Record of the invasive alien ladybird Harmonia axyridis (Coleoptera, Coccinellidae) from Kenya

The biological control agent and alien invasive ladybird Harmonia axyridis (Pallas, 1773) was recorded for the first time in Kenya, and in equatorial Africa, in 2010.     

Honeydew volatile emission acts as a kairomonal message for the Asian lady beetle Harmonia axyridis (Coleoptera: Coccinellidae)

Abstract The Asian lady beetle Harmonia axyridis Pallas is considered as an invasive species in most territories where it has been introduced. Because aphid honeydew acts as an attractant for many aphid predators and parasitoids, the objectives of this work were to collect and identify the volatile compounds released from the aphid excretory product to evaluate how these semiochemicals could affect the H. axyridis foraging behavior. Twelve volatile chemicals were identified from the Megoura viciae Buckton honeydew including four alcohols, three ketones, three aldehydes, a pyrazine and a monoterpene. The volatiles 3‐methyl‐1‐butanol and 3‐methyl‐butanal were highlighted as the two most abundant semiochemicals released from the M. viciae honeydew. Vicia faba L. plants treated with crude honeydew attracted more than 80% of the tested individuals with 40% of attracted beetles located on the plant. Four volatile compounds (3‐hydroxy‐2‐butanone, 3‐methyl‐butanal, 3‐methyl‐1‐butanol and limonene) were also highlighted to attract more than 75% of the coccinellids toward the odor source and to locate about 35% of them on the plants. Limonene was the most efficient attractant since 89% of the H. axyridis responded to this odor. The use of the identified semiochemicals as well as the composition of an artificial honeydew could certainly be helpful to control the dispersal of the Asian lady beetle H. axyridis.     

Differences in biological and behavioural characteristics of Harmonia axyridis (Coleoptera: Coccinellidae) according to colour patterns of elytra

Samples of overwintering populations of Harmonia axyridis were classified into five colour patterns, succinea 1, succinea 2, conspicua, spectabilis and axyridis. Generally, the succinea 1 colour pattern was dominant at six collecting sites in Daejon and Chungnam provinces of Korea and their occurrence proportion was over 50%. Non‐melanic forms such as succinea 1 and 2 were noted in about 90% of collected individuals. On the other hand, conspicua and spectabilis as melanic forms were dominant in the laboratory. One hypothesis is that the difference between field and laboratory populations is related to non‐random mating and environmental conditions. Practically, in mate‐choice experiments, most colour pattern ladybirds seemed to mate with the melanic form, especially the conspicua form. The body size (length) of H. axyridis in colour patterns was slightly different, but in females, there was no significant difference among colour patterns. However, both lengths of males were significantly different among colour patterns. In reproductive ability (fecundity), there was no difference among colour patterns.     

异色瓢虫不同色斑型成虫的耐寒性研究

【目的】研究异色瓢虫Harmonia axyridi不同色斑型成虫的耐寒能力,以明确其色斑型比例所呈现的季节性变化的适应意义。【方法】本文测定了异色瓢虫实验种群和自然种群不同色斑型成虫的过冷却点(Supercooling point,SCP)及低温存活率。【结果】同一发育温度下异色瓢虫实验种群不同色斑型成虫的SCP和低温存活率均没有显著性差异(P>0.05),但是低温暴露中饲养在25℃下成虫的存活率下降更为剧烈。异色瓢虫自然种群黑底型与黄底型成虫的SCP呈现出明显的季节性变化,其SCP在7月份最高,分别为﹣8.2℃和﹣7.5℃;在1月份达到最低,分别为﹣16.8℃和﹣18.2℃。越冬开始(11月)黑底型成虫的SCP低于黄底型成虫的,整个越冬期间黄底型成虫的SCP一直低于黑底型成虫的,但是异色瓢虫黑底型与黄底型越冬成虫的低温存活率却没有显著性差异(P>0.05)。【结论】越冬期间异色瓢虫黄底型成虫的SCP低于黑底型的,这样可以通过体液过冷却的方式来避免结冰造成的伤害,以增强其越冬种群的耐寒能力,这也可能是越冬期间异色瓢虫黄底型成虫数量显著上升的一个原因。     

