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(-1) 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(-1) , respectively). However, the facultative aphid symbionts Hamiltonella 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.

The boll weevil, Anthonomus grandis, has been monitored through deployment of traps baited with aggregation pheromone components. However, field studies have shown that the number of insects caught in these traps is significantly reduced during cotton squaring, suggesting that volatiles produced by plants at this phenological stage may be involved in attraction. Here, we evaluated the chemical profile of volatile organic compounds (VOCs) emitted by undamaged or damaged cotton plants at different phenological stages, under different infestation conditions, and determined the attractiveness of these VOCs to adults of A. grandis. In addition, we investigated whether or not VOCs released by cotton plants enhanced the attractiveness of the aggregation pheromone emitted by male boll weevils. Behavioral responses of A. grandis to VOCs from conspecific-damaged, heterospecific-damaged (Spodoptera frugiperda and Euschistus heros) and undamaged cotton plants, at different phenological stages, were assessed in Y-tube olfactometers. The results showed that volatiles emitted from reproductive cotton plants damaged by conspecifics were attractive to adults boll weevils, whereas volatiles induced by heterospecific herbivores were not as attractive. Additionally, addition of boll weevil-induced volatiles from reproductive cotton plants to aggregation pheromone gave increased attraction, relative to the pheromone alone. The VOC profiles of undamaged and mechanically damaged cotton plants, in both phenological stages, were not different. Chemical analysis showed that cotton plants produced qualitatively similar volatile profiles regardless of damage type, but the quantities produced differed according to the plant's phenological stage and the herbivore species. Notably, vegetative cotton plants released higher amounts of VOCs compared to reproductive plants. At both stages, the highest rate of VOC release was observed in A. grandis-damaged plants. Results show that A. grandis uses conspecific herbivore-induced volatiles in host location, and that homoterpene compounds, such as (E)-4,8-dimethylnona-1,3,7-triene and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene and the monoterpene (E)-ocimene, may be involved in preference for host plants at the reproductive stage.

Characterizing trophic networks is fundamental to many questions in ecology, but this typically requires painstaking efforts, especially to identify the diet of small generalist predators. Several attempts have been devoted to develop suitable molecular tools to determine predatory trophic interactions through gut content analysis, and the challenge has been to achieve simultaneously high taxonomic breadth and resolution. General and practical methods are still needed, preferably independent of PCR amplification of barcodes, to recover a broader range of interactions. Here we applied shotgun-sequencing of the DNA from arthropod predator gut contents, extracted from four common coccinellid and dermapteran predators co-occurring in an agroecosystem in Brazil. By matching unassembled reads against six DNA reference databases obtained from public databases and newly assembled mitogenomes, and filtering for high overlap length and identity, we identified prey and other foreign DNA in the predator guts. Good taxonomic breadth and resolution was achieved (93% of prey identified to species or genus), but with low recovery of matching reads. Two to nine trophic interactions were found for these predators, some of which were only inferred by the presence of parasitoids and components of the microbiome known to be associated with aphid prey. Intraguild predation was also found, including among closely related ladybird species. Uncertainty arises from the lack of comprehensive reference databases and reliance on low numbers of matching reads accentuating the risk of false positives. We discuss caveats and some future prospects that could improve the use of direct DNA shotgun-sequencing to characterize arthropod trophic networks.

The silverleaf whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is one of the most important pest insects in tomato crop systems worldwide. It has been previously demonstrated that intercropping tomato [Solanum lycopersicum L. Mill. (Solanaceae)] with coriander [Coriandrum sativum L. (Apiaceae)] reduces the incidence and severity of damage caused by B. tabaci. However, it is not yet known how coriander affects the insect′s behaviour. We evaluated the attractiveness of tomato constitutive volatiles to B. tabaci and what effect coriander constitutive volatiles have on the insect′s behaviour. To this end, we conducted three bioassays in a multiple‐choice four‐arm olfactometer (‘×’ type), measuring B. tabaci behaviour when offered tomato and coriander constitutive volatiles presented alone as well as together. We also evaluated the colonisation and establishment of B. tabaci in experimental plots with only single tomato plants and tomatoes intercropped with coriander in a greenhouse. Bemisia tabaci males and females recognised tomato constitutive volatiles as a positive stimulus (kairomonal effect), indicating that semiochemicals from this plant can play an important role in the insect’s host plant selection. Coriander constitutive volatiles reduced the attractiveness of tomato volatiles but no repellency to these volatiles was observed. Greater numbers of adults and nymphs of B. tabaci per plant were observed in tomato monoculture plots than in tomato intercropped with coriander. We suggest that coriander constitutive volatiles have an odour masking effect on tomato volatiles, thus interfering in the host plant selection of B. tabaci.

