Harmonine, a defence compound from the harlequin ladybird, inhibits mycobacterial growth and demonstrates multi-stage antimalarial activity

The harlequin ladybird beetle Harmonia axyridis has been introduced in many countries as a biological control agent, but has become an invasive species threatening the biodiversity of native ladybirds. Its invasive success has been attributed to its vigorous resistance against diverse pathogens. This study demonstrates that harmonine ((17R,9Z)-1,17-diaminooctadec-9-ene), which is present in H. axyridis haemolymph, displays broad-spectrum antimicrobial activity that includes human pathogens. Antibacterial activity is most pronounced against fast-growing mycobacteria and Mycobacterium tuberculosis, and the growth of both chloroquine-sensitive and -resistant Plasmodium falciparum strains is inhibited. Harmonine displays gametocytocidal activity, and inhibits the exflagellation of microgametocytes and zygote formation. In an Anopheles stephensi mosquito feeding model, harmonine displays transmission-blocking activity.     

异色瓢虫对与本土蚜虫共存的外来扶桑绵粉蚧的搜寻和捕食行为

[目的]外来入侵物种作为猎物可能影响本土广谱捕食者的搜寻和捕食行为.本研究旨在揭示异色瓢虫Harmonia axyridis对与本土豌豆修尾蚜Megoura japonica混合发生的外来入侵扶桑绵粉蚧Phenacoccus solenopsis的搜寻和捕食行为.[方法]试验前对异色瓢虫设两个饲喂猎物种类处理:用扶桑绵...     

The potential impacts of the arrival of the harlequin ladybird, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), in Britain

Abstract.  1. The harlequin ladybird, Harmonia axyridis , has recently arrived in Britain.
2. This species has been introduced from Asia into many parts of the world for biological control purposes.
3. In many parts of North America it has become the predominant aphidophagous coccinellid in less than 20 years, and in north-western Europe it is spreading and increasing in number rapidly.
4. Since establishment in North America and continental Europe, reports of its effectiveness as a biological control agent of aphids and coccids have been accompanied by accounts of negative effects on other aphidophagous species and humans.
5. Here the potential impacts of the arrival of the harlequin ladybird in Britain are assessed.     

Effect of cannibalism and predation on the larval performance of two ladybird beetles

The relative impact of cannibalism and predation on the development and survival of fourth instar larvae was assessed in two species of aphidophagous ladybird beetles, Coccinella septempunctata and Harmonia axyridis. The effect of eating aphids, conspecific larvae or heterospecific larvae on larval performance differed in the two species: aphids were the best food for C. septempunctata and survival of C. septempunctata larvae was significantly lower when offered heterospecific larvae rather than conspecific larvae or aphids as food, indicating that H. axyridis larvae were not suitable food for C. septempunctata. However, as the different foods did not affect the larval performance of H. axyridis, this species appears to be more polyphagous. Both intraguild predation by the aggressive larvae of H. axyridis and the polyphagous food habit of this species may account for its dominance in ladybird assemblages and its displacing other ladybird beetles in several places in the world.     

Experimental evidence for the phenotypic impact of admixture between wild and biocontrol Asian ladybird (Harmonia axyridis) involved in the European invasion

Hybridization can fuel evolutionary processes during biological invasions. The harlequin ladybird Harmonia axyridis has long been used as a biocontrol agent before the species became invasive worldwide. Previous analysis based on microsatellite data has shown that European invasive populations bear traces of admixture between an eastern North American source, which is at the origin of the worldwide invasion, and biocontrol strains used in Europe. In this study, we tested the hypothesis that this early admixture event may have fostered the European invasion by impacting on the phenotypes of wild European populations. Mean life history traits of experimental F(1) hybrids are compared with pure parental sources and wild European crosses. Our results reveal a biased impact whereby North American beetles benefitted from being admixed with European biocontrol strains. Resemblance between experimental hybrids and wild European invasive crosses further suggests a long-lasting effect of admixture that may still be at work and fostering invasiveness.     

