Showing papers in "Journal of Insect Science in 2008"

Reference Gene Selection for Insect Expression Studies Using Quantitative Real-Time PCR: The Head of the Honeybee, Apis mellifera, After a Bacterial Challenge

Abstract: In this study an important and often neglected aspect of gene expression studies in insects, the validation of appropriate reference genes with stable expression levels between sample groups, is addressed. Although in this paper the reference gene selection for the honeybee, Apis mellifera L. (Hymenoptera: Apidae) head was tested in the context of bacterial challenge with Escherichia coli, this work can serve as a resource to help select and screen insect reference genes for gene expression studies in any tissue and under any experimental manipulation. Since it is recommended to use multiple reference genes for accurate normalization, we analyzed the expression of eleven candidate reference genes in the honeybee head, for their potential use in the analysis of differential gene expression following bacterial challenge. Three software programs, BestKeeper, Normfinder and geNorm, were used to assess candidate reference genes. GeNorm recommended the use of four reference genes. Both geNorm and Normfinder identified the genes GAPDH, RPS18, actin and RPL13a as the most stable ones, only differing in their ranking order. BestKeeper identified RPS18 as being the reference gene with the least overall variation, but also actin and GAPDH were found to be the second and third most stable expressed gene. By a combination of three software programs the genes actin, RPS18 and GAPDH were found suitable reference genes in the honeybee head in the context of bacterial infection.

Effects of Heat Shock on Survival and Reproduction of Two Whitefly Species, Trialeurodes vaporariorum and Bemisia tabaci Biotype B

Abstract: The effects of heat shock on survival and reproduction of two whitefly species, Trialeurodes vaporariorum (Westwood) and Bemisia tabaci (Gennadius) biotype B (Homoptera: Aleyrodidae), were compared in the laboratory. Whitefly adults were exposed to 26 (control), 37, 39, 41, 43 and 45°C for 1 hour, and were then maintained at 26°C. Adult survival was significantly affected when they were exposed at 41°C or higher for B. tabaci or 39°C or higher for T. vaporariorum. All males of T. vaporariorum were killed at 45°C. In both whitefly species, females were more tolerant to high temperatures at 39°C or higher than males. Female fecundity was not significantly different when B. tabaci adults were heat-shocked at all temperatures. In contrast, the fecundity of T. vaporariorum females declined with the increase of temperature, and only a few eggs were oviposited at 43°C. Survival or hatch rates of the F1 nymphs of both whitefly species declined as heat-shock temperature increased, and no T. vaporariorum nymphs were hatched at 43°C. Similarly, percentages of F1 offspring developing to adults for both whitefly species also declined as the heat-shock temperature increased. Sex ratios of the F1 offspring were not significantly affected for T. vaporariorum but were slightly affected for B. tabaci at 43 and 45°C. The significance of heat shock in relation to dispersal, distribution and population dynamics of the two whitefly species is discussed.

Genetic Variability of the European Corn Borer, Ostrinia nubilalis, Suggests Gene Flow Between Populations in the Midwestern United States

Abstract: The European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae), is a widely distributed and serious economic pest to corn production in the U.S. Genetic variability of O. nubilalis was studied in 18 sub-populations in the upper Midwestern United States using amplified fragment length polymorphism. The relatively low GST values indicate that more variation exists within populations than between populations. High gene flow (Nm) values were indicated across the entire O. nubilalis population; the lowest degree of gene flow was in the northern samples (Nm = 1.96) and the highest degree of gene flow was in the southern samples (Nm = 2.77). The differences observed in the respective regions (north vs. south) may be explained by the voltinism patterns (univoltine vs. multivoltine, respectively) of O. nubilalis: southern multivoltine populations have opportunities for multiple matings for the duration of the year, further mix alleles. AMOVA results also indicated that most of the genetic variation was within sub-populations (≈ 81% of total variation); less variation (≈ 13%) was detected among populations within each of the three regions as designated for this study. However, the most striking and unexpected result was the low percentage of variation between all groups (≈ 6%), further supporting implications of a high degree of gene flow. These results provide support for current requirements of refugia corn planting in Bt-corn management. These results also indicate that if resistance to Bt were to evolve in O. nubilalis, quick action would be necessary to deter the rapid spread of the gene for resistance.

