Parasitism and Predation on Sentinel Egg Masses of Three Stink Bug Species (Hemiptera: Pentatomidae) in Native and Exotic Ornamental Landscapes.

This study evaluated parasitism and predation on sentinel egg masses of three stink bug species, the spined soldier bug, Podisus maculiventris (Say), the brown stink bug, Euschistus servus (Say), and the invasive brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), in ornamental landscapes composed of either native or exotic plants. This study also compared the species composition of parasitoids attacking two native stink bug species (P. maculiventris and E. servus) with those attacking the invasive BMSB on the same tree species in the same habitat. Overall, egg parasitism and predation were much higher on the two native stink bug species compared with BMSB, with an average parasitism rate of 20.6% for E. servus, 12.7% for P. maculiventris, and only 4.2% for H. halys and an average predation rate of 8.2% for E. servus,17.7% for P. maculiventris, and 2.3% for H. halys. Egg predation was also significantly higher on P. maculiventris than on E. servus eggs. Eight parasitoid species attacked sentinel stink bug eggs in the ornamental landscaped plots. Trissolcus euschisti (Ashmead) (Hymenoptera: Scelionidae) was the predominant parasitoid for all three stink bug species. There were no significant differences in parasitism and predation rates on any of the stink bug species between native and exotic plots. Therefore, there is no evidence that ornamental landscapes composed of native plants increased parasitism or predation rates of sentinel egg masses of two native stink bug species or the invasive BMSB, compared with those composed entirely of exotic plants.

The detectability half-life in arthropod predator-prey research: what it is, why we need it, how to measure it, and how to use it.

Molecular gut-content analysis enables detection of arthropod predation with minimal disruption of ecosystem processes. Most assays produce only qualitative results, with each predator testing either positive or negative for target prey remains. Nevertheless, they have yielded important insights into community processes. For example, they have confirmed the long-hypothesized role of generalist predators in retarding early-season build-up of pest populations prior to the arrival of more specialized predators and parasitoids and documented the ubiquity of secondary and intraguild predation. However, raw qualitative gut-content data cannot be used to assess the relative impact of different predator taxa on prey population dynamics: they must first be weighted by the relative detectability periods for molecular prey remains for each predator-prey combination. If this is not carried out, interpretations of predator impact will be biased towards those with the longest detectabilities. We review the challenges in determining detectability half-lives, including unstated assumptions that have often been ignored in the performance of feeding trials. We also show how detectability half-lives can be used to properly weight assay data to rank predators by their importance in prey population suppression, and how sets of half-lives can be used to test hypotheses concerning predator ecology and physiology. We use data from 32 publications, comprising 97 half-lives, to generate and test hypotheses on taxonomic differences in detectability half-lives and discuss the possible role of the detectability half-life in interpreting qPCR and next-generation sequencing data.

Removing external DNA contamination from arthropod predators destined for molecular gut-content analysis.

Ecological research requires large samples for statistical validity, typically hundreds or thousands of individuals, which are most efficiently gathered by mass-collecting techniques. For the study of interspecific interactions, molecular gut-content analysis enables detection of arthropod predation with minimal disruption of community interactions. Field experiments have demonstrated that standard mass-collection methods, such as sweep netting, vacuum sampling and foliage beating, sometimes lead to contamination of predators with nontarget DNA, thereby compromising resultant gut-content data. We deliberately contaminated immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of Leptinotarsa decemlineata by topically applying homogenate of the alternate prey Leptinotarsa juncta. We then attempted to remove contaminating DNA by washing in ethanol or bleach. A 40-min wash with end-over-end rotation in 80% EtOH did not reliably reduce external DNA contamination. Identical treatment with 2.5% commercial bleach removed most externally contaminating DNA without affecting the detectability of the target prey DNA in the gut. Use of this bleaching protocol, perhaps with minor modifications tailored to different predator-prey systems, should reliably eliminate external DNA contamination, thereby alleviating concerns about this possible source of cross-contamination for mass-collected arthropod predators destined for molecular gut-content analysis.

Barcode haplotype variation in north American agroecosystem lady beetles (Coleoptera: Coccinellidae).

