Alkaloids provide evidence of intraguild predation on native coccinellids by <Emphasis Type="Italic">Harmonia axyridis</Emphasis> in the field

Over a period of less than 5 years, Belgium was thoroughly invaded by the multicolored Asian coccinellid, Harmonia axyridis. At the same time, a decline of some native coccinellid species was observed in tree habitats. One hypothesis about the cause of this decline was that it might have been due to intraguild predation (IGP) by H. axyridis. In natural conditions, IGP between coccinellids can be tracked by using defensive compounds. Exogenous alkaloids in H. axyridis were therefore examined by gas chromatography-mass spectrometry (GC–MS), using individuals sampled from lime trees that were also occupied by other species of coccinellids. Harmonia axyridis was the dominant species at all life stages, in terms of both numbers of sites occupied and local abundance. The GC–MS analysis of H. axyridis larvae revealed traces of exogenous alkaloids from 19 of the 20 sites and, in nine of those 19 sites, more than 30% of the larvae analyzed contained exogenous alkaloids. Three alkaloids were detected: adaline from Adalia spp., calvine from Calvia spp. and propyleine from Propylea quatuordecimpunctata. Predation by H. axyridis on two different coccinellid species was also detected in the same larva, reinforcing the status of H. axyridis as a top predator. A generalized linear model indicated that IGP frequency was positively influenced by two variables: the abundance of extraguild and intraguild prey; and the interaction between these two variables. Our results therefore support the hypothesis that IGP by H. axyridis on native coccinellids in tree habitats has led to the decline of several of these species.     

Determining the effects of life stage,shared prey density and host plant on intraguild predation of a native lacewing (<Emphasis Type="Italic">Chrysoperla carnea</Emphasis>) by an invasive coccinellid (<Emphasis Type="Italic">Harmonia axyridis</Emphasis>)

Negative impacts of non-native Harmonia axyridis (Pallas) on members of the native aphid enemy guild have been widely hypothesised but mainly only assessed with other coccinellid species, and mostly in small experimental arenas. Here we investigated the interactions between H. axyridis and Chrysoperla carnea Stephens larvae. In small-scale (Petri dish) arenas 2nd-instar C. carnea were at risk of predation from larval (2nd and 4th-instar) and adult (male and female) H. axyridis while 3rd-instar C. carnea were only at minimal risk from 4th-instar and adult female H. axyridis. Plant species, aphid species and aphid density did not affect intraguild predation of 2nd-instar C. carnea by 4th-instar and adult H. axyridis in mesocosm experiments. Chrysoperla carnea consumed similar numbers of Megoura viciae Buckton, Aphis fabae Scop. and Acyrthosiphon pisum Harris aphids while H. axyridis consumed fewer M. viciae than the other two species. The greatest suppression of A. pisum was achieved in treatments with both C. carnea and H. axyridis. Life stage and the sex of H. axyridis as well as the life stage of C. carnea are important variables affecting intraguild predation and these attributes should be considered when assessing the potential threat of other potentially invasive alien predators.     

Heritable bacterial endosymbionts in native and invasive populations of <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

Harmonia axyridis Pallas (1773) (Coleoptera: Coccinellidae) is the well-studied system of invasive insect species. Native and invasive parts of the area of H. axyridis are isolated geographically. We studied the species composition and the distribution of bacterial symbionts Spiroplasma and Rickettsia in seven localities of the native area and six localities of the invasive area of H. axyridis. Rickettsia was detected in H. axyridis populations for the first time. We found that the proportion of beetles infected with Rickettsia in native and invasive populations of H. axyridis is about 0.03. Spiroplasma was found only in native populations of H. axyridis. The proportion of infected individuals with Spiroplasma in native populations of H. axyridis is about 0.08. All studied native populations of H. axyridis are infected with Spiroplasma, while all invasive populations are not. We discuss the possible influence of Spiroplasma and Rickettsia in the formation of invasive populations of H. axyridis.     