Cloning and phylogenetic analysis of sid-1-like genes from aphids

Abstract: The sid-1 (systemic interference defective) gene encodes a transmembrane protein that is an important participator in the systemic RNAi pathway and has been reported in several organisms. In insects, sid-1-like genes were described from Tribolium castaneum, Apis mellifera, Bombyx mori and Schistocerca americana, but were not found in Drosophila melanogaster and Anopheles gambiae. To investigate whether this gene occurs in aphid species, RT-PCRs were performed using degenerate primers designed using the conserved motif of sid-1-like genes. An sid-1-like full-length transcript was amplified from the cotton/melon aphid, Aphis gossypii Glover (Homopera: Aphididae), and a fragment was amplified from the grain aphid, Sitobion avenae (F.). The trancript from A. gossypii was 3067 bp long, with an open reading frame encoding 766 amino acids. Sequence analysis indicated that this transcript shares highest similarity with the reported sid-1-like gene in Schistocerca americana (53%, fragment), followed by A. mellifera (44%), T. castaneum (32–44%), B. mori (38–42%) and Caenorhabditis elegans (25%). Analysis of the transmembrane protein topological structure indicated that the protein encoded by this gene has a similar structure to SID-1 of C. elegans. A phylogenetic tree with all available sid-1-like genes suggests that sid-1-like genes may have had a long evolutionary history. Considering its importance in the RNAi pathway, the absence of a sid-1-like gene in D. melanogaster and A. gambiae is worthy of further investigation.

Influence of temperature and humidity on the efficacy of spinosad against four stored-grain beetle species

Abstract: In the present work, we examined the insecticidal effect of spinosad, against adults of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), the rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) on wheat and the larger grain borer, Prostephanus truncatus (Horn) (Coleoptera: Bostrychidae) on maize. The dose rates used were 0.01, 0.1, 0.5 and 1 ppm. The bioassays were carried out at three temperatures, 20, 25 and 30°C and two relative humidity levels, 55 and 75%. Mortality of R. dominica and S. oryzae was high even at 0.01 ppm of spinosad, reaching 100% at 55% relative humidity and 30° after 21 days of exposure. Generally, mortality of R. dominica, increased with temperature while for S. oryzae mortality increased with temperature and with the decrease of relative humidity. Moreover, for S. oryzae, mortality was low at 20°C. In the case of T. confusum, mortality was low at doses between 0.01 and 0.5 ppm even after 21 days of exposure. At 1 ppm, mortality exceeded 90% only at 30°C and only after 21 days of exposure. Mortality of P. truncatus was low on maize treated with 0.01 ppm, but increasing the dose to 0.1 ppm resulted in > 87% mortality after 14 days of exposure. In several combinations tested, spinosad efficacy notably varied according to the temperature and humidity regimes. Of the species tested, R. dominica and P. truncatus were very susceptible to spinosad, followed by S. oryzae, while T. confusum was the least susceptible.

Diversity of forest litter-inhabiting ants along elevations in the Wayanad region of the Western Ghats

Abstract: Litter ant diversity and abundance in relation to biotic and abiotic factors were analyzed at five primary forest sites lying between 300 to 1650 meter above mean sea level in the Wayanad region of the Western Ghats in Kerala, southern India. Ant abundance and species richness peaked at mid-elevations influenced by the presence of favourable physical conditions and abundance of prey resources. Dominance of ants preferring termites and Collembola as prey at sites rich in their specific prey resources indicate the influence of local prey resource availability in determining ant distribution. Dominant species (Tapinoma sp. and Solenopsis sp.) had wider distributions, being present at all elevations. Physical factors (slope of the terrain, rainfall, moisture, humidity, temperature) and prey resource availability (insect larvae, termites, Collembola) influenced ant species abundance at a regional scale, whereas at local scales, site specific variations in the relationship between abundance of ants and prey-predators and physical factors were recorded. The present study highlights the need to consider site-specific abiotic and biotic factors while examining the distribution patterns of litter ants along altitudinal gradients in other regions of the Western Ghats, which is a recognised hot spot of biodiversity with wide regional variation in vegetation types and faunal distribution patterns.

Latitudinal Body Size Clines in the Butterfly Polyommatus icarus are Shaped by Gene-Environment Interactions

Abstract: The study of latitudinal body size clines can illuminate processes of local adaptation, but there is a need for an increased understanding of the relative roles of genetic variation, environmental effectstions or this reason, we combined an investigation of a museum collection of the common blue butterfly Polyommatus icarus (Rottemburg) (Lycaenidae: Polyommatini) from Sweden with a common-garden experiment in the laboratory, using strains reared from individuals collected from three different latitudes. Sizes of the field-collected butterflies tended to smoothly decrease northwards in a latitudinal cline, but suddenly increase at the latitude where the life cycle changes from two to one generations per year, hence allowing more time for this single generation. Further north, the size of the field-collected butterflies again decreased with latitude (with the exception of the northernmost collection sites). This is in accordance with the “converse Bergmann” pattern and with the “saw-tooth model” suggesting that insect size is shaped by season length and number of generations along latitudinal transects. In contrast, under laboratory conditions with a constant long day-length there was a different pattern, with the butterflies pupating at a higher mass when individuals originated from southern populations under time stress to achieve a second generation. This is indirect evidence for field patterns being shaped by end-of-season cues cutting development short, and also suggests counter-gradient variation, as butterflies from the time-stressed populations over-compensated for decreasing larval development time by increasing their growth rates, thus obtaining higher mass. Hence, we found support for both adaptive phenotypic plasticity and local genetic adaptation, with gene-environment interactions explaining the observed field patterns.

