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.

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.

Electroantennographic Responses of Wild and Laboratory-Reared Females of Xyleborus affinis Eichhoff and Xyleborus ferrugineus (Fabricius) (Coleoptera: Curculionidae: Scolytinae) to Ethanol and Bark Volatiles of Three Host-Plant Species.

Chemical ecology studies on ambrosia beetles are typically conducted with either wild or laboratory-reared specimens. Unlike laboratory-reared insects, important aspects that potentially influence behavioral responses, such as age, physiological state, and prior experience are unknown in wild specimens. In this study, we compared the electroantennographic (EAG) responses of laboratory-reared and wild X. affinis and X. ferrugineus to 70% ethanol and bark odors (host kairomones) of Bursera simaruba, Mangifera indica, and Persea schiedeana aged for 2, 24, and 48 h. Chemical analyses of each odor treatment (bark species x length of aging) were performed to determine their volatilome composition. EAG responses were different between laboratory-reared and wild X. ferrugineus when exposed to ethanol, whereas wild X. affinis exhibited similar EAG responses to the laboratory-reared insects. Ethanol elicited the strongest olfactory responses in both species. Among the bark-odors, the highest responses were triggered by B. simaruba at 48 h in X. affinis, and P. schiedeana at 24 and 48 h in X. ferrugineus. Volatile profiles varied among aged bark samples; 3-carene and limonene were predominant in B. simaruba, whereas α-copaene and α-cubebene were abundant in P. schiedeana. Further studies are needed to determine the biological function of B. simaruba and P. schiedeana terpenes on X. affinis and X. ferrugineus, and their potential application for the development of effective lures.

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.

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.

Efecto del aceite esencial de Chrysactinia mexicana A. Gray sobre aislados clínicos de Candida glabrata / Effect of the essential oil of Chrysactinia mexicana A. Gray on clinical isolates of Candida glabrata

El objetivo del trabajo fue evaluar el efecto del aceite esencial Chrysactinia mexicana A. Gray en aislados clínicos de C. glabrata. Se obtuvo aceite esencial por arrastre de vapor de las flores, hojas y ramas secundarias de C. mexicana. Se realizó cromatografía de gases y espectrometría de masas para conocer la composición del aceite. Se hicieron ensayos de microdilución en fase logarítmica y fase estacionaria con Saccharomyces cerevisiae y aislados clínicos de C. glabrata para determinar la Concentración Mínima Inhibitoria (CMI) en presencia del aceite esencial. Los principales compuestos del aceite esencial fueron la piperitona (29.57 %), eucaliptol (26.86 %), a-terpineol (14.65 %), delta-3-careno (12.37 %) y linalool (3.56 %). Se determinaron las CMI del aceite esencial, para las cinco cepas utilizadas en las dos fases de crecimiento, siendo menor la CMI para S. cerevisiae que para los diferentes aislados clínicos de C. glabrata. El aceite esencial de C. mexicana tiene efecto fungicida en S. cerevisiae y los aislados clínicos de C. glabrata.

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 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.

Agar and Carrageenan as Cost-Effective Gelling Agents in Yeast-Reduced Artificial Diets for Mass-Rearing Fruit Flies and Their Parasitoids.

The development of cost-effective diets for mass-rearing fruit flies (Diptera: Tephritidae) and their parasitoids in pest control programs based on the Sterile Insect Technique is a high priority worldwide. To this end, we tested carrageenan, agar, gelatin and two types of pregelatinized corn starches as gelling agents at varying percentages in a yeast-reduced liquid larval diet for rearing the Mexfly, Anastrepha ludens. Only diets with 0.234% (w/w) agar or 0.424% carrageenan were identified as diets with potential for mass-rearing A. ludens in terms of the number of pupae recovered from the diet, pupal weight, adult emergence, flight ability and diet cost. Comparative experiments showed that yeast-reduced agar and carrageenan gel diets produced heavier pupae and higher proportions of flying adults than the standard mass-rearing diet. The gel-agar and mass-rearing diets produced more pupae than the gel-carrageenan diet, but the latter produced more suitable larvae as hosts for rearing of Diachasmimorpha longicaudata (Hymenoptera: Braconidae) females, a widely used fruit fly biocontrol agent. Yeast-reduced agar and carrageenan gel diets could represent cost-effective fruit fly mass-rearing diets if a practical system for gel diet preparation and dispensation at fruit fly mass-rearing facilities is developed.

Physalis peruviana L. (Solanaceae) Is Not a Host of Ceratitis capitata (Diptera: Tephritidae): Evidence from Multi-Year Field and Laboratory Studies in Colombia.