Relationships Between Attack and Escape Rates, Cannibalism, and Intraguild Predation in Larvae of Two Predatory Ladybirds

We compared incidences of cannibalism and intraguild (IG) predation (IGP) and quantified attack and escape rates—mechanisms which possibly account for the difference in incidences of these interactions—in laboratory experiments with Harmonia axyridis and Coccinella septempunctata. There was a tendency for H. axyridis to act as an IG predator and C. septempunctata as an IG prey. Cannibalism was also often observed in both species. The incidences of both IGP and cannibalism were different between the species. The average attack rates of C. septempunctata were less than 20%, but those of the more aggressive H. axyridis exceeded 50%. Larvae of both species attacked conspecifics and heterospecifics H. axyridis larvae successfully escaped when attacked by both conspecifics and by heterospecifics, while larvae of C. septempunctata escaped from attacks of conspecifics but not from those of heterospecifics. Thus the aggressive behavior of H. axyridis, in particular, of the third and fourth instars, negatively affects the larval survival of C. septempunctata. It may contribute to the dominance of H. axyridis in ladybird assemblages and its displacement of other ladybird species in several places in the world.     

Sequence Polymorphism of the Mitochondrial DNA Control Region in Native and Invasive Populations of Harmonia axyridis (Coleoptera,Coccinellidae)

Russian Journal of Genetics - The harlequin ladybird Harmonia axyridis forms stable natural populations in East Asia and southern Siberia and spreading invasive populations in Europe, Africa, North...     

Intraguild predation behaviour of ladybirds in semi-field experiments explains invasion success of Harmonia axyridis

Harmonia axyridis has been introduced as a biological control agent in Europe and the USA. Since its introduction, it has established and spread, and it is now regarded as an invasive alien species. It has been suggested that intraguild predation is especially important for the invasion success of H. axyridis. The aim of this study was to compare the intraguild predation behaviour of three ladybird species (Coccinella septempunctata, Adalia bipunctata, and H. axyridis). Predation behaviour was investigated in semi-field experiments on small lime trees (Tilia platyphyllos). Two fourth-instar larvae placed on a tree rarely made contact during 3-hour observations. When placed together on a single leaf in 23%-43% of the observations at least one contact was made. Of those contacts 0%-27% resulted in an attack. Harmonia axyridis attacked mostly heterospecifics, while A. bipunctata and C. septempunctata attacked heterospecifics as often as conspecifics. In comparison with A. bipunctata and C. septempunctata, H. axyridis was the most successful intraguild predator as it won 86% and 44% of heterospecific battles against A. bipunctata and C. septempunctata respectively, whilst A. bipunctata won none of the heterospecific battles and C. septempunctata won only the heterospecific battles against A. bipunctata. Coccinella septempunctata dropped from a leaf earlier and more often than the other two species but was in some cases able to return to the tree, especially under cloudy conditions. The frequency with which a species dropped did not depend on the species the larva was paired with. The results of these semi-field experiments confirm that H. axyridis is a strong intraguild predator as a consequence of its aggressiveness and good defence against predation from heterospecific species. The fact that H. axyridis is such a strong intraguild predator helps to explain its successful establishment as invasive alien species in Europe and the USA.     

Assessing the ecological risk posed by a recently established invasive alien predator: <Emphasis Type="Italic">Harmonia axyridis</Emphasis> as a case study

Invasive alien predators are a serious threat to biodiversity worldwide. However, there is no generic method for assessing which local species are most at risk following the invasion of a new predator. The harlequin ladybird, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), is an alien in Europe and many other parts of the world where it affects other species of ladybirds through competition for food and intra-guild predation (IGP). Here, we describe a method developed to assess which European ladybird species are most at risk following the invasion of H. axyridis. The three components of the risk assessment are: the likelihood that the assessed native species encounters H. axyridis in the field, the hazard of competition for food, and the IGP hazard. Thirty native European ladybird species were assessed through data obtained from field observations, laboratory experiments and literature reviews. The species that are considered most at risk are found on deciduous trees, have immature stages which are highly vulnerable to IGP by H. axyridis, and are primarily aphidophagous. These species should be the focus of specific studies and possibly conservation actions. The risk assessment method proposed here could be applied to other alien predators which are considered a threat to native species through competition and predation.     

