Anomala Cuprea Research Articles

Corazonin and corazonin-like substances in the central nervous system of the Pterygote and Apterygote insects.

Antisera against corazonin were used to investigate distribution of immunoreactive cells in the central nervous system (CNS) of representatives of six insect orders: Ctenolepisma lineata (Zygentoma), Locusta migratoria (Orthoptera), Oxya yezoensis (Orthoptera), Gryllus bimaculatus (Orthoptera), Pyrrhocoris apterus (Hemiptera), Arge nigrinodosa (Hymenoptera), Athalia rosae (Hymenoptera), Bombyx mori (Lepidoptera) and Anomala cuprea (Coleoptera). Corazonin-like immunoreactive (CLI) cells were detected in the brain and ventral ganglia of all insects studied except for the albino strain of L. migratoria and the beetle A. cuprea. Implantation of the brain or different ganglia from insects with detected immunoreactivity induced dark coloration in the albino locust, providing further evidence for the presence of authentic corazonins [His(7)- and Arg(7)-isoforms] in these insects. The protocerebral lateral neurosecretory cells projecting into the ipsilateral retrocerebral neurohemal organs and bilateral longitudinal tracts extending and branching throughout the entire CNS seem to be a well-conserved part of the corazonin system in insects. The bilateral longitudinal tracts were formed by species-specific numbers of bilateral interneurons segmentally distributed in the ventral ganglia. Additional immunoreactive somata, mostly interneurons, were detected in the CNS of various insects. The distribution of corazonin in the cephalic neurosecretory system and in the bilateral interneurons suggests that corazonin acts as a hormone as well as a neurotransmitter or a neuromodulator. An ancient origin of corazonin is suggested by the presence of a corazonin-like substance in the primitive insect, C. lineata. These results support previous findings on the common occurrence of corazonin among insects, except for the albino strain of L. migratoria and the Coleoptera.

A new strain of Paenibacillus lentimorbus isolated from larvae of the oriental beetle, Blitopertha orientalis (Coleoptera: Scarabaeidae), in Chiba Prefecture, Japan

In the summer of 1997, larvae of the oriental beetle, Blitopertha orientalis, with milky disease were collected from a turf field in Chiba Prefecture, Japan. Characteristic footprint-shaped sporangia containing a spore and a parasporal body were isolated from the larval hemolymph. Examination of the bacteriological characteristics of this strain revealed that it was unable to grow in 2% NaCl, was susceptible to vancomycin, and had two plasmids, of about 9.8 and 5.5 kb. The sequence of the 16S ribosomal RNA gene (16S rDNA) of this strain was more homologous with that of Paenibacillus (Bacillus) lentimorbus ATCC 14707T than those of the most famous, traditional American culture, Paenibacillus (Bacillus) popilliae ATCC 14706T and NRRL B-4081. From homology of the 16S rDNA, this strain was identified as P. lentimorbus, and named strain ‘Semadara’. The sequence of strain Semadara was highly homologous with those of three Japanese isolates of B. popilliae. Strain Semadara also infected four other scarabaeids: Anomala cuprea, Anomala rufocuprea, Phyllopertha diversa, and Popillia japonica. When second or third stadium larvae of each of the four scarabaeids were inoculated with sporangia of strain Semadara at a concentration of 1.5×108 sporangia/cup, most of the larvae died 2–4 weeks after inoculation.

Occurrence of Damaged Fruits Caused by Cupreous Chafer, Anomala cuprea HOPE and Control Methods in Peach Orchards

Occurrence of Damaged Fruits Caused by Cupreous Chafer, <I>Anomala cuprea</I> HOPE and Control Methods in Peach Orchards

A continuous cell line from the cupreous chafer, Anomala cuprea Hope (Insecta, Coleoptera, Scarabaeidae).

A continuous cell line was obtained from the culture of embryonic cells of the cupreous chafer, Anomala cuprea Hope. The cells showed substrate-dependent growth and formed loose networks. Population doubling time was about 4.5 d. The mode of chromosome number was about 32 (4n). The cell line was designated FRI-AnCu-35.

