Effects of Aphid Prey on Larval Development and Mortality of Adalia bipunctata and Coccinella septempunctata (Coleoptera: Coccinellidae)

Developmental time and mortality rate of Adalia bipunctata (L.) and Coccinella septempunctata L. (Coleoptera: Coccinellidae) were determined when feeding on five aphid species. Metopolophium dirhodum (Walker), Sitobion avenae (F.), Rhopalosiphum padi (L.), Hyalopterus pruni (Geoffr.) and Myzus cerasi F. (Homoptera: Aphididae) are widespread in Tekirda?, Turkey. Tests were carried out in a controlled environmental chamber (25±1°C temperature, 65±5% relative humidity and 16 h light:8 h dark period). Developmental times for A. bipunctata and C. septempunctata larvae varied significantly depending on species of aphid prey (P<0.05). Development time (±S.E.) varied from 17.50±0.84 to 20.83±1.60 days for C. septempunctata and 16.7±0.76 to 20.7±1.03 days for A. bipunctata. Mortality of A. bipunctata (50%) and C. septempunctata (63%) were highest on H. pruni.     

Laboratory studies on the reproduction of Adalia bipunctata (Coleoptera, Coccinellidae)

The pre-oviposition period, but not the longevity, of adult A. bipunctata was significantly shorter when they had been reared to the adult stage on Microlophium evansii and then fed on Acyrthosiphon pisum, than when fed in the adult stage on Aphis fabae. When adult beetles, reared as larvae on M. evansii, were fed on A. pisum they laid twice as many eggs as similarly reared adults fed on A. fabae. The species of aphid fed to the insects during their adult life (but not their larval life) greatly affected fecundity, and A. pisum is perhaps a more nutritious food for A. bipunctata than A. fabae is. Virgin females laid infertile eggs but only half as many as those laid by mated females.

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Laboratoriumsuntersuchungen über die vermehrung von Adalia Bipunctata (Coleoptera, Coccinellidae)

Zusammenfassung Die Praeovipositionszeit, jedoch nicht die Lebensdauer, von erwachsenen A. bipunctata war signifikant kürzer, wenn sie nach Aufzucht mit Microlophium evansii im Adultstadium mit Acyrthosiphum pisum statt mit Aphis fabae gefüttert wurden. Wenn erwachsene Käfer, die als Larven mit M. evansii aufgezogen worden waren, mit A. pisum gefüttert wurden, legten sie doppelt so viele Eier als entsprechend aufgezogene, aber mit A. fabae gefütterte Imagines. Die Blattlausart, welche den Käfern während ihres Imaginallebens-aber nicht während der Larvalentwicklung-als Nahrung geboten wird, beeinflußt ihre Fruchtbarkeit stark; und A. pisum ist vielleicht ein nahrhafteres Futter für A. bipunctata als A. fabae. Unbegattete Weibchen legten unbefruchtete Eier, aber nur halb so viele als begattete.

     

The Immediate Source of the Oviposition-Deterring Pheromone Produced by Larvae of Adalia bipunctata (L.) (Coleoptera, Coccinellidae)

As is the case for other insects ovipositing on or in resources that are limited in time and/or space, the two-spot ladybird beetle, Adalia bipunctata (L.) produces an oviposition-deterring pheromone (ODP), which is produced by the larval stages. Foraging larvae touch the substrate with their tarsi and the anal disk on the tenth abdominal segment. The aim of this paper was to determine whether the ODP produced by larvae was deposited by the tarsi or the anal disk. Fourth instar larvae either had their anal disk and tarsi, or anal disk, or tarsi coated with a water-soluble mounting medium. Larvae so treated were allowed to walk on filter paper that was subsequently presented to gravid females. The tracks of larvae that had both their tarsi and anal disk masked did not inhibit oviposition. However, the tracks of larvae that had only their tarsi masked significantly inhibited oviposition but those of larvae that had only their anal disk masked did not. It is concluded that the ODP is deposited on the substrate by the anal disk on the tenth abdominal segment of larvae.     