Extensive natural intraspecific variation in stoichiometric (C:N:p) composition in two terrestrial insect species.

Abstract: Heterotrophic organisms must obtain essential elements in sufficient quantities from their food. Because plants naturally exhibit extensive variation in their elemental content, it is important to quantify the within-species stoichiometric variation of consumers. If extensive stoichiometric variation exists, it may help explain consumer variation in life-history strategy and fitness. To date, however, research on stoichiometric variation has focused on interspecific differences and assumed minimal intraspecific differences. Here this assumption is tested. Natural variation is quantified in body stoichiometry of two terrestrial insects: the generalist field cricket, Gryllus texensis Cade and Otte (Orthoptera: Gryllidae) and a specialist curculionid weevil, Sabinia setosa (Le Conte) (Coleoptera: Curculionidae). Both species exhibited extensive intraspecific stoichiometric variation. Cricket body nitrogen content ranged from 8–12% and there was a four-fold difference in body phosphorus content, ranging from 0.32–1.27%. Body size explained half this stoichiometric variation, with larger individuals containing less nitrogen and phosphorus. Weevils exhibited an almost three-fold difference in body phosphorus content, ranging from 0.38–0.97%. Overall, the variation observed within each of these species is comparable to the variation previously observed across almost all terrestrial insect species.

Alkalinization in the Isolated and Perfused Anterior Midgut of the Larval Mosquito, Aedes aegypti

Abstract: In the present study, isolated midguts of larval Aedes aegypti L. (Diptera: Culicidae) were mounted on perfusion pipettes and bathed in high buffer mosquito saline. With low buffer perfusion saline, containing m-cresol purple, transepithelial voltage was monitored and luminal alkalinization became visible through color changes of m-cresol purple after perfusion stop. Lumen negative voltage and alkalinization depended on metabolic energy and were stimulated in the presence of serotonin (0.2 micromol l(-1)). In some experiments a pH microelectrode in the lumen recorded pH values up to 10 within minutes after perfusion stop. The V-ATPase inhibitor concanamycin (50 micromol l(-1)) on the hemolymph side almost abolished V(te) and inhibited luminal alkalinization. The carbonic anhydrase inhibitor, methazolamide (50 micromol l(-1)), on either the luminal or hemolymph-side, or the inhibitor of anion transport, DIDS (1 mmol l(-1)) on the luminal side, had no effect on V(te) or alkalinization. Cl(-) substitution in the lumen or on both sides of the tissue affected V(te), but the color change of m-cresol purple was unchanged from control conditions. Hemolymph-side Na(+) substitution or addition of the Na(+)/H(+) exchange inhibitor, amiloride (200 micromol l(-1)), reduced V(te) and luminal alkalinization. Luminal amiloride (200 micromol l(-1)) was without effects on V(te) or alkalinization. High K(+) (60 mmol l(-1)) in the lumen reduced V(te) without affecting alkalinization. These results indicate that strong luminal alkalinization in isolated and perfused anterior midgut of larval A. aegypti depends on basolateral V-ATPase, but is apparently independent of carbonic anhydrase, apical Cl(-)/HCO(3)(-) exchange or apical K(+)/2H(+) antiport.

Life History, Aggregation and Dormancy of the Rubber Plantation Litter Beetle, Luprops tristis, from the Rubber Plantations of Moist South Western Ghats

Abstract: Life history, aggregation and dormancy of rubber plantation litter beetle Luprops tristis Fabricius, (Tenebrionidae: Coleoptera) is described from rubber plantation belts in the western slopes of Western Ghats from the south Indian state of Kerala. The life cycle lasted 12 months, including the 5 larval instars lasting 1 month, the 3 day pupal stage, and the adult stage that can last 11 months. The adult stage includes an inactive dormancy phase of 9 months in shelters and 1 month each of active pre-dormancy (feeding) and post-dormancy (feeding and reproduction) phases that occur in rubber plantation litter. Reproductive activities are confined to the post-dormancy phase. With the onset of summer rains, huge aggregations of adults invade residential buildings and enter into a state of dormancy for 9 months. Beetle aggregations were in the range of 0.5 million to 4.5 million individuals per residential building. Dormancy in L. tristis is best classified as oligopause, which is intermediate between quiescence and diapause. Adults and larvae feed preferentially on wilted rubber tree leaves. Age-specific variation in mortality during dormancy is distinct with higher survivability for adults that have a longer pre-dormancy period. Generations are non-overlapping.

