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The genus Niphadomimus Zherikhin, 1987 is taxonomically revised herein. In addition to the two recorded Nepalese species, N. nigriventris Zherikhin and N. niger Zherikhin, known only from the holotypes, two additional specimens of N. nigriventris are reported and six new species from China represented by 96 specimens are described and illustrated. These are: N. alcyone sp. n. (Sichuan), N. celaeno sp. n. (Yunnan), N. electra sp. n. (Yunnan), N. maia sp. n. (Yunnan), N. merope sp. n. (Shaanxi) and N. sterope sp. n. (Sichuan). All known Niphadomimus species are apterous inhabitants of the leaf litter in the upper Rhododendron-dominated forest zone between 2000 and 4114 m. Phylogenetic analyses using DNA barcodes of six new species and representatives of 13 other Molytinae genera with available DNA data (A.) corroborates Niphadomimus monophyly; (B.) strongly argues for the sister-group relationship between N. merope sp. n. from the Qinling Mt. Range and the rest of the species distributed in the Hengduan mountains; (C.) in two among four analyses weakly relates the genus with the East Palaearctic Leiosoma. The tribe Typoderini could not be shown as monophyletic, which may be due to insufficient signal content of the cox1 marker at the tribal level. The detected phylogeographic pattern of Niphadomimus is compared with that of similarly distributed or closely related clades. Temporal DNA analysis estimates the N. merope sp. n. split at 6-11 MY, while the diversification of the Hengduan clade dates between 5.5 MY and 3.6 MY, i.e. well before the onset of the Quaternary climate fluctuations.

Trapping systems for the red palm weevil, Rhynchophorus ferrugineus Olivier, rely on the use of natural plant odor sources to boost the attractiveness of the aggregation pheromone. The identification of the key odorants involved in attraction is essential in the development of a synthetic pheromone synergist to replace the nonstandardized use of plant material in traps. Canary Islands date palms (Phoenix canariensis) have become preferred hosts for R. ferrugineus in Europe; thus, the volatile profile of different P. canariensis plant materials, including healthy and infested tissues, is investigated in the present work by means of solid phase microextraction (SPME-GC-MS), aimed to identify pheromone synergists. The electroantennography (EAG) response of the compounds identified was recorded, as well as the preliminary field response of several EAG-active compounds. The so-called "palm esters" (ethyl acetate, ethyl propionate, ethyl butyrate, and propyl butyrate) elicit the strongest EAG responses but performed poorly in the field. Mixtures of esters and alcohols give evidence of better performance, but release rates need further optimization.

Genotypes that hedge their bets can be favored by selection in an unpredictably varying environment. Bet hedging can be achieved by systematically expressing several phenotypes, such as one that readily attempts to reproduce and one that procrastinates in a dormant stage. But how much of each phenotype should a genotype express? Theory predicts that evolving bet-hedging strategies depend on local environmental variation, on how the population is regulated, and on exchanges with neighboring populations. Empirically, however, it remains unknown whether bet hedging can evolve to cope with the ecological conditions experienced by populations. Here we study the evolution of bet-hedging dormancy frequencies in two neighboring populations of the chestnut weevil, Curculio elephas. We estimate the temporal distribution of demographic parameters together with the form of the relationship between fecundity and population density and use both to parameterize models that predict the bet-hedging dormancy frequency expected to evolve in each population. Strikingly, the observed dormancy frequencies closely match predictions in their respective localities. We also found that dormancy frequencies vary randomly across generations, likely due to environmental perturbations of the underlying physiological mechanism. Using a model that includes these constraints, we predict the whole distribution of dormancy frequencies whose mean and shape agree with our observed data. Overall, our results suggest that dormancy frequencies have evolved according to local ecological conditions and physiological constraints.

Background: Root-feeding beetles, particularly Hylastes spp., Hylobius pales Herbst and Pachylobius picivorus Germar, increase in abundance in stressed forest stands and vector Grosmannia and Leptographium spp. fungi, which contribute to southern pine decline (SPD) in the southeastern United States. This study examined changes in the relative abundance of root-feeding beetles in response to mechanical thinning and clear-cut of even-age loblolly pine (Pinus taeda L.) stands in central Alabama and Georgia every 2 weeks during a 30 month study in 2009-2012., Results: The most abundant bark beetles were Hylastes salebrosus Eichhoff, H. porculus Erichson and H. tenuis Eichhoff. The relative abundance of the Hylastes spp. significantly increased after thinning treatments at all five sites. An initial decrease in Hylastes spp. occurred in response to clear-cut in some plots, but they typically recovered 2 months later and were stable for the remainder of the study., Conclusion: This study reports on the relative abundance responses of pathogen-vectoring root-feeding beetles to a thinning and clear-cut treatment in P. taeda stands. Thinning treatments conducted during the summer and winter may increase the relative abundance of Hylastes spp., vectors of Leptographium and Grosmannia spp., which are known to contribute to SPD by triggering plants to release defensive volatile compounds., (© 2013 Society of Chemical Industry.)

Chitinases have been implicated in the defence of conifers against insects and pathogens. cDNA for six chitinases were cloned from interior spruce (Picea glauca x engelmannii) and four from lodgepole pine (Pinus contorta). The cloned interior spruce chitinases were annotated class I PgeChia1-1 and PgeChia1-2, class II PgeChia2-1, class IV PgeChia4-1, and class VII PgeChia7-1 and PgeChia7-2; lodgepole pine chitinases were annotated class I PcChia1-1, class IV PcChia4-1, and class VII PcChia7-1 and PcChia7-2. Chitinases were expressed in Escherichia coli with maltose-binding-protein tags and soluble proteins purified. Functional characterization demonstrated chitinolytic activity for the three class I chitinases PgeChia1-1, PgeChia1-2 and PcChia1-1. Transcript analysis established strong induction of most of the tested chitinases, including all three class I chitinases, in interior spruce and lodgepole pine in response to inoculation with bark beetle associated fungi (Leptographium abietinum and Grosmannia clavigera) and in interior spruce in response to weevil (Pissodes strobi) feeding. Evidence of chitinolytic activity and inducibility by fungal and insect attack support the involvement of these chitinases in conifer defense., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: The systemic insecticides chlorantraniliprole (CAP) and thiamethoxam (TMX), applied to rice as seed treatments, may affect multiple life stages of the rice water weevil, Lissorhoptrus oryzophilus. Effects of CAP and TMX on adult survival, egg-laying and first- and late-instar survivals were determined by infesting plants treated as seeds with different rates of insecticides. The biological activity was related to insecticidal concentrations in leaves, shoots and roots., Results: CAP did not affect adult survival but decreased egg numbers and reduced the survival of the first and late instars. The greatest reduction in weevil population occurred in late instars feeding on roots. In contrast, TMX reduced adult survival and egg and larval numbers. The high biological activity of CAP on root-feeding stages was consistent with the accumulation of CAP in roots, whereas in TMX-treated plants the high activity on adults correlated with high concentrations of TMX in leaves and stems., Conclusions: The differential activity of insecticides on adults suggests poor inherent potency of CAP as an adulticide and/or its limited systemicity in foliage. The distribution of insecticide in specific plant parts can be attributed to the different physicochemical properties of CAP and TMX. The field implications of this research on management of L. oryzophilus are discussed., (© 2013 Society of Chemical Industry.)