Abstract.The synthetic racemic mixture of methyl 2,6,10‐trimethyltridecanoate, a component of the male produced pheromone of Euschistus heros (F.) (Heteroptera, Pentatomidae), was attractive to pentatomid species in a field test, using homemade pheromone trap designs. The pentatomid Piezodorus guildinii was caught in high numbers in field traps, during two field experiments, indicating a consistent response of this species to the E. heros pheromone. A correlation was found between the range of insects caught in the pheromone‐baited traps and a random sampling method. The synthetic stereoisomeric mixture of methyl 2,6,10‐trimethyldodecanoate, a minor component of E. heros pheromone, was also field tested and caught no pentatomids. Egg parasitoids were caught in traps baited with E. heros pheromone, indicating that this pheromone can be exploited as a kairomone. A synchrony in the periodicity of trap catch, between the egg parasitoids and their host, was also recorded.

SummaryThe egg parasitoid, Telenomus podisi, was shown to recognize its host, Euschistus heros, through both chemical and physical cues. These were determined in short‐range bioassays. The cues comprised fertile and infertile host eggs, egg extracts, crude extracts of adult males and females and successive dilutions of the synthetic racemic mixture of methyl 2,6,10‐trimethyltridecanoate, a male‐produced pheromone of E. heros.Some of the treatments used induced a selection process in T. podisi, through host recognition behaviour, indicating a consistent response of this species to E. heros. The possibility that some of these compounds could be exploited as long‐range kairomones, by T. podisi, is discussed.Using GC analysis, an unidentified compound (Rt 24.8 min), possibly from the egg adhesive material, was detected from E. heros egg extracts. Its retention time compared to the methyl 2,6,10‐trimethyltridecanoate showed a lower volatility, suggesting that the egg adhesive may act as a short‐range kairomone.Physical cues appeared to be an important component in the host selection process by T. podisi. The parasitoid discriminated infertile eggs which are differently shaped and smaller than fertile eggs.

-The boll weevil causes serious damage to the cotton crop in South America. Several studies have been published on this pest, but its phenology and behavior under the tropical conditions prevailing in Brazil are not well-known. In this study the feeding behavior and main food sources of adult boll weevils throughout the year in Central Brazil was investigated. The digestive tract contents of insects captured in pheromone traps in two cotton fields and two areas of native vegetation (gallery forest and cerrado sensu stricto) were analyzed. The insect was captured all through the year only in the cerrado. It fed on pollen of 19 different plant families, on Pteridophyta and fungi spores and algae cysts. Simpson Index test showed that the cerrado provided greater diversity of pollen sources. In the beginning of the cotton cycle, the plant families used for pollen feeding were varied: in cotton area 1, the weevil fed on Poaceae (50%), Malvaceae and Smilacaceae (25% each); in cotton area 2 the pollen sources were Malvaceae (50%), Asteraceae (25%) and Fabaceae and Clusiaceae (25% each); in the cerrado they were Chenopodiaceae (67%) and Scheuchzeriaceae (33%). No weevils were collected in the gallery forest in this period. After cotton was harvested, the family Smilacaceae was predominant among the food plants exploited in all the study areas. These results help to explain the survivorship of adult boll weevil during cotton fallow season in Central Brazil and they are discussed in the context of behavioral adaptations to the prevailing tropical environmental conditions.

-The objective of this research was to evaluate the parasitism behavior of Telenomus podisi Ashmead, Trissolcus basalis (Wollaston) e Trissolcus urichi Crawford (Hymenoptera: Scelionidae) on eggs of Nezara viridula L., Euschistus heros F., Piezodorus guildinii Westwood and Acrosternum aseadum Rolston (Heteroptera: Pentatomidae), in no choice and multiple choice experiments. For all parasitoid species, the results demonstrated the existence of a main host species that maximizes the reproductive success. The competitive interactions among the parasitoid species were investigated in experiments of sequential and simultaneous release of different combinations of parasitoid pairs on the hosts N. viridula, E. heros and A. aseadum. Exploitative competition was observed for egg batches at the genus level (Telenomus vs. Trissolcus) and interference competition at the species level (T. basalis vs. T. urichi). Trissolcus urichi was the most aggressive species, interfering with the parasitism of T. basalis. Generally, T. basalis showed an opportunistic behavior trying to parasitise eggs after T. urichi had abandoned the egg batch. The selection of parasitoid species for use in augmentative biological control programs should take into account the diversity of pentatomids present in soybean in addition to the interactions among the different species of parasitoids.Index terms: parasitism, host-parasite relations, biological competition, pest control methods. Termos para indexação: parasitismo, relação parasita-hospedeiro, competição biológica, métodos de combate às pragas. Interações inter-guilda e intra-guilda entre espécies de parasitóides de ovos que atacam o complexo de percevejos da soja