The influence of aphids on the behaviour of adults of the ladybird beetle,Harmonia axyridis (Col.: Coccinellidae)

The mechanism of prey finding by adultHarmonia axyridis (Pallas) (Coleoptera, Coccinellidae) was investigated in the laboratory by offering beetles small gauze or polyethylene bags containing either aphid-infested leaves or uninfested leaves along with empty control bags. The beetles were attracted to bags containing aphid-infested leaves. It is suggested that adults ofH. axyridis use olfactory and visual cues to detect prey. Mate-searching by males and the mating receptivity of females were enhanced in the presence of an abundance of aphids. The presence of aphids, perceived by the sensory organs of the beetles, possibly influence behaviour other than prey-searching.     

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.     

Individual variation in two host plants of the ladybird beetle,Epilachna pustulosa (Coleoptera: Coccinellidae)

Individual variation in two species of host plants (thistle,Cirsium kamtschaticum, and blue cohosh,Caulophyllum robustum) of the herbivorous ladybird beetleEpilachna pustulosa was examined under laboratory conditions for their acceptability to adult beetles as a food resource, for adult preference and for larval performance. When clones of these plants were subjected to non-choice tests using posthibernating female beetles, there was found to be significant intraspecific variation among clones in terms of their acceptability, but interspecific variation was not detected. Significant intraspecific as well as interspecific variation were frequently detected in the two host plants when clones of these plants were subjected to choice tests using posthibernating female beetles; the magnitude of interspecific plant variation for beetle preference is not necessarily larger than that of intraspecific plant variation. Individual variation across plant species with respect to beetle larval performance was also significant. A positive correlation between adult preference and larval performance is suggested across the two taxonomically remote host plant species, thistle and blue cohosh, although this needs further investigation.     

Role of photoperiod in the development and reproduction of Harmonia axyridis (Coleoptera,Coccinellidae)

To improve the laboratory mass rearing and the winter greenhouse use of the multicoloured Asian lady beetle Harmonia axyridis (Pallas) (Coleoptera, Coccinellidae), the effects of photoperiod on the development and fecundity of H. axyridis were investigated at 20°C and photoperiods ranging from 0L:24D to 24L:0D. We foun'd photoperiod to significantly affect the pre-imaginal development and reproduction of H. axyridis. The development of the pre-imaginal stage was significantly faster at shorter photoperiods (0–14?h) than at longer ones (16–24?h). The proportion of females laying eggs within the first 30 days after eclosion, the mean number of eggs per ovipositing female, and the long axial length of the first oocyte were all lower at shorter photoperiods (6–12?h) than at longer ones (14–24?h). These results suggest that adult H. axyridis may encounter problems in preying and reproducing in the winter conditions of unheated and unilluminated greenhouses in areas with temperatures lower than 20°C and day lengths shorter than 12?h.     

Larval Starvation Resistance in Invasive and Native Populations of the Multicolored Asian Ladybird,Harmonia axyridis (Pallas) (Coleoptera,Coccinellidae)

Entomological Review - Resistance of Harmonia axyridis (Pallas) (Coleoptera, Coccinellidae) larvae to long-term (up to 8 days) starvation was studied under laboratory conditions. The experiments...     

Trehalase in Harmonia axyridis (Coleoptera: Coccinellidae): effects on beetle locomotory activity and the correlation with trehalose metabolism under starvation conditions

Trehalose is important in the energy metabolism of insects and protects them from extreme environmental conditions. To study the synthesis and degradation of trehalose, which is hydrolyzed to glucose by trehalase, two soluble trehalase cDNAs of the beetle Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) (HaTreh1-1 and HaTreh1-2) were cloned. The HaTreh mRNA expression levels were higher during the larval than adult stages. The locomotory behavior of the beetle and its mRNA expression of HaTreh1-1 and HaTreh1-2 under starvation were then investigated. Sprint speed and maximum moving distance were increased, and the pause frequency decreased in H. axyridis adults starved for 8 h compared with control (0 h) adults. In contrast, the mRNA expression of both HaTrehs increased quickly in adults starved from 8 to 18 h, with HaTreh1-1 in particular being expressed 289-fold in adults starved for 18 h versus control. These results show that stored food reserves can provide energy to sustain vital activities for 8 h in H. axyridis adults. However, the beetles require trehalose and other energy sources to provide energy to find food and for vital activities when starved for more than 8 h.     

