Materials and methods.

Large numbers of dacine specimens were collected throughout Papua New Guinea by trapping and host fruit sampling. Steinertype fruit fly traps, baited with cue lure, methyl eugenol or vanillylacetone (zingerone), were set in many localities over a wide range of ecosystems. In most cases, the traps were serviced on 2-week cycles for at least 1 year. Samples of rainforest and cultivated fruits were collected in some provinces. All specimens collected were preserved in a dry state and sent to R.A.I. Drew at Griffith University, Brisbane, Australia, for microscopic identification and curation. Data and photographs of Bactrocera longicornis were received from the Museum Nationale d'Histoire Naturelle, Paris, France. The subgeneric classification used herein follows Drew and Hancock (2016) and Hancock and Drew (2006, 2015, 2016, 2017a,b,c,d,e, 2018a,b,c, 2019).

Species and speciation.

This chapter discusses two species models, which are diametrically opposed. The first, often called the 'biological species concept', defines species in terms of 'reproductive isolation', convinced that species arise when subsets of a population are split off and remain geographically isolated over evolutionary time. If and when such new species are reunited with their founder population, interbreeding does not occur, or if it does, infertile progeny result. Hence, from the biological species concept, natural selection is a primary agent of change and directly selects for new species. In this sense, species are the direct products of natural selection and they are therefore 'adaptive devices'. When applying this species concept, it has been impossible to separate some sibling species of fruit flies in the genus Bactrocera where distinct morphological species can be similar in molecular analyses of certain DNA sequences, while similar species morphologically are distinct in the same molecular characters. A radically different model, the 'recognition concept of species', relies heavily on a knowledge of species ecology and behaviour, particularly in their natural habitat. The principal points in this concept are given. In contrast to the now-outdated biological species concept that leads one to depend on laboratory-based research to define species, the recognition concept requires workers to undertake extensive field research in the habitat of the taxon under investigation. In translating this approach to research in the insect family Tephritidae, particularly the Dacinae, some 35 years of field surveys have been undertaken throughout the Indian subcontinent, South-east Asia and the South Pacific region. These surveys included trapping using male lure traps and host fruit collections of commercial/edible fruits. The results of this work have included the provision of specimens of almost all known species for morphological descriptions (c.800 species), material for male pheromone chemistry, and data on host fruit relationships and biogeographical studies.

Introduction.

Given the rich rainforest flora of Papua New Guinea, which includes some 8000 known plant species, it is understandable why this land mass contains such a rich dacine fauna, with the largest number of species of any land mass across the entire Asian/Pacific region. Major collections of Dacini have been obtained, over the past two decades, by male lure trapping and host fruit sampling across large areas of Papua New Guinea. These surveys have provided the specimens for the descriptions of the new species in this book. In particular, the use of vanillylacetone has resulted in the collection of a number of previously unknown species.

Preferential Attraction of Oviposition-Ready Oriental Fruit Flies to Host Fruit Odor over Protein Food Odor

Olfaction plays a key role in the location of food and oviposition resources by tephritid fruit flies. Adult females, including oriental fruit fly, Bactrocera dorsalis, can sustain egg production throughout their lives provided they obtain sufficient protein. Thus, preferential attraction to food or oviposition sites (host fruit) will depend on a fly’s particular physiological state. In this study, laboratory bioassays were conducted with mature, mated B. dorsalis (provisioned protein and sugar ad libitum) to evaluate attraction to traps baited with torula yeast versus six host fruit sources (guava, guava juice, mango, orange, Surinam cherry, or white sapote). Females that preferred fruit laid a significant number of eggs around the trap entrance (average 405 eggs/fly), while almost no eggs were laid by females that preferred yeast (0.5 and 1.3 eggs/fly on two occasions). Similar results were observed in a bioassay using headspace extracts of guava juice and torula yeast, supporting olfactory-mediated responses. When individual females were allowed to oviposit in guava juice traps 0–24 h after a choice test, 45.8% of females that chose guava juice laid eggs (average 14.7 eggs/fly), compared with 27.5% that chose yeast (average 6.5 eggs/fly). Dissections indicated that females with a preference for guava juice had an egg load 2.4 times greater than females that preferred yeast. These results suggest there is an olfactory-based behavioral switch in preference from protein to host odors when female B. dorsalis are oviposition-ready. We discuss the implications of our findings for practical behavioral management and detection programs for B. dorsalis.

