Pseudacteon notocaudatus and Pseudacteon obtusitus (Diptera: Phoridae), two new species of fire ant parasitoids from South America.

Ongoing studies in South America of phorid flies of the genus Pseudacteon Coquillett 1907 have revealed two further new species in this genus that are described here: P. obtusitus and P. notocaudatus. Both species are parasitoids of Solenopsis (F.) fire ants.

Pseudacteon tricuspis: its behavior and development according to the social form of its host and the role of interference competition among females.

We studied how the behavior and performance of Pseudacteon tricuspis Borgmeier varies with the social form of its host Solenopsis invicta Buren, in its native range in Argentina where monogyne colonies are more abundant than polygynes (approximately 75 vs. 25%). Female, P. tricuspis took 44% less time (50 vs. 89 s) to attack monogyne than polygyne ants, but oviposition attempts were similar (23 vs. 18 attacks). The presence of the parasitoid affected the average size of foragers on the trail, with the proportion of minor workers increasing on both social forms. In the laboratory, P. tricuspis selected similar host sizes, although pupal survival was 25% higher on monogynes than on polygynes. Developmental times of both genders were similar (33-35 d), although larger females emerged from bigger hosts. The sex ratio of P. tricuspis was more male biased when exploiting polygyne ants. Intraspecific competition significantly affected parasitoid reproductive success, being significantly higher for a solitary female than when three females were present, although the size of workers selected did not vary. The male:female ratio also changed, being 1:1 without competition but 2:1 with competition. We demonstrated for the first time the consequences of interference competition among P. tricuspis females, a common behavior observed in others parasitoids. We discuss why P. tricuspis sex ratios are always biased toward males in both social forms and suggest that similar studies of interference competition within and between already naturalized Pseudacteon species in the United States could help predict establishment patterns.

Behavior and development of Pseudacteon curvatus (Diptera: Phoridae) varies according to the social form of its host Solenopsis invicta (Hymenoptera: Formicidae) in its native range.

We assessed the performance of Pseudacteon curvatus Borgmeier with respect to the social form of Solenopsis invicta Buren in Argentina In the field, we studied the effect the parasitoid on size and proportion of ant foragers. In the laboratory, we evaluated P. curvatus oviposition preferences; host size elected; developmental periods; and sexual size dimorphism, sex ratio, and parasitoid survivorship. P. curvatus affected the average size of foraging workers on both social forms diminishing the proportion of big and increasing the proportion of minor workers. P. curvatus required a shorter orientation time and exhibited a greater number of attacks when ovipositing on monogynes workers. In the laboratory, host sizes elected by P. curvatus were similar between social forms. However, attacks on polygyne colonies were more variable, increasing the number of unviable offspring. Developmental times of females and males of P. curvatus were similar for both social forms, but total developmental periods were shorter for males from monogyne colonies. We did not find differences between sexes in emerging adults' size by social form and the female: male sex ratio was 1:1 for both social forms. P. curvatus pupae survival and adult emergence per trial from monogyne colonies were greater than from polygyne colonies. The rarity of polygyne S. invicta in its native range may prevent this phorid from adjusting its life history to that social form. Consequences of applying this phorid in biological control are discussed.

Cytogenetic analysis of three species of Pseudacteon (Diptera, Phoridae) parasitoids of the fire ants using standard and molecular techniques.

Pseudacteon flies, parasitoids of worker ants, are being intensively studied as potentially effective agents in the biological control of the invasive pest fire ant genus Solenopsis (Hymenoptera: Formicidae). This is the first attempt to describe the karyotype of P. curvatus Borgmeier, P. nocens Borgmeier and P. tricuspis Borgmeier. The three species possess 2n = 6; chromosomes I and II were metacentric in the three species, but chromosome pair III was subtelocentric in P. curvatus and P. tricuspis, and telocentric in P. nocens. All three species possess a C positive band in chromosome II, lack C positive heterochromatin on chromosome I, and are mostly differentiated with respect to chromosome III. P. curvatus and P. tricuspis possess a C positive band, but at different locations, whereas this band is absent in P. nocens. Heterochromatic bands are neither AT nor GC rich as revealed by fluorescent banding. In situ hybridization with an 18S rDNA probe revealed a signal on chromosome II in a similar location to the C positive band in the three species. The apparent lack of morphologically distinct sex chromosomes is consistent with proposals of environmental sex determination in the genus. Small differences detected in chromosome length and morphology suggests that chromosomes have been highly conserved during the evolutionary radiation of Pseudacteon. Possible mechanisms of karyotype evolution in the three species are suggested.

Distributional patterns of Pseudacteon associated with the Solenopsis saevissima complex in South America.

