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Feeding stimulative activity of steroidal and secoiridoid glucosides and their hydrolysed derivatives toward the olive weevil (Dyscerus perforatus).

Beta-sitosteryl-D-glucoside and oleuropein isolated from the olive tree (Olea europaea) and their hydrolysed derivatives were tested by a feeding stimulative activity bioassay using the olive weevil (Dyscerus perforatus). Although the steroidal glucoside showed potent feeding stimulative activity, the activity of the aglycone (beta-sitosterol) was significantly lower than that of the glucoside. On the other hand, the difference in the activity between oleuropein, a secoiridoid glucoside, and the hydrolysed derivatives was not significant.

Animals↗

Lima bean (Phaseolus lunatus) seed coat phaseolin is detrimental to the cowpea weevil (Callosobruchus maculatus).

The presence of phaseolin (a vicilin-like 7S storage globulin) peptides in the seed coat of the legume Phaseolus lunatus L. (lima bean) was demonstrated by N-terminal amino acid sequencing. Utilizing an artificial seed system assay we showed that phaseolin, isolated from both cotyledon and testa tissues of P. lunatus, is detrimental to the nonhost bruchid Callosobruchus maculatus (F) (cowpea weevil) with ED50 of 1.7 and 3.5%, respectively. The level of phaseolin in the seed coat (16.7%) was found to be sufficient to deter larval development of this bruchid. The expression of a C. maculatus-detrimental protein in the testa of nonhost seeds suggests that the protein may have played a significant role in the evolutionary adaptation of bruchids to legume seeds.

Amino Acid Sequence↗

Lethal and sublethal effects of selected insecticides and an insect growth regulator on the boll weevil (Coleoptera: Curculionidae) ectoparasitoid Catolaccus grandis (Hymenoptera: Pteromalidae).

A laboratory culture of Catolaccus grandis (Burks), an ectoparasitoid of the boll weevil, Anthonomus grandis grandis Boheman, was exposed to lethal and sublethal doses of insecticides and an insect growth regulator using a spray chamber bioassay. Materials tested were azinphos-methyl, endosulfan, fipronil, malathion, cyfluthrin, dimethoate, spinosad, methyl parathion, acephate, oxamyl, and tebufenozide. At full rates, spinosad was significantly less toxic to female C. grandis than other treatments except endosulfan. Fipronil and malathion were significantly more toxic to females than other treatments. Most of the chemicals tested were highly toxic to male C. grandis; spinosad was least toxic. At reduced rates, most of 4 selected chemicals tested were low in toxicity to C. grandis; however, a reduced rate of malathion was significantly more toxic to females than other treatments. No C. grandis pupae developed from parasitism during a 24-h treatment period with malathion or spinosad. The sex ratio of progeny from sprayed adults appeared to be unaffected by the treatments.

Animals↗

Alfalfa weevil (Coleoptera: Curculionidae) larval sampling: comparison of shake-bucket and sweep-net methods and effect of training.

Two field-sampling methods, shake-bucket and sweep-net, were compared for use in monitoring alfalfa weevil, Hypera postica (Gyllenhal), larvae in alfalfa hay of the high plains and intermountain region of North America. In this region, alfalfa grows to sufficient height to use both methods before the more damaging late instars peak in abundance. Both methods also were compared with extracting larvae by using Berlese funnels in the laboratory. The shake-bucket method was more sensitive in detecting small larvae (first and second instars) than large larvae (third and fourth instars), and the sweep-net method detected a lower proportion of small larvae. The number of larvae collected with the shake-bucket method was strongly correlated with number of larvae recovered from Berlese funnels (total larval counts, R2 = 0.85). Correlation of the sweep-net samples with the Berlese extraction was also significant but less strong (R2 = 0.56). In addition, sampler instruction was evaluated to determine whether demonstration training affects performance of inexperienced samplers using the two field-sampling methods. Training did not significantly change sampler performance in using the shake-bucket but did increase the number of large larvae when using the sweep-net. In addition, less variability was associated with the shake-bucket sampling method than with the sweep-net method for samplers who only had access to written sampling instructions. Therefore, when estimation of small larval abundance is desired for economic decision-making and sampling is performed by people with little or infrequent sampling experience, such as growers, the shake-bucket method is the preferred field-sampling method in the high plains and intermountain region of North America Sweep-net sampling is more variable than shake-bucket sampling, although demonstration training improves the usefulness of the sweep-net.

