Insights into the differences related to the resistance mechanisms to the highly toxic fruit Hippomane mancinella (Malpighiales: Euphorbiaceae) between the larvae of the sister species Anastrepha acris and Anastrepha ludens (Diptera: Tephritidae) through comparative transcriptomics.

The Manchineel, Hippomane mancinella ("Death Apple Tree") is one of the most toxic fruits worldwide and nevertheless is the host plant of the monophagous fruit fly species Anastrepha acris (Diptera: Tephritidae). Here we aimed at elucidating the detoxification mechanisms in larvae of A. acris reared on a diet enriched with the toxic fruit (6% lyophilizate) through comparative transcriptomics. We compared the performance of A. acris larvae with that of the sister species A. ludens, a highly polyphagous pest species that is unable to infest H. mancinella in nature. The transcriptional alterations in A. ludens were significantly greater than in A. acris. We mainly found two resistance mechanisms in both species: structural, activating cuticle protein biosynthesis (chitin-binding proteins likely reducing permeability to toxic compounds in the intestine), and metabolic, triggering biosynthesis of serine proteases and xenobiotic metabolism activation by glutathione-S-transferases and cytochrome P450 oxidoreductase. Some cuticle proteins and serine proteases were not orthologous between both species, suggesting that in A. acris, a structural resistance mechanism has been selected allowing specialization to the highly toxic host plant. Our results represent a nice example of how two phylogenetically close species diverged over recent evolutionary time related to resistance mechanisms to plant secondary metabolites.

Larval Rearing and Nutrition of the Polyphagous Tephritid Pest Anastrepha ludens on Artificial Diets with Calcium Alginate, Agar, or Carrageenan as Gelling Agents at Various Concentrations and across Extreme Larval Density Conditions.

Research on larval rearing and nutrition of tephritid flies on artificial diets is key for the sterile insect technique. Here, we examined the effects of the type of gel (calcium alginate, agar, or carrageenan), at varying percentages in artificial diets for the polyphagous pest Anastrepha ludens, on the physicochemical and nutritional traits of the diets, and the effects of the type of gel, the gel content and the larval density (larvae/g of diet) used in production, quality parameters for mass-reared tephritids, diet removal (an indirect estimation of diet consumption), and nutritional traits of flies. Regardless of the gel content, calcium alginate diets were firmer and more resistant to penetration than the agar and carrageenan diets. The larval recovery, pupation, pupal weight, and flight ability of A. ludens were lower in calcium alginate diets than in agar and carrageenan diets. Diet removal was higher in calcium alginate diets; however, low levels of ammonium and high levels of uric acid in excretions from larvae on these diets suggest an alteration in protein metabolism. The firmness and penetration resistance characteristics of calcium alginate diets may have limited movement and feeding of larvae, but this could be overcome by the collective feeding of large groups of larvae. Our findings provide insights into the mechanism governing gel-diet rearing systems for A. ludens.

New Insights on Antennal Sensilla of Anastrepha ludens (Diptera: Tephritidae) Using Advanced Microscopy Techniques.

Using light, transmission, scanning electron, and confocal microscopy, we carried out a morphological study of antennal sensilla and their ultrastructures of the Mexican Fruit Fly Anastrepha ludens (Loew), an economically important species that is a pest of mangos and citrus in Mexico and Central America. Our goal was to update the known information on the various sensilla in the antennae of A. ludens, involved in the perception of odors, temperature, humidity, and movement. Based on their external shape, size, cuticle-thickness, and presence of pores, we identified six types of sensilla with 16 subtypes (one chaetica in the pedicel, four clavate, two trichoid, four basiconic, one styloconic, and one campaniform-like in the flagellum, and three additional ones in the two chambers of the sensory pit (pit-basiconic I and II, and pit-styloconic)), some of them described for the first time in A. ludens. We also report, for the first time, two types of pores in the sensilla (hourglass and wedge shapes) that helped classify the sensilla. Additionally, we report a campaniform-like sensillum only observed by transmission electronic microscopy on the flagellum, styloconic and basiconic variants inside the sensory pit, and an "hourglass-shaped" pore in six sensilla types. We discuss and suggest the possible function of each sensillum according to their characteristics and unify previously used criteria in the only previous study on the topic.