The coffee berry borer Hypothenemus hampei is a pest that causes great economic damage to coffee grains worldwide. Because the proteins consumed are digested by aspartic proteases in the insect's midgut, the inhibition of these proteases by transferring a gene encoding an aspartic protease inhibitor from Lupinus bogotensis Benth. to coffee plants could provide a promising strategy to control this pest. Five aspartic protease inhibitors from L. bogotensis (LbAPI) were accordingly purified and characterized. The gene encoding the L. bogotensis aspartic protease inhibitor (LbAPI), with the highest inhibitory activity against H. hampei, was expressed in Escherichia coli and the purified recombinant protein (rLbAPI), with a molecular mass of 15 kDa, was subsequently assessed for its ability to inhibit the aspartic protease activity present in the H. hampei midgut in vitro, as well as its effects on the growth and development of H. hampei in vivo. The in vitro experiments showed that rLbAPI was highly effective against aspartic proteases from H. hampei guts, with a half maximal inhibitory concentration (IC50) of 2.9 μg. The in vivo experiments showed that the concentration of rLbAPI (w/w) in the artificial diet necessary to cause 50% mortality (LD50) of the larvae was 0.91%. The amino acid sequence of LbAPI had high homology (52-80%) to the seed storage proteins, vicilin and β-conglutin, suggesting that this protein was generated by evolutionary events from a β-conglutin precursor. Based on these results, LbAPI may have a dual function as storage protein, and as defense protein against H. hampei. These results provide a promising alternative to obtain a coffee plant resistant to H. hampei., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Bruchid beetles infest various seeds. The seed coat is the first protective barrier against bruchid infestation. Although non-host seed coats often impair the oviposition, eclosion and survival of the bruchid Callosobruchus maculatus larvae, morphological and biochemical aspects of this phenomenon remain unclear. Here we show that Phaseolus vulgaris (non-host) seed coat reduced C. maculatus female oviposition about 48%, increased 83% the seed penetration time, reduced larval mass and survival about 62 % and 40 % respectively. Interestingly, we found no visible effect on the major events of insect embryogenesis, namely the formation of the cellular blastoderm, germ band extension/retraction, embryo segmentation, appendage formation and dorsal closure. Larvae fed on P. vulgaris seed coat have greater FITC fluorescence signal in the midgut than in the feces, as opposed to what is observed in control larvae fed on Vigna unguiculata. Cysteine protease, α-amylase and α-glucosidase activities were reduced in larvae fed on P. vulgaris natural seed coat. Taken together, our results suggest that although P. vulgaris seed coat does not interfere with C. maculatus embryonic development, food digestion was clearly compromised, impacting larval fitness (e.g. body mass and survivability)., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Anthonomus grandis Boheman is a key pest in cotton crops in the New World. Its larval stage develops within the flower bud using it as food and as protection against its predators. This behavior limits the effectiveness of its control using conventional insecticide applications and biocontrol techniques. In spite of its importance, little is known about its genome sequence and, more important, its specific expression in key organs like the midgut. Total mRNA isolated from larval midguts was used for pyrosequencing. Sequence reads were assembled and annotated to generate a unigene data set. In total, 400,000 reads from A. grandis midgut with an average length of 237 bp were assembled and combined into 20,915 contigs. The assembled reads fell into 6,621 genes models. BlastX search using the NCBI-NR database showed that 3,006 unigenes had significant matches to known sequences. Gene Ontology (GO) mapping analysis evidenced that A. grandis is able to transcripts coding for proteins involved in catalytic processing of macromolecules that allows its adaptation to very different feeding source scenarios. Furthermore, transcripts encoding for proteins involved in detoxification mechanisms such as p450 genes, glutathione-S-transferase, and carboxylesterases are also expressed. This is the first report of a transcriptomic study in A. grandis and the largest set of sequence data reported for this species. These data are valuable resources to expand the knowledge of this insect group and could be used in the design of new control strategies based in molecular information., (© The Author 2014. Published by Oxford University Press on behalf of the Entomological Society of America.)

The entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, and Isaria fumosorosea were tested against the stored-grain pest Sitophilus oryzae. The fungi were isolated from the soil (from three locations in Attica, Greece: B. bassiana from Tatoion, M. anisopliae from Marathon, and I. fumosorosea from Aghios Stefanos) using larvae of Galleria mellonella as bait. Suspensions of 2.11 × 10(7) and 2.11 × 10(8), 1.77 × 10(7) and 1.77 × 10(8), and 1.81 × 10(7) and 1.81 × 10(8) conidia per ml of B. bassiana, M. anisopliae, and I. fumosorosea, respectively, were applied by three treatments: (i) sprayed on food and set in petri dishes with adults of S. oryzae, (ii) sprayed on adults of S. oryzae and set in petri dishes without food, and (iii) sprayed on adults of S. oryzae and set in petri dishes with food. The observed mortality of S. oryzae adults during the overall exposure period for the lowest, as well as for the highest, concentrations of B. bassiana, M. anisopliae, and I. fumosorosea ranged from 0 to 100%. Concentration was, in most of the cases tested, a critical parameter that determined the "speed of kill" of the exposed insect species for B. bassiana and M. anisopliae. Conversely, concentration was not that critical for I. fumosorosea, and survival was high in some of the combinations tested, even after 14 days of exposure. Both in the highest and the lowest concentrations of fungi, the mortality of S. oryzae adults was higher when the fungi were applied on adults than when they were applied on food. Higher mortality was observed when food was absent than when food was present, in most of the cases tested. The high efficacy levels recorded in the current study indicate that the tested fungi could be effective biocontrol agents against S. oryzae.

The red palm weevil (RPW; Rhynchophorus ferrugineus) is a devastating pest of palms, prevalent in the Middle East as well as many other regions of the world. Here, we report a large-scale de novo complementary DNA (cDNA) sequencing effort that acquired ∼5 million reads and assembled them into 26 765 contigs from 12 libraries made from samples of different RPW developmental stages based on the Roche/454 GS FLX platform. We annotated these contigs based on the publically available known insect genes and the Tribolium castaneum genome assembly. We find that over 80% of coding sequences (CDS) from the RPW contigs have high-identity homologs to known proteins with complete CDS. Gene expression analysis shows that the pupa and larval stages have the highest and lowest expression levels, respectively. In addition, we also identified more than 60 000 single nucleotide polymorphisms and 1 200 simple sequence repeat markers. This study provides the first large-scale cDNA dataset for RPW, a much-needed resource for future molecular studies., (© 2012 Institute of Zoology, Chinese Academy of Sciences.)

Sitophilus zeamais (Coleoptera: Curculionidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae) are important economic pests of corn, Zea mays (Poaceae). Psychotria spp. (Rubiaceae) plants are rich in secondary metabolites that could be toxic against such pests. We have screened extracts from four species of Psychotria abundant in the Cerrado (Savannah-type) biome of Brazil for the toxicity to two insects. We found that extracts from leaves and stems had significant effects on the hatching rate, parameters of caterpillar body (weight and length and width of head capsule), repellency, and mortality of these two pests, although the effects varied according to the Psychotria species and plant source (stem or leaf). Extracts of the stems of Psychotria hoffmannseggiana and of Psychotria capitata were more toxic to S. zeamais and the stems of Psychotria goyazensis to S. frugiperda; therefore, such extracts could have the potential for use in integrated pest management schemes.