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Toxicity and repellency of Tephrosia candida to larval and adult Diaprepes root weevil (Coleoptera: Curculionidae).

Leaves of the tropical legume Tephrosia candida DC deterred feeding by adults of the Diaprepes root weevil, Diaprepes abbreviatus (L.), compared with leaves of Citrus macrophylla Wester, a common citrus rootstock, or T. vogelii Hook. f. When larvae were placed in pots containing plants of the three species for 28 d in a growth chamber, larval survival and weight gain were significantly reduced in pots containing plants of T. candida compared with larvae in pots with C. macrophylla or T. vogelii. Diet incorporation of lyophilized roots of T. candida into an artificial diet increasingly inhibited the growth of larvae and increased larval mortality with increased concentration of roots, whereas roots from C. macrophylla or T. vogelii had no effect compared with the diet-only control. T. candida, but not T. vogelii, contains at least one constituent that acts as an antifeedant toward adult D. abbreviatus and as a toxicant toward larvae. No antifeedant effect of roots of T. candida toward larvae was observed in no-choice pot tests or in a diet incorporation bioassay. In pots, larval feeding damage to roots of T. candida was evident. In the diet incorporation assay, 97% of larvae survived 29 d on a diet of cellulose powder (a nutritionally inert filler) despite losing weight. We conclude that decreased survival and weight gain of larvae-fed fresh or lyophilized roots of T. candida were the result of ingestion of a toxicant and not deterrence from feeding.

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Effects of nitrogen fertilizer applied before permanent flood on the interaction between rice and rice water weevil (Coleoptera: Curculionidae).

Nitrogen fertilizer was applied to rice, Oryza sativa L., before permanent flood to determine the interaction between rice and the rice water weevil, Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae), through a series of experiments conducted over a 3-yr period in Texas and Louisiana. Both absolute and relative percentage of yield loss because of L. oryzophilus feeding was not affected by fertilizer rates in the Texas experiment. Absolute yield loss increased with nitrogen rates in 2001 and 2002 in the Louisiana experiment; however, percentage of yield loss was not affected. This finding suggests that nitrogen rate does not affect tolerance of rice to L. oryzophilus injury. In the Texas experiment, differences were detected for ratoon crop yield among nitrogen rates and insecticide that were applied at preflood on the main crop, indicating a carryover of fertilizer and insecticide effects from the main to the ratoon crop. L. oryzophilus populations tended to increase with nitrogen fertilizer in the Louisiana experiment. Our results show that farmers should not increase preflood nitrogen fertilizer to increase tolerance of rice to L. oryzophilus injury.

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A synergistic aggregation pheromone component in the banana weevil Cosmopolites sordidus Germar 1824 (Coleoptera: Curculionidae).

Cosmopolites sordidus is an important pest on banana plantations worldwide. The chemistry of the aggregation pheromone of this insect has been recently resolved and here we present the first evidence from field trails that sordidin, a compound from the male released aggregation pheromone, attracts significant number of weevils only if host plant odors are also present. Sordidin attracts few insects when it is presented without the host plant tissue. However, the attractiveness of host plant tissue increases more than tenfold when it is presented simultaneously with sordidin in field traps. We confirm experimentally that sordidin may be used as part of a system for mass trapping and monitoring this insect.

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Karyological notes on three weevil species from Armenia (Coleoptera, Curculionidae, Cleoninae).