Phylogenetic Relationships of Tribes Within Harpalinae (Coleoptera: Carabidae) as Inferred from 28S Ribosomal DNA and the Wingless Gene

Abstract: Harpalinae is a large, monophyletic subfamily of carabid ground beetles containing more than 19,000 species in approximately 40 tribes. The higher level phylogenetic relationships within harpalines were investigated based on nucleotide data from two nuclear genes, wingless and 28S rDNA. Phylogenetic analyses of combined data indicate that many harpaline tribes are monophyletic, however the reconstructed trees showed little support for deeper nodes. In addition, our results suggest that the Lebiomorph Assemblage (tribes Lebiini, Cyclosomini, Graphipterini, Perigonini, Odacanthini, Lachnophorini, Pentagonicini, Catapiesini and Calophaenini), which is united by a morphological synapomorphy, is not monophyletic, and the tribe Lebiini is paraphyletic with respect to members of Cyclosomini. Two unexpected clades of tribes were supported: the Zuphiitae, comprised of Anthiini, Zuphiini, Helluonini, Dryptini, Galeritini, and Physocrotaphini; and a clade comprised of Orthogoniini, Pseudomorphini, and Graphipterini. The data presented in this study represent a dense sample of taxa to examine the molecular phylogeny of Harpalinae and provide a useful framework to examine the origin and evolution of morphological and ecological diversity in this group.

Conglobation in the Pill Bug, Armadillidium vulgare, as a Water Conservation Mechanism

Abstract: Water balance of the terrestrial isopod, Armadillidium vulgare, was investigated during conglobation (rolling-up behavior). Water loss and metabolic rates were measured at 18 ± 1°C in dry air using flow-through respirometry. Water-loss rates decreased 34.8% when specimens were in their conglobated form, while CO2 release decreased by 37.1%. Water loss was also measured gravimetrically at humidities ranging from 6 to 75 %RH. Conglobation was associated with a decrease in water-loss rates up to 53 %RH, but no significant differences were observed at higher humidities. Our findings suggest that conglobation behavior may help to conserve water, in addition to its demonstrated role in protection from predation.

Molecular cloning and expression of three polygalacturonase cDNAs from the tarnished plant bug, Lygus lineolaris.

Abstract: Three unique cDNAs encoding putative polygalacturonase enzymes were isolated from the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae). The three nucleotide sequences were dissimilar to one another, but the deduced amino acid sequences were similar to each other and to other polygalacturonases from insects, fungi, plants, and bacteria. Four conserved segments characteristic of polygalacturonases were present, but with some notable semiconservative substitutions. Two of four expected disulfide bridge—forming cysteine pairs were present. All three inferred protein translations included predicted signal sequences of 17 to 20 amino acids. Amplification of genomic DNA identified an intron in one of the genes, Llpg1, in the 5′ untranslated region. Semiquantitative RT-PCR revealed expression in all stages of the insect except the eggs. Expression in adults, male and female, was highly variable, indicating a family of highly inducible and diverse enzymes adapted to the generalist polyphagous nature of this important pest.

Methodology for Assessing Rice Varieties for Resistance to the Lesser Grain Borer, Rhyzopertha dominica

Abstract: Several physical and chemical attributes of rice were evaluated to determine which character would be best to use to assess multiple rice varieties for resistance to the lesser grain borer, Rhyzopertha dominica (F.). Laboratory tests were conducted on single varieties of long-, short-, and medium grain-rice to develop procedures and methodologies that could be used for large-scale screening studies. Progeny production of R. dominica was positively correlated with the percentage of broken hulls. Although kernel hardness, amylose content, neonate preference for brown rice, and adult emergence from neonates varied among the three rice varieties tested they did not appear to be valid indicators of eventual progeny production, and may not be useful predictors of resistance or susceptibility. Soundness and integrity seem to be the best characters to use for varietal screening studies with R. dominica.

Production of Beauveria bassiana fungal spores on rice to control the coffee berry borer, Hypothenemus hampei, in Colombia

Abstract: Two isolates of fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) were grown on cooked rice using diphasic liquid-solid fermentation in plastic bags to produce and harvest spore powder. The cultures were dried and significant differences were found for isolates and time of harvest. The spores were harvested manually and mechanically and after the cultures were dried for nine days, when moisture content was near 10%. After harvesting, spores were submitted to quality control to assess concentration, germination, purity, moisture content, particle size and pathogenicity to the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae). Spore productivity on cooked rice was less than 1×1010 spores/g using both manually and mechanically harvesting methodologies. Germination at 24 hours was over 75% and pathogenicity against H. hampei was over 92.5%. This methodology is suitable for laboratory and field studies, but not for industrial production when a high concentration of spores are required for formulation and field applications.