Modified atmosphere based on lack of O2 offers a safe, residue-free alternative to chemical fumigants for pest control in stored grains. In this study, we intended to determine whether elevated CO2 (at a biologically achievable level) has an enhanced suppressive effect over low O2 atmosphere alone on the cowpea bruchid (Callosobruchus maculatus), a storage pest of cowpea and other legumes. Experiments were performed under two modified atmospheric conditions, (1) 2% O2+18% CO2+80% N2 and (2) 2% O2+98% N2. Both hypoxic environments significantly affected the development and survival of all insect developmental stages. Eggs were most vulnerable to hypoxia, particularly at the early stage (4-6h old), surviving only up to a maximum of 2 days in both treatments. These were followed by adults, pupae and larvae, in order of decreasing susceptibility. The 3rd and 4th instar larvae were most resilient to hypoxia and could survive up to 20 days of low O2. The presence of 18% CO2 significantly increased the mortality of adults, the later stage of eggs, as well as 1st and 4th instar larvae caused by hypoxia. However, the surviving insects exhibited faster development, evidenced by their earlier emergence from cowpea seeds compared to those without CO2. One interesting observation was the frequent, premature opening of the emergence windows in the 4th instar larvae when CO2 was involved. This phenomenon was not observed at all in insects stressed by low O2 alone. Differential expression profiling of metabolic genes and proteolytic activity of midgut digestive enzymes suggested that the rate of metabolic activity could contribute in part to the difference in insect development and survival under hypoxia in the presence and absence of CO2., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The South American weevil Listronotus bonariensis (Kuschel) is an important pest of pastures in New Zealand. As a component of management strategies for this pest, the South American parasitoid Microctonus hyperodae Loan (Hymenoptera: Braconidae) was released in northern New Zealand during 1991 as a biological control agent. Over the subsequent 5-6 yr, the reproductive biology of M. hyperodae and its relationship to, and effects on, the reproductive phenology and fitness of L. bonariensis were studied at three sites. M. hyperodae was first recovered in the field in December 1991. Subsequently, the incidence of parasitism in L. bonariensis increased to seasonal maxima of 75-90%. There was variable synchrony between parasitoid generations and the two generations of its host, leading to marked seasonal variation in rates of parasitism and parasitoid abundance. Despite marked inter-year variation, abundance of host adult and egg populations declined in the presence of parasitoids. Parasitized host females had lower ovarian maturity scores, had lower egg loads, and exhibited less investment in wing muscle development than females that had escaped parasitism. There was almost complete elimination of egg maturation in parasitized females and these hosts contributed little to population recruitment. Rate of buildup and seasonal maxima in parasitism, frequency of superparasitism, adult abundances, and wing muscle development in adult L. bonariensis varied among the three sites in a manner that was only partially related to climate differences across the 1.83° gradient of latitude. Site effects were weak to absent in measures of reproductive condition in L. bonariensis females.

The walnut twig beetle (Pityophthorus juglandis Blackman) vectors Geosmithia morbida, the causal agent of thousand cankers disease in Juglans, and is particularly damaging to Juglans nigra L. (black walnut). Native hosts of P. juglandis are distributed in the southwestern United States where winter temperatures tend to be higher than those found within the native range of black walnut. To better understand temperature effects on survival of P. juglandis, we initiated studies to determine: 1) seasonal variations in cold tolerance, as measured by the supercooling point (SCP), and 2) upper and lower lethal temperatures (LT). In the lower LT study, Xyleborinus saxeseni (Ratzeberg) was tested for comparison. Insects were either exposed to increasing or decreasing temperatures and then checked for survival. Upper and lower LTs were estimated using a logistic model. For the SCP study, data were grouped into seasons. Seasonal mean SCPs were highest in summer (-15.4°C) and lowest in fall (-18.1°C). The upper lethal limit estimations required to kill 99% of the population (LT99) for adults and larvae were 52.7 and 48.1°C, respectively, and lower limit LT99 estimations for adults and larvae were -18.1 and -18.7°C, respectively. The lower median LT (LT50) of X. saxeseni was -24.7°C. These studies, as well as beetle survival in infested Colorado trees where temperatures reached -29°C in February 2011, suggest P. juglandis could survive the winter in much of the native range of black walnut, but may be limited in trees where temperatures regularly exceed the lower LT.

There is empirical evidence of many diversified ways for energy to be acquired and allocated to reproduction, notably with strategies ranging from strict income breeding (females fueling their gametes with energy gained concomitantly during reproduction) to strict capital breeding (females storing nutrients prior reproduction). Until now, the question of whether diversification of these strategies might impact the way communities are organized has not been considered. Here, we suggest that diversified resource allocation strategies among competing species may contribute to their coexistence. We examined this hypothesis by focusing on communities composed of four phytophagous insect species that coexist and compete for egg-laying sites. From wild-caught females, we determined precisely the breeding period of each species and we described their resource acquisition and allocation to reproduction dynamics. We quantified in each species the total amount of larval energy stored by newly-emerging females and then monitored the total energy budget of females caught in the field before and throughout their breeding period. We found that the four sibling weevil species are markedly segregated along the income-capital-breeding continuum, which is correlated with clear time partitioning in their laying activity. Our results suggest that diversified resource allocation strategies might contribute to time partitioning of plant resources exploitation and thus indirectly to their coexistence. This work should further encourage studies examining the extent to which competitive coexistence might be affected by diversification of income-capital breeding strategies together with the intensity of interspecific competition, and considering the divergent evolution of these strategies.

The principal component analysis (PCA), non-metric multidimensional scaling (MDS) and analysis of similarity (ANOSIM) are multivariate statistical techniques that graphically represent numerical measures of several factors and display multiple relationships that may exist between them. In this study, we evaluated the applicability of these techniques to analyze the nutritional quality of diet, using as model, the bioassay rice weevil. The diets tested were: corn starch, potato starch, 5% glucose, peas, starved and starved with water supply. The variables studied were: survival, weight change and body composition. The PCA and MDS showed positive relationships of survival and weight change with body fat and carbohydrate parameters. Fat and carbohydrates were greater in starches diets, similar to the positive control. The PCA showed differences between populations fed with different diets, whereas the MDS showed similarity between diets. Both studies defined a gradient of the nutritive value of diets in the x-axis. The ANOSIM indicate significant (p < 0.05) differences between groups. This test is necessary to support the results obtained in the PCA and MDS. The application of these statistical tools is promising to analyze complex processes such as interaction of differents variables to measure the nutritional quality of diets.

Seven resistant varieties of sweetpotato were compared with three susceptible varieties in field trials and laboratory bioassays and showed that resistance was an active process rather than an escape mechanism, as field resistant varieties also had reduced root damage and oviposition compared with susceptible varieties in the laboratory. Liquid chromatography-mass spectrometry (LC-MS) of root surface and epidermal extracts showed significant variation in the concentration of hexadecyl, heptadecyl, octadecyl, and quinic acid esters of caffeic and coumaric acid, with higher concentrations correlated with resistance. All compounds were synthesized to enable their positive identification. Octadecyl coumarate and octadecyl caffeate applied to the surface of susceptible varieties in laboratory bioassays reduced feeding and oviposition, as observed on roots of resistant varieties, and therefore are implicated in weevil resistance. Segregating populations from breeding programs can use these compounds to identify trait loci for resistance and enable the development of resistant varieties.

Weevils of the genus Cionus (Curculionidae, Mecininae) sequester the iridoid glycosides (IGs) aucubin and catalpol from their host plants Scrophularia or Verbascum (Scrophulariaceae). Cionus hortulanus is the only member of the genus that feeds on both plant genera. We previously showed that sequestration patterns in C. hortulanus depend on the local host. To investigate whether IG patterns are driven by their availability in the hosts or genetic differences between populations, we collected C. hortulanus from S. nodosa in the field and reared them either on S. nodosa or on V. nigrum. The differences in IG concentrations were specific for the host plant upon which the weevils developed. Similar to monophagous species of the Cionini, individuals from S. nodosa had more aucubin than catalpol and mirrored the concentrations of their host plants. Specimens from V. nigrum, on the other hand, had higher concentrations of aucubin and of catalpol than their host. On V. nigrum, the ratio of catalpol to aucubin differed significantly between plant and beetle samples due to much higher catalpol concentrations in the weevils. Our data thus contradict genetically fixed differences between populations living on either plant but rather document the host plants' influence on the beetles' metabolism.

