. These data suggest that this MAb can be used as a diagnostic probe for gut content analysis of potential predators of pink bollworm eggs under field conditions.
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ABSTRACT: We extend detection of arthropod predator gut contents by polymerase chain reaction (PCR), heretofore restricted to insect predators, to spiders. Single individuals of the corn lead aphid, Rhopalosiphum maidis, were detected in the guts of spiderlings of Oxyopes salticus up to 12 h after feeding; individuals of the congeneric bird cherry oat aphid, R. padi, were not detected. Unfed O. salticus and Misumenops sp. were also negative.Journal of Arachnology 01/2009; · 0.73 Impact Factor
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ABSTRACT: Molecular analysis of predation enables accurate and reliable elucidation of trophic linkages in complex food webs, but identifying the strength of such interactions can be subject to error. Currently two techniques dominate: monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Although the optimization and characterization of these systems ensures their sensitivity and specificity, predator collection protocols such as sweep-netting and vacuum sampling could overestimate feeding rates because of surface-level contamination, yielding positive reactivity or predation within the sampling device. Therefore, two sampling techniques (sweep-net sampling and hand collection) were compared within an alfalfa agroecosystem using a monoclonal antibody-based ELISA to test the hypothesis that cross-contamination is a source of error, i.e., significantly more predators (linyphiid spiders) would test positive for prey (Diptera) proteins. A concurrent study examining the viability of trapping predators into saline solution was also undertaken. No significant differences were found between the proportions of spiders screening positive for Diptera when collected by sweep-net versus hand collection, rejecting the hypothesis that sweep-netting predators for subsequent molecular gut content analysis overestimates predation frequency. ELISA was also capable of detecting prey proteins in predator guts from pitfall traps containing phosphate-buffered saline, indicating the suitability of this approach for the collection and analysis of epigeal predators. Although these results indicate that sweep netting and pitfall trapping into solution is appropriate in this predator-prey and ELISA analysis system, caution should be exercised with other interactions and PCR-based analysis. The likelihood for false-positive reactivity should therefore be considered on a case-by-case basis.Environmental Entomology 09/2008; 37(4):990-5. · 1.31 Impact Factor
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ABSTRACT: The pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae), and related psyllids are important pests of pear (Pyrus spp.) worldwide. Many of these pests are thought to be partially controlled by predatory insects. To improve our understanding of the predator species that attack pear psylla, we developed monoclonal antibodies (MAbs) against this pest for predator gut content studies. Mice were immunized with homogenates of nymphal, adult, and egg stages of pear psylla. A mouse immunized with nymph homogenate showed high activity against all three antigen types and was used for MAb development. From 952 hybridomas screened, 35 showed good activity to pear psylla and low activity against nontarget arthropods. Four MAbs were retained: two from immunoglobulin M (IgM)-secreting hybridomas, both with high activity against all stages of psylla except young eggs, and two immunoglobin G-secreting hybridomas, both with high activity against psylla eggs and gravid adult females. Using one of the IgM-MAbs, pear psylla remains were detected in the predatory bugs Anthocoris tomentosus Péricart (Heteroptera: Anthocoridae) and Deraeocoris brevis (Uhler) (Heteroptera: Miridae) in laboratory feeding trials. Digestion half lives typically exceeded 24 h and were dependent on meal size and predator life stage. Gut content analysis of 970 field-collected D. brevis and Anthocoris spp. showed that the proportion which fed on psylla averaged 59% and that percentage closely tracked the density of pear psylla nymphs during three seasons. The utility of these antibodies for the study of trophic interactions and habitat management in relation to biological control of pear psylla is discussed.Annals of the Entomological Society of America 01/2009; · 1.20 Impact Factor