Ovicidal Activity of Organophosphate, Oxadiazine, Neonicotinoid and Insect Growth Regulator Chemistries on Northern Strain Plum Curculio, Conotrachelus nenuphar

Abstract: An in vitro method was developed for assessing ovicidal effects of the organophosphate azinphos-methyl, the neonicotioids thiacloprid, thiamethoxam and clothianidin, the oxadiazine indoxacarb and the insect growth regulators novaluron and pyriproxifen on the plum curculio, Conotrachelus nenuphar (Herbst)(Coleoptera: Curculionidae). The baseline survivorship of this method was 88 percent. Plum curculio eggs were most sensitive to azinphos-methyl. Thiacloprid, clothianidin and the chitin synthesis inhibitor, novaluron, had LC50 values below 100 ppm. Thiamethoxam, indoxacarb and pyriproxifen were not ovicidal at 100 ppm. Octanol-water partitioning coefficients, log Kow, appeared to be an important indicator of ovicidal activity within the neonicotinoids. This new bioassay method eliminates the confounding of the insect-chemical and plant-chemical interactions and the results highlight the utility of a post-infestation curative approach to plum curculio management.

Repellent Properties of δ-Octalactone Against the Tsetse Fly, Glossina morsitans Morsitans

Abstract: δ-octalactone, produced by several Bovidae, has been suggested as a potential repellant of tsetse fly attack. Racemic δ-octalactone was synthesized via an abbreviated route. The product was assayed against 3-day old starved teneral female tsetse flies, Glossina morsitans morsitans Wiedemann (Diptera: Glossinidae), in a choice wind tunnel and found to be a potent tsetse repellent at doses ≥0.05 mg in 200 µl of paraffin oil (0.05 >p >0.01).

The effects of hydration on growth of the house cricket, Acheta domesticus

Abstract: Maintenance of biochemical gradients, membrane fluidity, and sustained periods of activity are key physiological and behavioral functions of water for animals living in desiccating environments. Water stress may reduce the organism's ability to maintain these functions and as such, may reduce an organism's growth. However, few studies have examined this potential effect. The effects of altered hydration state of the house cricket, Acheta domesticus L. (Orthoptera: Gryllidae) on individual growth were studied under laboratory conditions. Crickets were permitted access to water for three different durations each day, resulting in significant differences in hydration state (32% greater hydration for maximum than minimum duration of water availability). Growth was 59% and 72% greater in dry mass and length, respectively, between the lowest and highest hydration state treatments. These findings may be representative for a variety of animal species and environments and could have important ecological implications.

A rapid decision sampling plan for implementing area–wide management of the red palm weevil, Rhynchophorus ferrugineus , in coconut plantations of India

Abstract: The red palm weevil Rhynchophorus ferrugineus Olivier (Curculionidae/Rhynchophoridae/Dryophthoridae) is a lethal pest of young coconut palms, Cocos nucifera L. (Arecales: Arecaceae), with a highly aggregated population distribution pattern. R. ferrugineus is managed in several coconut growing countries using area-wide pheromone based programmes that need a substantial commitment of funds over a period of time. Often, decisions to implement area-wide management of R. ferrugineus are based on pheromone trap captures in surveillance traps and or infestation reports. Implementing area-wide management of this pest on the basis of such data can be inaccurate, as it may either under or over estimate the pest intensity in the field. This study presents sampling plans for rapid and accurate classification of R. ferrugineus infestation in coconut plantations of India by inspecting palms to detect infestation in a sequence until a decision to either implement or not to initiate area-wide management of R. ferrugineus can be made. The sampling plans are based on a common aggregation index of 3.45, assumed action threshold values of either 1.0 (plan A) or 0.5 (plan B) per cent infested palms and a risk factor of making the wrong decision set at 0.05. Using plans A and B, if the cummulative number of infested palms in a young 1 hectare coconut plantation is zero out of 150 palms for both plans, then area-wide management is not required, while on the other hand, if the cummulative number of infested palms for the same area is 6 (plan A), or 5 (plan B), then area-wide management of R. ferrugineus is essential. The proposed sampling plans are efficient tools in decision making, particularly at very low and high levels of infestation and can also be used to assess the performance of R. ferrugineus IPM programmes that are in progress. These plans not only save time and money as only a small area needs to be sampled to arrive at a correct decision, but are also efficient in rating the infestation level accurately.