Feeding on the Fruit Waste Orange Bagasse Modifies Immature Protein Content, Body Weight, Scent Bouquet Composition, and Copula Duration in Males of a Tephritid Frugivorous Fly.

Anastrepha ludens is a polyphagous frugivorous tephritid that infests citrus and mango. Here, we report the establishment of a laboratory colony of A. ludens reared on a larval medium that is a waste for the citrus industry, specifically, orange (Citrus × sinensis) fruit bagasse. After 24 generations of rearing on a nutritionally poor orange bagasse diet, pupae weighed 41.1% less than pupae from a colony reared on a nutritionally rich artificial diet. Larvae from the orange bagasse diet had 6.94% less protein content than larvae from the artificial diet, although their pupation rate was similar. Males from the orange bagasse diet produced a scent bouquet with 21 chemical compounds and were sexually competitive, but they had significantly shorter copulations when compared to males from the artificial diet and from the wild host, Casimiroa edulis, which had relatively simple scent bouquets. The chemical complexity in the odors of males from the orange bagasse diet might initially have attracted females to novel scent combinations, but, once in the copula, they may have been able to sense negative characteristics in males, leading them to terminate copulations soon after they began. We conclude that A. ludens can adjust morphological, life history, nutritional, and chemical traits when adapted to a larval environment consisting of fruit bagasse.

Assessment of the Molecular Responses of an Ancient Angiosperm against Atypical Insect Oviposition: The Case of Hass Avocados and the Tephritid Fly Anastrepha ludens.

Anastrepha spp. (Diptera: Tephritidae) infestations cause significant economic losses in commercial fruit production worldwide. However, some plants quickly counteract the insertion of eggs by females by generating neoplasia and hindering eclosion, as is the case for Persea americana Mill., cv. Hass (Hass avocados). We followed a combined transcriptomics/metabolomics approach to identify the molecular mechanisms triggered by Hass avocados to detect and react to the oviposition of the pestiferous Anastrepha ludens (Loew). We evaluated two conditions: fruit damaged using a sterile pin (pin) and fruit oviposited by A. ludens females (ovi). We evaluated both of the conditions in a time course experiment covering five sampling points: without treatment (day 0), 20 min after the treatment (day 1), and days 3, 6, and 9 after the treatment. We identified 288 differentially expressed genes related to the treatments. Oviposition (and possibly bacteria on the eggs' surface) induces a plant hypersensitive response (HR), triggering a chitin receptor, producing an oxidative burst, and synthesizing phytoalexins. We also observed a process of cell wall modification and polyphenols biosynthesis, which could lead to polymerization in the neoplastic tissue surrounding the eggs.

Safety Assessment of the Potential Probiotic Bacterium Limosilactobacillus fermentum J23 Using the Mexican Fruit Fly (Anastrepha ludens Loew, Diptera: Tephritidae) as a Novel In Vivo Model.

Safety assessment of probiotics is difficult but essential. In this work, the Mexican fruit fly, Anastrepha ludens (Loew) (Diptera: Tephritidae), was used as in vivo model to assess the biosafety of Limosilactobacillus fermentum J23. In the first set of experiments, the strain was orally administered to adult flies through direct feeding, whereas in the second set of experiments, it was supplemented through the larval rearing medium. Data showed that L. fermentum J23 did not lead to increased mortality or treatment-related toxicity signs in adult female and male flies. Ingestion of L. fermentum J23 by adult female flies led to a statistically significant improvement in locomotor activity compared to the control groups (ca. 59% decrease in climbing time, p < 0.0001). A positive trend in lifespan extension under stress (maximum lifespan = 144 h) was also observed. When L. fermentum J23 was administered to the larvae, the adult emergence (p = 0.0099), sex ratio (p = 0.0043), and flight ability (p = 0.0009) increased significantly by 7%, 31%, and 8%, respectively, compared to the control diet. No statistical effect between the control diet and the L. fermentum J23-based diet for the number of pupae recovered, pupal weight, duration of the pupal stage, lifespan under stress, and morphological development was observed. We conclude that feeding L. fermentum J23 to the novel experimental model A. ludens had no toxic effects and could be safely considered a potential probiotic for food supplements; however, further studies are still needed to establish its biosafety in humans.

Bitter friends are not always toxic: The loss of acetic acid bacteria and the absence of Komagataeibacter in the gut microbiota of the polyphagous fly Anastrepha ludens could inhibit its development in Psidium guajava in contrast to A. striata and A. fraterculus that flourish in this host.