Rhynchophorus ferrugineus (Olivier 1790) (Coleoptera: Curculionidae) is becoming a serious problem in Mediterranean areas where it is well-adapted, and now is present even in the United States (California). The infestations are primarily in urban areas where chemical control is not advisable and million of Euros are spent to control it. The effects of the entomopathogenic nematode Steinernema carpocapsae (Nematoda: Steinernematidae) on mortality, growth, as well as the immune activity of R. ferrugineus larvae, were investigated. R. ferrugineus mortality exhibited a positive trend with the dosage and duration of exposure to S. carpocapsae. The median lethal dose and median lethal time, important to optimize the treatments, were calculated. S. carpocapsae also had a detrimental effect on R. ferrugineus weight. In vivo and in vitro effects of S. carpocapsae on the phagocytic responses of R. ferrugineus hemocytes also were recorded. S. carpocapsae was not encapsulated by R.ferrugineus hemocytes. After 24 h, the number of hemocytes recorded in treated larvae was reduced. To investigate the defensive abilities of R. ferrugineus humoral and cellular immune systems, specifically against the bacterium Xenorhabdus nematophila (Enterobacteraceae), the minimum inhibitory concentration that inhibits bacterial growth was measured. This is the first time that this technique is applied to entomopathogenic bacteria.

Ambrosia beetle fungiculture represents one of the most ecologically and evolutionarily successful symbioses, as evidenced by the 11 independent origins and 3500 species of ambrosia beetles. Here we document the evolution of a clade within Fusarium associated with ambrosia beetles in the genus Euwallacea (Coleoptera: Scolytinae). Ambrosia Fusarium Clade (AFC) symbionts are unusual in that some are plant pathogens that cause significant damage in naïve natural and cultivated ecosystems, and currently threaten avocado production in the United States, Israel and Australia. Most AFC fusaria produce unusual clavate macroconidia that serve as a putative food source for their insect mutualists. AFC symbionts were abundant in the heads of four Euwallacea spp., which suggests that they are transported within and from the natal gallery in mandibular mycangia. In a four-locus phylogenetic analysis, the AFC was resolved in a strongly supported monophyletic group within the previously described Clade 3 of the Fusarium solani species complex (FSSC). Divergence-time estimates place the origin of the AFC in the early Miocene ∼21.2 Mya, which coincides with the hypothesized adaptive radiation of the Xyleborini. Two strongly supported clades within the AFC (Clades A and B) were identified that include nine species lineages associated with ambrosia beetles, eight with Euwallacea spp. and one reportedly with Xyleborus ferrugineus, and two lineages with no known beetle association. More derived lineages within the AFC showed fixation of the clavate (club-shaped) macroconidial trait, while basal lineages showed a mix of clavate and more typical fusiform macroconidia. AFC lineages consisted mostly of genetically identical individuals associated with specific insect hosts in defined geographic locations, with at least three interspecific hybridization events inferred based on discordant placement in individual gene genealogies and detection of recombinant loci. Overall, these data are consistent with a strong evolutionary trend toward obligate symbiosis coupled with secondary contact and interspecific hybridization., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: The effect of transgenic maize (Zea mays L.) containing a lepidopteran-specific Bt toxin on a stored-product pest, Sitophilus zeamais Motschulsky, and its parasitoid, Lariophagus distinguendus Förster, was examined in the laboratory to test the impact of transgenic maize on stored-product pests and their biological control., Results: Weevils were not harmfully affected by transgenic Bt maize in their development characteristics (development time, body mass), and females emerging from transgenic maize kernels were larger. However, significantly fewer parasitoid females emerged from weevils that developed in transgenic kernels, although parasitoids did not develop more slowly and were not different in size or mass from their conspecifics emerging from hosts in non-transgenic maize kernels., Conclusion: The emergence of female parasitoids was reduced in transgenic Bt maize, and this effect cannot be explained by the known lepidopteran-specific toxicity of Bt Cry1Ab toxin., (© 2012 Society of Chemical Industry.)

The pecan weevil, Curculio caryae (Horn), is a key pest of pecan, Carya illinoinensis (Wangenh.) K. Koch. Prior research indicated the potential for use of Hypocreales fungi to suppress C. caryae. We compared the efficacy of two fungal spp., Beauveria bassiana (GHA strain) and Metarhizium brunneum (F52), in their ability to cause C. caryae mortality. The fungus, B. bassiana, was applied to trunks of pecan trees (a method previously shown to be effective in C. caryae suppression) and efficacy was compared with M. brunneum applied to the ground or to the trunk with or without SoyScreen Oil as an ultraviolet protecting agent. Results indicated B. bassiana to be superior to M. brunneum regardless of application method; consequently, the potential for applying B. bassiana to control C. caryae was explored further. Specifically, the impact of different fertilizer regimes (as used by pecan growers) on the persistence of B. bassiana (GHA) in soil was determined. B. bassiana was applied to soil in a pecan orchard after one of several fertilizer treatments--i.e., ammonium nitrate, crimson clover, poultry litter, clover plus poultry litter, and a no-fertilizer control. B. bassiana persistence up to 49 d in 2009 and 2010 was assessed by plating soil onto selective media and determining the number of colony forming units, and by baiting soil with a susceptible host, Galleria mellonella (L.). Fertilizer treatments did not impact B. bassiana persistence. We conclude that standard fertilizers for nitrogen management, when applied according to recommended practices, are unlikely to negatively impact survival of B. bassiana in pecan orchards when the fungus is applied for C. caryae suppression during weevil emergence. Additional research on interactions between entomopathogenic fungi and fertilizer amendments (or other tree nutrition or soil management practices) is merited.

Increasing empirical evidence has documented variability in elemental composition within species. However, the extent, causes, and pattern of variability in consumer stoichiometry across a large geographical scale are not well understood. Here, we investigated this issue using a holometabolous insect, weevils (Curculio davidi Fairmaire). Larvae of this species store energy needed for diapause, and variable energy requirements across the geographic range of this species could lead to differences in body elemental composition. Our results showed that variability was high (assessed as the coefficient of variation (CV)) in larval body nitrogen (N) and phosphorus (P) (CV, 10% for N and 13% for P) compared to emerging adults (CV, 5% for N and 8% for P). Temperature-related factors explained more variation than other climatic factors and food for carbon (C), N and P in weevil. In warmer regions, larval C concentration was higher, while N and P were lower. The high C content of weevil larvae relative to both their food source and their adult stage was attributed to energy storage. Across the climatic gradient of its geographic range, larval body C content increased with mean annual temperature and decreased with average diurnal temperature range. This finding implies that temperature-related C storage drives the high variability in elemental composition of larvae across the climate gradient, and also effectively dampens the stoichiometric imbalance between consumers and food resources while serving as an energy reservoir for overwintering and metamorphosis., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Coffee berry borer, Hypothenemus hampei (Ferrari) is the most serious insect pest of coffee around the world. Although it is already present in most of the world's major coffee growing regions, it is important to delay further spread and to prevent reintroductions that might include hyperparasites or improve the genetic base of existing populations. Green coffee is shipped around the world for custom blending and roasting and such shipments carry the risk of spreading H. hampei. We used heavily infested coffee berries as a surrogate for green coffee to test the freezing tolerance of H. hampei. After freezing, all life stages of H. hampei were dissected from coffee berries and mortality was assessed. Counting all life stages, > 15,000 insects were measured in this study. A temperature of approximately -15 degrees C (range, -13.9 to -15.5) for 48 h provided 100% control of all life stages. A logit regression model predicted < or = 1 survivor in a million for treatments of -20 degrees C for 5 d or -15 degrees C for 6 d. A freezing treatment for green coffee might be more economical and acceptable compared with fumigation with methyl bromide, especially for small-scale and organic growers and millers in Hawaii who ship green coffee beans to other islands for custom roasting. Freezing treatments could also be used to kill H. hampei in coffee seeds before export with minimal effects on seed germination if coffee seeds are first dried to critical water content levels in accordance with published methods.