Assessment of geographic and host-associated population variations of the carob moth, Ectomyelois ceratoniae, on pomegranate, fig, pistachio, and walnut, using AFLP markers

Abstract: The carob moth, Ectomyelois ceratoniae (Zeller 1839) (Lepidoptera: Pyralidae) is the most important pest of pomegranate, Punica granatum L. (Myrtales: Ponicaceae), in Iran. In this study, 6 amplified fragment length polymorphism primer combinations were used to survey the genetic structure of the geographic and putative host-associated populations of this pest in Iran. An AMOVA was performed on test populations. Pairwise differences, Mantel test, multidimensional analysis, cluster analysis and migration rate were calculated for 5 geographic populations of E. ceratoniae sharing the same host, pomegranate. In another part of the study, 3 comparisons were performed on pairwise populations that were collected on different hosts (pomegranate, fig, pistachio and walnut) in same geographic regions. The results showed high within population variation (85.51% of total variation), however geographic populations differed significantly. The Mantel test did not show correlations between genetic and geographic distances. The probable factors that affect genetic distances are discussed. Multidimensional scaling analysis, migration rate and cluster analysis on geographic populations showed that the Arsanjan population was the most different from the others while the Saveh population was more similar to the Sabzevar population. The comparisons didn't show any host fidelity in test populations. It seems that the ability of E. ceratoniae to broaden its host range with no fidelity to hosts can decrease the efficiency of common control methods that are used on pomegranate. The results of this study suggest that in spite of the effects of geographic barriers, high within-population genetic variation, migration rate and gene flow can provide the opportunity for emerging new phenotypes or behaviors in pest populations, such as broadening host range, changing egg lying places, or changing over-wintering sites to adapt to difficult conditions such as those caused by intensive control methods.

The basic helix-loop-helix transcription factor family in the honey bee, Apis mellifera.

Abstract: The basic helix-loop-helix (bHLH) transcription factors play important roles in a wide range of developmental processes in higher organisms. bHLH family members have been identified in a dozen of organisms including fruit fly, mouse and human. In this study, we identified 51 bHLH sequences in silico in the honey bee, Apis mellifera L. (Hymenoptera: Apidae), genome. Phylogenetic analyses revealed that they belong to 38 bHLH families with 21, 11, 9, 1, 8 and 1 members in high-order groups A, B, C, D, E and F, respectively. Using phylogenetic analyses, all of the 51 bHLH sequences were assigned to their corresponding families. Genes that encode ASCb, NeuroD, Oligo, Delilah, MyoRb, Figa and Mad were not found in the honey bee genome. The present study provides useful background information for future studies using the honey bee as a model system for insect development.

Diaspididae (Hemiptera: Coccoidea) of Espírito Santo, Brazil.

Abstract: Twenty-seven species of armored scale insects (Hemiptera: Diaspididae) are newly recorded from Espirito Santo, Brazil, and information on the host plants and geographic distribution of the 31 species of Diaspididae that have been identified in the State is provided. New plant host records are reported for 11 of the diaspidid species studied and results are discussed with respect to development of agriculture in this and similar areas with objectives of modernization and diversification.

Effect of plant structure on searching strategy and searching efficiency of Trichogramma turkestanica.

Abstract: When searching for hosts on a plant, female parasitoids use strategies to maximize efficiency. Searching strategies include the expressed behaviors, the time budget associated with each behavior, the time allocated to the different plant parts and the exploration sequence of plant parts. Searching efficiency refers to the time taken to find the first egg, the number of eggs found per foraging time unit and the re-encountering frequency of eggs during a foraging period. This study examines the effect of artificial simple (few leaves and connections) and complex plant structures (more leaves and connections) on searching strategy and searching efficiency of the egg parasitoid Trichogramma turkestanica Meyer (Hymenoptera: Trichogrammatidae). Analyses of frequency and duration of behaviors associated with searching on artificial plants of different complexities were performed. Plant structure had no effect on time associated with locomotion behaviors such as walking, standing and flying. However, it had an impact on the area searched, which was significantly greater on simple plant structure. Also, time spent on a leaf without encountering an egg was greater on complex plant structure compared to simple one. No significant differences were found between simple and complex plant structures regarding time spent walking on the different plant parts such as twigs, limbs, leaf perimeters, and limbs of inferior and superior leaf sides. Results showed that female parasitoids spent less time actively exploring complex than simple plants. Encountering and re-encountering frequencies of eggs were significantly greater on simple than on complex plant structure. Plant structure had no effect on handling time of eggs. This study demonstrates that plant structure can modulate activities inherent to searching and ovipositing, which in turn affects area searched per foraging time unit and therefore host finding success.