Glutathione S-transferase isoenzymes in the two-spot ladybird, Adalia bipunctata (Coleoptera: Coccinellidae)

Isoenzymes of glutathione S-transferase (GST) in adult Adalia bipunctata, an aphidophagous predator, were studied. Cytosolic GST activity was studied in each beetle developmental stage. The highest activities towards both 1-chloro-2,4-dinitrobenzene (CDNB) and 2,4-dinitro-1-iodobenzene (DNIB) occurred in adults. The enzyme distribution was investigated in adults. While most of the enzymatic activity was found in the abdomen (40-50 and 34-63% respectively) using several concentrations of both CDNB and DNIB, significant differences were observed for the head and the thorax depending on the substrate. Activities were more abundant in the thorax with DNIB (37-47%) compared to the 13-19% obtained with CDNB. Some GST activity was also detected in the elytra. GSTs were purified by epoxy-activated Sepharose 6B affinity chromatography and applied to an HPLC column to determine the native molecular weight (69 kDa). Three isoenzymes were separated by chromatofocusing at pH ranges 7-4. Three bands with molecular mass from 23 to 26 kDa were visualised on SDS-PAGE. Their isoelectric points were 6.66, 6.36, and 6.21. The substrate specificities and the kinetic parameters (Vm and Km) of the isoenzymes showed large differences depending on the isoenzyme. Arch.     

The Stockholm populations of Adalia bipunctata (L) (Coleoptera: Coccinellidae)--a case of extreme female-biased population sex ratio

The genetic composition and sex ratio in the Stockholm populations of Adalia bipunctata have been studied. The overall frequency of melanics is 3.2%, which is significantly lower than in the populations of St. Petersburg and other large cities along the Baltic Sea. The secondary sex ratio in the Stockholm populations is female-biased 82:18. More than half of A. bipunctata females are infected with the male-killing Spiroplasma bacterium. Beetles of the co-existing species Adalia decempunctata are infected with a different bacterium belonging to the genus Rickettsia.     

The effects of two microsporidian pathogens on the two-spotted lady beetle, Adalia bipunctata L. (Coleoptera: Coccinellidae)

Two-spotted lady beetles, Adalia bipunctata L. are available for biological pest control in North America. Lady beetles are known to host microsporidia and although these pathogens are able to infect more than one host under laboratory conditions, little is known regarding the effects of more than one microsporidian pathogen on host fitness. In this study, egg cannibalism was used to examine the effects of the microsporidium Tubulinosema hippodamiae from Hippodamia convergens and an undescribed microsporidium from A. bipunctata (alone and in combination) on A. bipunctata host fitness (larval development and mortality, sex ratio, adult fecundity and longevity). Development was prolonged significantly for larvae that were infected with the undescribed microsporidium but T. hippodamiae had no effect and as a result, conclusions could not be made regarding the effects of both pathogens on larval development. The two microsporidia had no effect on sex ratios (1♀:1♂) or on adult fecundity and longevity. Spores were detected in the majority of smear preparations of individuals that were fed microsporidia-infected eggs and molecular analysis confirmed the identity of both pathogens in sampled individuals. T. hippodamiae spores were smaller than spores of the undescribed microsporidium (3.76±0.03×2.32±0.02 μm and 5.43±0.06×2.75±0.03 μm, respectively) and although the former stained less intensely than did those of latter, spores of the two pathogens are difficult to differentiate when examined by light microscopy alone. The ability of some microsporidia to infect more than one lady beetle host makes it difficult to conclude with certainty as to the number of species that are present in infected Adalia when specimens are examined solely by light microscopy.     