The Western Ghats of Karnataka natural ecosystem are among the most diverse and is one of the eight hottest hotspots of biological diversity in the world, that runs along the western part of India through four states including Karnataka. Bacillus thuringiensis (Bt) strains were isolated from soils of Western Ghats of Karnataka and characterized by molecular and analytical methods as a result of which 28 new Bt-like isolates were identified. Bt strains were isolated from soil samples using sodium acetate selection method. The morphology of crystals was studied using light and phase contrast microscopy. Isolates were further characterized for insecticidal cry gene by PCR, composition of toxins in bacterial crystals by SDS-PAGE cloning, sequencing and evaluation of toxicity was done. As a result 28 new Bt-like isolates were identified. Majority of the isolates showed the presence of a 55 kDa protein bands on SDS-PAGE while the rest showed 130, 73, 34, and 25 kDa bands. PCR analysis revealed predominance of Coleopteran-active cry genes in these isolates. The variations in the nucleotide sequences, crystal morphology, and mass of crystal protein(s) purified from the Bt isolates revealed genetic and molecular diversity. Three strains containing Coleopteran-active cry genes showed higher activity against larvae Myllocerus undecimpustulatus undatus Marshall (Coleoptera: Curculionidae) than B. thuringiensis subsp. Morrisoni. Results indicated that Bt isolates could be utilized for bioinsecticide production, aiming to reduce the use of chemical insecticide which could be useful to use in integrated pest management to control agriculturally important pests for sustainable crop production.

Bark beetles are among the most destructive of pine forest pests and they form close symbiotic relationships with ophiostomatoid fungi. Although some fungi are considered to be mutualistic symbionts of bark beetles with respect to the supply of nutrients, detrimental effects of fungal symbionts on larval growth have also been frequently reported. The mechanisms of such antagonistic effects are hypothesized to be a decrease in nutritional resources caused by competition for saccharides by the fungi. Here, we provide experimental evidence that three beetle-associated fungi modify the nutritional content of an artificial phloem diet, leading to a detrimental effect on the growth of Dendroctonus valens larvae. When larvae were fed a diet of pine phloem in agar medium colonized with any of these fungi, feeding activity was not affected but weight significantly decreased. Additional analysis showed that fungi depleted the fructose and glucose concentrations in the phloem media. Furthermore, these detrimental effects were neutralized by supplementing the media with fructose or glucose, suggesting that fungi may affect larval growth by modifying diet saccharide contents. These data indicate that fungus-induced nutritional changes in bark beetle diet can affect larval growth, and that the mechanism involves fungus-induced saccharide depletion from the larval diet.

Background: Feeding assays using adult rice water weevils and foliage of plants treated as seeds with chlorantraniliprole and thiamethoxam at different rates were conducted to evaluate the systemic adulticidal and feeding effects. Dose-mortality relationships were determined for thiamethoxam seed treatments by combining leaf area lost due to feeding and insecticide residues analyzed by LC/MS/MS. Changes in adulticidal activity of thiamethoxam were also investigated by contrasting adult mortalities at the 5-6-leaf and tillering stages of rice., Results: Adult weevil mortalities and leaf consumption rates on foliage were affected in thiamethoxam but not in chlorantraniliprole treatments when rice was at the 6-7-leaf stage. The LD(50) for weevils feeding on thiamethoxam-treated rice at the 2-3-leaf stage was 447 pg insecticide weevil(-1) (95% CL: 25-830 pg weevil(-1)) but was lower (142 pg weevil(-1); 95% CL: 102-180 pg weevil(-1)) in experiments with 3-4-leaf-stage plants. Mortalities on leaves from 5-6-leaf-stage plants were consistently higher than on leaves from tillering plants. Thiamethoxam residues measured by ELISA increased with seed treatment rate and differed between plant stages., Conclusion: The LD(50) values developed in this study are the first values for leaf-feeding insects on foliage of plants treated as seeds with thiamethoxam. The attrition of adulticidal activity of thiamethoxam in foliage of older plants may help to explain the reduced effectiveness of seed treatments against rice water larvae that is seen at later stages of rice growth in field studies. The differential activity of these two seed treatments on adults suggests that adult mortality contributes to the field efficacy of thiamethoxam but not to that of chlorantraniliprole., (Copyright © 2012 Society of Chemical Industry.)

Larva and pupa of Loncophorus pustulatus (Champion, 1903) (Curculionidae: Curculioninae: Anthonomini) are described, illustrated and compared with descriptions of immatures of two other species of Loncophorus. Weevil larvae were found inside aborted flowers on the ground under Ceiba speciosa (A. St.-Hil.) Ravenna (Malvaceae), in the city of Sgo Paulo, State of S5o Paulo, and reared to adults in laboratory. Data obtained in the field and under laboratory conditions are presented. Parasitoidism of weevil larvae by wasps of the genus Catolaccus (Hymenoptera: Pteromalidae) is reported.

Anchylorhynchus vanini sp. nov. from the Amazon is described, including the mouthparts and male genitalia. The new species is compared with similar species within the genus and the key to the species of Anchylorhynchus provided by Vaurie (1954) is modified to include the new species. Adult specimens were collected in flowers of the palm Syagrus vermicularis Noblick and additional collections in other palms species suggest that this association is specific. This is the first record of the palm Syagrus vermicularis as host for a species of Anchylorhynchus.

The size of adult parasitoids is influenced by the quantity and quality of resources available during immature development. Gregarious development of endoparasitoids results in scramble competition where the resources are shared among individuals developing into the same host. Individuals that developed gregariously are therefore smaller and that reduced size generally translates into lower fitness due to reductions in several life history traits including longevity, mobility and traits linked to reproduction. We measured the reproductive constraints induced by size in Anaphes listronoti (Hymenoptera: Mymaridae), a facultative gregarious egg parasitoid of Curculionidae. Size decreased with number of immatures developing in a host egg for both male and female A. listronoti. This reduction in size induced a reproductive cost as both males and females produced fewer gametes when developing gregariously. Contrarily to other egg parasitoids, A. listronoti males are not prospermatogenic and produced some sperm during their adult life. Their spermatogeny index is estimated at 0.6-0.7 that places this species as moderately synspermatogenic. Female A. listronoti have an ovigeny index of 0.70 and are therefore moderately synovigenic. Large females that developed singly received a full sperm complement only when mated by a large male that also developed singly. When mating with a small triplet male, a large female received less than half her sperm complement. Large males were able to mate three females before the number of sperm transferred started to decrease., (Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.)