Plant host shift: The viability and persistence of a polyphagous non-native insect

Drosophila suzukii (Diptera: Drosophilidae) has become a pervasive pest in several countries around the world. Many studies have investigated the preference and attractiveness of potential hosts on this invasive, polyphagous drosophilid. Thus far, no studies have investigated whether a shift of fruit host could affect its ecological viability or spatiotemporal persistence. In this study, we analysed the fecundity and oviposition period jointly with the survival time of D. suzukii subject to a shift in host fruit, using a joint modelling method for longitudinal outcomes and time-until-event outcomes. The number of eggs laid by females was higher in raspberry than in strawberry and when setting adults of F1 generation underwent a first host shift. The joint modelling also suggested that insects reared on raspberry survived longer. We then combined experimental results with a two-patch dispersal model to investigate how host shift in a species that exhibits both passive and density-dependent dispersal may affect its asymptotic dynamics. In line with empirical evidence, we found that a shift in host choice can significantly affect the growth potential and fecundity of a species such as D. suzukii, which ultimately could aid such invasive populations in their ability to persist within a changing environment.

Comparative Transcriptomic Analyses of Antibiotic-Treated and Normally Reared Bactrocera dorsalis Reveals a Possible Gut Self-Immunity Mechanism

Bactrocera dorsalis (Hendel) is a notorious agricultural pest worldwide, and its prevention and control have been widely studied. Bacteria in the midgut of B. dorsalis help improve host insecticide resistance and environmental adaption, regulate growth and development, and affect male mating selection, among other functions. Insects have an effective gut defense system that maintains self-immunity and the balance among microorganisms in the gut, in addition to stabilizing the diversity among the gut symbiotic bacteria. However, the detailed regulatory mechanisms governing the gut bacteria and self-immunity are still unclear in oriental fruit flies. In this study, the diversity of the gut symbiotic bacteria in B. dorsalis was altered by feeding host fruit flies antibiotics, and the function of the gut bacteria was predicted. Then, a database of the intestinal transcriptome of the host fruit fly was established and analyzed using the Illumina HiSeq Platform. The gut bacteria shifted from Gram negative to Gram positive after antibiotic feeding. Antibiotics lead to a reduction in gut bacteria, particularly Gram-positive bacteria, which ultimately reduced the reproduction of the host flies. Ten immunity-related genes that were differentially expressed in the response to intestinal bacterial community changes were selected for qRT-PCR validation. Peptidoglycan-recognition protein SC2 gene (PGRP-SC2) was one of the 10 immunity-related genes analyzed. The differential expression of PGRP-SC2 was the most significant, which confirms that PGRP-SC2 may affect immunity of B. dorsalis toward gut bacteria.

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.

A Minor Role of Host Fruit on the Parasitic Performance of Aganaspis daci (Hymenoptera: Figitidae) on Medfly Larvae

Host fruit is known to strongly affect the performance of both fruit pests and their potential natural enemies. This is particularly important in the control of tephritid fruit flies, whose larvae develop inside the fruit and thus create a set of foraging problems for parasitoids. In the present study, we assessed the response of female Aganaspis daci (Weld) (Hymenoptera: Figitidae), one of the most promising parasitoids for tephritid biocontrol in the Mediterranean Basin, to different potential host fruit species. We measured the olfactory response to medfly-infested and uninfested fruits, and several biological parameters of A. daci when different infested fruits were offered under both laboratory and greenhouse conditions. Our results showed that this parasitoid was significantly more attracted to apples and uninfested fruit. Moreover, parasitic activity was similar among the tested fruits under both conditions, showing very high values in the laboratory and a much poorer performance when conditions were variable. This suggests that A. daci may be a good candidate to be included in mass releases against the medfly regardless of the affected crop, but only when climate conditions are not expected to hinder its normal activity.

Predation on Drosophila suzukii within Hedges in the Agricultural Landscape

The invasive Drosophila suzukii feeds and reproduces on various cultivated and wild fruits and moves between agricultural and semi-natural habitats. Hedges in agricultural landscapes play a vital role in the population development of D. suzukii, but also harbor a diverse community of natural enemies. We investigated predation by repeatedly exposing cohorts of D. suzukii pupae between June and October in dry and humid hedges at five different locations in Switzerland. We sampled predator communities and analyzed their gut content for the presence of D. suzukii DNA based on the COI marker. On average, 44% of the exposed pupae were predated. Predation was higher in dry than humid hedges, but did not differ significantly between pupae exposed on the ground or on branches and among sampling periods. Earwigs, spiders, and ants were the dominant predators. Predator communities did not vary significantly between hedge types or sampling periods. DNA of D. suzukii was detected in 3.4% of the earwigs, 1.8% of the spiders, and in one predatory bug (1.6%). While the molecular gut content analysis detected only a small proportion of predators that had fed on D. suzukii, overall predation seemed sufficient to reduce D. suzukii populations, in particular in hedges that provide few host fruit resources.