The development and characterization of an anti‐haemolymph antiserum for the detection of mollusc remains within carabid beetles

Abstract

Molluscan haemolymph was evaluated as an antigen for the detection, by enzyme-linked immunosorbent assay (ELISA), of mollusc proteins within the crop contents of carabid beetles, using an anti-haemolymph antiserum. The value of using a narrow range of prey-specific immunogens, rather than whole-body macerates, was discussed. Predators and potential prey were tested by a quantitative indirect ELISA, with separate evaluation of antigen reactions with non-specific and specific antibodies. The assay proved to be capable of detecting less than 1 ng of haemolymph protein and the antiserum reacted strongly with all molluscs tested. Initial cross-reactions with earthworms were effectively eliminated by absorption. A range of invertebrates was tested, and a significance level established relative to the invertebrates giving the strongest cross-reaction, which proved to be wolf spiders, Pardosa sp. Harvestmen (Opiliones), that initially gave strong reactions to the antiserum, were shown to have fed upon molluscs. Laboratory tests on the crop contents of slug-fed beetles demonstrated clear detection for at least 4 days after feeding.

Identification of predator–prey relationships between coccinellids and Saissetia oleae (Hemiptera: Coccidae), in olive groves, using an enzyme-linked immunosorbent assay

“…After 30 min in the dark, the reaction was stopped with 50 lL of sulfuric acid 2.5 M and the absorbance of each well was measured in a microplate reader Multiskan Ex (Labsystems, Finland) at 492 nm. The detection limit was conventionally established at AS C 2 9 NRS (Symondson and Liddell 1993).…”

“…The detection of S. oleae antigens was performed by indirect ELISA following the general protocol described by Symondson and Liddell (1993).…”

“…Consequently, analytical methods must often be used to assess prey consumption (Sopp et al 1992). Serological arthropod gut analyses, such as the enzyme-linked immunosorbent assay (ELISA), have been widely used to investigate predation on invertebrates offering a rapid identification of the remains of specific prey in the guts of predators (Fichter and Stephen 1981;Sunderland et al 1987;Sopp et al 1992;McIver and Tempelis 1993;Symondson and Liddell 1993;Symondson et al 1996;Morris et al 1999). Serological tests were successfully used in olive groves to study the natural enemies of the olive moth, Prays oleae (Bernard), whose main predators were found to be Formicidae, Heteroptera and Coleoptera (Morris et al 1999;Lozano et al 2002).…”

Detection of predation on Euzophera pingüis (Lepidoptera: Pyralidae) using an enzyme-linked immunosorbent assay

“…To protect against cross-reactions Lozano et al (1999) applied the antiserum to 27 different arthropod species. All predators tested with the polyclonal antiserum which gave a calculated concentration equivalent (CE) greater than 2.0, the mean CE for the highest cross-reacting species (Prays oleae) plus 2.5 S.D., were considered positive sensu Symondson & Liddell (1993a).…”

“…Various factors should be considered (Morris et al, 1999a), including meal size, mixed feeding and hunger level (Lövei et al, 1985(Lövei et al, , 1987(Lövei et al, , 1990, time that has elapsed since consumption (Symondson & Liddell, 1993a), temperature (Hagler & Cohen, 1990) and the species in question (Symondson & Liddell, 1993b). In 1998, the highest number of positives was recorded for spiders, and this may be due to the slower rate of digestion in these arthropods (Greenstone, 1983;.…”