Scientifically-based, tephritid fly host status determination lies at the heart of strategic regulatory decisions impinging on international fruit trade. Here we conducted intensive field and laboratory studies with peaches as controls, to determine the host status of Physalis peruviana for the Medfly-Ceratitis capitata, as this fruit is experiencing a consumption boom worldwide. A total of 98,132 Uchuvas (local name), collected in Colombia from the plant or the ground over a three-year period (2016-2018) did not yield a single C. capitata larva or pupa, thus reaching a Probit 9 level with 99.9968% efficacy and 96% confidence level. Field-cage studies with enclosed fruit-bearing Uchuva plants, exposing fruit with an intact, damaged or totally removed husk to the attack of C. capitata, also failed to yield infestations. Highly artificial choice experiments, exposing gravid females to unripe and fully ripe fruit, resulted in an absence of infestations, even when overripe Uchuvas were artificially damaged. The husk and surface resins/waxes inhibit fly landings on fruit and oviposition activity. Considering our results and the fact that the foliage, husk and fruit of P. peruviana are repellent/toxic to insects, we conclude that this plant should be treated as a non-natural and non-conditional host of C. capitata.

Structural Differences in the Digestive Tract Between Females and Males Could Modulate Regurgitation Behavior in Anastrepha ludens (Diptera: Tephritidae).

With the aim of understanding the mechanisms involved in the regurgitation behavior of tephritid flies, we performed a structural study of the digestive system of the economically important fruit-fly pest, Anastrepha ludens (Loew) using optical, scanning electronic microscopy (SEM) and transmission electron microscopy (TEM), plus a feeding assay. Most structures studied are similar to those previously reported in other adult dipterans, but, importantly, we found sexual differences in some structures that apparently affect regurgitation. We report for the first time sexual differences in the crop duct nerve and large numbers of dense core vesicles within the nerve bundle. Male nerve bundles are bigger and have more secretory vesicles than female ones. The close proximity to the muscles of both the crop lobes and duct suggest that these vesicles (i.e., possibly neurosecretions) might help modulate the muscles regulating regurgitation. The salivary glands are connected to the crop via tracheae, however, SEM/TEM studies failed to find any direct structural connection. Results of the feeding assay indicate that, independently of food type (sucrose or protein) and age, males regurgitate significantly more than females. Regurgitation behavior may also play an important role in capturing bacteria in the environment, and possibly help adults eliminate ingested toxicants such as insecticides. Our findings shed light on an interesting phenomenon that has important practical implications.

The Effect of Winter Length on Duration of Dormancy and Survival of Rhagoletis completa (Diptera: Tephritidae) and Associated Parasitoids From Northeastern Mexico.

The walnut husk fly Rhagoletis completa (Cresson), native to the Midwestern United States and Mexico, is invasive in California and Europe. It is one of the most important pests of walnuts in areas gathering 30% of the world production. Knowledge of life-history regulation is important for the design of management strategies. Research on dormancy has been performed on invasive populations, and not on populations at the southern extreme of its native range. Here, we examined the effect of winter length on fly and parasitoid emergence, survival, and duration of dormancy. Percent emergence was higher for chill periods at 5°C ranging from 8 to 20 wk. No or insufficient chill resulted in low emergence and a significant proportion of individuals in prolonged dormancy (>1 yr). Duration of dormancy was longer for pupae at constant temperatures and a 4-wk chill period than longer winter durations. Dormancy was longer for Mexican than that reported for U.S. populations, suggesting the existence of a latitudinal cline where populations at southern latitudes have evolved slower metabolic rates. Three parasitoid species were found associated with R. completa (Aganaspis alujai (Wharton and Ovruski) (Hymenoptera: Figitidae), Diachasmimorpha juglandis Muesebeck, and Diachasmimorpha mellea Gahan) (Hymenoptera: Braconidae). Results suggest that rearing of R. completa is possible by subjecting pupae to chill periods between 8 and 20 wk. Overwintering mortality of flies and A. alujai could be further reduced above 5°C. Our findings can contribute for the accurate development of predictive models on invasion potential, development, fly and parasitoid rearing, and biological control.

Effects of Larval Density and Support Substrate in Liquid Diet on Productivity and Quality of Artificially Reared Anastrepha ludens (Diptera: Tephritidae).

The larval developmental environment can affect the quality of artificially reared insects used in pest control programs that apply the sterile insect technique. We used a liquid larval diet lacking corncob fractions (an ingredient which may be contaminated by toxins that inhibit larval development and drastically reduce insect production in mass-rearing facilities) to examine quality indicators of Anastrepha ludens (Loew) (Diptera: Tephritidae) as a function of larval density (1-5 larvae per ml of diet) and on two types of supporting materials (synthetic sponge and carpet felt) compared with a standard solid diet with corncob fractions inoculated with a fixed quantity of A. ludens eggs. Pupal weight and adult emergence of A. ludens reared on a liquid artificial diet were negatively and positively density dependent, respectively. One tray of solid diet produced an average (95% CI) of 689.5 (594.4, 794.3) pupae, whereas one tray of liquid diet with synthetic sponge and carpet felt produced an average of 266.0 (208.7, 332.9) and 43.7 (23.2, 73.9) pupae, respectively. Pupal weight and adult emergence of flies reared on the solid diet were atypically low probably due to the type of rearing tray used, whereas pupal weight and adult emergence on liquid diet were equal to, or exceeded 20 mg and a proportion of 0.93, respectively. The highest proportion of adults capable of flight was observed in the liquid diet with carpet felt. The potential causes by which larval density and support substrate affected productivity and quality of A. ludens reared on liquid diet are discussed.

