Gabriele Rondoni

The harlequin ladybird, Harmonia axyridis: global perspectives on invasion history and ecology

The harlequin ladybird, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), is native to Asia but has been intentionally introduced to many countries as a biological control agent of pest insects. In numerous countries, however, it has been introduced unintentionally. The dramatic spread of H. axyridis within many countries has been met with considerable trepidation. It is a generalist top predator, able to thrive in many habitats and across wide climatic conditions. It poses a threat to biodiversity, particularly aphidophagous insects, through competition and predation, and in many countries adverse effects have been reported on other species, particularly coccinellids. However, the patterns are not consistent around the world and seem to be affected by many factors including landscape and climate. Research on H. axyridis has provided detailed insights into invasion biology from broad patterns and processes to approaches in surveillance and monitoring. An impressive number of studies on this alien species have provided mechanistic evidence alongside models explaining large-scale patterns and processes. The involvement of citizens in monitoring this species in a number of countries around the world is inspiring and has provided data on scales that would be otherwise unachievable. Harmonia axyridis has successfully been used as a model invasive alien species and has been the inspiration for global collaborations at various scales. There is considerable scope to expand the research and associated collaborations, particularly to increase the breadth of parallel studies conducted in the native and invaded regions. Indeed a qualitative comparison of biological traits across the native and invaded range suggests that there are differences which ultimately could influence the population dynamics of this invader. Here we provide an overview of the invasion history and ecology of H. axyridis globally with consideration of future research perspectives. We reflect broadly on the contributions of such research to our understanding of invasion biology while also informing policy and people.

Escape from parasitism by the invasive alien ladybird, Harmonia axyridis

Alien species are often reported to perform better than functionally similar species native to the invaded range, resulting in high population densities, and a tendency to become invasive. The enemy release hypothesis (ERH) explains the success of invasive alien species (IAS) as a consequence of reduced mortality from natural enemies (predators, parasites and pathogens) compared with native species. The harlequin ladybird, Harmonia axyridis, a species alien to Britain, provides a model system for testing the ERH. Pupae of H. axyridis and the native ladybird Coccinella septempunctata were monitored for parasitism between 2008 and 2011, from populations across southern England in areas first invaded by H. axyridis between 2004 and 2009. In addition, a semi‐field experiment was established to investigate the incidence of parasitism of adult H. axyridis and C. septempunctata by Dinocampus coccinellae. Harmonia axyridis pupae were parasitised at a much lower rate than conspecifics in the native range, and both pupae and adults were parasitised at a considerably lower rate than C. septempunctata populations from the same place and time (H. axyridis: 1.67%; C. septempunctata: 18.02%) or in previous studies on Asian H. axyridis (2–7%). We found no evidence that the presence of H. axyridis affected the parasitism rate of C. septempunctata by D. coccinellae. Our results are consistent with the general prediction that the prevalence of natural enemies is lower for introduced species than for native species at early stages of invasion. This may partly explain why H. axyridis is such a successful IAS.

Development and application of molecular gut-content analysis to detect aphid and coccinellid predation by Harmonia axyridis (Coleoptera: Coccinellidae) in Italy.

Despite their positive effect in reducing pest populations, exotic generalist predators sometimes become invasive and contribute to the displacement of indigenous species in the same trophic level. Although laboratory experiments have linked intraguild predation (IGP) to these interactions, field evidence and quantification of IGP are still lacking for most systems. The recent establishment of the exotic Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in Italy raises concern about the detrimental effect that the ladybird could have on native coccinellids. Here we assessed, under laboratory conditions, the acceptability and suitability of eggs of 2 native ladybirds, Adalia bipunctata L. and Oenopia conglobata (L.), as prey items for H. axyridis larvae. Then we developed primers for molecular gut‐content analysis to detect predation by H. axyridis on the 2 ladybirds and on the aphid Eucallipterus tiliae L. Species‐specific 16S primers were developed for the 3 species and laboratory feeding trials were conducted to quantify the rate of prey DNA breakdown in the gut of H. axyridis. Moreover, to field evaluate primers, H. axyridis 4th instars (n = 132) were systematically collected from linden trees in northern Italy and screened for the presence of prey DNA. Seventy‐three percent and 7% of field collected H. axyridis were positive for aphid and coccinellid DNA, respectively. Predation upon aphid and A. bipunctata was lower than predicted if density dependent consumption was expected, while predation upon O. conglobata was significantly higher. Here, we provided the first evidence of IGP among feral populations of H. axyridis and indigenous ladybird beetles, occurring in Italy.