Classical biological control efforts against imported fire ants have largely involved the use of Pseudacteon parasitoids. To facilitate further exploration for species and population biotypes a database of collection records for Pseudacteon species was organized, including those from the literature and other sources. These data were then used to map the geographical ranges of species associated with the imported fire ants in their native range in South America. In addition, we found geographical range metrics for all species in the genus and related these metrics to latitude and host use. Approximately equal numbers of Pseudacteon species were found in temperate and tropical regions, though the majority of taxa found only in temperate areas were found in the Northern Hemisphere. No significant differences in sizes of geographical ranges were found between Pseudacteon associated with the different host complexes of fire ants despite the much larger and systemic collection effort associated with the S. saevissima host group. The geographical range of the flies was loosely associated with both the number of hosts and the geographical range of their hosts. Pseudacteon with the most extensive ranges had either multiple hosts or hosts with broad distributions. Mean species richnesses of Pseudacteon in locality species assemblages associated with S. saevissima complex ants was 2.8 species, but intensively sampled locations were usually much higher. Possible factors are discussed related to variation in the size of geographical range, and areas in southern South America are outlined that are likely to have been under-explored for Pseudacteon associated with imported fire ants.

Cytogenetic analysis of three species of Pseudacteon (Diptera, Phoridae) parasitoids of the fire ants using standard and molecular techniques

Pseudacteon flies, parasitoids of worker ants, are being intensively studied as potentially effective agents in the biological control of the invasive pest fire ant genus Solenopsis (Hymenoptera: Formicidae). This is the first attempt to describe the karyotype of P. curvatus Borgmeier, P. nocens Borgmeier and P. tricuspis Borgmeier. The three species possess 2n = 6; chromosomes I and II were metacentric in the three species, but chromosome pair III was subtelocentric in P. curvatus and P. tricuspis, and telocentric in P. nocens. All three species possess a C positive band in chromosome II, lack C positive heterochromatin on chromosome I, and are mostly differentiated with respect to chromosome III. P. curvatus and P. tricuspis possess a C positive band, but at different locations, whereas this band is absent in P. nocens. Heterochromatic bands are neither AT nor GC rich as revealed by fluorescent banding. In situ hybridization with an 18S rDNA probe revealed a signal on chromosome II in a similar location to the C positive band in the three species. The apparent lack of morphologically distinct sex chromosomes is consistent with proposals of environmental sex determination in the genus. Small differences detected in chromosome length and morphology suggests that chromosomes have been highly conserved during the evolutionary radiation of Pseudacteon. Possible mechanisms of karyotype evolution in the three species are suggested.

Geographic variation in resource dominance-discovery in Brazilian ant communities.

A predictive framework for the ecology of species invasions requires that we learn what limits successful invaders in their native range. The red imported fire ant (Solenopsis invicta) is invasive in the United States, Puerto Rico, Australia, New Zealand, and China. Solenopsis invicta appears to be a superior competitor in its introduced range, where it can cause the local extirpation of native species, but little is known about its competitive ability in its native range in South America. Here we examine the competitive ability of S. invicta for food resources in three widely separated Brazilian ant communities. Each of these communities contains 20-40 ant species, 8-10 of which were common and frequently interacted with S. invicta. S. invicta at all three sites was attacked by several species-specific phorid parasitoids, and at one site, two other species were attacked by their own specialized parasitoids. We examined interactions in these local communities for evidence that trade-offs among ant species between resource dominance and resource discovery, and between resource dominance and parasitoid vulnerability facilitate local coexistence. The trade-off between resource dominance and resource discovery was strong and significant only at Santa Genebra, where parasitoids had no effect on the outcome of confrontations at resources. At Bonito, parasitoids significantly reduced the ability of S. invicta, which was the top-ranked behavioral dominant, from defending and usurping food resources from subordinate species. In the Pantanal, S. invicta ranked behind three other ant species in a linear hierarchy of behavioral dominance, and lost the majority of its interactions with a fourth more subordinate species, Paratrechina fulva, another invasive species. Parasitoids of S. invicta were uncommon in the Pantanal, and did not affect its low position in the hierarchy relative to the other two sites. Parasitoids, however, did affect the ability of Linepithema angulatum, the top-ranked behavioral dominant in this community, from defending and usurping resources from behavioral subordinates. These results indicate that both interspecific competition and trait-mediated indirect effects of phorid parasitoids affect the ecological success of the red imported fire ant in its native range, but that the relative importance of these factors varies geographically.

The population genetics of mimetic diversity in Heliconius butterflies.