Hidden costs to an invasive intraguild predator from chemically defended native prey

Studies of invasive species tend to concentrate on the reasons for their success, rather than on the potential costs that they face. Invasive predators are potentially vulnerable to chemical defences of native prey that they have not previously encountered, and evolved to tolerate, within their natural range. While novel chemical defences have been cited as a factor facilitating invasion by exotic prey, the potential costs of chemical novelty to invasive predators have been ignored or remained hidden. We here consider one such example. Although a dominant intraguild predator in interactions with native ladybird beetles, the invasive ladybird Harmonia axyridis finds the defensive alkaloids of certain native ladybirds either distasteful or toxic, although laboratory and field evidence suggests it readily attacks these prey; these natives appear to possess novel alkaloids not previously encountered by H. axyridis . This effect has hitherto remained hidden, due to the methodological approaches used in earlier studies.     

Asymmetric reproductive interference between specialist and generalist predatory ladybirds

1.?Closely related species often differ greatly in the quality and breadth of resources exploited, but the actual mechanisms causing these differences are poorly understood. Because in the laboratory specialized species often survive and perform as well or better on host species that are never utilized in nature, negative ecological interactions restricting host range must exist. Here, we focused on reproductive interference, which has been theoretically predicted to drive niche separation between closely related species with overlapping mating signals. 2.?We examined the interspecific sexual interactions in relation to ecological specialization and generalization in two sibling ladybird species, Harmonia yedoensis and Harmonia axyridis. Harmonia yedoensis is a specialist predator that preys only on pine aphids, which are highly elusive prey for ladybird hatchlings, whereas H.?axyridis is a generalist predator with a broad prey and habitat range. 3.?We experimentally showed that conspecific sperm fertilized the vast majority of eggs regardless of mating order (i.e. conspecific sperm precedence) when a female of H.?yedoensis or H.?axyridis mated with both a conspecific and a heterospecific male. Moreover, we demonstrated that mating opportunities of H.?yedoensis females strongly decreased as heterospecific density increased relative to conspecific density. In contrast, in H.?axyridis, female mating success was high regardless of conspecific or heterospecific density. 4.?Our results suggest that the generalist H.?axyridis should be dominant to the specialist H.?yedoensis in terms of reproductive interference. Our results support the hypothesis that asymmetric reproductive interference from the dominant species may force the non-dominant species to become a specialist predator that exclusively utilizes less preferred prey in nature.     

Isolation and characterization of microsatellites in the harlequin ladybird, Harmonia axyridis (Coleoptera, Coccinellidae), and cross-species amplification within the family Coccinellidae

A total of 18 microsatellite DNA loci were isolated and characterized from the harlequin ladybird, Harmonia axyridis (Coleoptera: Coccinellidae). We optimized a multiplex panel consisting of two polymerase chain reactions, allowing the genotyping of all loci. The number of alleles and heterozygosity observed at each locus ranged from 1 to 12 and from 0 to 100%, respectively. After Bonferroni correction for multiple tests, none of the loci deviated significantly from Hardy-Weinberg equilibrium and there was no indication of significant linkage disequilibrium among pairs of loci. Successful cross-species amplification was obtained for only three of the seven tested species of Coccinellidae.     