Effects of Vegetated Field Borders on Arthropods in Cotton Fields in Eastern North Carolina

Abstract: The influence, if any, of 5m wide, feral, herbaceous field borders on pest and beneficial arthropods in commercial cotton, Gossypium hirsutum (L.) (Malvales: Malvaceae), fields was measured through a variety of sampling techniques over three years. In each year, 5 fields with managed, feral vegetation borders and five fields without such borders were examined. Sampling was stratified from the field border or edge in each field in an attempt to elucidate any edge effects that might have occurred. Early season thrips populations appeared to be unaffected by the presence of a border. Pitfall sampling disclosed no differences in ground-dwelling predaceous arthropods but did detect increased populations of crickets around fields with borders. Cotton aphid (Aphis gossypii Glover) (Hemiptera: Aphididae) populations were too low during the study to adequately assess border effects. Heliothines, Heliothis virescens (F.) and Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), egg numbers and damage rates were largely unaffected by the presence or absence of a border, although in one instance egg numbers were significantly lower in fields with borders. Overall, foliage-dwelling predaceous arthropods were somewhat more abundant in fields with borders than in fields without borders. Tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois) (Heteroptera: Miridae) were significantly more abundant in fields with borders, but stink bugs, Acrosternum hilare (Say), and Euschistus servus (Say) (Hemiptera: Pentatomidae) numbers appeared to be largely unaffected by border treatment. Few taxa clearly exhibited distributional edge effects relative to the presence or absence of border vegetation. Field borders like those examined in this study likely will have little impact on insect pest management in cotton under current insect management regimens.

Status of insecticide susceptibility in Anopheles gambiae sensu lato and Anopheles funestus mosquitoes from western Kenya.

Abstract: The status of resistance was investigated in Anopheles gambiae sensu lato and An. funestus (Diptera: Culicidae) mosquitoes from western Kenya to four classes of insecticides approved by World Health Organization for indoor residual spraying. The prevalence of the knockdown-resistance (kdr) mutation associated with resistance to pyrethroids and DDT was determined in An. gambiae s.l.. Standard World Health Organization diagnostic bioassay kits for DDT (an organochlorine), fenitrothion (an organophosphate), bendiocarb (a carbamate), and the pyrethoirds, lambdacyhalothrin and permethrin, were used. Knockdown every 10 min and mortality 24 h after exposure were noted. Controls not treated with insecticides and with the susceptible An. gambiae KISUMU strain were included in the bioassays. The presence of the kdr gene was determined using a standard diagnostic polymerase chain reaction assay. Over 98% mortality was observed for tests with all insecticides for both An. gambiae s.l. and An. funestus. Knockdown rates were not significantly different between An. gambiae s.l. and the KISUMU strain control. 50% and 95% knockdown times were either slightly lower than those for the KISUMU strain or higher by factors of less than 1.6. The mean frequency of the East African kdr mutation was 24.7% in An. gambiae sensu strictu. Based on conventional criteria where susceptibility is defined by mortality rates >98% 24 h after exposure, no evidence for resistance was found, implying that vector control measures employing any of the insecticides tested would be unhampered by resistance. The observed frequencies of the kdr mutation do not appear to compromise the effectiveness of the insecticides. The need for continuous monitoring of the status of insecticide resistance and of the impact of any observed resistance on the efficacy of vector control programs employing insecticides is apparent.

Molecular phylogenetic relationships of flightless beetles belonging to the genus Mesechthistatus Breuning, (Coleoptera: Cerambycidae) inferred from mitochondrial COI gene sequences.

Abstract: The longicorn beetles belonging to the genus MesechthistatusBreuning., 1950 (Coleoptera: Cerambycidae) cannot fly since their hindwings are atrophied. This slows down gene flow between local populations. Currently, it is considered that the genus contains four endemic species from the eastern Honshu Is., Japan, M. binodosus, M. furciferus, M. taniguchii and M. fujisanus, that are distributed parapatrically. Sequence analyses of the cytochrome oxidase subunit I gene suggests that lineages of mitochondrial haplotypes split approximately in the same era. However, this result is not consistent with the monophyly of morphological species. The estimated evolutionary rate of the COI gene in other insects suggests that mitochondrial haplotypes of Mesechthistatus differentiated at the end of the Pliocene epoch during the Tertiary era.