Searching by Adalia bipunctata (L.) (Coleoptera: Coccinellidae) and escape behaviour of its aphid and cicadellid prey on lime (Tilia×vulgaris Hayne)

• 1 The paper describes the behavioural interactions between searching larvae of Adalia bipunctata and its aphid and cicadellid prey on lime (Tilia×vulgaris Hayne).
• 2 The frequency with which the predator achieves contact with cicadellid nymphs of differing ages is related to direction of predator approach and predator perception by the cicadellid.
• 3 The success of a coccinellid in contacting and capturing its aphid prey is related to the predator's size and direction of approach, the perception of the predator by an aphid and the latter's ability to escape, the effectiveness of which is discussed.

     

Programmed cell death of the ovarian nurse cells during oogenesis of the ladybird beetle Adalia bipunctata (Coleoptera: Coccinellidae)

Programmed cell death (PCD) is an evolutionary conserved and genetically regulated form of cell death, in which the cell plays an active role in its own demise. It is widely recognized that PCD can be morphologically classified into three major types: type I, known as apoptosis, type II, called autophagy, and type III, specified as cytoplasmic cell death. So far, PCD has been morphologically analyzed in certain model insect species of the meroistic polytrophic ovary-type, but has never been examined before in insects carrying meroistic telotrophic ovaries. In the present study, we attempted to thoroughly describe the three different types (I, II and III) of PCD occurring during oogenesis in the meroistic telotrophic ovary of the Coleoptera species Adalia bipunctata, at different developmental ages of the adult female insects. We reveal that in the ladybird beetle A. bipunctata, the ovarian tropharia undergo age-dependent forms of apoptotic, autophagic and cytoplasmic (paraptotic-like) cell death, which seem to operate in a rather synergistic fashion, in accordance with previous observations in Diptera and Lepidoptera species. Furthermore, we herein demonstrate the occurrence of morphogenetically abnormal ovarioles in A. bipunctata female insects. These atretic ovarioles collapse and die through a PCD-mediated process that is characterized by the combined activation of all three types of PCD. Conclusively, the distinct cell death programs (I, II and III) specifically engaged during oogenesis of A. bipunctata provide strong evidence for the structural and functional conserved nature of PCD during insect evolution among meroistic telotrophic and meroistic polytrophic ovary-type insects.     

Characterization of the alimentary canal of the aphidophagous ladybird,Adalia bipunctata (Coleoptera: Coccinellidae): anatomical and histological approaches

The alimentary canal of the two‐spot ladybird Adalia bipunctata (Linnaeus) (Coleoptera: Coccinellidae) presents the foregut (stomodeum), the midgut (mesenteron) and the hindgut (proctodeum). The shortest region is the foregut and the longest is the midgut. The relative proportions of the main regions were found to be similar for males and females. In the foregut it was possible to distinguish the pharynx, the esophagus and the proventriculus but no crop. The hindgut is composed of the ileum, rectum and rectal canal. Generally the organ width is similar for males and females, but females presented a wider proventriculus. The epithelium of the foregut varied from squamous to simple cuboidal and columnar. In the midgut the epithelium is simple columnar with goblet and regenerative cells. The epithelium of the hindgut varied from simple cuboidal to squamous. Females presented thicker midgut epithelium whereas males presented thicker epithelium in the esophagus. The anatomy of the alimentary canal of A. bipunctata seems to conform to its carnivorous and recent phylogenetic status within the family Coccinellidae.     

The effect of female mating history on sperm precedence in the two-spot ladybird, Adalia bipunctata (Coleoptera, Coccinellidae)

Effects of two different mating regimes on sperm precedencein the two-spot ladybird, Adalia bipunctata, were studied usingthe polymorphic gene for melanism as a marker for paternity.Virgin nonmelanic females (homozygous recessive) were matedto nonmelanic male(s) and then, after laying fertilized eggs,were mated to a melanic male of known genotype. The resultsafter the two successive single matings showed a highly variabledegree of paternity of the second male. Initial multiple matingwith nonmelanic males did not alter the pattern of paternityafter the subsequent single mating with a melanic male, butit had two other effects: (1) the female showed an increasein rejection behavior, and (2) a longer copulation was requiredfor high success of the melanic male. Additional observationsin which families were reared from beetles collected in copulain the field demonstrated that sperm competition also occursunder natural conditions. The outcome of the competition wasvariable with frequent sperm mixing.     

