Seasonal and annual abundance of Ephuta wasp (Hymenoptera: Mutillidae) in Panama

Introduction: Ephuta Say is a widespread New World genus of mutillid wasp whose ecology is poorly understood. Objective: The objective of this study was to determine how Ephuta species abundance varies annually and seasonally driven by weather conditions and hosts. Methods: Ephuta specimens, located in the "old forest" at Barro Colorado Island, were examined from weekly samples (2001-2006) of ten Malaise traps. The monthly abundance of each Ephuta species was compared with monthly average humidity, solar radiation, temperature and rainfall. Results: Nine species and ten morphospecies were identified. Most specimens were collected from March to June. April was the month reported with the greatest abundance. Of the four abiotic variables measured, only average monthly temperature was correlated with Ephuta abundance. Months with the highest number of Ephuta specimens were correlated with suspected host abundance, as indicated by the monthly abundance of Pompilidae (Hymenoptera) captured during the year 2007. Conclusions: We concluded that Ephuta display strong seasonal variation in abundance, with the peaks occurring during the end of the dry season and beginning of the rainy season, which correlate broadly with temperature and the abundance of their pompilid hosts.

Introducción: Ephuta es un género de avispas mutílidas ampliamente distribuido en el Nuevo Mundo y cuya ecología es poco conocida. Objetivo: El objetivo de este trabajo fue determinar cómo varía la abundancia de especies de Ephuta anualmente y estacionalmente debido a las condiciones climáticas y sus hospederos. Métodos: Se examinaron especímenes de Ephuta tomados de muestras semanales (2001-2006) de diez trampas Malaise, ubicadas en el "bosque viejo" de Isla Barro Colorado. La abundancia mensual de cada especie de Ephuta se comparó con el promedio mensual de la humedad, la radiación solar, la temperatura y las precipitaciones. Resultados: Se identificaron nueve especies y diez morfoespecies. La mayoría de los especímenes fueron recolectados de marzo a junio. La mayor abundancia mensual promedio se dio en abril. De las cuatro variables abióticas medidas, solo la temperatura mensual promedio se correlacionó con la abundancia de Ephuta. Los meses con el mayor número de especímenes de Ephuta se correlacionan con la sospechosa abundancia del huésped, como lo indica la abundancia mensual de Pompilidae (Hymenoptera) capturada durante el 2007. Conclusiones: Ephuta muestra una fuerte variación estacional en la abundancia, con picos durante el final de la estación seca y el comienzo de la temporada de lluvias. Esta se correlaciona ampliamente con la temperatura y la abundancia de sus huéspedes pompílidos.

Variation in Host Plant Usage and Diet Breadth Predict Sibling Preference and Performance in the Neotropical Tortoise Beetle Chelymorpha alternans (Coleoptera: Chrysomelidae: Cassidinae).

Specialized interactions between insects and the plants that they consume are one of the most ubiquitous and consequential ecological associations on the plant. Decades of investigation suggest that a narrow diet favors an individual phytophagous insect's performance relative to a dietary generalist. However, this body of research has tended to approach questions of diet breadth and host usage from the perspective of temperate plant-insect associations. Relationships between diet breadth, host usage, and variation in tropical insect preference and performance remain largely uninvestigated. Here we characterize how variation in diet breadth and host usage affect oviposition preference, development, survival, and gain in mass of a Neotropical tortoise beetle Chelymorpha alternans Boheman 1854 (Coleoptera: Chrysomelidae), using a split-brood, sibling experimental design. Host performance was measured after splitting broods among four no-choice host diets. Groups consuming single hosts varied among themselves in developmental time and survival from larva to adult. Performance did not vary among groups consuming multiple and single hosts. Oviposition preference was measured in choice and no-choice tests. Females displayed preference for the original host in both experiments. Developmental time and survival of offspring sourced from the no-choice experiment was measured for two complete generations to explore correlations with female oviposition preference. Preference for the original host correlated with high survivorship and an intermediate developmental time. Survivorship and time to develop were also high on an alternative host that was less preferred. Departures from predictions of prevailing preference-performance hypotheses suggest that host usage presents C. alternans with fitness trade-offs.