First report of <Emphasis Type="Italic">Harmonia axyridis</Emphasis> Pallas being attacked by <Emphasis Type="Italic">Dinocampus coccinellae</Emphasis> Schrank in Canada

Field parasitism of two lady beetles, a recently introduced species, Harmonia axyridis Pallas, and an indigenous species, Coleomegilla maculata lengi Timberlake, by Dinocampus coccinellae Schrank (Hymenoptera: Braconidae) was investigated in southwestern Québec. Adult coccinellids were sampled from June to September 2001 in alfalfa and corn fields. Parasitism rates were recorded by dissection and rearing of coccinellids in controlled condition. The average proportions of coccinellids with a parasitoid larva were 4.6%for H. axyridis and 32%for C. maculata but 0%of the H. axyridis and 5.9%of theC. maculatawere successfully parasitized. In July, more C. maculata than H. axyridiswere found to be parasitized and among all individuals dissected, C. maculata were more frequently found superparasitized than H. axyridis. Our results suggest that D. coccinellae is not well adapted to H. axyridis in Quebec. This is the first mention of H. axyridis being attacked by D. coccinellae in Canada.     

Intraguild predation on the aphid parasitoid <Emphasis Type="Italic">Aphelinus asychis</Emphasis> by the ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

We investigated intraguild predation (IGP) on an aphid parasitoid, Aphelinus asychis Walker (Hymenoptera: Aphelinidae), by the multicolored Asian ladybird, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), and used the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae) as the prey/host in the laboratory. The ladybirds reared on artificial diet and on aphids consumed more aphids than mummies, while those reared on parasitized aphids consumed similar numbers of aphids and mummies. The ladybirds chose more mummies in treatments when mummies were more abundant, and more aphids when numbers of aphids and mummies were equal, or when aphids were more abundant. However, at all density treatments, rejection rates of mummies (36%) were much greater than of aphids (2%). H. axyridis prey on more aphids than A. asychis mummies, which enhances biological control by the two species. However, prior feeding experience affected subsequent choice, increasing the competition between natural enemies which would reduce their combined effectiveness for biological control.     

The chemical ecology of <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

We review the chemical ecology of the ladybird beetle Harmonia axyridis from the perspective of its invasiveness and the deleterious effects it exerts in the regions it has colonised. We outline the nature and quantification of its chemical defence, and discuss the protection this provides against natural enemies, particularly intraguild predators. We consider the role of infochemicals in location of prey, intraspecific communication and intraguild interactions. We also discuss the role of prey allelochemicals in relation to H. axyridis extreme dietary generalism. Harmonia axyridis poses a number of practical problems for human health and well-being, including “ladybug taint” wine contamination and problems resulting from large aggregations overwintering in buildings. We consider chemical insights into these issues and, in particular, how attractants and repellents might help manage H. axyridis populations through a push–pull strategy. We conclude by discussing future perspectives for research.     

Population variation of elytral ridge occurrence in ladybirds <Emphasis Type="Italic">Harmonia axyridis</Emphasis> Pallas

Intra- and interpopulation variation of occurrence of elytral ridge (neutral morphological character with the known genetic determination) in some populations of Harmonia axyridis from the Russian part of the species area was studied for the first time. Comparative analysis of the frequencies of the recessive allele r of a corresponding gene and the frequencies of recessive homozygotes in 32 samples from 16 localities has shown the lack of temporal and microgeographic variations of this trait. The character of its geographic variation con-firms the hypothesis advanced earlier on the basis of analysis of elytral pattern variation about the existence of two subspecies of H. axyridis and suggests the existence of a zone of secondary hybridization between them in the central part of the area.     

Life cycle of <Emphasis Type="Italic">Harmonia axyridis</Emphasis> in central Europe

The development and reproduction, and maximum number of generations across seasons were ascertained for populations of Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in Central Europe. Number of generations was calculated using the thermal characteristics of development and summation of daily average air temperatures, in 2007–2016. This species can produce 2–3 generations per year. However, there is insufficient time for it to achieve its full reproductive potential. Delayed emigration from hibernacula (late April) shortened the period of seasonal activity by 1–6%. Breeding occurred from May to mid September. Oviposition continued into late summer, which left insufficient time for the late-hatching immatures to complete their development before the onset of winter. A high mobility, opportunistic use of prey patches and weak tendency to enter dormancy provide the potential for population increase by extending the period of development of immatures at the risk of failure if the cold period comes early.     