Effects of food and temperature on development,fecundity and life‐table parameters of Adalia bipunctata (Coleoptera: Coccinellidae)

Development, reproduction and life tables of Adalia bipunctata (L.) were studied at three temperatures (19, 23 and 27°C) on a mixture of frozen pollen and Ephestia kuehniella Zeller eggs as a factitious food and on the aphids Myzus persicae (Sulzer) and Acyrthosiphon pisum (Harris) as natural foods. Development time of A. bipunctata on all tested diets decreased with increasing temperature. Mortality was lowest at 23°C, averaging 44.5%, 42.6% and 24.3% on factitious food, A. pisum and M. persicae respectively. The shortest developmental time from egg to adult at this temperature was observed on factitious food (18.55 days). However, the factitious food was inferior to the aphid diets in terms of reproduction, yielding the longest pre‐oviposition period, shortest oviposition period and lowest fecundity. The mean oviposition rate at 23°C varied from 19.94 to 25.03 eggs day^?1 on factitious food and M. persicae respectively. The intrinsic rate of increase (r_m) on different foods increased with increasing temperature and ranged from a minimum of 0.08 females/female/day on factitious food (19°C) to a maximum of 0.18 females/female/day on A. pisum (27°C). The results suggest that a mixture of E. kuehniella eggs and pollen fully support development of A. bipunctata larvae and can be used as an alternative to live aphids in the mass rearing of the pre‐imaginal stages of the predator. However, reproductive performance of a laboratory population may be better on aphids than on the factitious food.     

Bacteria in ovarioles of females from maleless families of ladybird beetles Adalia bipunctata L. (Coleoptera: Coccinellidae) naturally infected with Rickettsia,Wolbachia, and Spiroplasma

Ovarioles were found to be infected with Spiroplasma, Wolbachia, and Rickettsia in Adalia bipunctata females with maleless progeny in different natural populations. Ooplasm was infected with few Wolbachia bacteria. In ooplasm infected by Rickettsia, bacteria were present in small foci. Spiroplasmas were found encapsulated into ooplasm from the wider intercellular spaces between epithelial and oocyte cells. The cytoplasm of follicular epithelia infected with Rickettsia was heavily destroyed, but the nucleus was intact and free from bacteria. The essential feature of follicular epithelium cells from Spiroplasma and Wolbachia infected A. bipunctata females was inclusions of three types: crystalline, filaments, and concentric myelin-like lamellae. Observations of smears prepared from ovaries of A. bipunctata from natural populations revealed a low concentration of bacteria within a microscopy field (less 10 bacteria) in more than 90% of specimens, and only a few ovaries were heavily infected. Two different ways of bacterial invasion of the oocyte are suggested: Spiroplasma-like, through the intercellular spaces in the epithelium and Rickettsia-like, through the cytoplasm of follicular epithelium cells. Bacteria were not found in germarium zones and we suggest that each follicle is infected from haemolymph.     

Heterodynamic processes in Coccinella septempunctata L. (Coccinellidae: Coleoptera): a mini review

Among ladybirds, diapause has evidently been most studied in Coccinella septempunctata L. adults. Coccinella septempunctata (C‐7), the seven spot, has attracted attention both in Europe and the USA for being heterogeneous as to the induction of diapause. A polyphenic character of diapause is a prominent feature in C. septempunctata and this phenomenon generally and often generates voltinism heterogeneity within populations. A greater part of the C‐7 population shows an obligatory univoltine cycle, whereas a smaller proportion is facultatively polyvoltine. Coccinella s. bruckii in Japan is bivoltine, in which the first generation of adults aestivate while the second generation hibernates. This paper reviews this heterodynamic cycle and its importance in the life history of C‐7, as reported from different regions of the world. Heterogeneous voltinism and diapause smooth the progress of a portion of C‐7 populations to take benefit of aphids well into late summer and autumn. Understanding such variation and plasticity in the life history of this species can help in organizing proper biological control attempts using it as a biocontrol agent.     