Karyological studies were carried out on three Armenian weevil species from the subfamily Cleoninae. The following chromosome numbers were found in individual species: 2n = 38, n Male = 18 + Xyp in Menecleonus anxius (Gyllenhal, 1824), 2n = 40, n Male = 19 + Xyp in Conorhynchus nigrivittis (Pallas, 1781) and 2n = 44, n Male = 21 + Xyp in Lixus iridis Olivier, 1807. The heterochromosomes of all the examined species form, in the first meiotic metaphase, a typical parachute bivalent.

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Karyological investigations on seven weevil species (Coleoptera, Curculionidae).

Karyological studies were carried out on seven Palaearctic weevils. The following chromosome numbers were found in individual species, i.e. Otiorhynchus niger (F.), Phyllobius viridearis (Laich.), Phyllobius scutellaris Redt., Phyllobius calcaratus (F.), Polydrusus cervinus (L.), and Brachyderes incanus (L.) 2n = 22, n Male = 10 + Xyp, in Lixus elegantulus (Boh.) 2n = 22, n Male = 21 + Xyp. The heterochromosomes of all the examined species form, in the first meiotic metaphase, a typical parachute bivalent.

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Cytogenetic investigations on seven Palaearctic weevil species (Coleoptera, Curculionidae).

Cytogenetic investigations were carried out on seven species of bisexual weevils from five subfamilies. The following numbers of chromosomes were found in individual species: 2n = 22, n male = 10 + Xyp in Ptochus porcellus Boh. (Otiorhynchinae); 2n = 40, n male = 19 + Xyp in Larinodontes turbinatus Gyll. (Cleoninae); 2n = 26, n male = 12 + Xyp in Curculio pellitus (Boh.) (Curculioninae); 2n = 30, n male = 14 + Xyp in Acalles camelus (F.), Acalles echinatus (Germ.), Acalles hypocrita Boh. (Cryptorhynchinae), and 2n = 40, n male = 19 + Xyp in Cionus hortulanus (Foucr.) (Mecininae). The heterochromosomes of all the examined species form, in the first meiotic metaphase, a parachute bivalent. The chromosome number and sex determining system of six species and two genera are described for the first time.

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[Life table of natural population of banana pseudostem weevil, Odoiporus longicollis Oliver, in spring].

The active factor life tables of natural population of banana pseudostem weevil were built, and the controlling effect of active factors was evaluated by using exclusion index of population control (EIPC). The results showed that the population trend indices in growing and harvested banana trunks were 4,3481 and 3,6628, and the EIPC of predation and others were 1,9256 and 2,3577, respectively. The EIPC of natural death were 1,3607 and 1,3554, and that of parasitism and disease in growing and harvested banana trunks were 1,1988 and 1,1668, respectively. The factor of predation and others was important.

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Purification of a lectin from the marine red alga Gracilaria ornata and its effect on the development of the cowpea weevil Callosobruchus maculatus (Coleoptera: Bruchidae).

A lectin from the marine red alga Gracilaria ornata (Gracilariaceae, Rodophyta) was purified and characterized. The purification procedure consisted of extracting soluble proteins in 0.025 M Tris-HCl buffer, pH 7.5, followed by ammonium sulfate precipitation (70% saturation), ion exchange chromatography on DEAE-cellulose and affinity chromatography on mucin-Sepharose 4B. The purified G. ornata lectin (GOL) showed a single protein band with an apparent molecular mass of 17 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing conditions. The native molecular mass of GOL determined by gel filtration on a Sephadex G-100 column was 17.4 kDa and its carbohydrate content was estimated to be 2.9%. Therefore, GOL is a monomeric glycoprotein. The purified lectin agglutinated trypsin-treated erythrocytes from rabbit and chicken but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested but by the complex glycoproteins porcine stomach mucin, lactotransferrin, asialofetuin and bovine and porcine thyroglobulins. Isoelectric focusing showed that GOL is an acidic protein with a pI of 5.4 with analysis of its amino acid composition revealing high contents of Asx, Glx, Ser, Glu, Ala and Cys. When incorporated in artificial seeds, GOL significantly affected the development of Callosobruchus maculatus larvae, indicating the possibility of using this lectin in a biotechnological strategy for insect management of stored cowpea seeds.