DNA barcodes have proven invaluable in identifying and distinguishing insect pests, most notably for determining the provenance of exotic invasives, but relatively few insect natural enemies have been barcoded. We used Folmer et al.'s (1994) universal invertebrate primers and Hebert et al.'s (2004) for Lepidoptera, to amplify 658 bp at the 5' end of the mitochondrial cytochrome oxidase c subunit I (COI) gene in five species of lady beetles from crop fields in six states in the US Mid-Atlantic, Plains and Midwest: three native species, Hippodamia convergens Guérin-Méneville, H. parenthesis (Say) and Coleomegilla maculata (De Geer); and two exotic species, Harmonia axyridis (Pallas) and Coccinella septempunctata Linnaeus. Sequence divergences within species were low, never exceeding 0.9% (Kimura 2-parameter distances). Sequence divergences between the two Hippodamia species ranged from 14.7 to 16.4%, mirroring the relationships found for other arthropod taxa. Among the exotic species, C. septempunctata sequences were as variable as those of the three native species, while H. axyridis populations comprised a single haplotype. Limited data on two Coleomegilla subspecies, C. m. lengi Timberlake and C. m. fuscilabris (Mulsant), are consistent with their belonging to the same species, although morphological and reproductive data indicate that they represent separate species. Our results support the general utility of COI barcodes for distinguishing and diagnosing coccinellid species, but point to possible limitations in the use of barcodes to resolve species assignments in recently divergent sibling species.

Unnecessary roughness? Testing the hypothesis that predators destined for molecular gut-content analysis must be hand-collected to avoid cross-contamination.

Molecular gut-content analysis enables detection of arthropod predation with minimal disruption of ecosystem processes. Mass-collection methods, such as sweep-netting, vacuum sampling and foliage beating, could lead to regurgitation or rupturing of predators along with uneaten prey, thereby contaminating specimens and compromising resultant gut-content data. Proponents of this 'cross-contamination hypothesis' advocate hand-collection as the best way to avoid cross-contamination. However, hand-collection is inefficient when large samples are needed, as with most ecological research. We tested the cross-contamination hypothesis by setting out onto potato plants immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of either Leptinotarsa decemlineata or Leptinotarsa juncta, or unfed individuals of these predator species along with L. decemlineata larvae. The animals were then immediately re-collected, either by knocking them vigorously off the plants onto a beat cloth and capturing them en masse with an aspirator ('rough' treatment) or by hand-searching and collection with a brush ('best practice'). Collected predators were transferred in the field to individual vials of chilled ethanol and subsequently assayed by PCR for fragments of cytochrome oxidase I of L. decemlineata and L. juncta. Ten to 39 per cent of re-collected fed predators tested positive by PCR for DNA of both Leptinotarsa species, and 14-38% of re-collected unfed predators contained L. decemlineata DNA. Overall levels of cross-contamination in the rough (31%) and best-practice (11%) samples were statistically different and supported the cross-contamination hypothesis. A pilot study on eliminating external DNA contamination with bleach prior to DNA extraction and amplification gave promising results.

Tracking the role of alternative prey in soybean aphid predation by Orius insidiosus: a molecular approach.

The soybean aphid, Aphis glycines (Hemiptera: Aphididae), is a pest of soybeans in Asia, and in recent years has caused extensive damage to soybeans in North America. Within these agroecosystems, generalist predators form an important component of the assemblage of natural enemies, and can exert significant pressure on prey populations. These food webs are complex and molecular gut-content analyses offer nondisruptive approaches for examining trophic linkages in the field. We describe the development of a molecular detection system to examine the feeding behaviour of Orius insidiosus (Hemiptera: Anthocoridae) upon soybean aphids, an alternative prey item, Neohydatothrips variabilis (Thysanoptera: Thripidae), and an intraguild prey species, Harmonia axyridis (Coleoptera: Coccinellidae). Specific primer pairs were designed to target prey and were used to examine key trophic connections within this soybean food web. In total, 32% of O. insidiosus were found to have preyed upon A. glycines, but disproportionately high consumption occurred early in the season, when aphid densities were low. The intensity of early season predation indicates that O. insidiosus are important biological control agents of A. glycines, although data suggest that N. variabilis constitute a significant proportion of the diet of these generalist predators. No Orius were found to contain DNA of H. axyridis, suggesting intraguild predation upon these important late-season predators during 2005 was low. In their entirety, these results implicate O. insidiosus as a valuable natural enemy of A. glycines in this soybean agroecosystem.

Molecular markers for identification of the hyperparasitoids Dendrocerus carpenteri and Alloxysta xanthopsis in Lysiphlebus testaceipes parasitizing cereal aphids.

