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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"One disadvantage of PCR in comparison with monoclonal antibody technology is the inability to achieve stage or instar level specificity (cf. Greenstone & Morgan 1989; Hagler et al. 1994; Greenstone 1995) because of the presence of DNA in all tissues of all life stages. Such specificity might be achieved by reverse-transcriptase PCR, enabling the detection of mRNAs expressed at different developmental periods in the life of the insect. "
[Show abstract][Hide abstract] ABSTRACT: We describe polymerase chain reaction (PCR) primers for gut analysis of aphid predators. The primers amplify aphid mitochondrial COII fragments ranging in size from 77 to 386 bp. Using these primers, we were able to distinguish six species of US Great Plains cereal aphids, including two congeners, Rhopalosiphum maidis (Fitch) and R. padi (L.), and to detect them in extracts of coccinellid and chrysopid predators. We devised a protocol for deriving half-lives of detectability for the DNA of a single aphid consumed by predators maintained under simulated field dietary and temperature conditions. Using this protocol and primers that amplify a 198-bp fragment, we determined statistically different half-lives of detectability for a single R. maidis of 3.95 h in Chrysoperla plorabunda (Fitch) and 8. 78 h in Hippodamia convergens Guerin. The detectability half-life for a 339-bp R. maidis fragment was statistically longer in C. plorabunda but not in H. convergens. The sensitivity of the assay for the 198-bp fragment is 10-7 aphid equivalents. For species-specific predator gut analysis, PCR is superior to monoclonal antibody technology, giving comparable detectability half-lives with lower expense, much shorter development times, and greater certainty of a successful outcome.
"This study was designed to evaluate the efficacy of several gut content immunoassays on whole-body homogenized adult lady beetles, Hippodamia convergens Guérin-Meneville that had consumed either one or five pink bollworm, Pectinophora gossypiella (Saunders), eggs. An indirect ELISA and a dot blot designed specifically to detect pink bollworm egg remains in arthropod guts have been described (Hagler et al., 1994; 1995). "
[Show abstract][Hide abstract] ABSTRACT: Five different immunoassay formats were examined for their ability to detect a minute quantity of prey remains in predator guts. The convergent lady beetle, Hippodamia convergens Guerin-Meneville, that had consumed either one or five pink bollworm, Pectinophora gossypiella (Saunders), eggs was evaluated by the following immunoassays: three variations of enzyme-linked immunosorbent assay (ELISA), a dot blot, and a Western blot. Sandwich ELISA, dot blot, and Western blot were the most sensitive immunoassays based on the proportion of individual predators scoring positive for prey remains. The direct ELISA and indirect ELISA were ineffective at detecting prey in the predators. The advantages and disadvantages of each immunoassay format are discussed.
Biological Control 05/1998; 12(1). DOI:10.1006/bcon.1998.0612 · 1.64 Impact Factor
"Predators were removed from the environmental chambers at 12, 24, 48, or 72 h after feeding. Each individual was homogenized in 500 µl of PBS and assayed for the pink bollworm egg antigen by the indirect ELISA described by Hagler et al. (1994). We fitted negative exponential equations to describe the decay in the proportion of positive responses with time after feeding for each predator species and holding temperature (SAS Institute, 1990). "
[Show abstract][Hide abstract] ABSTRACT: The gut contents of three species of insect predators that were fed either a variable or a fixed number of pink bollworm eggs but held at variable time and temperature regimes were assayed by an indirect enzyme-linked immunosorbent assay (ELISA). The sensitivity and efficacy of the monoclonal antibody-based ELISA was dependent on the predator species examined. Small predators were more immunoresponsive to the ELISA than large predators. Furthermore, the assay sensitivity was dependent on the number of prey consumed, elapsed time after feeding, and temperature at which the predators were held. The smaller predator species retained recognizable traces of prey remains for longer periods than larger predator species. The ELISA efficacy decreased with increasing ambient temperature. A series of regression equations have been developed to estimate the median detection interval of prey in a predator's gut that takes into account the predator species examined, the quantity of prey consumed, and ambient after-meal temperature.
Biological Control 06/1997; 9(2). DOI:10.1006/bcon.1997.0521 · 1.64 Impact Factor