Effect of Resin Ducts and Sap Content on Infestation and Development of Immature Stages of Anastrepha obliqua and Anastrepha ludens (Diptera: Tephritidae) in Four Mango (Sapindales: Anacardiaceae) Cultivars.

We determined the influence of resin ducts, sap content, and fruit physicochemical features of four mango cultivars (Criollo, Manila, Ataulfo, and Tommy Atkins) on their susceptibility to the attack of the two most pestiferous fruit fly species infesting mangoes in Mexico: Anastrepha ludens (Loew) and Anastrepha obliqua (Macquart). We performed three studies: 1) analysis of resin ducts in mango fruit exocarp to determine the density and area occupied by resin ducts in each mango cultivar, 2) assessment of mango physicochemical features including fruit sap content, and 3) a forced infestation trial under field conditions using enclosed fruit-bearing branches to expose mangoes to gravid A. ludens or A. obliqua females. Infestation rates, development time from egg to prepupae and pupae, pupal weight, and percent of adult emergence, were assessed. 'Ataulfo' and 'Tommy Atkins' cultivars exhibited the highest resin duct density and sap content, the lowest infestation rate, and had a negative effect on immature development and pupal weight. In sharp contrast, 'Manila' and 'Criollo' cultivars, with the lowest resin duct density and sap content, were highly susceptible to A. ludens and A. obliqua attack. We conclude that sap content and the number, size, and distribution of resin ducts as well as firmness in mango fruit exocarp are all involved in the resistance of mango to A. ludens and A. obliqua attack.

Developing diagnostic SNP panels for the identification of true fruit flies (Diptera: Tephritidae) within the limits of COI-based species delimitation.

BACKGROUND: Rapid and reliable identification of quarantine pests is essential for plant inspection services to prevent introduction of invasive species. For insects, this may be a serious problem when dealing with morphologically similar cryptic species complexes and early developmental stages that lack distinctive characters useful for taxonomic identification. DNA based barcoding could solve many of these problems. The standard barcode fragment, an approx. 650 base pairs long sequence of the 5'end of the mitochondrial cytochrome oxidase I (COI), enables differentiation of a very wide range of arthropods. However, problems remain in some taxa, such as Tephritidae, where recent genetic differentiation among some of the described species hinders accurate molecular discrimination. RESULTS: In order to explore the full species discrimination potential of COI, we sequenced the barcoding region of the COI gene of a range of economically important Tephritid species and complemented these data with all GenBank and BOLD entries for the systematic group available as of January 2012. We explored the limits of species delimitation of this barcode fragment among 193 putative Tephritid species and established operational taxonomic units (OTUs), between which discrimination is reliably possible. Furthermore, to enable future development of rapid diagnostic assays based on this sequence information, we characterized all single nucleotide polymorphisms (SNPs) and established "near-minimal" sets of SNPs that differentiate among all included OTUs with at least three and four SNPs, respectively. CONCLUSIONS: We found that although several species cannot be differentiated based on the genetic diversity observed in COI and hence form composite OTUs, 85% of all OTUs correspond to described species. Because our SNP panels are developed based on all currently available sequence information and rely on a minimal pairwise difference of three SNPs, they are highly reliable and hence represent an important resource for developing taxon-specific diagnostic assays. For selected cases, possible explanations that may cause composite OTUs are discussed.

Understanding long-term fruit fly (Diptera: Tephritidae) population dynamics: implications for areawide management.

Fruit flies (Diptera: Tephritidae) are devastating agricultural pests worldwide but studies on their long-term population dynamics are sparse. Our aim was to determine the mechanisms driving long-term population dynamics as a prerequisite for ecologically based areawide pest management. The population density of three pestiferous Anastrepha species [Anastrepha ludens (Loew), Anastrepha obliqua (Macquart), and Anastrepha serpentina (Wiedemann)] was determined in grapefruit (Citrus x paradisi Macfad.), mango (Mangifera indica L.), and sapodilla [Manilkara zapota (L.) P. Royen] orchards in central Veracruz, México, on a weekly basis over an 11-yr period. Fly populations exhibited relatively stable dynamics over time. Population dynamics were mainly driven by a direct density-dependent effect and a seasonal feedback process. We discovered direct and delayed influences that were correlated with both local (rainfall and air temperature) and global climatic variation (El Niño Southern Oscillation [ENSO] and North Atlantic Oscillation [NAO]), and detected differences among species and location of orchards with respect to the magnitude and nature (linear or nonlinear) of the observed effects, suggesting that highly mobile pest outbreaks become uncertain in response to significant climatic events at both global and local levels. That both NAO and ENSO affected Anastrepha population dynamics, coupled with the high mobility of Anastrepha adults and the discovery that when measured as rate of population change, local population fluctuations exhibited stable dynamics over time, suggests potential management scenarios for the species studied lie beyond the local scale and should be approached from an areawide perspective. Localized efforts, from individual growers will probably prove ineffective, and nonsustainable.