Nutrient intake in the third instar larvae of Anomala cuprea and Protaetia orientalis submarmorea (Coleoptera: Scarabaeidae) from a mixture of cow dung and wood chips: Results from stable isotope analyses of nitrogen and carbon

The nutrient intake of the third instar larvae of Anomala cuprea (Ac) and Protaetia orientalis submarmorea (Pos) (Coleoptera: Scarabaeidae) from a mixture of cow dung and wood chips of two wood species (Cryptomeria japonica and Quercus mongolica), was assayed by the stable isotope technique. The values of δ15N and δ13C (deviations of 15N/14N and 13C/12C from the standard materials) were measured in the following samples: (1) the integument plus the head of the 3rd instar larvae of both species reared in 1999 autumn fed with a mixture of cow dung and wood chips or with leaf mold, (2) cow dung, (3) wood chips, and (4) leaf mold as the larval food before onset of rearing. The δ values of the bodies of larvae fed with leaf mold differed with the beetle species. The δ15N values of the larval bodies of both species fed with the mixture were close to those of cow dung, whereas the δ13C values were in between those of cow dung and wood chips. Therefore, both species were presumed to obtain almost all their nitrogen from cow dung, and their carbon from both cow dung and wood chips.

Individual variation in pheromone emission and termination patterns in female Anomala cuprea

Individual variations in pheromone emission patterns were examined in a scarab beetle, Anomala cuprea Hope (Coleoptera: Scarabaeidae), by headspace collection of airborne volatiles from individual females. The amount of pheromone obtained varied among virgin females, and about 16% of these females (“silent” females) did not emit detectable amount of pheromone throughout the experimental period. There was no clear temporal pattern of peak pheromone emission for 19 days after the onset. More than half of the laboratory mated females completely stopped releasing pheromone after the first mating, while the rest of them continued releasing pheromone, frequently followed by additional mating.

Unisex Pheromone Detectors and Pheromone-binding Proteins in Scarab Beetles

Olfaction was studied in two species of scarab beetle, Anomala octiescostata and Anomala cuprea (Coleoptera: Scarabaeidae: Rutelinae), which are temporarily isolated and use the same sex pheromone compounds, (R)-buibuilactone and (R)-japonilure. Single sensillum recordings in A. octiescostata revealed highly sensitive olfactory receptor neurons (ORNs) (threshold <1 pg) that were tuned to the detection of the green leaf volatile compound (Z)-3-hexenyl acetate. As opposed to similar ORNs in another scarab species, Phyllopertha diversa, in A. octiescostata a diazo analogue elicited much lower neuronal responses than the natural ligand. Detectors for other floral and leaf compounds were also characterized. Extremely stereoselective ORNs tuned to sex pheromone were identified in male and female antennae. Biochemical investigations showed that, in A. octiescostata and A. cuprea, the pheromone-binding proteins (PBPs) isolated from male antennae were identical to PBPs obtained from female antennae. AoctPBP and AcupPBP had seven different amino acid residues. Binding of AoctPBP to (R)-japonilure is shown. PdivOBP1, which is also known to bind to (R)-japonilure, differed from AcupPBP in only two amino acid residues, one at the N-terminus and the other near the C-terminus. The structural features of the Bombyx mori PBP are compared with the sequences of eight known scarab odorant-binding proteins.

Two novel insect defensins from larvae of the cupreous chafer, Anomala cuprea: purification, amino acid sequences and antibacterial activity

A humoral immune response in larvae of the coleopteran insect, Anomala cuprea has been examined for exploring the molecular basis of host–pathogen interactions. The antibacterial activity against the Gram-positive strain, Micrococcus luteus was detected at a low level in absence of injection. The activity increased strikingly in the hemolymph of the larvae challenged with Escherichia coli, showing the fluctuating profile through a time course, which consists of the static induction phase, the production phase rising to a maximum level, and the reduction phase extending over a long duration. Two peptides were purified and characterized by reverse-phase HPLC, Edman degradation and mass spectrometry. They were isoforms, composed of similar sequences with two amino acid substitutions in 43 residues, and novel members of the insect defensins, cysteine-rich antibacterial peptides. Anomala defensins A and B showed potent activity against Gram-positive bacteria, with slight differences in activity against a few strains of tested bacteria. Anomala defensin B was active at high concentration of 40 μM against the Gram-negative strain, Xenorhabdus japonicus, a pathogen toward the host, A. cuprea larvae.

Olfactory receptor neurons detecting plant odours and male volatiles in Anomala cuprea beetles (Coleoptera: Scarabaeidae)

We have identified several types of olfactory receptor neurons in male and female Anomala cuprea beetles. The receptor neurons were sensitive to female sex pheromone components, flower volatiles, green leaf volatiles and unknown volatiles from males. Olfactory sensilla were located on three lamellae forming the antennal club. There was a clear spatial separation between some types of sensilla on each lamella. Receptor neurons for the two sex pheromone components were situated in sensilla placodea covering a specific area on each lamella in both males and females. All sex pheromone receptor neurons were found in these sensilla. Most other receptor neurons were located in a longitudinal, heterogeneous streak formed by various types of sensilla. Receptor neurons for plant-derived compounds appeared to be specialists with a high sensitivity to their respective key compound. The most remarkable among these are the green leaf volatile-specific receptor neurons, which were both sensitive and selective, with the key compound being at least 1000 times as effective as any other compound. These green leaf volatile detectors are apparently homologous to detectors recently found in the scarab Phyllopertha diversa. Our results emphasize the role of single-sensillum recordings as a tool in the identification of biologically active odours.