The gut microbiota is key for the homeostasis of many phytophagous insects, but there are few studies comparing its role on host use by stenophagous or polyphagous frugivores. Guava (Psidium guajava) is a fruit infested in nature by the tephritids Anastrepha striata and A. fraterculus. In contrast, the extremely polyphagous A. ludens infests guava only under artificial conditions, but unlike A. striata and the Mexican A. fraterculus, it infests bitter oranges (Citrus x aurantium). We used these models to analyze whether the gut microbiota could explain the differences in host use observed in these flies. We compared the gut microbiota of the larvae of the three species when they developed in guava and the microbiota of the fruit pulp larvae fed on. We also compared the gut microbiota of A. ludens developing in C. x aurantium with the pulp microbiota of this widely used host. The three flies modified the composition of the host pulp microbiota (i.e., pulp the larvae fed on). We observed a depletion of Acetic Acid Bacteria (AAB) associated with a deleterious phenotype in A. ludens when infesting P. guajava. In contrast, the ability of A. striata and A. fraterculus to infest this fruit is likely associated to a symbiotic interaction with species of the Komagataeibacter genus, which are known to degrade a wide spectrum of tannins and polyphenols. The three flies establish genera specific symbiotic associations with AABs. In the case of A. ludens, the association is with Gluconobacter and Acetobacter, but importantly, it cannot be colonized by Komagataeibacter, a factor likely inhibiting its development in guava.

Coping with global warming: Adult thermal thresholds in four pestiferous Anastrepha species determined under experimental laboratory conditions and development/survival times of immatures and adults under natural field conditions.

Climate change, particularly global warming, is disturbing biological processes in unexpected ways and forcing us to re-study/reanalyze the effects of varying temperatures, among them extreme ones, on insect functional traits such as lifespan and fecundity/fertility. Here we experimentally tested, under both laboratory and field conditions, the effects of an extreme range of temperatures (5, 10, 15, 20, 30, 40, and 45 °C, and the naturally varying conditions experienced in the field), on survivorship/lifespan, fecundity, and fertility of four pestiferous fruit fly species exhibiting contrasting life histories and belonging to two phylogenetic groups within the genus Anastrepha: A. ludens, A. obliqua, A. striata, and A. serpentina. In the field, we also measured the length of the entire life cycle (egg to adult), and in one species (A. ludens), the effect on the latter of the host plant (mango and grapefruit). Under laboratory conditions, none of the adults, independent of species, could survive a single day when exposed to a constant temperature of 45 °C, but A. striata and A. serpentina females/males survived at the highly contrasting temperatures of 5 and 40 °C at least 7 days. Maximum longevity was achieved in all species at 15 °C (375, 225, 175 and 160 days in A. ludens, A. serpentina, A. striata and A. obliqua females, respectively). Anastrepha ludens layed many eggs until late in life (368 days) at 15 °C, but none eclosed. Eclosion was only observed in all species at 20 and 30 °C. Under natural conditions, flies lived ca. 100 days less than in the laboratory at 15 °C, likely due to the physiological cost of dealing with the highly varying environmental patterns over 24 h (minimum and maximum temperatures and relative humidity of ca. 10-40 °C, and 22-100%, respectively). In the case of A. ludens, the immature's developmental time was shorter in mango, but adult survival was longer than in grapefruit. We discuss our results considering the physiological processes regulating the traits measured and tie them to the increasing problem of global warming and its hidden effects on the physiology of insects, as well as the ecological and pest management implications.

Characterization of reproductive proteins in the Mexican fruit fly points towards the evolution of novel functions.

Seminal fluid proteins (Sfps) modify female phenotypes and have wide-ranging evolutionary implications on fitness in many insects. However, in the Mexican fruit fly, Anastrepha ludens, a highly destructive agricultural pest, the functions of Sfps are still largely unknown. To gain insights into female phenotypes regulated by Sfps, we used nano-liquid chromatography mass spectrometry to conduct a proteomic analysis of the soluble proteins from reproductive organs of A. ludens. The proteins predicted to be transferred from males to females during copulation were 100 proteins from the accessory glands, 69 from the testes and 20 from the ejaculatory bulb, resulting in 141 unique proteins after accounting for redundancies from multiple tissues. These 141 included orthologues to Drosophila melanogaster proteins involved mainly in oogenesis, spermatogenesis, immune response, lifespan and fecundity. In particular, we found one protein associated with female olfactory response to repellent stimuli (Scribble), and two related to memory formation (aPKC and Shibire). Together, these results raise the possibility that A. ludens Sfps could play a role in regulating female olfactory responses and memory formation and could be indicative of novel evolutionary functions in this important agricultural pest.