Susceptibility of Eggs and Adult Fecundity of the Lesser Grain Borer, Rhyzopertha dominca, Exposed to Methoprene

Abstract: A series of tests were conducted to determine the susceptibility of eggs and neonates of the lesser grain borer Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae = Bostrychidae), exposed to the insect growth regulator, methoprene, on filter paper and on rough rice. In the first test, the hatch rate of eggs exposed on filter paper treated with methoprene at the label rate of 0.003 mg [AI]/cm2 when used as a surface treatment in structures was 52.0 ± 7.3% compared to 93.0 ± 3.3% on untreated controls. In the second test, eggs were exposed to a dose-response series of 0.00003 to 0.03 mg[AI]/cm2. Egg hatch was directly proportional to concentration and ranged from 85.0 ± 2.0% on untreated controls to 26.7 ± 8.3% at the highest concentration tested. In the third test, 1 ppm of methoprene was sprayed on long grain rough rice (paddy) (Cocodrie variety), and then individual kernels were cracked and an egg of R. dominica was placed directly on the kernel. On untreated rice kernels, 67.5 ± 11.6% of the eggs hatched and were able to bore inside, and all of these larvae emerged as adults. In contrast, 40.0 ± 5.3% of the eggs placed on treated cracked kernels were able to develop to where the larvae were visible through X-ray detection, but none emerged as adults. In the final test, newly-emerged adults were exposed on rough rice treated with 1 ppm methoprene. The number of eggs from adults on untreated rice was 52.1 ± 4.3 eggs per female, and on treated rice the average egg production was 12.5 ± 1.1 eggs per female. Methoprene applied on a surface or on rough rice affected development of egg hatch also reduced fecundity of parent adults exposed on the treated rough rice.

Habitat Preferences of Butterflies in the Bumbuna Forest, Northern Sierra Leone

Abstract: The habitat preferences of the butterfly fauna were studied in the Bumbuna Forest Reserve in northern Sierra Leone. The intact forest reserve and a secondary forest regrowth, disturbed as a result of slash-and-burn agriculture, were compared to savanna habitats. Of the 290 specimens collected, 195 butterfly species were included, of which significant proportion were Nymphalidae. Of the 147 forest species, 111 (75.5%) showed preferences for the forest habitats, while 70 (47.6%) and 34 (23.1%) preferred disturbed and savannah habitats, respectively. Numerically, a comparable proportion of savannah species were recorded in the 18 disturbed (73.9%) and 16 savannah habitats (63.2%). Accumulated species richness and diversity indices were lower in the disturbed habitats compared to the forest reserve, but lowest in the savanna habitats. However, a large proportion of forest species, especially those with either a more restricted geographic range or species for which no information on geographic distribution was available, were exclusively captured in the forest patches. The survey indicated the presence of a rich butterfly fauna, which should be systematically collected for further research and study in order to build a good taxonomic database for Sierra Leone.

Odorant Receptor C-Terminal Motifs in Divergent Insect Species

Abstract: Insect odorant receptors are a large family of seven transmembrane proteins believed to be G-protein coupled receptors. The peptide sequences of two odorant receptors within a given species may share as little as 17% identity, and there is limited similarity between receptors of divergent species. One exception is DmOr83b, which is found in Drosophila melanogaster and is highly conserved in at least ten other insect species. DmOr83b is broadly expressed in most of the olfactory sensory neurons of D. melanogaster at most developmental stages, while other odorant receptors tend to have more restricted and specific expression patterns. DmOr83b is critical for D. melanogaster olfaction, and it is involved in properly localizing other odorant receptors possibly by forming heterodimers with these receptors. The C-terminal region has been implicated as sites for such heterodimer formation. Multiple em for motif elicitation (MEME), a hidden markov model based program, was used to uncover three conserved motifs in the C-termini of a vast majority of the odorant receptor peptides from Anopheles gambiae, D. melanogaster, and Apis mellifera. These motifs are also found in DmOr83b and its orthologs and the order of these motifs is conserved as well. The conservation of these motifs among divergent odorant receptors in divergent species suggests functional importance. We propose that these motifs are involved in receptor- receptor protein interactions, contributing to the heterodimer formation between DmOr83b (or its orthologs) and other odorant receptors.

Magnetic resonance microscopy of flows and compressions of the circulatory, respiratory, and digestive systems in pupae of the tobacco hornworm, Manduca sexta

Abstract: Circulatory, respiratory, and digestive motions in Manduca sexta pupae were observed using proton-density weighted and fast-imaging with steady-state free procession magnetic resonance microscopy. Proton-density weighted images clearly differentiated pupal air sacs from the hemolymph and organs because, as expected, the air sacs appeared dark in these images. Steady-state free procession imaging allowed real-time monitoring of respiration and circulation, creating movies of hemolymph circulation. Some of the movies show compression and inflation of the air sacs as well as abdominal movements consistent with previously reported ceolopulses. To our knowledge, this is the first magnetic resonance microscopy study of insect circulation and respiration and these preliminary results demonstrate the potential of magnetic resonance microscopy for studying in vivo dynamic processes in insects.