Theory predicts strong stabilizing selection on warning patterns within species and convergent evolution among species in Müllerian mimicry systems yet Heliconius butterflies exhibit extreme wing pattern diversity. One potential explanation for the evolution of this diversity is that genetic drift occasionally allows novel warning patterns to reach the frequency threshold at which they gain protection. This idea is controversial, however, because Heliconius butterflies are unlikely to experience pronounced population subdivision and local genetic drift. To examine the fine-scale population genetic structure of Heliconius butterflies we genotyped 316 individuals from eight Costa Rican Heliconius species with 1428 AFLP markers. Six species exhibited evidence of population subdivision and/or isolation by distance indicating genetic differentiation among populations. Across species, variation in the extent of local genetic drift correlated with the roles different species have played in generating pattern diversity: species that originally generated the diversity of warning patterns exhibited striking population subdivision while species that later radiated onto these patterns had intermediate levels of genetic diversity and less genetic differentiation among populations. These data reveal that Heliconius butterflies possess the coarse population genetic structure necessary for local populations to experience pronounced genetic drift which, in turn, could explain the origin of mimetic diversity.

Macrolides from the scent glands of the tropical butterflies Heliconius cydno and Heliconius pachinus.

The four major components present in scent gland extracts of the male Costa Rica longwing butterflies Heliconius cydno and Heliconius pachinus were identified as 12- and 14-membered macrolides containing a C(18)-carbon skeleton. By use of micro-reactions and spectrometric examinations, structural proposals were made and subsequently proven by synthesis, using ring-closing-metathesis as the key steps. These macrolides, (9Z,11E,13S)-octadeca-9,11-dien-13-olide (5, S-coriolide), (9Z,11E,13S,15Z)-octadeca-9,11,15-trien-13-olide (6), (9Z,13S)-octadec-9-en-13-olide (13), and (9Z,11S)-octadec-9-en-11-olide (25), are biosynthetically obviously derived from oleic, linoleic, and linolenic acids. Their absolute configurations were determined by gas chromatographic investigations on chiral phases, showing all to possess (S)-configuration.

Associations of fire ant phorids and microhabitats.

We examined flight activity patterns for a guild of fire ant parasitoids in western Argentina in relationship to their host's location (mound/foraging trail) and light condition (full sun/partial sun/full shade) at different scales, from the individually sampled mound to the full day's summation for each species. We asked first whether taxa showed preferences among these conditions, and second, whether certain species and sexes might be found together more frequently than expected to by chance. All species, except the P. obtusus species complex, were significantly more likely to be found attacking ants at disturbed mounds than at paired foraging trails. The P. nocens complex and P. litoralis were more likely to be in the shade when temperatures were above the overall mean of the study (28.3 degrees C), whereas others, such as the P. obtusus complex and P. tricuspis, were more likely to be in full sun under these same conditions. Our analyses indicated that a limited set of species, particularly P. nocens with P. litoralis, and males with female P. obtusus and P. tricuspis, were more likely to be found together than expected. We also found decreasing proportions of males with increasing time of analysis. We discuss the implications of host location, metereological conditions, and sex ratios in relationship to ongoing classical biological control efforts using species of these phorids.

No genomic mosaicism in a putative hybrid butterfly species.

Recent descriptions of hybrid animal species have spurred interest in this phenomenon, but little genomic data exist to support it. Here, we use frequency variation for 657 amplified fragment length polymorphism (AFLP) markers and DNA sequence variation from 16 genes to determine whether the genome of Heliconius pachinus, a suspected hybrid butterfly species, is a mixture of the putative parental species, Heliconius cydno and Heliconius melpomene. Despite substantial shared genetic variation among all three species, we show that the genome of H. pachinus is not a mosaic; both AFLP and DNA sequence data overwhelmingly associate H. pachinus with just one of the potential parents, H. cydno. This pattern also applies to the gene wingless, which is tightly linked to the locus that determines forewing colour-one specific H. pachinus trait that has been hypothesized to have originated from H. melpomene. As a whole, the data support a traditional, bifurcating model of speciation in which H. pachinus split from a common ancestor with H. cydno without a genetic contribution from H. melpomene. However, comparison of our data to DNA sequence data for another putative hybrid Heliconius species, Heliconius heurippa, suggests that the H. heurippa genome may be a mosaic.

A male gift to its partner? Cyanogenic glycosides in the spermatophore of longwing butterflies (Heliconius).

Males of several insect species transfer nuptial gifts to females during mating, typically in the form of a protein-rich spermatophore. In chemically defended species, males could potentially enhance such a gift with chemicals that help protect the female, her eggs, or both. This was shown for lepidopteran species that accumulate pyrrolizidine alkaloids. Most Heliconius butterflies are presumably protected from predators by virtue of de novo synthesized and/or sequestered cyanogenic glycosides. Males of Heliconius species are known to transfer nutritional gifts to the females but whether defensive chemicals could also be transferred is not known. To ascertain whether transfer of cyanogens occurs, we dissected freshly mated females from nine different Heliconius species and analyzed spermatophores for cyanogenic glycosides. We found cyanogens in the spermatophores of all nine species. This is the first time cyanogenic glycosides are reported in the spermatophores of arthropods. We discuss the implications of these findings for Heliconius biology and for other cyanogenic insects as well. We suggest that chemically defended species commonly lace their nuptial gifts with defensive chemicals to improve gift quality.

