Response of two parasitoid species (Hymenoptera: Braconidae,Figitidae) to tephritid host and host food substrate cues

The Neotropical‐native figitid Aganaspis pelleranoi (Brèthes) and the Asian braconid Diachasmimorpha longicaudata (Ashmead) are two parasitoids of Tephritidae fruit flies with long and recent, respectively, evolutionary histories in the Neotropics. Both species experienced a recent range of overlap. In Argentina, A. pelleranoi is a potential species in biological control programs against the pestiferous tephritid species, Anastrepha fraterculus (Wiedemann) and Ceratitis capitata (Wiedemann), whereas D. longicaudata is already used in open‐field releases against Medfly in central‐western Argentina. To characterize the host‐foraging strategies of A. pelleranoi and D. longicaudata, olfactometer experiments were conducted comparing responses to C. capitata and A. fraterculus larvae, in two kinds of food substrate: fruit and artificial larval medium. To control the possible influence of host larvae used for parasitoid rearing on olfactory response, two strains of both parasitoid species, reared on both tephrtid species, were studied. Volatiles directly emanating either from A. fraterculus or C. capitata larvae may be detected by both A. pelleranoi and D. longicaudata, although chemical stimuli originating from the combination of host larvae and the habitat of the host were preferred. However, olfactory cues associated with host larvae probably play a relevant role in host searching behaviour of A. pelleranoi, whereas for D. longicaudata, the host‐habitat olfactory stimuli would be highly essential in short‐range host location. The strain of the parasitoids did not affect host search ability on the two tephritid species evaluated. These evidences are relevant for mass production of both parasitoids and their impact following open‐field augmentative releases.     

Susceptibility of selected apple cultivars to the Mediterranean fruit fly

Ceratitis capitata (Wiedemann), the Mediterranean fruit fly, is one of the key pest species affecting deciduous fruit orchards along the Mediterranean coasts. Because of global warming, C. capitata is gradually spreading north and is becoming a major pest of apples. Determining the susceptibility of the main apple varieties grown in the region will serve as a cornerstone to the management of this pest. In this study, we show the results of a field and laboratory no‐choice test conducted to determine the Medfly preferences on different apple cultivars. The seven main varieties of apples (Gala, Red Delicious, Golden Delicious, Granny Smith, Kanzi, Morgen Dallago and Fuji) were tested. The results demonstrate that C. capitata lays eggs on all apple cultivars in both field and laboratory conditions. The Granny Smith, Red Delicious and Morgen Dallago varieties showed the lowest susceptibility in laboratory conditions, (0.75, 1.55, 2 oviposition punctures/fruit, respectively), with significant differences in oviposition compared to the Golden Delicious, Kanzi and Fuji (3.27, 3.31, 3.1 oviposition punctures/fruit, respectively) varieties, which were shown to be the most susceptible to Medfly attack in laboratory conditions. On the other hand, only slight and not statistically significant differences emerged from the field trials. In relation to the physico‐chemical characteristics, the apple cultivars showing the lowest susceptibility (Granny Smith, Red Delicious and Morgen Dallago) had harder peels and pulps and lower sugar contents than the most susceptible cultivars (Golden Delicious, Fuji and Kanzi). These results were also confirmed through evaluation of larval development on different varieties. In fact, Granny Smith, Red Delicious and Morgen Dallago were the three varieties that did not allow adequate larval and adult development and reduced the possibility of the emergence of a new generation.     

Response of the fruit fly parasitoid Diachasmimorpha longicaudata to host and host‐habitat volatile cues

Chemical information is crucial to insect parasitoids for successful host location. Here, we evaluated the innate response of Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), a fruit fly larval parasitoid, to cues from host and host habitat (i.e., fruit infested with host larvae). We first assessed the preference of female parasitoids between oranges infested with Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) and non‐infested fruit. Females were highly attracted towards infested oranges on the basis of volatile chemical cues. After this initial experiment, we aimed at revealing the potential sources of volatile cues present in an infested fruit. To this end, we considered five potential sources: (1) punctured fruit; (2) fly feeding, frass, or host‐marking pheromone deposited on the orange surface; (3) larval activity inside the fruit; (4) the larvae themselves; and (5) fungi associated with infestation of oranges. Habitat cues associated with host activity and those produced by rotten oranges or oranges colonized by fungi were highly attractive for female wasps, whereas odours associated with the activity of the adults on the surface of the fruit, and those released by the fruit after being damaged (as happens during fruit fly egg‐laying) were not used as cues by female parasitoids. Once the female had landed on the fruit, direct cues associated with larval activity became important although some indirect signals (e.g., products derived from larval activity inside the fruit) also increased host searching activity. Our findings indicate that naïve D. longicaudata uses chemical cues during host habitat searching and that these cues are produced both by the habitat and by the host larvae.     