Impact of oilseed rape expressing the insecticidal cysteine protease inhibitor oryzacystatin on the beneficial predator Harmonia axyridis (multicoloured Asian ladybeetle)

Insect-resistant transgenic plants have been suggested to have deleterious effects on beneficial predators through transmission of the transgene product by the pest to the predator. To test this hypothesis, effects of oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) on the predatory ladybird Harmonia axyridis were investigated using diamondback moth Plutella xylostella as the pest species. As expected, oilseed rape expressing OC-1 had no effects on either development or survival of the pest, which utilizes serine digestive proteases. Immunoassays confirmed accumulation of the transgene product in pest larval tissues at levels of up to 3 ng per gut. Characterization of proteolytic digestive enzymes of H. axyridis demonstrated that larvae and adults utilize cysteine and aspartic proteases; the former activity was completely inhibited by oryzacystatin in vitro. However, when H. axyridis larvae consumed prey reared on OC-1 expressing plants over their entire life cycle, no significant effects upon survival or overall development were observed. The inhibitor initially stimulated development, with a shortening of the developmental period of the second instar by 27% (P < 0.0001) accompanied by a 36% increase in weight of second instar larvae (P = 0.007). OC-1 had no detrimental effects on reproductive fitness of adult H. axyridis. Interestingly there was a significant increase in consumption of OC-1 dosed prey. The results show that prey reared on transgenic plants expressing a protein which inhibited ladybird digestive enzymes in vitro had no effects in vivo; the ladybird was able to up-regulate digestive proteases in response to the inhibitor.     

Intraguild predation of the aphid pathogenic fungus Pandora neoaphidis by the invasive coccinellid Harmonia axyridis

Abstract.  1. The ladybird Harmonia axyridis is an invasive alien species in many countries and is predicted to have a negative impact on native biodiversity. However, little is known on the status of this aphidophage as an intraguild predator of natural enemies of aphids such as insect-pathogenic fungi.
2. The study assessed the predation of the aphid-specific pathogenic fungus Pandora neoaphidis by adult and larval H. axyridis collected from the U.K. (an invasive population) and Japan (a native population) relative to that of the ladybird Coccinella septempunctata (native to the U.K.) and the non-U.K. C. septempunctata subspecies brucki that were either starved or unstarved.
3. Overall, predation of uninfected aphids was greater than infected aphids and, when given a choice, a preference for aphids was shown. However, the extent of this preference was dependent on the species and origin of the coccinellid. Harmonia axyridis (U.K.) consumed the greatest quantity of fungal cadavers and showed little preference for uninfected aphids over infected aphids. In contrast, C. septempunctata rarely consumed infected aphids. Life stage had no direct effects on predation but starved coccinellids consumed more uninfected aphids than infected aphids.
4.  Harmonia axyridis (U.K.) is a stronger intraguild predator of P. neoaphidis cadavers than the native species C. septempunctata and, therefore, may have an impact on the occurrence and persistence of P. neoaphidis . The differences in intraguild predation by H. axyridis collected in the U.K. and those from Japan suggests that individuals that invaded the U.K. could have a different genetic profile to those in its native range.     

Interactions between introduced and native predatory ladybirds (Coleoptera, Coccinellidae): factors influencing the success of species introductions

Abstract.  1. Why some exotic species invade successfully while others do not is poorly understood. This study focuses on the inability of the exotic predatory ladybird Adalia bipunctata (L.) (Coleoptera, Coccinellidae) to establish and spread in Japan. In particular, this study considers the interactions between A. bipunctata and three native Japanese ladybird species, Harmonia axyridis Pallas, Coccinella septempunctata Linn., and Propylea japonica Mulsant.
2. The number of aphids occurring on the host plants declined dramatically over the course of larval development in the presence of H. axyridis or C. septempunctata . In contrast, many aphids remained at the end of the experiment when only A. bipunctata or P. japonica was present.
3. The survivorship of A. bipunctata from second instar to adult was significantly reduced in the presence of H. axyridis , but not in the presence of C. septempunctata or P. japonica . Most deaths were the result of intra-guild predation when H. axyridis or C. septempunctata larvae were present. Larvae of H. axyridis and C. septempunctata suffered very high mortality in this experiment, but survived better when co-occurring with A. bipunctata than when occurring only with conspecifics. In contrast, P. japonica survived better when occurring with conspecifics than with A. bipunctata .
4. The finding that larvae of the exotic species A. bipunctata died at high rates from intra-guild predation by the native Japanese ladybirds H. axyridis and C. septempunctata may bear on the question of why A. bipunctata has not established more successfully in Japan whereas H. axyridis and C. septempunctata have been very successful in establishing themselves in North America.     