Peroral infectivity of non-occluded viruses of Bombyx mori nucleopolyhedrovirus and polyhedrin-negative recombinant baculoviruses to silkworm larvae is drastically enhanced when administered with Anomala cuprea entomopoxvirus spindles.

Non-occluded viruses (NOVs) of Bombyx mori nucleopolyhedrovirus (BmNPV) are poorly infectious to silkworm larvae when administered by peroral inoculation, although they are highly infectious when injected into the insect haemocoel. In the present study, it is demonstrated that NOVs of BmNPV became highly infectious even through peroral inoculation when administered with spindles (proteinaceous structures) of Anomala cuprea entomopoxvirus (AcEPV). Marked enhancement of peroral infectivity of NOVs by AcEPV spindles (nearly 1000-fold higher in the strongest case) was observed in all growth stages of silkworm larvae tested (2nd to 5th instar). Similarly, peroral infectivity of polyhedrin-negative recombinants of BmNPV, which do not produce polyhedra, was also enhanced remarkably by AcEPV spindles. In contrast, spheroids (proteinaceous structures containing AcEPV virions) did not enhance the peroral infectivity of either NOVs or the recombinant BmNPV in silkworm larvae.

Factor analysis of synergistic effect between the entomopathogenic fungus Metarhizium anisopliae and synthetic insecticides.

Anomala cuprea larvae were inoculated with Metarhizium anisopliae after topical application of synthetic insecticides, and the blood cells in the larval bodies were counted. There were fewer granular cells in A. cuprea after a mixed application of M. anisopliae and fenitrothion or teflubenzuron than after a single application of M. anisopliae. The phenoloxidase activity of larval hemolymph declined with time after the treatment of M. anisopliae and insecticides. One of the mechanisms of this synergistic effect is attributed to be weakening of the immune system of A. cuprea larvae by insecticidal stress, which facilitates infection of M. anisopliae to larvae.

Involvement of spindles of an entomopoxvirus (EPV) in infectivity of the EPVs to their host insect.

The biological function of entomopoxvirus (EPV) spindles (inclusion bodies that lack virions), has not been elucidated. We characterized the function of EPV spindles in the cupreous chafer, Anomala cuprea (Coleoptera: Scarabaeidae). Spheroids or spheroids mixed with spindles of Anomala cuprea EPV were administered per os to the A. cuprea larvae. A significant increase in infectivity was induced by the addition of spindles to the spheroids, strongly suggesting that the spindles play an important role in the infection of the host insect.

Leaf-Mold Trap Attractiveness to the Larvae Cupreous Chafer, Anomala cuprea (Coleoptera: Scarabaeidae), and the Role of the Antenna for Food Searching Behavior

Leaf-Mold Trap Attractiveness to the Larvae Cupreous Chafer, <I>Anomala cuprea</I> (Coleoptera: Scarabaeidae), and the Role of the Antenna for Food Searching Behavior

Control of cupreous chafer, Anomala cuprea Hope, by treatment of &ldquo;Diazinon S L so1&reg;&rdquo; microencapsulated Diazinon formulation, at &ldquo;coring&rdquo; of lawn field

Control of cupreous chafer, <I>Anomala cuprea</I> Hope, by treatment of &ldquo;Diazinon S L so1&reg;&rdquo; microencapsulated Diazinon formulation, at &ldquo;coring&rdquo; of lawn field

Olfactory receptor neurons specific to chiral sex pheromone components in male and female Anomala cuprea beetles (Coleoptera: Scarabaeidae)

Chemical communication in scarab beetles involves female-released long-distance sex pheromones. Electrophysiological recordings using tungsten microelectrodes demonstrated two types of olfactory receptor neurons in the scarab beetle Anomala cuprea, each specific for one of the two pheromone components (R)-buibuilactone and (R)-japonilure, respectively. No neurons were found that responded specifically to enantiomers of the pheromone compounds, i.e. (S)-buibuilactone and (S)-japonilure. Pheromone receptor neurons are present in high numbers on both the male and the female antenna, with a lower sensitivity in the females. As in bark beetles and moths, the pheromone receptor neurons in A. cuprea are very sensitive and selective. The difference in response thresholds between (R)- and (S)-enantiomers is almost three orders of magnitude. Pheromone receptor neurons are found in sensilla placodea located in a defined area on each lamella in the antennal club. (R)-buibuilactone and (R)-japonilure neurons are always found in different sensilla. Both types of sensilla contain two neurons, with the pheromone-sensitive neuron displaying a high spike amplitude and the second neuron, not responding to any of the tested compounds, always with a lower spike amplitude.