Toxicity of two ecdysone agonists, halofenozide and methoxyfenozide, against the multicoloured Asian lady beetle Harmonia axyridis (Col., Coccinellidae)

Abstract: The insecticidal activity of two ecdysone agonists, methoxyfenozide (RH-2485) and halofenozide (RH-0345), was tested against last-instar larvae of the natural predator Harmonia axyridis (Col., Coccinellidae). In addition, the relative weight gain of the larvae after application was followed. Both products proved to be equally toxic at concentrations of 25, 50 and 100 mg/l. The ecdysteroidal activity of the compounds caused premature induction of larval moulting, cessation of feeding and incomplete pupation in affected larvae. Although, compared with previous results with methoxyfenozide and halofenozide in target pests such as the Colorado potato beetle, Leptinotarsa decemlineata , these compounds caused mortality only after application at relatively high concentrations.     

Mating refusal and its significance in females of the ladybird beetle, Harmonia axyridis

ABSTRACT. . Mating receptivity in females of the ladybird, Harmonia axyridis Pallas (Coleoptera: Coccinellidae), was studied with reference to internal and environmental conditions. Females sometimes reject copulation: they run away, or shake off the male and bend the abdomen upwards. Newly emerged females show a greater likelihood of accepting copulation with an advance in the stage of ovarian development, this being related to age. Females usually mate repeatedly, but frequently show refusal behaviour under unfavourable food conditions. Dissections reveal that females which accept copulation have a greater number of mature eggs in the ovary than do rejectors. Results also suggest that the food conditions are a major environmental factor, directly controlling mating receptivity.     

Selection of reference genes for Harmonia axyridis (Coleoptera: Coccinellidae) feeding on different diets

The selection of a stable reference gene is vital to gene expression studies and to improving the accuracy of RT-qPCR data. With the deep research on the artificial feed of the Harmonia axyridis, there is an increasing need to evaluate the effects of feed on insects at the molecular level. To identify a reference gene to assess the expression of related genes in Harmonia axyridis (Pallas), ensure the reliability of target gene expression analysis using real-time PCR. Especially for H. axyridis were fed Rhopalosiphum padi (L.) or an artificial diet. In this study, the expression profiles of nine candidate reference genes, including 28SrRNA, 18SrRNA, RPS23, EF1, Actin, ATPase, GAPDH, UBI, RPL13 from different H. axyridis tissues (head, thorax, wing, leg, ovary, and fat body) were investigated. The stability of the nine candidate genes was assessed using geNorm, NormFinder, BestKeeper and RefFinder software, and a comprehensive analysis showed that EF1 is a suitable reference gene for eating different diets of different organizations from H. axyridis. 28SrRNA, 18SrRNA, and RPS23 can also be used as reference genes, but Actin, ATPase, RPL13 are not suitable as an internal reference gene.     

Walking activity of flightless Harmonia axyridis (Coleoptera: Coccinellidae) as a biological control agent

The use of flightless strains of the multicolored Asian lady beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), established via artificial selection, can be highly effective as a biological control agent for aphids. However, flightless H. axyridis must depend on walking for dispersion. Therefore, data on the walking activity levels in flightless strains are important for the development of effective methods when releasing these agents in the field. Results of measurement of walking activity levels using an infrared actograph showed that walking activity levels during the daytime (but not nighttime) in both sexes of pure flightless strains tended to be lower than those of control strains. We also found that walking activity levels during the daytime for the F1 generation of hybrid strains, produced by reciprocal crossing between two pure flightless strains, were approximately equal to those of pure strains; the reduction in walking activity levels was not recovered by hybrid vigor. Our results indicate that the reduction in walking activity levels in the pure flightless strains was not caused merely by inbreeding depression stemming from the artificial selection process. Instead, potentially flight ability and walking activity levels in this species may be controlled by the pleiotropic effect of a gene.     