Influence of Sunlight Incidence and Fruit Chemical Features on Oviposition Site Selection in Mango by Anastrepha obliqua: Implications for Management.

With the aim of identifying key factors that determine oviposition decisions by Anastrepha obliqua for management purposes, we conducted a behavioral study under natural/semi-natural field conditions to identify where exactly in the fruit (upper, middle, or lower sections) females preferred to lay eggs in a highly susceptible mango cultivar ("Criollo"), and whether sunlight incidence and fruit chemical compounds influenced oviposition site selection by this pestiferous fly. Females oviposited in shaded, upper fruit sections where pulp had higher total carbohydrate concentrations but similar total protein, lipid, and polyphenol concentrations than non-oviposited sections. Peel had higher overall nutrient and mangiferin/quercetin-3-D-galactoside (polyphenols) concentrations. An untargeted metabolomic analysis of oviposited and non-oviposited fruit sections identified abscisic acid (ABA) and dihydrophaseic acid glucoside, a by-product of ABA catabolism, as potential chemical markers that could play a role in fruit acceptance behaviors by female flies. We conclude that females preferentially oviposit in fruit sections with optimal chemical and environmental conditions for larval development: more carbohydrates and antioxidants such as mangiferin and ferulic acid and lesser sunlight exposure to avoid lethal egg/larval desiccation/overheating. We make specific recommendations for A. obliqua management based on female host selection behavior, a tree pruning scheme exposing fruit to direct sunlight, application of a host marking pheromone, and the use of egg sinks in the orchard.

Integrating proteomics and metabolomics approaches to elucidate the ripening process in white Psidium guajava.

Psidium guajava (guava) exhibits a high content of biomolecules with nutraceutical properties. However, the biochemistry and molecular foundation of guava ripening is unknown. We performed comparative proteomics and metabolomics studies in different fruit tissues at two ripening stages to understand this process in white guava. Our results, suggest the positive contribution of ethylene and abscisic acid (ABA) signaling to the regulation of biochemical changes during guava ripening. We characterized the modulation of several metabolic pathways, including those of sugar and chlorophyll metabolism, abiotic and biotic stress responses, and biosynthesis of carotenoids and secondary metabolites, among others. In addition to ethylene and ABA, we also found a differential accumulation of other growth regulators such as brassinosteroids, cytokinin, methyl-jasmonate, gibberellins and proteins, and discuss their possible implications in the intricate biochemical network associated with guava ripening process. This integrative approach represents a global overview of the metabolic pathway dynamics during guava ripening.

Testing the potential contribution of Wolbachia to speciation when cytoplasmic incompatibility becomes associated with host-related reproductive isolation.

Endosymbiont-induced cytoplasmic incompatibility (CI) may play an important role in arthropod speciation. However, whether CI consistently becomes associated or coupled with other host-related forms of reproductive isolation (RI) to impede the transfer of endosymbionts between hybridizing populations and further the divergence process remains an open question. Here, we show that varying degrees of pre- and postmating RI exist among allopatric populations of two interbreeding cherry-infesting tephritid fruit flies (Rhagoletis cingulata and R. indifferens) across North America. These flies display allochronic and sexual isolation among populations, as well as unidirectional reductions in egg hatch in hybrid crosses involving southwestern USA males. All populations are infected by a Wolbachia strain, wCin2, whereas a second strain, wCin3, only co-infects flies from the southwest USA and Mexico. Strain wCin3 is associated with a unique mitochondrial DNA haplotype and unidirectional postmating RI, implicating the strain as the cause of CI. When coupled with nonendosymbiont RI barriers, we estimate the strength of CI associated with wCin3 would not prevent the strain from introgressing from infected southwestern to uninfected populations elsewhere in the USA if populations were to come into secondary contact and hybridize. In contrast, cytoplasmic-nuclear coupling may impede the transfer of wCin3 if Mexican and USA populations were to come into contact. We discuss our results in the context of the general paucity of examples demonstrating stable Wolbachia hybrid zones and whether the spread of Wolbachia among taxa can be constrained in natural hybrid zones long enough for the endosymbiont to participate in speciation.