转cry1Ab基因玉米对瓢虫科天敌种群动态的影响

【背景】瓢虫科天敌昆虫是玉米田中重要的天敌类群,可捕食多种农业害虫,对害虫有很好的控制作用。为了明确转基因玉米对瓢虫科天敌的潜在效应。【方法】在田间条件下,采用系统调查结合吸虫器方法,研究了cry1Ab基因玉米对玉米田主要瓢虫科天敌种类丰富度和种群动态的影响。【结果】转cry1Ab基因玉米田与其对照田瓢虫类群和优势种群组成相同。春播时Bt玉米田与对照田以龟纹瓢虫为优势种,仅在玉米生长后期Bt玉米田中的龟纹瓢虫数量显著高于对照田,其他时期2种田间龟纹瓢虫的种群动态没有显著差异。夏播玉米田以龟纹瓢虫和异色瓢虫为主,在8月17日Bt玉米田中异色瓢虫数量显著高于对照田,在9月27日其异色瓢虫数量显著低于对照田,而其他时期Bt玉米田和对照田中异色瓢虫优势度以及田间种群动态没有显著差异。【结论与意义】转cry1Ab基因玉米对玉米田重要的瓢虫科天敌丰富度、优势度和种群数量没有不良影响。因此,该转基因玉米可用于农田生态系统的有害生物管理。     

Dropping behaviour of larvae of aphidophagous ladybirds and its effects on incidence of intraguild predation: interactions between the intraguild prey, Adalia bipunctata (L.) and Coccinella septempunctata (L.), and the intraguild predator, Harmonia axyridis Pallas

Abstract.  1. Two experiments were performed in the laboratory to assess the behaviour of dropping from a host plant as a defence against intraguild predation in aphidophagous ladybird larvae.
2. In the first experiment, encounters were observed on bean plants between fourth instars of the intraguild predator species, Harmonia axyridis , and first instars of two other ladybird species, Adalia bipunctata (L.) and Coccinella septempunctata (L.). The percentages of first instars of the latter two species that dropped from the plant in response to attack differed dramatically, with 47.5% of C. septempunctata first instars dropping vs. 0% of A. bipunctata .
3. In the second experiment, first instars of A. bipunctata or C. septempunctata and a fourth instar of H. axyridis were allowed to forage together on bean plants for 3 h. During this time, 44.3% of C. septempunctata larvae dropped from the plant, but less than 2% of A. bipunctata larvae did so. In contrast, 95.0% of A. bipunctata larvae fell victim to intraguild predation by H. axyridis vs. only 54.5% of C. septempunctata larvae.
4. The significance of dropping behaviour of ladybird larvae as a defence against intraguild predation, and the relationship of dropping behaviour to species-specific habitat affinity of ladybirds, is discussed.     

