Genome divergence and reproductive incompatibility among populations of Ganaspis near brasiliensis.

During the last decade, the spotted wing drosophila, Drosophila suzukii, has spread from eastern Asia to the Americas, Europe, and Africa. This fly attacks many species of cultivated and wild fruits with soft, thin skins, where its serrated ovipositor allows it to lay eggs in undamaged fruit. Parasitoids from the native range of D. suzukii may provide sustainable management of this polyphagous pest. Among these parasitoids, host-specificity testing has revealed a lineage of Ganaspis near brasiliensis, referred to in this paper as G1, that appears to be a cryptic species more host-specific to D. suzukii than other parasitoids. Differentiation among cryptic species is critical for introduction and subsequent evaluation of their impact on D. suzukii. Here, we present results on divergence in genomic sequences and architecture and reproductive isolation between lineages of Ganaspis near brasiliensis that appear to be cryptic species. We studied five populations, two from China, two from Japan, and one from Canada, identified as the G1 vs G3 lineages based on differences in cytochrome oxidase l sequences. We assembled and annotated the genomes of these populations and analyzed divergences in sequence and genome architecture between them. We also report results from crosses to test reproductive compatibility between the G3 lineage from China and the G1 lineage from Japan. The combined results on sequence divergence, differences in genome architectures, ortholog divergence, reproductive incompatibility, differences in host ranges and microhabitat preferences, and differences in morphology show that these lineages are different species. Thus, the decision to evaluate the lineages separately and only import and introduce the more host-specific lineage to North America and Europe was appropriate.

Discrimination Abilities and Parasitism Success of Pupal Parasitoids Towards Spotted-Wing Drosophila Pupae Previously Parasitized by the Larval Parasitoid Ganaspis brasiliensis (Hymenoptera: Figitidae).

Pachycrepoideus vindemiae (Rondani) (Hymenoptera: Pteromalidae) and Trichopria drosophilae (Perkins) (Hymenoptera: Diapriidae) are two cosmopolitan and generalist pupal parasitoids that are among a few of the resident parasitoids in North America capable of attacking Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), an invasive pest of small and soft fruit crops worldwide. Ganaspis brasiliensis (Ihering) is a specialist larval parasitoid of D. suzukii that was recently approved for biological control introduction against D. suzukii in the USA. As a solitary koinobiont species, G. brasiliensis oviposits in the host larva but emerges as an adult from the host puparium. This study investigated the discrimination ability and parasitism success by the pupal parasitoids towards D. suzukii pupae previously parasitized by G. brasiliensis, to examine whether interactions with resident parasitoids will affect G. brasiliensis after it is released in the USA. We found preliminary evidence that neither pupal parasitoid could discriminate towards D. suzukii pupae parasitized by early instars of G. brasiliensis. Pachycrepoideus vindemiae was able to successfully develop on D. suzukii pupae containing all preimaginal stages of G. brasiliensis, although parasitism success was significantly higher on those bearing later rather than early stages of G. brasiliensis. Trichopria drosophilae was only able to successfully develop on D. suzukii puparia containing early instars of G. brasiliensis. These results suggest that D. suzukii parasitized by the larval parasitoid could be subsequently attacked by the pupal parasitoids, possibly affecting the success of G. brasiliensis releases.

Methods for Rearing the Parasitoid Ganaspis brasiliensis, a Promising Biological Control Agent for the Invasive Drosophila suzukii.

Native to East Asia, the spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), has established widely in the Americas, Europe, and parts of Africa over the last decade, becoming a devastating pest of various soft-skinned fruits in its invaded regions. Biological control, especially by means of self-perpetuating and specialized parasitoids, is expected to be a viable option for sustainable area-wide management of this highly mobile and polyphagous pest. Ganaspis brasiliensis Ihering (Hymenoptera: Figitidae) is a larval parasitoid that is widely distributed in East Asia, and has been found to be one of the most effective parasitoids of D. suzukii. Following rigorous pre-introduction evaluations of its efficacy and potential non-target risks, one of the more host-specific genetic groups of this species (G1 G. brasiliensis) has been approved recently for introduction and field release in the United States and Italy. Another genetic group (G3 G. brasiliensis), which was also commonly found to attack D. suzukii in East Asia, may be considered for introduction in the near future. There is currently enormous interest in rearing G. brasiliensis for research or in mass-production for field release against D. suzukii. This protocol and associated video article describe effective rearing methods for this parasitoid, both on a small scale for research and a large scale for mass-production and field release. These methods may benefit further long-term research and use of this Asian-native parasitoid as a promising biological control agent for this global invasive pest.

Foraging Distance of the Argentine Ant in California Vineyards.

Argentine ants, Linepithema humile (Mayr) (Hymenoptera: Formicidae), form mutualisms with hemipteran pests in crop systems. In vineyards, they feed on honeydew produced by mealybugs and soft scales, which they tend and protect from natural enemies. Few options for controlling Argentine ants are available; one of the more effective approaches is to use liquid baits containing a low dose of an insecticide. Knowledge of ant foraging patterns is required to estimate how many bait stations to deploy per unit area. To measure how far ants move liquid bait in vineyards, we placed bait stations containing sugar water and a protein marker in plots for 6 d, and then collected ants along transects extending away from bait stations. The ants moved an average of 16.08 m and 12.21 m from bait stations in the first and second years of the study, respectively. Marked ants were found up to 63 m from bait stations; however, proportions of marked ants decreased exponentially as distance from the bait station increased. Results indicate that Argentine ants generally forage at distances <36 m in California vineyards, thus suggesting that insecticide bait stations must be deployed at intervals of 36 m or less to control ants. We found no effect of insecticide on distances that ants moved the liquid bait, but this may have been because bait station densities were too low to affect the high numbers of Argentine ants that were present at the study sites.