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Antennal transcriptome analysis of chemosensory proteins in the raspberry weevil, Aegorhinus superciliosus (Coleoptera: Curculionidae).

Aegorhinus superciliosus (Coleoptera: Curculionidae) is a polyphagous pest of economic importance in southern Chile, the chemical ecology of which remains poorly characterized. Across insect species, chemosensory proteins, including odorant receptors (ORs), gustatory receptors (GRs), ionotropic receptors (IRs), odorant-binding proteins (OBPs), chemosensory proteins (CSPs), and sensory neuron membrane proteins (SNMPs), mediate the detection of chemical cues involved in host selection, reproduction, and other ecologically relevant behaviors. In this study, the antennal transcriptome of adult A. superciliosus was sequenced and analyzed using a de novo RNA-seq approach. Three independent biological replicates per sex were used for RNA-seq, and the same number of independent biological replicates was used for RT-qPCR validation; sequencing yielded 147,409,936 high-quality reads after quality filtering. A total of 112 candidate chemosensory genes were identified, comprising 43 ORs, 34 OBPs, 10 CSPs, 18 IRs, 5 GRs, and 2 SNMPs. Phylogenetic analyses assigned these candidate proteins to established clades, providing a comparative framework for functional inference for ORs and OBPs. Sex- and tissue-biased expression analyses revealed that several ORs, including AsupOR4, AsupOR19, and AsupOBP13, exhibit antennal enrichment and sex-specific expression patterns. Notably, AsupOR19 and AsupOBP13 displayed strong female-biased expression. In addition, transcripts of selected ORs and OBPs were detected in non-antennal tissues, such as the rostrum and legs, suggesting potential functional versatility beyond canonical olfaction. Together, these findings represent the first molecular identification of the chemosensory repertoire of A. superciliosus. This study establishes a foundation for reverse chemical ecology approaches aimed at identifying behaviorally active volatile organic compounds (VOCs) toward environmentally sustainable strategies for integrated pest management.

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Temperature- and body mass-related variation in cyclic gas exchange characteristics and metabolic rate of seven weevil species: Broader implications.

The influence of temperature on metabolic rate and characteristics of the gas exchange patterns of flightless, sub-Antarctic Ectemnorhinus-group species from Heard and Marion islands was investigated. All of the species showed cyclic gas exchange with no Flutter period, indicating that these species are not characterized by discontinuous gas exchange cycles. Metabolic rate estimates were substantially lower in this study than in a previous one of a subset of the species, demonstrating that open-system respirometry methods provide more representative estimates of standard metabolic rate than do many closed-system methods. We recommend that the latter, and especially constant-pressure methods, either be abandoned for estimates of standard metabolic rate in insects, or have their outputs subject to careful scrutiny, given the wide availability of the former. V(.)CO(2) increase with an increase in temperature (range: 0-15 degrees C) was modulated by an increase in cycle frequency, but typically not by an increase in burst volume. Previous investigations of temperature-related changes in cyclic gas exchange (both cyclic and discontinuous) in several other insect species were therefore substantiated. Interspecific mass-scaling of metabolic rate (ca. 0.466-0.573, excluding and including phylogenetic non-independence, respectively) produced an exponent lower than 0.75 (but not distinguishable from it or from 0.67). The increase of metabolic rate with mass was modulated by an increase in burst volume and not by a change in cycle frequency, in keeping with investigations of species showing discontinuous gas exchange. These findings are discussed in the context of the emerging macrophysiological metabolic theory of ecology.