Rearing of the fruit fly parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae) on X-ray irradiated larvae of Ceratitis capitata (Diptera: Tephritidae)

We evaluated the effects of X-ray irradiation on larvae of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), with the aim of finding a treatment that prevented adult fly emergence, yet did not adversely affect larval quality as rearing hosts for the parasitoid Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Ichneumonidae). Two experiments were carried out. In the first experiment, small numbers of larvae were held in Petri dishes with and without small amounts of rearing media and then irradiated with doses of X-rays ranging from 0 to 8333.6 R (equivalent to 80 Gy). In the second experiment, higher numbers of larvae were held in a manner resembling mass-rearing conditions, and were then irradiated with X-rays ranging from 0 to 10,417 R (equivalent to 100 Gy). In both experiments, the only factor that significantly affected fly emergence was irradiation of larvae. Fly emergence decreased markedly as the irradiation dose increased, and complete suppression of fly emergence was achieved at 6250.2 R (equivalent to 60 Gy) when larvae were irradiated in small batches with or without rearing media. Irradiation also affected the fertility of those flies that did emerge following treatment. In the second experiment, we found the parasitoids reared from irradiated larvae produced a higher parasitism rate and a higher number of female offspring than did parasitoids reared from control (non-irradiated) larvae. Mean fecundity of F1 parasitoids reared from irradiated larvae were affected positively by irradiation only at the 8333.6 R (80 Gy) dose. Our results show that X-ray irradiation can be used to inhibit fruit fly adult emergence and that irradiated larvae are at least as good a rearing substrate as non-irradiated larvae. Future studies should focus on the adjustment of our findings to a mass-rearing scale.     

Genome size estimation with quantitative real‐time PCR in two Tephritidae species: Ceratitis capitata and Bactrocera oleae

The medfly Ceratitis capitata and the olive fruit fly Bactrocera oleae belong to the Tephritidae family of Diptera, a family whose members cause severe damages in agriculture worldwide. For such insect pests, the utmost concern is their population control. The sterile insect technique (SIT) has been used in the Tephritidae family with varying degrees of success. Its efficient use usually depends on the development of genetic sexing strains and the release of only male flies. However, such advances are based on modern genetic, molecular and genomic tools. The medfly is clearly the prototype of the family, since such tools have advanced considerably, which has resulted in effective SIT efforts around the world. A whole‐genome sequencing project of this insect is already underway. In contrast, similar tools in the olive fly lag behind, even though the insect is considered a promising candidate for a next SIT target. An accurate estimate of genome size provides a preliminary view of genome complexity and indicates possible difficulties in genome assembly in whole‐genome projects. Taking advantage of a quantitative real‐time PCR approach, we determined the genome size of these two species C. capitata and B. oleae as 591 Mb (CI range: 577–605 Mb) and 322 Mb (CI range: 310–334 Mb) respectively.     

Novel bait stations for attract-and-kill of pestiferous fruit flies

A novel, visually-attractive bait station was developed in Hawaii for application of insecticidal baits against oriental fruit fly, Bactrocera dorsalis (Hendel), melon fly, Bactrocera cucurbitae (Coquillett), and Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (all Diptera: Tephritidae). The bait station developed represents a supernormal visual stimulus of papaya foliage and takes advantage of the flies' strong response to the high light-reflecting properties of yellow color and of their need for shelter, while fully protecting the bait against rainfall. Field studies revealed that the behavioral response of female fruit flies, in particular C. capitata and B. cucurbitae , to yellow-painted bait stations sprayed with GF-120 NF Naturalyte Fruit Fly Bait was significantly enhanced compared with similarly sprayed bait stations that mimicked the green color of fully grown papaya leaves. Field studies conducted with B. cucurbitae indicated that the period of bait attractiveness can be extended for at least 1 week after bait application due to the rain-fastness properties of the bait stations and the use of a visually-attractive color. Our studies provide the behavioral basis for the development of improved attract-and-kill bait stations for fruit flies in Hawaii. These devices also provide a standardized way of evaluating bait spray formulations, thus allowing for proper comparisons over time, across species, and among geographical areas.     