Paternity following multiple mating in ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

We investigated factors which may determine realised paternity following multiple mating in a promiscuous ladybird Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), namely the mating order and ability of males of different morphs to fertilize females. In experiments using males of two colour morphs, we found that the first male had a reproductive advantage over the second male (sperm precedence P2 = 0.38). Males of the melanic morph spectabilis had a reproductive advantage over males of the semi-melanic morph axyridis (P2 = 0.35). Analysis of paternity using microsatellites applied to young larvae, which avoids possible differential mortality of progeny, confirmed the precedence of the first male and higher paternity rate of colour morphs that are more frequent in the wild. Hatchability of eggs increased from 65% after the first mating to 85% after the second mating delayed by five days. Average duration of mating was 157 min and did not differ between the first and second mating and between the colour morphs of males.     

Effect of long-term cold storage on the fitness of pre-wintering <Emphasis Type="Italic">Harmonia axyridis</Emphasis> (Pallas)

The multicolored Asian ladybird beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), is considered an important generalist predator that can be used as a biological control agent against Hemiptera Sternorrhyncha, Thysanoptera, and the eggs and larvae of Lepidoptera, Coleoptera and Diptera. There are currently abundant natural resources of overwintering H. axyridis in Asia and North America. Given its potential as a biological control agent, methods can be developed to increase its effectiveness for pest control. The availability of an adequate cold storage method would enable the use of field-collected pre-wintering ladybirds for pest suppression in the following season. We studied the effect of cold storage (30, 60, 90, 120 and 150 days stored at −3, 0, 3 and 6°C) on survival, fecundity and predation in field-collected populations. The survival of both female and male ladybirds decreased significantly as storage duration increased at −3°C and 0°C. The ladybirds showed more than 80% survival when they were stored for 150 days at 3°C and 6°C. Long-term cold storage had different effects on the fecundity of H. axyridis at different temperatures. Prolonged cold storage at both 3°C and 6°C shortened pre-oviposition duration and had no adverse effect on reproductive capacity as compared to that of unstored individuals. The adults that experienced 90-day storage at 0°C had the shortest pre-oviposition duration and the largest reproductive capacity. The individuals that were stored for 150 days at 3°C consumed significantly more aphids than the unstored ones. Generally, 3–6°C is a suitable temperature for cold storage of the ladybird without any reduction in fitness. This study will help the exploitation and application of pre-wintering H. axyridis for the biological control of insect pests.     

Parasites of <Emphasis Type="Italic">Harmonia axyridis</Emphasis>: current research and perspectives

Harmonia axyridis (Coleoptera: Coccinellidae) has been introduced widely for biological control of agricultural pests. Harmonia axyridis has established in four continents outside of its native range in Asia and it is considered an invasive alien species (IAS). Despite a large body of work on invasion ecology, establishment mechanisms of IAS and their interactions with natural enemies remain open questions. Parasites, defined as multicellular organisms that do not directly kill the host, could potentially play an important role in regulating host populations. This study presents a review of the parasites of H. axyridis, discussing their distributions and effects on host populations across the host’s native and invasive range. These parasites are: Hesperomyces virescens Thaxt. fungi, Coccipolipus hippodamiae (McDaniel and Morrill) mites, and Parasitylenchus bifurcatus Poinar and Steenberg nematodes.     

The aggregation behavior of <Emphasis Type="Italic">Harmonia axyridis</Emphasis> in its native range in Northeast China