Performance of Psyttalia humilis (Hymenoptera: Braconidae) reared from irradiated host on olive fruit fly (Diptera: Tephritidae) in California.

The parasitoid Psyttalia humilis (Silvestri) was reared on Mediterranean fruit fly, Ceratitis capitata (Wiedemann), third instars irradiated at 0-70 Gy at the USDA, APHIS, PPQ, Moscamed biological control laboratory in San Miguel Petapa, Guatemala, and shipped to the USDA, ARS, Parlier, CA. Irradiation dose did not affect the parasitoid's offspring sex ratio (53-62% females), percentage of unemerged adults (12-34%), number of progeny produced per female (1.4-1.8), and parasitism (19-24%). Host irradiation dose had no significant effect on the forewing length of female P. humilis and its parasitism on olive fruit fly, Bactrocera oleae (Rossi) and offspring sex ratio, but dissection of 1-wk-old female parasitoids reared from hosts irradiated with 70 Gy had a significantly lower number of mature eggs than females from nonirradiated hosts. Longevity of P. humilis adults decreased with increased temperature from 15 to 35°C, regardless of food provisions, gender, and host irradiation dose. Females survived 37-49 d at 15°C with water and food, and only 1-2 d at 35°C without food, whereas males lived shorter than females at all temperatures and food combinations tested. Adult P. humilis reared from fertile C. capitata and aspirated for dispensing in cups lived significantly longer after shipment than those specimens chilled and dispensed by weight. At 21 and 32°C, 50% of parasitoids departed release cages after 180 and 30 min, respectively, but none departed at 12°C. Thirteen shipments of P. humilis (2,980-21,922 parasitoids per shipment) were received between September and December 2009, and seven shipments (7,502-22,560 parasitoids per shipment) were received between October and December 2010 from San Miguel Petapa, Guatemala. Daily number of olive fruit fly adult and percentage female trap captures ranged <1-19 and 8-58% in 2009, and <1-11 and 0-42% in 2010, respectively. The number of parasitoids released ranged 848-12,257 in 2009 and 3,675-11,154 in 2010. Percentage parasitism of olive fruit fly third instars at all locations ranged 0-9% in 2009 and 0-36% in 2010.

Testing baits to control Argentine ants (Hymenoptera: Formicidae) in vineyards.

Liquid baits were evaluated for control of the Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae), and associated mealybug and soft scale pests in California vineyards. In 2003, liquid baits with small doses ofimidacloprid, boric acid, or thiamethoxam dissolved in 25% sucrose water resulted in lower ant and mealybug densities and fruit damage, compared with an untreated control. Similar treatments in a soft scale-infested vineyard showed only a reduction of ant density and fruit infestation in only the boric acid and thiamethoxam treatments. In 2004, commercial and noncommercial formulations of liquid baits reduced ant densities in three separate trials, but they had inconsistent effects on mealybug densities and fruit infestation; granular protein bait had no effect. Using large plots and commercial application methodologies, liquid bait deployed in June resulted in lower ant density and fruit infestation, but it had no effect on mealybug density. Across all trials, liquid bait treatments resulted in lower ant density (12 of 14 trials) and fruit damage (11 of 14 sites), presenting the first report of liquid baits applied using commercial methodologies that resulted in a reduction of ants and their associated hemipteran crop damage. For commercialization of liquid baits, we showed that any of the tested insecticides can suppress Argentine ants when properly delivered in the crop system. For imidacloprid, bait dispensers must be protected from sunlight to reduce photodegradation. Results suggest that incomplete ant suppression can suppress mealybug densities. However, after ant populations are suppressed, there may be a longer period before hemipteran populations are effectively suppressed. Therefore, liquid baits should be considered part of a multiseason program rather than a direct, in-season control of hemipteran pest populations.

Improving liquid bait programs for Argentine ant control: bait station density.

Argentine ants, Linepithema humile (Mayr), have a positive effect on populations of mealybugs (Pseudococcus spp.) in California vineyards. Previous studies have shown reductions in both ant activity and mealybug numbers after liquid ant baits were deployed in vineyards at densities of 85-620 bait stations/ha. However, bait station densities may need to be <85 bait stations/ha before bait-based strategies for ant control are economically comparable to spray-based insecticide treatments-a condition that, if met, will encourage the commercial adoption of liquid baits for ant control. This research assessed the effectiveness of baits deployed at lower densities. Two field experiments were conducted in commercial vineyards. In experiment 1, baits were deployed at 54-225 bait stations/ha in 2005 and 2006. In experiment 2, baits were deployed at 34-205 bait stations/ha in 2006 only. In both experiments, ant activity and the density of mealybugs in grape fruit clusters at harvest time declined with increasing bait station density. In 2005 only, European fruit lecanium scale [Parthenolecanium corni (Bouché)] were also present in fruit clusters, and scale densities were negatively related to bait station density. The results indicate that the amount of ant and mealybug control achieved by an incremental increase in the number of bait stations per hectare is constant across a broad range of bait station densities. The results are discussed in the context of commercializing liquid ant baits to provide a more sustainable Argentine ant control strategy.