In this study, we describe and present original data on the bionomics and ecology of Tachygonus erythroxyli Hespenheide, New Species, on Erythroxylum subsessile (Mart.). Thirty individuals of E. subsessile were analysed every two months, from October, 2006 to October, 2007 at the National Park of Restinga de Jurubatiba (PNRJ) Carapebus, Rio de Janeiro, Brazil. Females of T. erythroxyli lay their eggs singly on the upper surface of the host's leaves. The mine of T. erythroxyli is expanded with three bubbles, each one representing one larval instar. These mines occur throughout the year in at least half of the host plants and are markedly more abundant in summer. We obtained three parasitoid species, making a total of 53% of the rate of parasitism. Cirrospilus sp. (Eulophidae) was the most prevalent and represented 88% of the parasitoids. The great abundance of Tachygonus mines, their high frequency in their host plant, and local monophagy of this leaf miner weevil suggest a close association between T. erythroxyli and Erythroxylum subsessile.

Understanding how the population dynamics of insect pests are affected by environmental factors and agricultural practices is important for pest management. To investigate how the abundance of the banana weevil, Cosmopolites sordidus (Coleoptera: Curculionidae), is related to environmental factors and the length of the fallow period in Martinique, we developed an extensive data set (18,130 observations of weevil abundance obtained with pheromone traps plus associated environmental data) and analysed it with generalized mixed-effects models. At the island scale, C. sordidus abundance was positively related to mean temperature and negatively related to mean rainfall but was not related to soil type. The number of insects trapped was highest during the driest months of the year. Abundance of C. sordidus decreased as the duration of the preceding fallow period increased. The latter finding is inconsistent with the view that fallow-generated decomposing banana tissue is an important resource for larvae that leads to an increase in the pest population. The results are consistent with the view that fallows, in association with pheromone traps, are effective for the control of the banana weevil.

The large pine weevil, Hylobius abietis, is a major pest in European conifer forests causing millions of Euros of damage annually. Larvae develop in the stumps of recently felled trees; the emerging adults feed on the bark of seedlings and may kill them. This study investigated the susceptibility of different developmental stages of H. abietis to commercial and commercially viable isolates of entomopathogenic fungi, Metarhizium and Beauveria. All the developmental stages of H. abietis can be killed by Metarhizium robertsii, Metarhizium brunneum, and Beauveria bassiana. The most virulent isolate of M. robertsii ARSEF4556 caused 100% mortality of pupae, larvae and adults on day 4, 6 and 12, respectively. This strain was further tested against adult weevils in different concentrations (10(5)-10(8) conidia cm(-2) or ml(-1)) using two types of fungal formulation: 'dry' conidia and 'wet' conidia (suspended in 0.03% aq. Tween 80) applied on different substrates (tissue paper, peat and Sitka spruce seedlings). 'Dry' conidia were more effective than 'wet' conidia on tissue paper and on spruce or 'dry' conidia premixed in peat. The LC(50) value for 'dry' conidia of isolate ARSEF4556 was three folds lower than 'wet' conidia on tissue paper. This study showed that 'dry' conidia are more effective than 'wet' conidia, causing 100% adult mortality within 12 days. Possible strategies for fungal applications are discussed in light of the high susceptibility of larvae and pupae to fungal pathogen., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Dendroctonus rhizophagus Thomas and Bright (Curculionidae: Scolytinae) is an endemic economically important insect of the Sierra Madre Occidental in Mexico. This bark beetle has an atypical behavior within the genus because just one beetle couple colonizes and kills seedlings and young trees of 11 pine species. In this work, the bacteria associated with the Dendroctonus rhizophagus gut were analyzed by culture-dependent and culture-independent methods. Analysis of 16S rRNA sequences amplified directly from isolates of gut bacteria suggests that the bacterial community associated with Dendroctonus rhizophagus, like that of other Dendroctonus spp. and Ips pini, is limited in number. Nine bacterial genera of γ-Proteobacteria and Actinobacteria classes were detected in the gut of Dendroctonus rhizophagus. Stenotrophomonas and Rahnella genera were the most frequently found bacteria from Dendroctonus rhizophagus gut throughout their life cycle. Stenotrophomonas maltophilia, Ponticoccus gilvus, and Kocuria marina showed cellulolytic activity in vitro. Stenotrophomonas maltophilia, Rahnella aquatilis, Raoultella terrigena, Ponticoccus gilvus, and Kocuria marina associated with larvae or adults of Dendroctonus rhizophagus could be implicated in nitrogen fixation and cellulose breakdown, important roles associated to insect development and fitness, especially under the particularly difficult life conditions of this beetle.

Insecticidal potential of natural zeolites and diatomaceous earths originating from Serbia against Sitophilus oryzae (L.) and Tribolium castaneum (Herbst) was evaluated. Two natural zeolite formulations (NZ and NZ Modified) were applied to wheat at rates of 0.50, 0.75, and 1.0 g/kg, while two diatomaceous earth (DE) formulations (DE S-1 and DE S-2) were applied at rates of 0.25, 0.50, 0.75, and 1.0 g/kg. A bioassay was conducted under laboratory conditions: temperature of 24 +/- 1 degrees C, relative humidity in the range 50-55%, in tests with natural zeolites, and 60-65%, in tests with DEs, and in all combinations for progeny production. Mortality was assessed after 7, 14, and 21 d of insect contact with treated wheat, and the total mortality after an additional 7-d recovery on untreated broken wheat. Progeny production was also assessed after 8 wk for S. oryzae and 12 wk for T. castaneum. The highest mortality for S. oryzae and T. castaneum was found after the longest exposure period and 7 d of recovery, on wheat treated with NZ at the highest rate and DEs at rates of 0.50 -1.0 g/kg. Progeny reduction higher than 90% was achieved after 14 and 21 d of contact of both beetle pests with wheat treated with DE S-1 at 0.50-1.0 g/kg and DE S-2 at 0.75-1.0 g/kg, while the same level of reduction was achieved only for T. castaneum after its contact with the highest rate of NZ formulation. NZ Modified, applied even at the highest rate, revealed much lower insecticidal potential.

Background: The granary weevil, Sitophilus granarius (L.), is one of the most damaging pests of stored grains, causing severe quantitative and qualitative losses. Sustainable control means, alternative to the commonly used fumigants and broad-spectrum contact insecticides, are urgently needed owing to legislative limits, the development of resistant insect strains and increasing consumer demand for safe food. Short-chain aliphatic ketones, known to be emitted by cereal grains and previously identified as repellents to adult granary weevils, were evaluated for their ability to disrupt insect orientation towards wheat grains and as possible natural fumigants., Results: In behavioural bioassays, 2-pentanone, 2-hexanone, 2-heptanone and 2,3-butanedione significantly reduced insect orientation towards odours of wheat grains, with 2-hexanone and 2-heptanone being the most active. In fumigation tests, all compounds were effective in killing weevil adults, but they performed differently according to chemical structure, speed of action and presence of wheat grains. In the presence of grains, the highest fumigant toxicity was shown by 2-pentanone (LC(50) = 8.4 ± 1.0 mg L(-1)) after 24 h exposure, and by 2-pentanone (LC(50) = 4.5 ± 0.3 mg L(-1)), 2-heptanone (LC(50) = 7.1 ± 0.3 mg L(-1) ) and 2-hexanone (LC(50) = 8.1 ± 0.6 mg L(-1)) 1 week after the treatment end., Conclusion: Short-chain aliphatic ketones have potential for applications in IPM programmes for the granary weevil because of their behaviour-altering activity and fumigant toxicity., (Copyright © 2011 Society of Chemical Industry.)