Structural and physical evidence for an endocuticular gold reflector in the tortoise beetle, Charidotella ambita.

Charidotella ambita offers a unique opportunity for unambiguously locating its gold reflector by comparing the structure of reflecting and non-reflecting cuticle of the elytron and pronotum. Using light microscopy and TEM, the reflector was located underneath the macrofiber endocuticle just above the epidermis. The reflector is a multilayer comprising up to 50 bilayers alternating high and low density layers parallel to the surface of the cuticle. It is chirped, i.e., showing a progressive decrease in layer thickness from approximately 150 nm-100 nm across its depth. The high density layers in contact with the endocuticle fuse to the last macrofiber when the reflector is interrupted by a trabecula, demonstrating their cuticular nature. Simulated reflectance spectra from models of the multilayer matched the reflection spectra measured on the major gold patch of the elytron of living specimens. Previous reports in adult insects exhibiting metallic colors located their reflector in the upper strata and structures of the cuticle, i.e., epicuticle, exocuticle, scales and hairs. Thus, the endocuticular location of the reflector in C. ambita (and other tortoise beetles) appears unique for adult insects. Gold reflection appears in C. ambita only when the synthesis of the macrolayer endocuticle is complete, which may take up to 2 weeks. The development of the gold reflector coincides with the start of mating behavior, possibly suggesting a signaling function in conspecific recognition once sexual maturity has been reached.

Cuticular extracts from Acromis sparsa (Coleoptera: Cassidinae) mediate arrestment behavior of the commensal canestriniid mite Grandiella rugosita.

Astigmatid mites in the family Canestriniidae are often closely associated with tortoise leaf beetles (Chrysomelidae: Cassidinae). For example, the survival of the commensal canestriniid mite Grandiella rugosita depends on dispersal to the cassidine beetle Acromis sparsa. Here, we tested whether the beetle cuticle provides chemical cues for host recognition for G. rugosita. In two-choice assays with cuticular extracts from A. sparsa and the co-occurring, non-host cassidine Chelymorpha alternans offered simultaneously, mites clearly preferred the area treated with extract from their host. In no-choice assays, G. rugosita spent three times longer and moved three times slower on host cuticular extracts compared to non-host extracts and the solvent control. Analyses of the chemical composition of cuticular extracts from males and females of A. sparsa and C. alternans revealed complex mixtures of mainly methyl branched hydrocarbons, which clearly separated both species in a principal component analysis. We found no qualitative difference between males and females of either species, but in C. alternans quantitative differences between males and females were detected. Our results demonstrate that G. rugosita is able to discriminate between cuticular extracts from its host A. sparsa and the non-host C. alternans. The components eliciting the observed arrestment behavior remain to be determined.

Insect leaf-chewing damage tracks herbivore richness in modern and ancient forests.

The fossil record demonstrates that past climate changes and extinctions significantly affected the diversity of insect leaf-feeding damage, implying that the richness of damage types reflects that of the unsampled damage makers, and that the two are correlated through time. However, this relationship has not been quantified for living leaf-chewing insects, whose richness and mouthpart convergence have obscured their value for understanding past and present herbivore diversity. We hypothesized that the correlation of leaf-chewing damage types (DTs) and damage maker richness is directly observable in living forests. Using canopy access cranes at two lowland tropical rainforest sites in Panamá to survey 24 host-plant species, we found significant correlations between the numbers of leaf chewing insect species collected and the numbers of DTs observed to be made by the same species in feeding experiments, strongly supporting our hypothesis. Damage type richness was largely driven by insect species that make multiple DTs. Also, the rank-order abundances of DTs recorded at the Panamá sites and across a set of latest Cretaceous to middle Eocene fossil floras were highly correlated, indicating remarkable consistency of feeding-mode distributions through time. Most fossil and modern host-plant pairs displayed high similarity indices for their leaf-chewing DTs, but informative differences and trends in fossil damage composition became apparent when endophytic damage was included. Our results greatly expand the potential of insect-mediated leaf damage for interpreting insect herbivore richness and compositional heterogeneity from fossil floras and, equally promisingly, in living forests.

