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Biomedical subjects

M H Greenstone

Publications and source records attributed to M H Greenstone.

4 recordsLinked to original sources

Molecular methods for assessing insect parasitism.

Determining insect parasitism rates is problematic due to the small size and lack of useful distinguishing morphological characters of many parasitoid taxa. To solve this problem, entomologists have employed one of four general methods to detect parasitoid protein or nucleic acid markers: serological assay; random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR); allozyme electrophoresis; or specific PCR. Serological methods, especially with monoclonal antibodies, are unrivalled for specificity, enabling discrimination at the stage as well as species level. However, they have not found favour with many workers, possibly due to complexity and expense. RAPD-PCR has been widely used, but can only be recommended for restricted applications because of its poor reproducibility. Allozyme electrophoresis provides reproducible detection and discrimination of closely related species. Specific-PCR is highly specific and reproducible, and also has the shortest latency for detection, usually 24 h or less after parasitization. The substantial existing literature on allozyme electrophoresis and specific PCR is used to support recommendations on what are apt to be fruitful enzyme systems or genomic regions for detecting and discriminating parasitoids in untried parasitoid-host assemblages.

Animals↗

Barcoding generalist predators by polymerase chain reaction: carabids and spiders.

Identification of arthropod predators is challenging when closely related species are found at a given locality. Identification of the immature stages is especially problematic, because distinguishing morphological features are difficult to use or have not been described. We used polymerase chain reaction (PCR) to distinguish closely related carabids and spiders, and to match eggs and larvae (or nymphs) with identified adult parents. Within the Carabidae, we amplified species-specific mitochondrial cytochrome oxidase I (COI) fragments for three species each in the genera Poecilus and Harpalus, and two each in Chlaenius and Bembidion. Within the Araneae, we amplified species-specific COI fragments for two Hibana species (Anyphaenidae), Pardosa milvina and Rabidosa rabida (Lycosidae), Frontinella communis and Grammonota texana (Linyphiidae), and Cheiracanthium inclusum (Miturgidae). We are able to correctly identify all immature stages tested--eggs, larvae (or nymphs) and pupae--by comparison of the amplified fragments with those of the adults. Using COI markers as species identifiers is a tenet of the Barcode of Life initiative, an international consortium to provide a molecular identifier for every animal species.

Animals↗

Can generalist predators be effective biocontrol agents?

Theoretical developments are helping us to comprehend the basic parameters governing the dynamics of the interactions between generalist predators and their many pest and nonpest prey. In practice, however, inter- and intraspecific interactions between generalist predators, and between the predators and their prey, within multispecies systems under the influence of rapidly changing biotic and abiotic variables are difficult to predict. We discuss trade-offs between the relative merits of specialists and generalists that allow both to be effective, and often complementary, under different circumstances. A review of manipulative field studies showed that in approximately 75% of cases, generalist predators, whether single species or species assemblages, reduced pest numbers significantly. Techniques for manipulating predator numbers to enhance pest control at different scales are discussed. We now need to find ways of disentangling the factors influencing positive and negative interactions within natural enemy communities in order to optimize beneficial synergies leading to pest control.

Animals↗

Identifying key cereal aphid predators by molecular gut analysis.

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.

Animals↗