Seasonal Occurence of the Adult Cupreous Chafer, Anomala cuprea (Coleoptera: Scarabaeidae), by a Light Trap and the Number of Eggs per Female Captured by a Light Trap

Seasonal Occurence of the Adult Cupreous Chafer, <I>Anomala cuprea</I> (Coleoptera: Scarabaeidae), by a Light Trap and the Number of Eggs per Female Captured by a Light Trap

Enantiomeric anosmia in scarab beetles

The sex pheromone for Anomala octiescostata has been previously identified as a 4:1 mixture of (R)-buibuilactone and (R)-japonilure. Field experiments showed that although not active per se, the secondary component increased the activity of major component, buibuilactone. The enantiomers of these semiochemicals, i.e., (S)-buibuilactone and (S)-japonilure are neither attractants nor behavioral antagonists to the pheromone system of A. octiescostata. Chiral GC-EAD experiments demonstrated that, in marked contrast to the Japanese beetle and Osaka beetle, which can detect a stereoisomer not produced by their conspecific females, the male antennae of A. octiescostata lack olfactory receptor neurons (ORNs) tuned to (S)-buibuilactone. The same enantiomeric anosmia (inability to smell a compound) was also observed in Anomala cuprea male antennae. Both in a Y-olfactometer and field experiments, it was clarified that (S)-buibuilactone was not a behavioral antagonist for A. cuprea. Contrary to male antennae of both A. octiescostata and A. cuprea, which lack ORNs tuned to the nonnatural (S)-buibuilactone, fragrance evaluation by skilled perfumers revealed that the human nose can discriminate (both in intensity and quality) enantiomerically pure samples of (R)- and (S)-buibuilactone and japonilure.

Biosynthesis of scarab beetle pheromones.

Chemical communication in scarab beetles (Coleoptera: Scarabaeidae) is achieved with a wide variety of pheromones, but one typical structure is the gamma-lactone having a long unsaturated hydrocarbon chain. Several species utilize (R, Z)-5-(-)-(oct-1-enyl)-oxacyclopentan-2-one (buibuilactone), (R, Z)-5-(-)-(dec-1-enyl)-oxacyclopentan-2-one and (S, Z)-5-(+)-(dec-1-enyl)-oxacyclopentan-2-one [(R)-japonilure and (S)-japonilure]. Using deuterated precursors, we have demonstrated that these compounds are biosynthesized from fatty acids. (9, 10-d4)-Palmitic acid, (9,10-d4)-stearic acid, (9,10-d2)-palmitoleic acid, (9,10-d2)-oleic acid, (9,10-d2)-8-hydroxypalmitoleic acid and (9,10-d2)-8-hydroxyoleic acid were readily incorporated by female Anomala cuprea into the pheromone molecules, while (Z)-(5, 6-d2)-5-dodecenoic acid and (Z)-(5,6-d2)-5-tetradecenoic acid were not. Therefore, the reaction pathway starts from saturated fatty acids, involves their desaturation, followed by 8-hydroxylation, chain shortening and cyclization. The products obtained from racemic (9,10-d2)-8-hydroxypalmitoleic acid and (9,10-d2)-8-hydroxyoleic acid were also racemic, implying that the steps following hydroxylation were not stereospecific. Perdeuterated palmitic acid was applied to disclose the mechanism of the unique hydroxylation reaction. Retention of all deuterium atoms implied that this reaction was a direct process mediated by a specific fatty acid hydroxylase, and preceding desaturation or epoxidation was not involved.

The Spindles of an Entomopoxvirus ofColeoptera(Anomala cuprea) Strongly Enhance the Infectivity of a Nucleopolyhedrovirus inLepidoptera(Bombyx mori)

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Variability of Susceptibility of Anomala cuprea against Metarhizium anisopliae, Using Synthetic Insecticides or Entomopathogenic Fungi as Stressor

Variability of Susceptibility of <I>Anomala cuprea against Metarhizium anisopliae</I>, Using Synthetic Insecticides or Entomopathogenic Fungi as Stressor