Metagenomic Survey of the Highly Polyphagous Anastrepha ludens Developing in Ancestral and Exotic Hosts Reveals the Lack of a Stable Microbiota in Larvae and the Strong Influence of Metamorphosis on Adult Gut Microbiota.

We studied the microbiota of a highly polyphagous insect, Anastrepha ludens (Diptera: Tephritidae), developing in six of its hosts, including two ancestral (Casimiroa edulis and C. greggii), three exotic (Mangifera indica cv. Ataulfo, Prunus persica cv. Criollo, and Citrus x aurantium) and one occasional host (Capsicum pubescens cv. Manzano), that is only used when extreme drought conditions limit fruiting by the common hosts. One of the exotic hosts ("criollo" peach) is rife with polyphenols and the occasional host with capsaicinoids exerting high fitness costs on the larvae. We pursued the following questions: (1) How is the microbial composition of the larval food related to the composition of the larval and adult microbiota, and what does this tell us about transience and stability of this species' gut microbiota? (2) How does metamorphosis affect the adult microbiota? We surveyed the microbiota of the pulp of each host fruit, as well as the gut microbiota of larvae and adult flies and found that the gut of A. ludens larvae lacks a stable microbiota, since it was invariably associated with the composition of the pulp microbiota of the host plant species studied and was also different from the microbiota of adult flies indicating that metamorphosis filters out much of the microbiota present in larvae. The microbiota of adult males and females was similar between them, independent of host plant and was dominated by bacteria within the Enterobacteriaceae. We found that in the case of the "toxic" occasional host C. pubescens the microbiota is enriched in potentially deleterious genera that were much less abundant in the other hosts. In contrast, the pulp of the ancestral host C. edulis is enriched in several bacterial groups that can be beneficial for larval development. We also report for the first time the presence of bacteria within the Arcobacteraceae family in the gut microbiota of A. ludens stemming from C. edulis. Based on our findings, we conclude that changes in the food-associated microbiota dictate major changes in the larval microbiota, suggesting that most larval gut microbiota is originated from the food.

Pupation Substrate Type and Volume Affect Pupation, Quality Parameters and Production Costs of a Reproductive Colony of Ceratitis capitata (Diptera: Tephritidae) VIENNA 8 Genetic Sexing Strain.

Adequate pupation substrates and substrate volume are critical factors in the mass-rearing of insects for Sterile Insect Technique (SIT) applications. To identify an ideal pupation substrate for a reproductive colony of Ceratitis capitata (Wiedemann) VIENNA 8 genetic sexing strain, we first examined pupation in cellulose from recycled paper (cellulose I), sawdust, fine wheat bran, vermiculite and coconut fiber using a volume of 2.5-12.5 mL of substrate for each 5 mL volume of fly larvae. We found a positive relationship between substrate volume and pupation, with cellulose I generating the highest proportions of pupation and coconut fiber the lowest. Higher proportions of female flies (white pupae) pupated in sawdust. The proportion of female fliers increased as substrate volume rose in sawdust and coconut fiber, whereas it decreased in vermiculite and cellulose. In a second experiment, we tested three types of cellulose differing in physicochemical characteristics (celluloses I, II and III), sawdust, and fine wheat bran using a substrate:larvae ratio of 1:1. The three types of cellulose produced the highest pupation levels. The highest proportions of female fliers were observed in sawdust, and cellulose types III and II. Cellulose III and sawdust at relatively low volumes were more cost-effective to produce one million pupae than other substrates, including fine wheat bran used in a mass-rearing facility in Mexico.

Chitosan coatings reduce fruit fly (Anastrepha obliqua) infestation and development of the fungus Colletotrichum gloeosporioides in Manila mangoes.