Polymerase chain reaction (PCR)-based molecular markers have been developed to detect the presence of primary parasitoids in cereal aphids and used to estimate primary parasitism rates. However, the presence of secondary parasitoids (hyperparasitoids) may lead to underestimates of primary parasitism rates based on PCR markers. This is because even though they kill the primary parasitoid, it's DNA can still be amplified, leading to an erroneous interpretation of a positive result. Another issue with secondary parasitoids is that adults are extremely difficult to identify using morphological characters. Therefore, we developed species-specific molecular markers to detect hyperparasitoids. A 16S ribosomal RNA mitochondrial gene fragment was amplified by PCR and sequenced from two secondary parasitoid species, Dendrocerus carpenteri (Curtis) (Hymenoptera: Megaspilidae) and Alloxysta xanthopsis (Ashmead) (Hymenoptera: Charipidae), four geographic isolates of the primary parasitoid, Lysiphlebus testaceipes (Cresson) (Hymenoptera: Braconidae), and six aphid species common to cereal crops. Species-specific PCR primers were designed for each insect on the basis of these 16S rRNA gene sequences. Amplification of template DNA, followed by agarose gel electrophoresis, successfully distinguished D. carpenteri and A. xanthopsis from all four isolates of L. testaceipes and all six cereal aphid species in this laboratory test.

Prey preference and host suitability of the predatory and parasitoid carabid beetle, Lebia grandis, for several species of Leptinotarsa beetles.

Lebia grandis (Coleoptera: Carabidae), recorded as a parasitoid only on Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae), is capable of parasitizing the false potato beetle, L. juncta, and also L. haldemani. Historical records show that L. decemlineata, while the only recorded host, was not present in much of the original range of L. grandis, and may not have been its host prior to its expansion into eastern North America, where L. juncta is endemic. Our laboratory comparisons suggest that L. juncta, the presumptive original host, best supports the development of the parasitoid larval L. grandis, based on 43.6% successful emergence of the adult carabid parasitoid, compared to 11.5% from the two other Leptinotarsa species. L. grandis adults accept eggs and larvae of all 3 Leptinotarsa species as adult food. Naive, newly-emerged adults show no preference when presented the 3 species of third-instar larvae, which they consume at a mean rate of 3.3 per day, a rate which does not differ significantly by sex, larval host, or weight at emergence. When presented with equal amounts by weight of the 3 species of Leptinotarsa eggs, such adults consume the equivalent of 23.0 L. decemlineata eggs per day, with consumption of L. juncta eggs 67% higher by weight than L. decemlineata consumption. Insight into the biotic and abiotic limitations on L. grandis should aid in determining its potential for suppression of Colorado potato beetle by biological control in diverse agroecosystems.

Overlapping confidence intervals or standard error intervals: what do they mean in terms of statistical significance?

We investigate the procedure of checking for overlap between confidence intervals or standard error intervals to draw conclusions regarding hypotheses about differences between population parameters. Mathematical expressions and algebraic manipulations are given, and computer simulations are performed to assess the usefulness of confidence and standard error intervals in this manner. We make recommendations for their use in situations in which standard tests of hypotheses do not exist. An example is given that tests this methodology for comparing effective dose levels in independent probit regressions, an application that is also pertinent to derivations of LC50s for insect pathogens and of detectability half-lives for prey proteins or DNA sequences in predator gut analysis.

Determinants of web spider species diversity: Vegetation structural diversity vs. prey availability.

The hypotheses that vegetation structural diversity and prey availability determine alpha diversity were examined for scrub- and meadow-inhabiting web spider species assemblages along elevational gradients in Costa Rica and California. Prey availabilities were estimated by sticky trap catches, using only the orders and size classes of insects actually captured by the spiders. The measured component of vegetation structural diversity was the maximum tip height in 20 cm increments from 0 to 2.0 m. Spider species diversity and vegetation tip height diversity were both expressed by the Inverse Simpson Index. Web spider species diversity is highly significantly correlated with vegetation tip height diversity. Prey availability is not a significant predictor of web spider species diversity in these habitats.

Site-specificity and site tenacity in a wolf spider: A serological dietary analysis.

The wolf spider Pardosa ramulosa is a dietary specialist on aquatic insects. Site tenacity of this species was determined by serological analysis of spiders collected on the margins of and at known distances from each of eight closely-spaced pools whose prey compositions were known. Remains of a mosquito species found uniquely in three of these pools were found only in spiders closely associated with them, despite the presence of otherwise suitable pools less than 10 m away. This indicates that spiders stay at specific pools for at least two weeks, the detectability period of this marker species in the assay.The only exception to this site tenacity, except for inadvertant rafting during high tides, is the movement of adult females with egg cases away from pools. This is attributed to selection for avoidance of spiderling cannibalism, rather than a search for sunning or foraging sites as in some other lycosids.The extreme site-tenacity of P. ramulosa is attributed to the high productivity of all pools, such that spiders from the most and least productive pools differ little if at all in fitness, and the high costs of moving to other pools, which are widely dispersed. The situation in crab spiders may be analogous. Conversely high and biologically significant variances in web site productivity, low distances between acceptable web sites, and low moving costs reduce site tenactiy in north temperate second growth orb weavers and a desert funnel web weaver.