Influence of some pesticides on mortality and fecundity of the aphidophagous coccinellid Adalia bipunctata L. (Col., Coccinellidae)

The influence of 30 pesticides (insecticides, acaricides and fungicides) on different stages of Adalia bipunctata was evaluated under laboratory conditions by: (1) immersing individuals for 5 s in the pesticide solution; (2) placing the second and fourth instar larvae on leaves picked from trees treated with the pesticide; and (3) feeding adult coccinellids with aphids contaminated by a recommended concentration of the pesticide. Fenpropathrin, alphacypermethrin, esfenvalerate, acrinathrin, phosalone and propoxur + methoxychlor caused high mortality (up to 100%) not only by direct contact but also as fresh residues on leaves, or even 28 days after application. The mortality also varied with stage and mode of treatment. Feeding with aphids contaminated by fenpropathrin, clofentezine, hexythiazox, brompropylate and vinclozolin decreased the coccinellid fecundity.     

Variability of the mitochondrial gene for cytochrome oxidase I within the Adalia bipunctata species and within species of ladybird beetles (Coleoptera: Coccinellidae)

Intergeneric, interspecific, and intraspecific genetic variation of the 310-bp 3'-end region of the mitochondrial gene of cytochrome oxidase I (COI) has been assessed in ladybirds (Coleoptera: Coccinnellidae). The phylogenetic distances between eight species of ladybirds have been determined. Mitochondrial DNA (mtDNA) nucleotide sequences have been compared in Adalia bipunctata L. differing in the elytron and pronotum colors that have been sampled from several geographically remote populations. The taxonomic statuses of two morphs from the genus Adalia, A. bipunctata bipunctata and A. bipunctata fasciatopunctata, have been identified.     

Functional significance of Micraspis discolor (F.) (Coccinellidae: Coleoptera) in rice ecosystem

Coccinellids are key predators that are conserved and augmented in agricultural ecosystems, to achieve biological control of pests. Actual quantification of field predation has not been attempted for many of the beneficial coccinellids. Numerous reports show coccinellids as opportunistic, feeding on a variety of food material in addition to their preferred prey. Micraspis discolor is the most abundant species of coccinellid in rice ecosystems and touted as a biocontrol option for brown planthopper (BPH), Nilaparvata lugens (Stal), a key pest of rice. However, it has been reported as both entomophagous and phytophagous. Native polyacrylamide gel electrophoresis (PAGE) was used to separate esterases from whole‐body homogenates of Micraspis and its prey viz., green leafhopper (GLH) Nephotettix virescens (Distant), BPH N. lugens, whitebacked planthopper (WBPH) Sogatella furcifera (Horvath), aphid Rhopalosiphum padi L., thrips, Haplothrips sp., and pollen. Field‐collected beetles showed a range of bands, some corresponding with pollen and GLH, while others were different from prey offered, indicating a wider range of prey spectrum than envisaged. Feeding preference studies confirmed a preference for pollen and GLH in no‐choice (H = 20.724; P = 0.001) and multiple‐choice tests (H = 20.52; P < 0.001) and a significant preference for pollen over all insects offered in the paired‐choice test (t = 5.099; P = 0.007). The abundance of adult M. discolor in rice at flowering phase does not correspond to prey abundance in the field but rather reflects an inclination to pollen feeding more than entomophagy.     