Larval feeding on Bt hybrid and non-Bt corn seedlings by Harmonia axyridis (Coleoptera: Coccinellidae) and Coleomegilla maculata (Coleoptera: Coccinellidae)

Zoophytophagy is an omnivorous activity that occurs when a primarily carnivorous species feeds on plant material. Plant feeding by beneficial predators may have negative consequences if the plant material has been chemically treated, contains toxins, or was transgenically altered. Although common in predaceous Hemiptera, zoophytophagy has been rarely studied in aphidophagous coccinellids. This study examined the likelihood of feeding on Bt and non-Bt corn seedlings by third- and fourth-instar coccinellid larvae, the regularity of feeding events by fourth instars, and the effect of leaf feeding on development time and adult size. Both third- and fourth-instar Harmonia axyridis Pallas and Coleomegilla maculata DeGeer consume leaf tissue, with fourth instars being significantly more likely to feed on corn seedlings. C. maculata larvae ingested leaf tissue more frequently than H. axyridis. Furthermore, when given access to corn seedlings daily, development time of fourth-instar C. maculata increased after Bt hybrid corn treatments compared with non-Bt corn treatments. Zoophytophagous feeding behavior is thought to sustain predators during times of low prey availability, and leaf tissue feeding by coccinellids has typically been attributed to their need for water. However, in this study, tissue feeding regularly occurred even though coccinellid larvae had constant access to water and a daily ad libitum supply of aphids. We suggest that, in addition to environmental conditions, the physiological state of the zoophytophagous species will influence the probability of plant feeding.     

Expansion of Harmonia axyridis Pallas (Coleoptera: Coccinellidae) to European Russia and adjacent regions

An invasive alien species, the harlequin ladybird Harmonia axyridis (Pallas, 1773), has quickly expanded its distribution in Eastern Europe. Records of H. axyridis from 31 localities in Lithuania, Latvia, the Ukraine, European Russia, and the Northern Caucasus are summarized and mapped. Within the last few years this species has established in south Latvia, on the Baltic Sea shore (Kaliningrad oblast and Lithuania), in the western and central Ukraine, Crimea, and in the Northern Caucasus. Besides that, individual specimens have been found in 4 more localities in European Russia. The species is recorded from Lipetsk oblast (European Russia), Crimea, and Nikolaev oblast (the Ukraine) for the first time.     

Role of long-chain hydrocarbons in the aggregation behaviour of Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae)

The multicoloured Asian ladybirds, Harmonia axyridis (Pallas), form large aggregations inside dwellings to survive cold winters, causing annoyance to householders due to their abundance, allergenic properties and problems consequent on reflex bleeding. Flight to overwintering sites and macrosite choice of this species is well documented. H. axyridis shows a hypsotactic behaviour and a clear preference for contrasting visual elements. However, how the microsite is selected remains undocumented, although a better understanding of the factors implicated could lead to the development of new control methods for this pest. In this work, we hypothesised that non-volatile compounds are involved in the microsite choice and aggregation of this beetle. Long chain hydrocarbons were identified at aggregation sites, comprising saturated and unsaturated homologues. An aggregation bioassay was then conducted on overwintering individuals, highlighting the retention capacity of the identified compounds on the tested ladybirds. Additional investigations showed that H. axyridis males and females, originating from overwintering sites, deposit a similar blend of molecules while walking. A Y-shaped tube assay revealed that this blend is used by male and female congeners as cue allowing individuals to orientate towards the treated side of the olfactometer. These results suggest the use of two different blends of long chain hydrocarbons by H. axyridis during its aggregative period, the first one to lead conspecifics towards aggregation sites (microsites) and the second to ensure the cohesion of the aggregation. These findings support the potential use of these blends, in association with volatiles, in the design of traps in order to control infestations of this species in dwellings.