Ailanthus altissima (Mill.) Swingle, tree-of-heaven, is an invasive species native to Asia. It first was introduced into the United States in the 1700 s and now is distributed throughout much of North America. Mechanical and chemical controls are current suppression tactics, however, implementation is costly. A weevil, Eucryptorrhynchus brandti (Harold), was identified in China and imported for quarantine testing in 2004 as a potential biological control agent. Host specificity tests on adult feeding, larval development, and oviposition of this weevil were conducted from 2007 to 2011 on A. altissima and 29 nontarget species. Eucryptorrhynchus brandti adults fed significantly more on A. altissima foliage when compared with all test species. Range of means for feeding on A. altissima was 32.5-106.5 mm(2)/adult/d. In no-choice tests, Simarouba glauca DC, Leitneria floridana Chapm., and Citrus limon (L.) Burm. F., had feeding rates of only 10, 49, and 10%, respectively, compared with the level of feeding on A. altissima. The mean range of adult feeding by E. brandti on all other test species was <7% of feeding on A. altissima (0.0-3.3 ± 5.0 mm(2)/adult/d). In the no-choice larval inoculation tests, larval development only occurred in two of 10 L. floridana seedlings compared with seven of 10 A. altissima seedlings. In the no-choice oviposition tests, oviposition and subsequent larval development did not occur in L. floridiana, whereas all seven A. altissima seedlings supported oviposition and subsequent larval development. The weevil did not appear to be a threat to L. floridana or any other nontarget species tested. Therefore, we conclude that Eucryptorrhynchus brandti is highly host specific to A. altissima.

Interactions between invasive insects and their fungal associates have important effects on the behavior, reproductive success, population dynamics and evolution of the organisms involved. The red turpentine beetle (RTB), Dendroctonus valens LeConte (Coleoptera: Scolytinae), an invasive forest pest in China, is closely associated with fungi. By carrying fungi on specialized structures in the exoskeleton, RTB inoculates fungi in the phloem of pines (when females dig galleries for egg laying and when males join them for mating). After eggs hatch, larvae gregariously feed on the phloem colonized by the fungi. We examined the effects of five isolates of RTB associated fungi (two from North America, Leptographium terebrantis and L. procerum, and three from China, Ophiostoma minus, L. sinoprocerum and L. procerum) on larval feeding activity, development and mortality. We also studied the effects of volatile chemicals produced in the beetle hindgut on fungal growth. Ophiostoma minus impaired feeding activity and reduced weight in RTB larvae. Leptographium sinoprocerum, L. terebrantis and L. procerum did not dramatically influence larval feeding and development compared to fungi-free controls. Larval mortality was not influenced by any of the tested fungi. Hindgut volatiles of RTB larvae, verbenol, myrtenol and myrtenal, inhibited growth rate of all the fungi. Our results not only show that D. valens associated fungus, O. minus, can be detrimental to its larvae; but, most importantly, they also show that these notorious beetles have an outstanding adaptive response evidenced by the ability to produce volatiles that inhibit growth of harmful fungus.

The preference-performance hypothesis has principally considered insect herbivores with aboveground lifecycles, although the hypothesis could be equally relevant to insects with life stages occurring both aboveground and belowground. Moreover, most studies have focussed on either laboratory or field experiments, with little attempt to relate the two. In this study, the preference-performance hypothesis was examined in an aboveground-belowground context in the laboratory using the vine weevil (Otiorhynchus sulcatus (F.)) (Coleoptera: Curculionidae) and two cultivars of red raspberry (Rubus idaeus), Glen Rosa and Glen Ample. A two-year field study (2008-2009) was also undertaken to characterise the population dynamics of adult weevils on the two raspberry cultivars. Larval performance (abundance and mass) differed significantly between Glen Rosa and Glen Ample, with Glen Rosa resulting in 26% larger but 56% fewer larvae compared to Glen Ample. Larval abundances were significantly and positively correlated with root nitrogen and magnesium concentrations, but negatively correlated with root iron. However, concentrations of these minerals were not significantly different in the two cultivars. Adult weevils did not preferentially select either of the two cultivars for egg laying (laying 3.08 and 2.80 eggs per day on Glen Ample and Glen Rosa, respectively), suggesting that there was no strong preference-performance relationship between adult vine weevils and their belowground offspring. Field populations of adult vine weevils were significantly higher on Glen Ample than Glen Rosa, which may reflect the higher larval survival on Glen Ample observed in laboratory experiments.

The red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae), is an economically important pest of palms worldwide. To better understand the phenology of R. ferrugineus populations in the Mediterranean basin, the objective of the present work has been to estimate the effect of temperatures within the range 10-25°C on the reproductive parameters of R. ferrugineus and to determine its lower temperature thresholds for oviposition and egg hatching. Our results confirm that oviposition in R. ferrugineus is strongly affected by temperature. Lower thresholds for oviposition and egg hatching (15.45° and 13.95°C, respectively) are below the mean monthly temperatures registered in winter in most of the northern shore of the Mediterranean basin. Under these circumstances, new palm infestations would be difficult during most of the winter. These results should be taken into account when planning some palm management practices, such as pruning or pesticide treatments, in areas under a Mediterranean climate.

Domestic sheep (Ovis spp.) grazing is emerging as an integrated pest management tactic for alfalfa weevil, Hypera postica (Gyllenhal), management and a degree-day model is needed as a decision and support tool. In response to this need, grazing exclosures with unique degree-days and stocking rates were established at weekly intervals in a central Montana alfalfa field during 2008 and 2009. Analyses indicate that increased stocking rates and grazing degree-days were associated with decreased crop levels of weevil larvae. Larval data collected from grazing treatments were regressed against on-site and near-site temperatures that produced the same accuracy. The near-site model was chosen to encourage producer acceptance. The regression slope differed from zero, had an r2 of 0.83, and a root mean square error of 0.2. Crop data were collected to achieve optimal weevil management with forage quality and yield. Differences were recorded in crude protein, acid and neutral detergent fibers, total digestible nutrients, and mean stage by weight. Stem heights differed with higher stocking rates and degree-days recording the shortest alfalfa canopy height at harvest. The degree-day model was validated at four sites during 2010 with a mean square prediction error of 0.74. The recommendation from this research is to stock alfalfa fields in the spring before 63 DD with rates between 251 and 583 sheep days per hectare (d/ha). Sheep should be allowed to graze to a minimum of 106 and maximum of 150 DD before removal. This model gives field entomologists a new method for implementing grazing in an integrated pest management program.

Dry artificial diet pellets prepared with maize, cassava chips, and amino acid supplements (lysine and methionine) were evaluated for mass culture of Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), a highly polyphagous pest of many stored grains. Evaluation was done in the laboratory at temperature 26 ± 2 °C, 60-70% RH, 12:12 L:D photoperiod. The artificial diet was compounded from different proportions of maize (M) variety TZPB-SW-R, cassava (C) variety TMS-2110, and amino acid supplements, and was pelletized into 6 mm diameter pellets on which five pairs of one-day-old S. zeamais were bioassayed. The diet M(9)C(1) (90% M and 10% C) was the most suitable diet with comparatively shorter developmental period (34.8 days) and the highest F(1) emergence of progeny (145.4) compared to the control, M(10)C(0) (100% M and 0% C).