Susceptibility of the Mediterranean fruit fly (Ceratitis capitata) and the Natal fruit fly (Ceratitis rosa) to entomopathogenic nematodes

The potential of entomopathogenic nematodes, Heterorhabditis bacteriophora, Heterorhabditis zealandica and Steinernema khoisanae, to infect pupariating larvae, pupae and adults of Ceratitis capitata and Ceratitis rosa was investigated in laboratory bioassays. Pupariating larvae and adult flies were susceptible to nematode infection, with no infection recorded for the pupae. Pupariating larvae of C. capitata were generally more susceptible to infection than those of C. rosa. Significantly more larvae of C. capitata were infected by H. bacteriophora. For C. rosa, highest infectivity of larvae was obtained with H. zealandica. In contrast, adults of both species were highly infected by S. khoisanae.     

Recovering full DNA barcodes from natural history collections of Tephritid fruitflies (Tephritidae, Diptera) using mini barcodes

The family of Tephritid fruit flies (Tephritidae, Diptera) is composed of more than 4000 species and more than 350 are of economic importance (EI). The Tephritid Barcoding Initiative (TBI) aims at obtaining DNA barcodes for all EI species and the majority of their congeners. Dry pinned specimens from natural history collections are an important resource for reference material, but were often collected decades ago. We observed a strong decrease in the success rate of obtaining a full COX1 DNA barcode (658 bp), with an increasing age of the specimens. Obtaining full barcodes is often not possible using standard protocols. We developed a universal Tephritid primer set for multiple overlapping mini-barcodes that allows reconstructing the full COX1 DNA barcode. These newly developed primers and the corresponding protocol will facilitate the utilization of the extensive natural history collection by the TBI consortium.     

High sexual signalling rates of young individuals predict extended life span in male Mediterranean fruit flies

In a laboratory study, we monitored the lifetime sexual signalling (advertisement) of wild male Mediterranean fruit flies, and we tested the hypothesis that high lifetime intensity of sexual signalling indicates high survival probabilities. Almost all males exhibited signalling and individual signalling rates were highly variable from the beginning of the adults maturity and throughout their life span (average life span 62.3 days). Sexual signalling rates after day 10 (peak maturity) were consistently high until about 1 week before death. There was a positive relationship between daily signalling rates and life span, and an increase in signalling level by one unit over all times was associated with an approximately 50% decrease in mortality rate. Signalling rates early in adult life (day 6–20) were higher in the longest-lived than in the shortest-lived flies. These results support the hypothesis that intense sexual signalling indicates longer life span. We discuss the importance of age-specific behavioural studies for understanding the evolution of male life histories.     

Biosynthetic maturation of the corpus allatum of the female adult medfly,Ceratitis capitata,and its putative control

The major juvenile hormone (JH) homolog synthesized in vitro by the adult female Medfly (Ceratitis capitata) corpus allatum (CA) is JHB(3), with JH-III the minor homolog. Methyl-incorporation in vitro in post-eclosion virgin females is age-dependent. Basal activity occurs during the first four days post-eclosion and increases significantly thereafter, peaking at five days. Biosynthetic maturation of the mated female CA is delayed by one day and reduced considerably. The delayed response may be due to direct cerebral or neural inhibition. Synthetic Drosophila melanogaster sex peptide depresses JH biosynthesis by the Medfly female CA in vitro. Male-derived accessory gland peptides of the Medfly are transferred to the female during mating and a Medfly SP-analog may be responsible for down-regulation of JH synthesis by the CA in mated Medfly females. Mevinolin, an inhibitor of the mevalonate pathway, significantly reduces the biosynthesis of JHB(3), while farnesoic acid, a proximate precursor of JHIII, significantly stimulates the biosynthesis of both JHB(3) and JHIII in vitro.