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Repeated cycles of chilling and warming effectively terminate prolonged larval diapause in the chestnut weevil, Curculio sikkimensis.

Curculio sikkimensis (Coleoptera: Curculionidae) requires one or more years to complete its life cycle, owing to prolonged larval diapause. To compare the effects of temperature cycles and total periods of chilling on the termination of prolonged diapause, larvae were subjected to different chilling (5 degrees C) and warming (20 degrees C) cycles ranging from 30 to 720 days, and all cycles were repeated until the sum of chilling and warming periods reached 720 days. The prolonged diapause of C. sikkimensis was more effectively terminated by repeated cycles of chilling and warming than by prolonging the continuous chilling period. However, extremely short temperature cycles were not highly effective in enhancing diapause termination, even when such cycles were repeated many times. To examine the role of warming periods on diapause termination, diapause larvae were subjected to a sequence of chilling (120 days at 5 degrees C) and warming (240 days at 20 degrees C) with a warming period (0-120 days at 20 degrees C) inserted in the chilling period. Diapause larvae that were not reactivated in the first chilling period required exposure to a certain period of warming before they were able to complete diapause development in the subsequent chilling. Thus, C. sikkimensis appears to spread its reactivation times over several years in response to seasonal temperature cycles.

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Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly).

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulfate precipitation, affinity chromatography on immobilized trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named CpaTI, had M(r) of 32.5 kDa as determined by SDS-PAGE and was composed of two subunits with 27.7 and 5.6 kDa linked by disulfide bridges. CpaTI was stable at 50 degrees C and lost 40% of activity at 100 degrees C. CpaTI was also stable from pH 2 to 12 at 37 degrees C. CpaTI weakly inhibited chymotrypsin and elastase and its inhibition of papain, a cysteine proteinase, were indicative of its bi-functionality. CpaTI inhibited, in different degrees, digestive enzymes from Spodoptera frugiperda, Alabama argillacea, Plodiainterpunctella, Anthonomus grandis and Zabrotes subfasciatus guts. In vitro and in vivo susceptibility of Callosobruchus maculatus and Ceratitis capitata to CpaTI was evaluated. C. maculatus and C. capitata enzymes were strongly susceptible, 74.4+/-15.8% and 100.0+/-7.3%, respectively, to CpaTI. When CpaTI was added to artificial diets and offered to both insect larvae, the results showed that C. maculatus was more susceptible to CpaTI with an LD(50) of 3.0 and ED(50) of 2.17%. C. capitata larvae were more resistant to CpaTI, in disagreement with the in vitro effects. The larvae were more affected at lower concentrations, causing 27% mortality and 44.4% mass decrease. The action was constant at 2-4% (w/w) with 15% mortality and 38% mass decrease.

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The phylogeny of acorn weevils (genus Curculio) from mitochondrial and nuclear DNA sequences: the problem of incomplete data.

We considered the contribution of two mitochondrial and two nuclear data sets for the phylogenetic reconstruction of 22 species of seed beetles in the genus Curculio (Coleoptera: Cuculionidae). A phylogenetic tree from representatives found on various hosts was inferred from a combined data set of mitochondrial DNA cytochrome oxidase subunit I, mitochondrial cytochrome b, nuclear elongation factor 1alpha, and nuclear phosphoglycerate mutase, used for the first time as a molecular marker. Separate parsimony analyses of each data set showed that individual gene trees were mainly congruent and often complementary in the support of clades but the analysis was complicated by failure of PCR amplification of nuclear genes for many taxa and hence missing data entries. When the four gene partitions were combined in a simultaneous analysis despite the missing data, this increased the resolution and taxonomic coverage compared to the individual source trees. Alternative approaches of combining the information via supertree methodology produced a comparatively less resolved tree, and hence seem inferior to combining data matrices even in cases where numerous taxa are missing. The molecular data suggest a classification of the European species into two species groups that are in accordance with morphological characteristics but the data do no support any of the previously recognised American species groups.

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