Using food for different purposes: female responses to prey in the predator Coccinella septempunctata L. (Coleoptera: Coccinellidae)

Abstract.  1. Although predatory insects often feed on diverse prey, their reproductive activity may be linked most strongly to a more restricted range of prey. The propensity of adult females of the ladybird beetle, Coccinella septempunctata L., to attack two natural prey species, pea aphids [ Acyrthosiphon pisum (Harris)] and alfalfa weevil larvae [ Hypera postica (Gyllenhal)], was compared, and the degree to which ladybird egg production depends on consumption of aphids vs. weevils was assessed.
2. Coccinella septempunctata females more readily attacked aphids than weevil larvae. This was true regardless of whether females had fed previously on aphids or on weevil larvae.
3. When females were provided with few to many aphids daily, or few aphids plus an excess number of weevil larvae, their rates of egg production depended primarily on the number of aphids consumed.
4. Addition of weevil larvae to diets of limited numbers of aphids increased egg production, but only modestly. Thus, consumption of weevil larvae may have served mostly for self-maintenance, thereby enabling females to use for egg production more of the nutrients and energy obtained from limited consumption of aphids.
5. The females' linkage of egg production primarily to aphid rather than weevil consumption may be adaptive, as their offspring are much less able as larvae to survive and mature on a diet of weevils rather than aphids.     

Incidence of male-killing Rickettsia spp. (alpha-proteobacteria) in the ten-spot ladybird beetle Adalia decempunctata L. (Coleoptera: Coccinellidae)

The diversity of endosymbiotic bacteria that kill male host offspring during embryogenesis and their frequencies in certain groups of host taxa suggest that the evolution of male killing and the subsequent spread of male-killing symbionts are primarily determined by host life history characteristics. We studied the 10-spot ladybird beetle, Adalia decempunctata L. (Coleoptera: Coccinellidae), in which male killing has not been recorded previously, to test this hypothesis, and we also assessed the evolution of the male killer identified by DNA sequence analysis. Our results show that A. decempunctata harbors male-killing Rickettsia (alpha-proteobacteria). Male-killing bacteria belonging to the genus Rickettsia have previously been reported only for the congeneric two-spot ladybird beetle, Adalia bipunctata L. Phylogenetic analysis of Rickettsia DNA sequences isolated from different populations of the two host species revealed a single origin of male killing in the genus Rickettsia. The data also indicated possible horizontal transfer of symbionts between host species. In addition, A. bipunctata is known to bear at least four different male-killing symbionts in its geographic range two of which coexist in the two locations from which A. decempunctata specimens were obtained for the present study. Since only a single male-killing taxon was found in A. decempunctata, we assume that the two closely related ladybird beetle species must differ in the number and/or geographic distribution of male killers. We discuss the importance of these findings to our understanding of the evolution and dynamics of symbiotic associations between male-killing bacteria and their insect hosts.     

Response to Contrast Objects of the Seven-Spot Ladybird Coccinella septempunctata (Coleoptera, Coccinellidae)

The goal of the work was to establish whether the seven-spot ladybird Coccinella septempunctata has a spatial constancy towards the shape of images. The beetles were presented with black figures on the white wall of the cylindrical arena. The beetles were walking with different orientation of the body relative to the force of gravity: in the horizontal plane, on a three-ray labyrinth or on the flat ring in the bottom of the arena; with an inclination of 90° on the cylindrical ring wall; upwards on the vertical labyrinth or upside down, under a thin wire labyrinth stretched over the arena. The beetles in the horizontal position did not discriminate from each other differently orientated images, except for two types of the figures: vertical or horizontal bands (the choice was 80:20%) and meanders orientated downwards and upwards (the choice was 56:44%). The bands oriented vertically towards the Earth were preferred by the beetles when observed from the inclination position, but not in the case of a vertical ascension. The meanders were not discriminated when observed from the upside down position. The ascending oblique bands were preferred over the descending ones. No ability to discriminate the shape of objects was found in the ladybirds, and, correspondingly, they have no spatial constancy. The discrimination of figures and ornaments has the simplest mechanism: detection of the movement (directional) of a contrast margin.