BACKGROUND: Mangoes are tropical fruits appreciated worldwide but are extremely perishable, being susceptible to decay, pest infestation and fungal diseases. Using the flavorful and highly valued 'Manila' cultivar, we examined the effect of second-generation chitosan coatings on shelf-life, phenolic compound variation, phytohormones, pest infestation by fruit flies (Anastrepha obliqua) and anthracnose disease caused by the fungus Colletotrichum gloeosporioides. RESULTS: We observed almost total elimination of A. obliqua eggs with 10 and 20 g L-1 chitosan in diluted acetic acid and a five- to sixfold reduction in anthracnose damage. Treatment with 20 g L-1 chitosan also extended the shelf-life. External (skin) and internal (pulp) discoloration processes were delayed. Fruit firmness was higher when compared with control and acetic acid treatments, and total soluble solids were lower in chitosan-treated fruit. Targeted and non-targeted metabolomics analyses on chitosan-coated fruit identified some phenolic compounds related to the tannin pathway. In addition, abscisic acid and jasmonic acid in the peel were downregulated in chitosan-coated mango peels. Both phytohormones and phenolic content may explain the reduced susceptibility of mangoes to anthracnose development and A. obliqua egg eclosion or larval development. CONCLUSIONS: We conclude that chitosan coatings represent an effective postharvest treatment that significantly reduces anthracnose disease, inhibits A. obliqua egg eclosion and significantly extends 'Manila' mango shelf-life, a key factor currently inhibiting large-scale commercialization of this valuable fruit. © 2020 Society of Chemical Industry.

Evidence for spatial clines and mixed geographic modes of speciation for North American cherry-infesting Rhagoletis (Diptera: Tephritidae) flies.

An important criterion for understanding speciation is the geographic context of population divergence. Three major modes of allopatric, parapatric, and sympatric speciation define the extent of spatial overlap and gene flow between diverging populations. However, mixed modes of speciation are also possible, whereby populations experience periods of allopatry, parapatry, and/or sympatry at different times as they diverge. Here, we report clinal patterns of variation for 21 nuclear-encoded microsatellites and a wing spot phenotype for cherry-infesting Rhagoletis (Diptera: Tephritidae) across North America consistent with these flies having initially diverged in parapatry followed by a period of allopatric differentiation in the early Holocene. However, mitochondrial DNA (mtDNA) displays a different pattern; cherry flies at the ends of the clines in the eastern USA and Pacific Northwest share identical haplotypes, while centrally located populations in the southwestern USA and Mexico possess a different haplotype. We hypothesize that the mitochondrial difference could be due to lineage sorting but more likely reflects a selective sweep of a favorable mtDNA variant or the spread of an endosymbiont. The estimated divergence time for mtDNA suggests possible past allopatry, secondary contact, and subsequent isolation between USA and Mexican fly populations initiated before the Wisconsin glaciation. Thus, the current genetics of cherry flies may involve different mixed modes of divergence occurring in different portions of the fly's range. We discuss the need for additional DNA sequencing and quantification of prezygotic and postzygotic reproductive isolation to verify the multiple mixed-mode hypothesis for cherry flies and draw parallels from other systems to assess the generality that speciation may commonly involve complex biogeographies of varying combinations of allopatric, parapatric, and sympatric divergence.

A First Glimpse of the Mexican Fruit Fly Anastrepha ludens (Diptera: Tephritidae) Antenna Morphology and Proteome in Response to a Proteinaceous Attractant.

Anastrepha ludens is a key pest of mangoes and citrus from Texas to Costa Rica but the mechanisms of odorant perception in this species are poorly understood. Detection of volatiles in insects occurs mainly in the antenna, where molecules penetrate sensillum pores and link to soluble proteins in the hemolymph until reaching specific odor receptors that trigger signal transduction and lead to behavioral responses. Scrutinizing the molecular foundation of odorant perception in A. ludens is necessary to improve biorational management strategies against this pest. After exposing adults of three maturity stages to a proteinaceous attractant, we studied antennal morphology and comparative proteomic profiles using nano-LC-MS/MS with tandem mass tags combined with synchronous precursor selection (SPS)-MS3. Antennas from newly emerged flies exhibited dense agglomerations of olfactory sensory neurons. We discovered 4618 unique proteins in the antennas of A. ludens and identified some associated with odor signaling, including odorant-binding and calcium signaling related proteins, the odorant receptor co-receptor (Orco), and putative odorant-degrading enzymes. Antennas of sexually immature flies exhibited the most upregulation of odor perception proteins compared to mature flies exposed to the attractant. This is the first report where critical molecular players are linked to the odor perception mechanism of A. ludens.