Trophic interactions can trigger the development of exaggerated specialized characters and promote morphological diversification. For example, acorn weevils (genus Curculio) present strikingly long rostrums, which are used by females to perforate oviposition holes through the seed coat. Species exhibiting longer rostrums are known to exploit larger acorns, and therefore rostrum length is thought to be subject to selection to match the preferred acorn type. However, rostrum length is strongly correlated with body size, and morphological divergence could result from either selection on rostrum length for optimal food exploitation or from other pressures acting on body size. We collected infested acorns at oak forests where the large Curculio elephas and the small-bodied Curculio glandium co-occur. There were no interspecific differences in adult female body size to rostrum length allometric relationships, and rostrum length is equally correlated with body size in either species. MtDNA-based species identification showed that C. glandium larvae were present within acorns of all sizes, whereas C. elephas larvae were restricted to acorns above a minimum size, irrespective of oak species. Hence, exploitation of large acorns can hardly have triggered rostrum enlargement, as the small sized C. glandium adults (with short rostrums) could perforate and oviposit in both small and large acorns. Rather, increased rostrum length is probably a by-product of the larger body sizes of individuals emerging from bigger acorns, which allow increased larval size and enhance larval survival likelihood. Summarizing, when exaggerated feeding traits co-vary with other body features, interspecific morphological variability may result from contrasting selective pressures acting on these correlated characters.

Widespread destruction of tallgrass prairies in the midwestern United States has fragmented plant communities with the result that populations of endemic animal species have become geographically isolated from one another. The goal of the research summarized here was to evaluate the potential for conserving endemic prairie species of herbivorous insects by managing their host plants. Our study species was the weevil Haplorhynchites aeneus (Boehman), adults of which feed on pollen of plants in the genus Silphium (Asteraceae: Heliantheae). The female weevils clip the peduncles of flower heads and oviposit into the heads, where the larvae feed on the ovules. The research was conducted in 12 prairie sites in eastern Illinois. An allozyme analysis revealed that most populations of H. aeneus at the various prairie sites were genetically differentiated from one another, but the degree of differentiation was not associated with geographic distance between sites. Adult H. aeneus fed and oviposited on the plant species Silphium laciniatum L., S. integrifolium Michx., and S. terebinthinaceum Jacq, which differ in bloom phenology. There was no evidence of genetic differentiation of weevil populations with respect to host plant species, and adult weevils strongly preferred S. terebinthinaceum. We conclude that the oligophagous nature of the weevil assures its survival in small prairie remnants even where some of the host plant species are absent. Although H. aeneus can have a significant impact on reproduction of host plants by clipping flower heads, the perennial nature of Silphium species prevents their local extinction.

A documentation and review of weevil rostrum growth is made through examination of the developmental life stages in the rice weevil Sitophilus oryzae (Linnaeus). Histological and morphological examinations are made utilizing light, fluorescent, and electron microscopy. In S. oryzae, rostral tissue begins proliferating in the late 4th instar larva and continues through to the pupal stage, with the majority of rostrum growth taking place in the prepupa. Adult cranial and rostral morphology is also reviewed, focusing on structures that may be pertinent to phylogeny reconstruction. The weevil rostrum is essentially an extension of various head sclerites that are basal to the mouthparts. Therefore, while the mouthparts are fairly similar to other Coleoptera in basic form, the head is markedly different due to its anterior extension. By understanding the more noticeable details of rostrum growth and structure, this study may serve as a foundation for comparative studies of a similar nature and as a basis for beginning research on the genetic nature of rostrum formation and evolution throughout the weevil clade., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The spruce beetle, Dendroctonus rufipennis (Kirby), is an important mortality agent of native spruces throughout North America. The life-cycle duration of this species varies from 1 to 3 years depending temperature. The univoltine cycle (one generation per year) is thought to maximize outbreak risk and accelerate host mortality in established outbreaks. Prepupal diapause is associated with the semivoltine cycle (one generation per 2 years) and we investigated thermal conditions that result in diapause induction. Preliminary experiments used respirometry in an attempt to distinguish the diapause state of experimental insects but the technique was apparently confounded by low respiration before and during pupation, regardless of diapause status. Therefore, diapause induction was deduced using developmental delays. The observed developmental response was not a "switch", with developmental delay either present or absent, but instead varied continuously. We found that temperatures <15°C from instar III through mid-instar IV were associated with developmental delays beyond that expected from cool temperatures. Moreover, the duration of exposure to cool temperatures was important in determining the degree of developmental delay. Small, if any, delays were observed if the cumulative exposure to <15°C was <20 d whereas >40 d cumulative exposure was associated with distinct developmental suppression. Intermediate exposure to cool temperatures resulted in minor developmental delays. We used our results to parameterize a maximum likelihood estimation model of temperature-dependent instar IV developmental rates, including the effect of diapause. This model can be included as part of a spruce beetle phenology model for predicting population dynamics., (Published by Elsevier Ltd.)

Serine peptidase inhibitors (serpins) form a superfamily of proteins covering abroad spectrum of different biological functions. Here we describe the inhibitory characterization of leviserpin, the first serpin from the sugar cane weevil Sphenophorus levis. Leviserpin was able to inhibit bovine trypsin by the formation of the covalent complex serpin-peptidase, demonstrated by SDS-PAGE and mass spectroscopy analysis. We also have determined the cleavage site at the reactive center loop, by the analysis of the polypeptides released from de C-terminus of leviserpin. Moreover we investigated the mRNA expression of leviserpin in different stages of S. levis development. Thus the specificity of leviserpin, in addition with its mRNA coding being transcribed through all lifecycle of the insect, can suggest a possible role in defense mechanism by regulating the action of prophenoloxidase (proPO) activating enzyme.

The invasive red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera, Curculionidae), is one of the most destructive pests of palms in the world. Since its detection in the Mediterranean Basin, the ornamental Phoenix canariensis Hort. ex Chabaud has become its main host. This study was aimed at determining the life cycle of R. ferrugineus in live P. canariensis palms. Egg lethal temperature threshold and thermal constant were determined in the laboratory and resulted in 13.1°C and 40.4 degree days (DD), respectively. A semi field assay was carried out in a mesh enclosure where living P. canariensis palms were artificially infested with neonate larvae at one-month intervals from June 2008 to May 2009 under natural conditions. Infested palms were dissected at different time intervals. Maximum mortality rates for R. ferrugineus were observed for palms infested either in December or January (100%), whereas those infested from April through September showed maximum survival rates. Mean monthly temperatures below 10.3°C were lethal for neonate larvae, as 4.5°C were for older immature stages. All recovered larvae could be classed according to one of 13 instars. A thermal constant of 666.5 DD was estimated for complete larval development. Pupal develoment required an additional 282.5 DD. Based on these results and on the temperatures from 46 climatic stations selected in the Iberian Peninsula, less than one generation per year can be expected in areas with mean annual temperature below 15°C and more than two where mean annual temperature is above 19°C.

Three Sitophilus species (S. granarius L., S. oryzae L., and S. zeamais Mots.) are closely related based on DNA analysis of their endosymbionts. All are seed parasites of cereal crops and important economic pest species in stored grain. The Sitophilus species that currently exist, including these three species, are generally believed to be endemic to Asia's forested areas, suggesting that the first infestations of stored grain must have taken place near the forested mountains of southwestern Asia. Previous archaeological data and historical records suggest that the three species may have been diffused by the spread of Neolithic agriculture, but this hypothesis has only been established for granary weevils in European and southwestern Asian archaeological records. There was little archeological evidence for grain pests in East Asia before the discovery of maize weevil impressions in Jomon pottery in 2004 using the "impression replica" method. Our research on Jomon agriculture based on seed and insect impressions in pottery continued to seek additional evidence. In 2010, we discovered older weevil impressions in Jomon pottery dating to ca. 10 500 BP. These specimens are the oldest harmful insects in the world discovered at archaeological sites. Our results provide evidence of harmful insects living in the villages from the Earliest Jomon, when no cereals were cultivated. This suggests we must reconsider previous scenarios for the evolution and propagation of grain pest weevils, especially in eastern Asia. Although details of their biology or the foods they infested remain unclear, we hope future interdisciplinary collaborations among geneticists, entomologists, and archaeologists will provide the missing details.