Histological investigation on gall development induced by a worldwide invasive pest, Dryocosmus kuriphilus, on Castanea sativa

The Asian chestnut gall wasp Dryocosmus kuriphilus (Hymenoptera: Cynipidae) is one of the most serious pests of chestnut trees worldwide. Wasps lay eggs into chestnut buds from mid Jane to late July, depending on latitude, and galls develop the following spring on new vegetation, reducing photosynthesis and nut production. We observed that modification of tissues surrounding D. kuriphilus eggs, which differentiate to form the larval chamber, started approximately 1 month after oviposition, shortly after eggs hatch. The larval chambers continued to increase slightly in size throughout the autumn months until January. After that, a period of stagnation, which corresponds to the plants dormancy, occurred, followed by rapid growth from March to May, during the period from bud swelling to bud break. Galls continued to grow during the leaf expansion after bud break and stopped when plant organs achieved their final size. Our results have implications for the management of the pest, providing a better understanding of the critical time periods for the effective control.

Differential susceptibility in a specialised aphidophagous ladybird, Platynaspis luteorubra (Coleoptera: Coccinellidae), facing intraguild predation by exotic and native generalist predators

Abstract The establishment of exotic generalist predators raises concern about possible domination of the invaded community and displacement of native species. In this respect, recent studies suggest that specialists are likely to be more at risk of niche displacement than generalists, thereby compromising biological control in agroecosystems. Under laboratory conditions, we examined the susceptibility of a specialised myrmecophilous ladybird, Platynaspis luteorubra Goeze, as intraguild (IG) prey of generalist native (Oenopia conglobata L. and Adalia bipunctata L.) and exotic (Harmonia axyridis Pallas) coccinellid species. We described parameters of predatory behaviour (for example, attack rate and effectiveness of antipredator defence) and their variation among predators. Furthermore, we estimated the effect of these parameters on the time to predation using a Cox proportional hazards model. We found that A. bipunctata was the most effective IG predator of P. luteorubra, with the highest rate of successful attacks; however, P. luteorubra was not very susceptibile to attack by O. conglobata or H. axyridis, and these species exhibited lower rates of successful attack and predation. Our results, together with field observations, suggest that IG predation is a factor that fosters niche specialization in P. luteorubra and decreases the realised niche of this species in central Italy. Furthermore the lower susceptibility of P. luteorubra to H. axyridis may be due to a lack of historical co-occurrence between preimmaginal stages, possibly arising from differental selection of oviposition sites.

An invasive insect herbivore disrupts plant volatile-mediated tritrophic signalling

Plants respond to insect attack by emission of volatile organic compounds, which recruit natural enemies of the attacking herbivore, constituting an indirect plant defence strategy. In this context, the egg parasitoid Trissolcus basalis is attracted by oviposition-induced plant volatiles emitted by Vicia faba plants as a consequence of feeding and oviposition by the pentatomid host Nezara viridula. However, this local tritrophic web could be affected by the recent invasion by the alien pentatomid bug Halyomorpha halys, an herbivore that shares the same environments as native pentatomid pests. Therefore, we investigated in laboratory conditions the possible impact of H. halys on the plant volatile-mediated signalling in the local tritrophic web V. faba–N. viridula–T. basalis. We found that T. basalis wasps were not attracted by volatiles induced in the plants by feeding and oviposition activities of H. halys, indicating specificity in the wasps’ response. However, the parasitoid attraction towards plant volatiles emitted as a consequence of feeding and oviposition by the associated host was disrupted when host, N. viridula, and non-associated host, H. halys, were concurrently present on the same plant, indicating that invasion by the alien herbivore interferes with established semiochemical webs. These outcomes are discussed in a context of multiple herbivory by evaluating the possible influences of alien insects on local parasitoid foraging behaviour.

Behavioural and physiological responses to prey-related cues reflect higher competitiveness of invasive vs. native ladybirds

Understanding the traits that might be linked with biological invasions represents a great challenge for preventing non-target effects on local biodiversity. In predatory insects, the ability to exploit habitats for oviposition and the physiological response to prey availability differs between species. Those species that respond more readily to environmental changes may confer to their offspring a competitive advantage over other species. Here, we tested the hypothesis that the invasive Harmonia axyridis (Coleoptera: Coccinellidae) makes better use of information from a plant-prey (Vicia faba - Aphis fabae) system compared to the native Oenopia conglobata. Y-tube olfactometer bioassays revealed that both species used olfactory cues from the system, but H. axyridis exhibited a more complete response. This species was also attracted by plants previously infested by aphids, indicating the capacity to exploit volatile synomones induced in plants by aphid attack. Oocyte resorption was investigated when different olfactory stimuli were provided under prey shortage and the readiness of new oogenesis was measured when prey was available again. H. axyridis exhibited higher plasticity in oogenesis related to the presence/absence of plant-aphid volatiles. Our results support the hypothesis that H. axyridis is more reactive than O. conglobata to olfactory cues from the plant-prey system.