Subsocial Neotropical Doryphorini (Chrysomelidae, Chrysomelinae): new observations on behavior, host plants and systematics.

A summary of literature, documented observations and field studies finds evidence that mothers actively defend offspring in at least eight species and three genera of Neotropical Chrysomelinae associated with two host plant families. Reports on three Doryphora species reveal that all are oviparous and feed on vines in the Apocyanaceae. Mothers in the two subsocial species defend eggs and larvae by straddling, blocking access at the petiole and greeting potential predators with leaf-shaking and jerky advances. A less aggressive form of maternal care is found in two Platyphora and four Proseicela species associated with Solanaceae, shrubs and small trees. For these and other morphologically similar taxa associated with Solanaceae, genetic distances support morphology-based taxonomy at the species level, reveal one new species, but raise questions regarding boundaries separating genera. We urge continued study of these magnificent insects, their enemies and their defenses, both behavioral and chemical, especially in forests along the eastern versant of the Central and South American cordillera.

Two genera of Aulacoscelinae beetles reflexively bleed azoxyglycosides found in their host cycads.

Aulacoscelinae beetles have an ancient relationship with cycads (Cycadophyta: Zamiaceae), which contain highly toxic azoxyglycoside (AZG) compounds. How these "primitive" leaf beetles deal with such host-derived compounds remains largely unknown. Collections were made of adult Aulacoscelis appendiculata from Zamia cf. elegantissima in Panama, A. vogti from Dioon edule in Mexico, and Janbechynea paradoxa from Zamia boliviana in Bolivia. Total AZG levels were quantified in both cycad leaves and adult beetles by high performance liquid chromatography (HPLC). On average, cycad leaves contained between 0.5-0.8% AZG (frozen weight, FW), while adult beetles feeding on the same leaves contained even higher levels of the compounds (average 0.9-1.5% FW). High AZG levels were isolated from reflex bleeding secreted at the leg joints when beetles were disturbed. Nuclear magnetic resonance and mass spectroscopy identified two AZGs, cycasin and macrozamin, in the reflex bleeding; this is the first account of potentially plant-derived compounds in secretions of the Aulacoscelinae. These data as well as the basal phylogenetic position of the Aulacoscelinae suggest that sequestration of plant secondary metabolites appeared early in leaf beetle evolution.

Reverse color sequence in the diffraction of white light by the wing of the male butterfly Pierella luna (Nymphalidae: Satyrinae).

The butterfly Pierella luna (Nymphalidae) shows an intriguing rainbow iridescence effect: the forewings of the male, when illuminated along the axis from the body to the wing tip, decompose a white light beam as a diffraction grating would do. Violet light, however, emerges along a grazing angle, near the wing surface, while the other colors, from blue to red, exit respectively at angles progressively closer to the direction perpendicular to the wing plane. This sequence is the reverse of the usual decomposition of light by a grating with a periodicity parallel to the wing surface. It is shown that this effect is produced by a macroscopic deformation of the entire scale, which curls in such a way that it forms a "vertical" grating, perpendicular to the wing surface, and functions in transmission instead of reflection.

Switchable reflector in the Panamanian tortoise beetle Charidotella egregia (Chrysomelidae: Cassidinae).

The tortoise beetle Charidotella egregia is able to modify the structural color of its cuticle reversibly, when disturbed by stressful external events. After field observations, measurements of the optical properties in the two main stable color states and scanning electron microscope and transmission electron microscope investigations, a physical mechanism is proposed to explain the color switching of this insect. It is shown that the gold coloration displayed by animals at rest arises from a chirped multilayer reflector maintained in a perfect coherent state by the presence of humidity in the porous patches within each layer, while the red color displayed by disturbed animals results from the destruction of this reflector by the expulsion of the liquid from the porous patches, turning the multilayer into a translucent slab that leaves an unobstructed view of the deeper-lying, pigmented red substrate. This mechanism not only explains the change of hue but also the change of scattering mode from specular to diffuse. Quantitative modeling is developed in support of this analysis.

Dietary specialization influences the efficacy of larval tortoise beetle shield defenses.