Host Suitability and Fitness-Related Parameters in Coptera haywardi (Hymenoptera: Diapriidae) Reared on Irradiated Ceratitis capitata (Diptera: Tephritidae) Pupae Stemming From the tsl Vienna-8 Genetic Sexing Strain.

Coptera haywardi (Ogloblin) is a pupal endoparasitoid of tephritid flies with great potential as a biological control agent worldwide as it does not attack other Diptera. To reach its full potential, its mass rearing needs to be enhanced lowering costs. Here, we focused on the use of irradiated pupae of Ceratitis capitata (Wiedemann) stemming from the temperature-sensitive lethal (tsl) Vienna-8 genetic sexing strain (= CcVienna-8), which is mass-produced in the San Juan Medfly and Parasitoid Mass Rearing Facility in Argentina. Exposure of 1- to 2-d-old CcVienna-8 pupae irradiated at 90 Gy to 6- to 8-d-old C. haywardi females at a 10:1 host/parasitoid ratio for 24 h turned out to be highly successful for the rearing of this parasitoid. High radiation doses (90-100 Gy) did not adversely influence fitness parameters of C. haywardi offspring F1, namely lifetime reproductive rates, adult life expectancy, and survival time. Demographic parameters in C. haywardi F1 from irradiated CcVienna-8 young pupae were improved compared to those values recorded from parasitoid originated from nonirradiated CcVienna-8 pupae. These findings will help to enhance parasitoid mass rearing for augmentative releases against medfly in Argentinean fruit-producing regions.

Host Plant and Antibiotic Effects on Scent Bouquet Composition of Anastrepha ludens and Anastrepha obliqua Calling Males, Two Polyphagous Tephritid Pests.

In insects, the quality of sex pheromones plays a critical role in mating success and can be determined by the ability of larvae/adults to accrue chemical precursors. We tested the host-quality-effect hypothesis by analyzing the chemical composition of scent bouquets emitted by calling males of two polyphagous tephritid species (Anastrepha ludens and A. obliqua) that originated from 13 fruit species representing diverse plant families. In A. ludens, we worked with an ancestral host (Rutaceae), nine exotic ones (Rutaceae, Anacardiaceae, Rosaceae, Solanaceae, Lythraceae), and two species never attacked in nature but that represent candidates for host-range expansion (Solanaceae, Myrtaceae). In A. obliqua, we tested an ancestral, a native, and an exotic host (Anacardiaceae), one occasional (Myrtaceae), and one fruit never attacked in nature (Solanaceae). We identified a core scent bouquet and significant variation in the bouquet's composition depending on the fruit the larvae developed in. We also tested the possible microbial role on the scent bouquet by treating adults with antibiotics, finding a significant effect on quantity but not composition. We dwell on plasticity to partially explain our results and discuss the influence hosts could have on male competitiveness driven by variations in scent bouquet composition and how this could impact insect sterile technique programs.

Insights into the Interaction between the Monophagous Tephritid Fly Anastrepha acris and its Highly Toxic Host Hippomane mancinella (Euphorbiaceae).

Despite their enormous economic importance and the fact that there are almost 5000 tephritid (Diptera) species, fruit fly - host plant interactions are poorly understood from a chemical perspective. We analyzed the interactions among Anastrepha acris (a little studied monophagous tephritid) and its highly toxic host plant Hippomane mancinella from chemical, ecological and experimental perspectives, and also searched for toxicants from H. mancinella in the larval-pupal endoparasitoid Doryctobracon areolatus. We identified 18 phenolic compounds from H. mancinella pulp belonging to different chemical groups including phenylpropanoids, flavonoids, chalcones and coumarins. No traces of Hippomanin A were detected in larvae, pupae or A. acris adults, or in D. areolatus adults, implying that A. acris larvae can metabolize this toxicant, that as a result does not reach the third trophic level. We tested the "behavioral preference - lack of larval specialization-hypothesis" via feeding experiments with a larval rearing medium containing H. mancinella fruit (skin + pulp or pulp alone). The high toxicity of H. mancinella was confirmed as only two (out of 2520 in three experiments) A. ludens larvae (a polyphagous pest species that preferentially feeds on plants within the Rutaceae) survived without reaching the adult stage when fed on media containing H. mancinella, whereas A. acris larvae developed well and produced healthy adults. Together, these findings open a window of opportunity to study the detoxification mechanisms used by tephritid fruit flies.