Harmonia axyridis has become notorious as an urban pest in many of the regions where it has been introduced, despite its numerous contributions to the biological control of insects injurious to agriculture and horticulture. Aggregative behavior prior to overwintering leads to invasions of human habitations as beetles seek refuge from freezing temperatures. Here we describe the aggregation behavior of native H. axyridis populations of northeast China that breed in agricultural fields (mostly corn and rice) and shrub/forest habitats and then migrate through rural villages in autumn. More than 140,000 beetles were collected during direct observations in 16 villages in five townships in Jilin Province. Beetles aggregated on dwellings shortly after agricultural harvests, favoring white walls with southern exposures, the largest aggregations occurring in villages in mountainous townships at higher elevations. The sex ratio was consistently female-biased and succinic phenotypes were more than twice as abundant as melanic phenotypes in all locations. A special trap compared the relative attractiveness of different surface colors (white > yellow = black > green > red = natural wood) and potential baits (corn pollen = honey > caramel = cocoa > milk = blank control). All aggregations disappeared abruptly just prior to the first frost, whereupon beetles were discovered sheltering in montane caves with southern aspects at higher elevations. Villagers reported substantial expenditures on pesticides in efforts to eliminate beetles from their homes every autumn. Invasion of human habitations appears to be an intrinsic tendency of native H. axyridis populations in China, which is the result of behavioral adaptations for cold-avoidance.     

Long term changes in the communities of native ladybirds in Northern Italy: impact of the invasive species <Emphasis Type="Italic">Harmonia axyridis</Emphasis> (Pallas)

Since the mid-2000s, the exotic coccinellid species Harmonia axyridis (Pallas) has established itself in Northern Italy, raising serious concerns about possible repercussions on native ladybirds. In this study we compared the ladybird assemblages in 2015–2016 with those sampled in 1995–1996, before the arrival of H. axyridis. Surveys were carried out in the same sites and with the same methods for both periods. Aphidophagous ladybirds were sampled in hedgerows and herbaceous habitats at field margins by mechanical knockdown and sweeping net, respectively. The changes in the structure of ladybird communities after the arrival of H. axyridis were significantly different between arboreal and herbaceous habitats. Harmonia axyridis is currently the dominant species in shrubs and trees, and all the native ladybird species taken together account for only approximately one third of the total individuals sampled in 2015–2016. On the other hand, the relative abundance of the exotic species in herbaceous habitats was low, and it has had a negligible relevance on the structure of ladybird communities on grasslands. Among native species, Adalia bipunctata (L.), whose ecological niche largely overlaps with that of H. axyridis, suffered the largest decline between 1995–1996 and 2015–2016. The co-occurrence of H. axyridis invasion and the regression of A. bipunctata suggest a direct impact of the exotic species, because no other major modifications occurred in the studied areas between sampling periods.     

Importance of early arrival of adult <Emphasis Type="Italic">Harmonia axyridis</Emphasis> for control of <Emphasis Type="Italic">Aphis spiraecola</Emphasis> on apple

The biological control of aphid populations may only be possible when natural enemies arrive soon after aphid colonization. This study was done to identify how quickly adult Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) need to arrive at newly established spirea aphid [Aphis spiraecola Patch (Homoptera: Aphididae)] colonies on apple (Malus domestica Borkh.) to provide population control. A total of 100 newly established spirea aphid colonies were caged in an experimental apple orchard in West Virginia, USA. A single adult H. axyridis was added to each of ten caged colonies at day 0, 5, 10, 15 and 20 days after caging. An additional ten caged colonies were opened for exposure to natural levels of predation at each of the treatment intervals as a control. The single H. axyridis eliminated the aphid colonies significantly more quickly than natural predation for up to ten days after colony establishment. The probability of an aphid colony producing alates was significantly lower in the presence of a single H. axyridis adult than when exposed to natural predation for the first ten days. Adult H. axyridis beetles are capable of completely controlling individual spirea aphid colonies on apple only if they are abundant enough to find colonies within one week of colony establishment.     

Effect of size,extraguild prey and habitat complexity on intraguild interactions: a case study with the invasive ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis> and the hoverfly <Emphasis Type="Italic">Episyrphus balteatus</Emphasis>

