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International audience; DNA methods are useful to identify ingested prey items from the gut of predators, but reliable detection is hampered by low amounts of degraded DNA. PCR-based methods can retrieve minute amounts of starting material but suffer from amplification biases and cross-reactions with the predator and related species genomes. Here, we use PCR-free direct shotgun sequencing of total DNA isolated from the gut of the harlequin ladybird Harmonia axyridis at five time points after feeding on a single pea aphid Acyrthosiphon pisum. Sequence reads were matched to three reference databases: Insecta mitogenomes of 587 species, including H. axyridis sequenced here; A. pisum nuclear genome scaffolds; and scaffolds and complete genomes of 13 potential bacterial symbionts. Immediately after feeding , multicopy mtDNA of A. pisum was detected in tens of reads, while hundreds of matches to nuclear scaffolds were detected. Aphid nuclear DNA and mtDNA decayed at similar rates (0.281 and 0.11 h À1 respectively), and the detectability periods were 32.7 and 23.1 h. Metagenomic sequencing also revealed thousands of reads of the obligate Buchnera aphidicola and facultative Regiella insecticola aphid symbionts, which showed exponential decay rates significantly faster than aphid DNA (0.694 and 0.80 h À1 , respectively). However, the facultative aphid symbionts Hamil-tonella defensa, Arsenophonus spp. and Serratia symbiotica showed an unexpected temporary increase in population size by 1–2 orders of magnitude in the predator guts before declining. Metagenomics is a powerful tool that can reveal complex relationships and the dynamics of interactions among predators, prey and their symbionts.

Insect predators are exposed to the Cry1Ac toxin in Bt cotton fields through several pathways. In this study, we investigated the effects of activated Cry1Ac added to a diet on Cycloneda sanguinea (L.) (Coleoptera: Coccinellidae), which is one of the main predators of non-target pests in Brazilian cotton. Direct bitrophic exposure of C. sanguinea to Cry1Ac was done by feeding beetles with Aphis gossypii (Glover) (Hemiptera: Aphidae) sprayed with 500 μg per ml Cry1Ac solution. Larval and pupal survival, development time, aphid consumption, and adult longevity were recorded daily. Couples within the same experimental treatment were paired and numbers of eggs laid and hatched per female were recorded daily. Net replacement rate was calculated for each female. During development, a C. sanguinea larva consumed on average 1.8 μg of activated Cry1Ac. No significant differences due to Cry1Ac were observed for any of the response variables, except aphid consumption. Larvae receiving Cry1Ac consumed more aphids than larvae receiving distilled water alone. Additional statistical analyses were conducted to evaluate independence of responses, and for the independent responses, a simple meta-analysis was conducted to test the null hypothesis that all responses were zero. Nearly all of the response variables were statistically independent. Two pairs of responses were not independent, but the associated multivariate tests were not significant. The meta-analysis suggested that all effects were not different from random variation around zero and no cumulative effects could be detected. Our results indicated that bitrophic exposure to activated Cry1Ac is likely to have little or no adverse ecological effect on C. sanguinea.

Herbivory is known to increase the emission of volatiles, which attract natural enemies to herbivore-damaged plants in laboratory and agricultural systems. We report on signalling through volatiles induced by Euschistus heros (F.) (Heteroptera: Pentatomidae) in two legumes that influence the attraction and retention of the egg parasitoid Telenomus podisi (Ashmead) (Hymenoptera: Scelionidae). Air-borne extracts obtained from two host plants of E. heros, soybean, Glycine max, and pigeon pea, Cajanus cajan (Leguminosae), produce a different blend of emitted volatiles when attacked by adult males or females and nymphs of the pest species, compared with the undamaged plants. The same results were obtained when the plants were treated with extracted saliva of E. heros which had been mechanically introduced into the plants. This indicates that some substance in the saliva contributed to the release of the volatiles. Bioassays in a Y-tube olfactometer with female T. podisi and treated plants confirmed the significant preference of the egg parasitoid for plants attacked by either males, females, or fourth instar nymphs of the pest species. On the other hand, volatile extracts obtained from soybean subjected to the velvetbean caterpillar, Anticarsia gemmatalis, a non-host species for T. podisi, showed a different blend of volatiles compared to those obtained from plants subjected to E. heros. Additionally, the volatiles obtained from this plant–host complex were not attractive to T. podisi. These results indicate that E. heros causes an increase in the emission of specific plant volatiles, and that the induction is possibly caused by an elicitor present in the pest saliva. The possibility that these plant volatiles play an important role in the attraction and retention of the egg parasitoid T. podisi is discussed.

Summary 1. Predictions from the Phylogenetic Constraints Hypothesis were tested for the first time in a tropical system using the pasture pest Deois flavopicta Stal, which oviposits into the ground and not into plant tissues. The prediction that there is no oviposition preference–larval performance linkage was supported. The absence of such a link provides an evolutionary basis for eruptive population dynamics. 2. The effects of host species and host plant quality on the preference of ovipositing females of D. flavopicta and performance of their offspring on the selected host plants were tested at the population level. 3. Female oviposition behaviour was affected by the presence of the host plant. Females of D. flavopicta showed a strong preference to oviposit close to host plants. The number of eggs was higher in pots containing Brachiaria ruziziensis (121.88 ± 13.70) than in pots containing only the wet oviposition substrate (5.2 ± 1.98) or dry oviposition substrate (0.067 ± 0.067). Ovipositing females did not, however, discriminate between plants of Brachiaria decumbens and Axonopus marginatus and did not show a strong oviposition preference in relation to differences in plant quality (protein and fibre content). They did show oviposition preference for plants under the high watering regime. The mean number of eggs collected from pots with non water-stressed plants was 60% higher than the mean number of eggs collected on pots with water-stressed plants. 4. Although females did not show ovipositional preference, spittlebug larval performance, measured as percentage survival and duration of nymphal period, was better on plants of high protein and low fibre content. These results indicate that there is not a linkage between female oviposition preference and subsequent nymphal performance in relation to differences in protein and fibre content in the host plants. There was, however, a limited linkage between oviposition preference and nymphal performance in relation to plant water content. Females showed preference for moist sites that have high survival of newly hatched nymphs. 5. Evidence indicates that for D. flavopicta, the influences of natality and female oviposition behaviour in response to plant quality are not the major factors driving population outbreaks, which is in accordance with the Phylogenetic Constraints Hypothesis.