Plant chemical defenses and escape from natural enemies have been postulated to select for dietary specialization in herbivorous insects. In field and laboratory bioassays, we evaluated the effectiveness of intact and chemically modified larval shield defenses of the generalist Chelymorpha alternans and the specialists Acromis sparsa and Stolas plagiata (Chrysomelidae: Cassidinae) against three natural predators, using larvae reared on two morning glory (Convolvulaceae) species. We assessed whether: (1) specialists were better defended than generalists when both were fed and assayed on the same plant; (2) larval shield defenses were chemical, physical, or both; and (3) specialists exploit chemistry better than generalists. Live specialist larvae survived at higher rates than did generalists in predator bioassays with the bug Montina nigripes (Reduviidae), but there were no differences among groups against two species of Azteca ants (Hymenoptera: Dolichoderinae). Solvent leaching by H(2)O or MeOH significantly reduced shield efficacy for all species compared to larvae with intact shields. In contrast, freshly killed specialist larvae exhibited significantly lower capture rates and frequencies than the generalists. Although solvent leaching significantly reduced overall shield efficacy for freshly killed larvae of all species, the pattern of leaching effects differed between specialists and generalists, with H(2)O-leaching having a greater impact on the specialists. The overall vulnerability of the generalists appears due to lower chemical protection, which is ameliorated by increased escape behaviors, suggesting a selective trade-off between these defensive components. These experiments indicate that shield defenses are essential for larval survival and that specialists are superior at exploiting plant compounds residing in the aqueous fraction. Our results support the hypothesis that diet-specialized herbivorous insects have more effective defenses than generalists when both feed on the same plant due to the differential ability to exploit defensive precursors obtained from the host. The evolution of dietary specialization may therefore confer the advantage of enhanced enemy-free space.

The distribution of Wolbachia in fig wasps: correlations with host phylogeny, ecology and population structure.

We surveyed for the presence and identity of Wolbachia in 44 species of chalcid wasps associated with 18 species of Panamanian figs. We used existing detailed knowledge of the population structures of the host wasps, as well as the ecological and evolutionary relationships among them, to explore the relevance of each of these factors to Wolbachia prevalence and mode of transmission. Fifty-nine per cent of these wasp species have Wolbachia infections, the highest proportion reported for any group of insects. Further, neither the presence nor the frequency of Wolbachia within hosts was correlated with the population structure of pollinator hosts. Phylogenetic analyses of wsp sequence data from 70 individuals representing 22 wasp species show that neither the close phylogenetic relationship nor close ecological association among host species is consistently linked to close phylogenetic affinities of the Wolbachia associated with them. Moreover, no genetic variation was detected within any Wolbachia strain from a given host species. Thus, the spread of Wolbachia within host species exceeds the rate of horizontal transmission among species and both exceed the rate of mutation of the wsp gene in Wolbachia. The presence and, in some cases, high frequency of Wolbachia infections within highly inbred species indicate that the Wolbachia either directly increase host fitness or are frequently horizontally transferred within these wasp species. However, the paucity of cospeciation of Wolbachia and their wasp hosts indicates that Wolbachia do not persist within a given host lineage for long time-periods relative to speciation times.

Dual chemical sequestration: a key mechanism in transitions among ecological specialization.

Platyphora leaf beetles form a vast group of tropical species each feeding on a restricted set of host plants and exhibiting bright coloration warning predators against their chemical protection. These beetles offer an exceptional opportunity for understanding the evolution of phytochemical sequestration. Indeed, qualitative studies of defensive secretions indicate that Platyphora species acquire toxicity via sequestration of plant secondary metabolites. All produce pentacyclic triterpene saponins from sequestered plant amyrins, but our analyses also indicate that many Platyphora species produce a dual chemical defence, that is, they are additionally protected by lycopsamine-type pyrolyzidine alkaloids that they also sequester from their host. This paper reports on the evolution of chemical defence and host affiliation in Platyphora leaf beetles as reconstructed on a molecular phylogeny of mitochondrial and nuclear DNA sequences. The analyses indicate that dual sequestration could be the key mechanistic means by which transitions among ecological specializations (i.e. restricted host-plant affiliations) are made possible.