Intraguild predation (IGP) is a widespread interaction between predatory arthropods, and is influenced by several factors. The harlequin ladybird, Harmonia axyridis (Coleoptera: Coccinellidae), has frequently been reported as an intraguild predator of other Coccinellidae, but little is known about its interactions with other aphidophagous predators, including syrphids. This study investigated the incidence of IGP between H. axyridis and Episyrphus balteatus (Diptera: Syrphidae), the most abundant hoverfly species in Europe and a commercially available aphid biocontrol agent. The influence of size, presence of extraguild prey and habitat complexity were investigated through laboratory experiments in Petri dishes and on potted broad bean plants. In both types of arenas, IGP between H. axyridis and E. balteatus was found to be asymmetric, with the coccinellid in the majority of cases being the intraguild predator. There was a significant effect of size on the frequency of IGP. The efficiency of H. axyridis as an intraguild predator increased with the developmental stage. Early instars of E. balteatus were the most vulnerable to IGP. Pupae of either species were not attacked. In the presence of extraguild prey, the frequency of IGP was substantially reduced. However, IGP still occurred, mainly in combinations of older larvae of H. axyridis with first or second instars of E. balteatus. The size of the arena affected the incidence of IGP in combinations with second instars of E. balteatus, but not in combinations with third instars. Field research is needed to elucidate the ecological relevance of IGP among these predators.     

Molecular evidence for hybridization between invasive <Emphasis Type="Italic">Solidago canadensis</Emphasis> and native <Emphasis Type="Italic">S</Emphasis>. <Emphasis Type="Italic">virgaurea</Emphasis>

Hybridization between alien and native species is biologically very important and could lead to genetic erosion of native taxa. Solidago × niederederi was discovered over a century ago in Austria and described by Khek as a natural hybrid between the alien (nowadays regarded also as invasive) S. canadensis and native S. virgaurea. Although interspecific hybridization in the genus Solidago is considered to be relatively common, hybrid nature of S. × niederederi has not been independently proven using molecular tools, to date. Because proper identification of the parentage for the hybrid Solidago individuals solely based on morphological features can be misleading, in this paper we report an additive polymorphism pattern expressed in the ITS sequences obtained from individuals representing S. × niederederi, and confirm the previous hypothesis that the parental species of this hybrid are S. canadensis and S. virgaurea. Additionally, based on variability at the cpDNA rpl32-trnL locus, we showed that in natural populations hybridization occurs in both directions.     

Reaping benefits from an invasive species: role of <Emphasis Type="Italic">Harmonia axyridis</Emphasis> in natural biological control of <Emphasis Type="Italic">Aphis glycines</Emphasis> in North America

Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is an invasive species present in numerous agroecosystems in North America. Despite adverse impacts as a threat to native biodiversity, a nuisance household invader and a pest in fruit production, H. axyridis also plays a beneficial role as a major component of assemblages of generalist predators in several agricultural crops. Here, we review the role of H. axyridis as a natural enemy of Aphis glycines Matsumura (Hemiptera: Aphididae), an invasive pest of soybean, Glycine max (L.) Merrill (Fabales: Fabaceae), in North America. Harmonia axyridis is often the most abundant predator species attacking A. glycines in soybean agroecosystems. This predator has the potential to both prevent and suppress A. glycines outbreaks. Further studies are needed to fully understand and utilize the potential of H. axyridis as a natural enemy in the management of A. glycines and other agricultural pests in agroecosystems worldwide.     

Genetic structure of native and invasive populations of <Emphasis Type="Italic">Harmonia axyridis</Emphasis> Pall. in the light of global invasion

The population genetic structure, possible causes, sources, and routes of global invasion of the ladybird Harmonia axyridis are discussed. Infection with the reproductive endosymbionts is considered in the light of possible influence on the formation of evolutionary preadaptations that contributed to the invasion.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

Spotlight on the positive effects of the ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis> on agriculture

In the midst of considerable negativity surrounding the ladybird Harmonia axyridis (Pallas), this paper sheds some light on the positive effects that this predator has had on agriculture. Using resources available at the USDA, National Agricultural Library (DigiTop literature database, Navigator platform), I searched the abstracts of published literature on H. axyridis using the search term “Harmonia axyridis” or “Leis axyridis.” After deleting duplicate records, there were 1193 total records on this ladybird in the database dating from 1961–2015. Of this total, 90 records (7.54 %) highlighted positive effects of H. axyridis on agriculture in terms of its spectacular voracity, predation capacity, and effectiveness in suppressing plant pests, primarily aphids and scales, in forests, orchards, crop fields, gardens, greenhouses, and in laboratory bioassays. More concerted research is necessary to develop methodologies to exploit the positive behaviors of H. axyridis, while managing its negative ones.