Intraguild Predation Responses in Two Aphidophagous Coccinellids Identify Differences among Juvenile Stages and Aphid Densities

(1) Intraguild predation (IGP) can occur among aphidophagous predators thus reducing their effectiveness in controlling crop pests. Among ladybirds, Coccinella septempunctata L. and Hippodamia variegata Goeze are the most effective predators upon Aphis gossypii Glov., which is an economically important pest of melon. Understanding their likelihood to engage in reciprocal predation is a key point for conservation of biological control. Here, we aim to investigate, under laboratory conditions, the level of IGP between the two above mentioned aphidophagous species. (2) Fourth-instars of the two species were isolated in petri dishes with combinations of different stages of the heterospecific ladybird and different densities of A. gossypii. The occurrence of IGP events was recorded after six hours. (3) C. septempunctata predated H. variegata at a higher rate than vice versa (70% vs. 43% overall). Higher density of the aphid or older juvenile stage of the IG-prey (22% of fourth instars vs. 74% of eggs and second instars) reduces the likelihood of predation. (4) To our knowledge, IGP between C. septempunctata and H. variegata was investigated for the first time. Results represent a baseline, necessary to predict the likelihood of IGP occurrence in the field.

Molecular detection of field predation among larvae of two ladybird beetles is partially predicted from laboratory experiments

Despite the fact that natural enemies can synergistically contribute to herbivore pest suppression, sometimes predators engage in intraguild predation (IGP) that might dampen trophic cascades. DNA-based gut-content analysis has become common in assessing trophic connections and biocontrol potential by predators in field systems. Here, we developed a molecular technique that can be used to unravel predation among two ladybirds, Coccinella septempunctata and Hippodamia variegata, and their shared prey, Aphis gossypii. Both ladybirds may provide effective control of the pest. Therefore, understanding their likelihood to engage in IGP is crucial for conservation biological control. Ladybird specimens were collected in melon crop. DNA extraction, primer design and evaluation were conducted. Detectability of prey DNA did not differ significantly between the two ladybirds. H. variegata exhibited higher predation on A. gossypii than C. septempunctata (90.6% vs. 70.9%) and data correction based on DNA detectability confirmed this ranking. IGP was similar among the two species, although corrected data might suggest a stronger predation by C. septempunctata. Intriguingly, IGP by C. septempunctata was lower than predicted by laboratory bioassays, possibly due to the high complexity that arises under field conditions. Implications of our results for biological control and perspectives for ecological network analysis are discussed.

Tracking seasonal emergence dynamics of an invasive gall wasp and its associated parasitoids with an open-source, microcontroller-based device

The invasive Asian chestnut gall wasp, Dryocosmus kuriphilus (Hymenoptera: Cynipidae), is one of the most economically important pests of chestnut forest systems. Control is either based on the use of pesticides or the release of the parasitoid Torymus sinensis (Torymidae) for classical biological control of the pest. Both approaches require frequent monitoring of wasp emergence patterns, usually by recording adult emergence from samples of galls stored in dark boxes. Here, we have improved this monitoring method by developing and evaluating a microcontroller-based device. Cages in which galls were placed were modified by insertion of a turntable, which automatically separates wasps emerging at different times. We used this device to monitor the seasonal emergence of D. kuriphilus, its coevolved parasitoid T. sinensis and newly associated native parasitoids from fresh or overwintered chestnut galls formed in 2013 and 2014. In summer, native parasitoids (3.6 and 5.8% of 2-year parasitism) associated with current-year chestnut galls started emerging before the gall wasp and only partially overlapped with D. kuriphilus emergence. After overwintering, in the spring of the following year, T. sinensis (4.3 and 5.5% of 2-year parasitism from overwintered galls) emerged earlier than indigenous parasitoids (1.6 and 4.8%) and indicated potential for D. kuriphilus control. Overall, 14 species of native parasitoids belonging to five families were recorded from fresh and overwintered galls. We provide the schematic and the programming codes for this new device and propose that this automatic device be used to better track the flight periods of D. kuriphilus and its parasitoids.