Multilocus analyses of admixture and introgression among hybridizing Heliconius butterflies.

Introgressive hybridization is an important evolutionary process and new analytical methods provide substantial power to detect and quantify it. In this study we use variation in the frequency of 657 AFLP fragments and DNA sequence variation from 15 genes to measure the extent of admixture and the direction of interspecific gene flow among three Heliconius butterfly species that diverged recently as a result of natural selection for Miillerian mimicry, and which continue to hybridize. Bayesian clustering based on AFLP genotypes correctly delineated the three species and identified four H. cydno, three H. pachinus, and three H. melpomene individuals that were of mixed ancestry. Gene genealogies revealed substantial shared DNA sequence variation among all three species and coalescent simulations based on the Isolation with Migration (IM) model pointed to interspecific gene flow as its cause. The IM simulations further indicated that interspecific gene flow was significantly asymmetrical, with greater gene flow from H. pachinus into H. cydno (2Nm = 4.326) than the reverse (2Nm = 0.502), and unidirectional gene flow from H. cydno and H. pachinus into H. melpomene (2Nm = 0.294 and 0.252, respectively). These asymmetries are in the directions expected based on the genetics of wing patterning and the probability that hybrids of various phenotypes will survive and reproduce in different mimetic environments. This empirical demonstration of extensive interspecific gene flow is in contrast to a previous study which found little evidence of gene flow between another pair of hybridizing Heliconius species, H. himera and H. erato, and it highlights the critical role of natural selection in maintaining species diversity. Furthermore, these results lend support to the hypotheses that phenotypic diversification in the genus Heliconius has been fueled by introgressive hybridization and that reinforcement has driven the evolution of assortative mate preferences.

Development of Pseudacteon nocens (Diptera: Phoridae) on Solenopsis invicta and Solenopsis richteri fire ants (Hymenoptera: Formicidae).

We studied the developmental performance of the large morph of Pseudacteon nocens Borgmeier (Diptera: Phoridae), a prospective biological control agent of imported fire ants (Hymenoptera: Formicidae). We measured selected life history traits of this parasitoid as a function of 1) host species (Solenopsis invicta Buren versus Solenopsis richteri Forel), 2) temperature (22 versus 28 degrees C), 3) source population of the fly (Corrientes and Santiago del Estero, Argentina), and 4) varied size distributions of offered host ants. Developmental periods were influenced by host species, although the populations responded in opposing manners. Developmental times, however, were most strongly influenced by temperature with total developmental periods lengthened by 17-32% at 22 degrees C. Pupal mortality was also significantly lower at this temperature. Although numbers of progeny per female were significantly higher for the Corrientes population, we found no significant differences in progeny per female according to host species. Interestingly, we found that females were larger than males, and flies from Corrientes were larger than those from Santiago del Estero, even after statistical adjustments for host size. The modal frequency of host size elected in all treatment combinations tested was identical (0.6 mm), a size that represented the apparent threshold for producing female progeny. These laboratory and additional field observations demonstrate considerable interpopulational variation in P. nocens and lend further support to the applied approach focusing at the population, as opposed to the species level, with respect to both source and target areas for classical biological control introductions of Pseudacteon flies.

Developmental rates and host specificity for Pseudacteon parasitoids (Diptera: Phoridae) of fire ants (Hymenoptera: Formicidae) in Argentina.

This study extends our comparative knowledge of Pseudacteon interactions with Solenopsis fire ant workers. Reported in this work are development times for seven Argentinean parasitoid species reared on two hosts, Solenopsis richteri Forel and Solenopsis invicta Buren, under laboratory temperature regimes comparable with those of the climatic zones occupied by these host species. Developmental times spanned 31-66 d across phorid species, and in general did not differ between genders or host species, but were longer at lower temperatures. The size distribution of flies reared was bimodal, with a group of large (Pseudacteon borgmeieri, Pseudacteon nocens, Pseudacteon obtusus and Pseudacteon tricuspis) and small (Pseudacteon cultellatus, Pseudacteon curvatus, and Pseudacteon nudicornis) species. P. borgmeieri was exceptional with respect to length of developmental time. Also reported are results of initial oviposition and developmental studies of some of these phorid species on other Argentinean Solenopsis ant species; P. curvatus was the only species able to complete its development on nonhost fire ants. These results support the concept of incorporating several complementary species of Pseudacteon in the biological control of pest fire ants.