Life history of the harlequin ladybird, <Emphasis Type="Italic">Harmonia axyridis</Emphasis>: a global meta-analysis

The harlequin ladybird, Harmonia axyridis, is an important natural enemy of aphids throughout the world, but is now also considered an invasive alien species. We performed a meta-analysis of published life history data to address the question whether invading populations in Europe and North America have life history parameters that differ from native populations in Asia, explaining the beetle’s invasion success in new territories. In this meta-analysis, we accounted for important covariables that are often reported in published studies such as temperature, food source (aphids or eggs of Ephestia kuehniella), strain (laboratory or field populations) and photoperiod. Temperature was a key factor having consistent large effects on development rate, survival and reproductive characteristics of H. axyridis. Food source, strain, and photoperiod had effects on some, but not all characteristics, and their overall effect across characteristics was minor. Individuals of invasive populations had a shorter pre-oviposition period and higher fecundity at low temperatures than those of native populations, and a greater longevity across all temperatures. No differences in survival were found between native and invasive populations, while differences in development rate were not consistent, with opposing results obtained according to the way development rate was measured in trials reported in the literature. Results of this meta-analysis support the hypothesis that the life history of the beetle has changed during its invasion into North America and Europe. Invasive populations had a shorter pre-oviposition period and higher fecundity at low temperatures, as well as a greater longevity across all temperatures than native populations. These differences may partially explain the invasive success of H. axyridis.     

Symbiont‐mediated chemical defense in the invasive ladybird Harmonia axyridis

The volatile alkylpyrazines methyl‐ and methoxypyrazines (MPs) present in the reflex bleeds of coccinellid beetles such as the harlequin ladybird beetle Harmonia axyridis are important semiochemicals that function in antipredatory defense behavior. Pyrazines have also been coadapted from a primarily defensive role into pheromones that function in intraspecific communication, attraction, and aggregation behavior. However, the biosynthesis of MPs in ladybird beetles is poorly understood. Here, we tested the hypothesis that MPs could be produced by microbial symbionts in H. axyridis, which generates four different MPs. The evaluation of tissue‐specific MP production showed that MP concentrations were highest in the gut tissue and hemolymph of the beetles rather than the fat body tissue as the presumed site of MP biosynthesis. Furthermore, manipulation of gut microbiota by antibiotic‐containing diets resulted in a lower MP content in adult beetles. The analysis of the bacterial community of the digestive tract revealed the presence of bacteria of the genera Serratia and Lactococcus which are reportedly able to produce MPs. In line with the known diet‐dependent production of MP in H. axyridis, we determined that the presence or relative abundance of some of the potential MP producers (Enterococcus and Staphylococcus) is also diet‐dependent. We hypothesize a potential role of the microbiota in MP production in H. axyridis as a possible example for outsourcing the synthesis of ecologically important semiochemicals to its gut bacteria.     

异色瓢虫与隐斑瓢虫的区别及其色斑型和横脊的频率

异色瓢虫Harmonia axyridis(Pallas)和隐斑瓢虫H.yedoensis(Takizawa)是亲缘关系很近的姐妹种,在中国(北京到南岭,西至甘肃南部和西藏东部)、日本(北海道以南)和南韩等地同域分布,常常在松树等植物上共存。长期以来,隐斑瓢虫被认为是异色瓢虫的一个异名。有时从外形上很难区分这2种瓢虫,因此对于依据野外数据,分析异色瓢虫色斑型和鞘翅横脊的发生频率及小进化会产生一些错误。本文从没有隐斑瓢虫分布的东北地区的材料及其他数据,报道异色瓢虫的色斑型及鞘翅横脊发生频率。异色瓢虫的花斑型在我国东部地区(从东北至广东北部,西至甘肃和云南)发生率很低,而在新疆西北部发生率较高。鞘翅横脊的发生率从东北佳木斯的98.85%降低到云南大理的78.26%。本文列出了区分这2种瓢虫的形态特征及自然分布。如果鞘翅具横脊,则属于异色瓢虫,但如果标本来自2种瓢虫的共存区,鞘翅没有横脊,鞘翅的斑纹呈花斑型、四窗型或二窗型,则很难从外部形态上对它们进行鉴定。但这2种瓢虫的幼虫很容易区分。本文还提供了2种瓢虫的雄性外生殖器形态图、幼虫和成虫。