Prevalence of Endosymbionts in Polish Populations of Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

Colorado potato beetle (CPB, Leptinotarsa decemlineata Say) (Coleoptera: Chrysomelidae) is one of the most serious insect pest feeding on wild and cultivated Solanaceae plants. This pest poses a significant threat to potato crops. CPB originated from North America but has become widespread and has adapted in new localizations. Currently, it is reported in many countries worldwide. Endosymbiotic bacteria might have an influence on insect adaptation to new conditions. They are known to play a role in invasiveness of insect hosts and to facilitate colonization of new niches; however, information on endosymbionts of the CPB is very limited. In this study, we screened CPB populations collected from 20 evenly distributed locations in Poland for the presence of Arsenophonus, Cardinium, Wolbachia, and Flavobacterium. We found the presence of Flavobacterium in the studied insects. Little is known about CPB–endosymbionts interactions, thus this study may provide a reference for future studies in this subject.     

The Glycolytic Enzymes Activity in the Midgut of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) adult and their Seasonal Changes

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) is an important pest of maize. The diet of the D. virgifera imago is rich in starch and other polysaccharides present in cereals such as maize. Therefore, knowledge about enzymes involved in digestion of such specific food of this pest seems to be important. The paper shows, for the first time, the activities of main glycolytic enzymes in the midgut of D. virgifera imago: endoglycosidases (α-amylase, cellulase, chitinase, licheninase, laminarinase); exoglycosidases (α- and β-glucosidases, α- and β-galactosidases) and disaccharidases (maltase, isomaltase, sucrase, trehalase, lactase, and cellobiase). Activities of α-amylase, α-glucosidase, and maltase were the highest among assayed endoglycosidases, exoglycosidases, and disaccharidases, respectively. This indicates that in the midgut of D. virgifera imago α-amylase, α-glucosidase and maltase are important enzymes in starch hydrolysis and products of its digestion. These results lead to conclusion that inhibition of most active glycolytic enzymes of D. virgifera imago may be another promising method for chemical control of this pest of maize.     

Efficacy of Plectranthus glandulosus (Lamiaceae) and Callistemon rigidus (Myrtaceae) Leaf Extract Fractions to Callosobruchus maculatus (Coleoptera: Bruchidae)

As part of on-going efforts to use eco-friendly alternatives to chemical pesticides, methanol crude extracts of Plectranthus glandulosus and Callistemon rigidus leaves were sequentially fractionated in hexane, chloroform, ethyl acetate, and methanol to establish the most active fraction(s) against Callosobruchus maculatus in cowpea. Cowpea seeds (25 g) were treated with 0.5, 1, 2, and 4 g/kg of extract to evaluate the contact toxicity and F₁ progeny production of the beetles in the laboratory. Mortality was recorded 1, 3, and 7 d postexposure. P. glandulosus hexane fraction was more toxic than the other fractions recording 100% mortality at 4 g/kg, within 7 d with LC₅₀ of 0.39 g/kg. Hexane fraction of C. rigidus showed superior toxicity, causing 100% mortality at 4 g/kg within only 1 d of exposure with LC₅₀ of 1.02 g/kg. All the fractions greatly reduced progeny emergence, with C. rigidus hexane fraction being the best progeny inhibitor. Fractions of P. glandulosus and C. rigidus leaves had sufficient efficacy to be a component of storage pest management package for C. maculatus.     

Population Dynamics of Empoasca fabae (Hemiptera: Cicadellidae) in Central Iowa Alfalfa Fields

Adults and nymphs of Empoasca fabae Harris (Hemiptera: Cicadellidae) and adults of predatory species in the families Coccinellidae, Anthocoridae, Nabidae, Chrysopidae, and Hemerobiidae were sampled in Iowa alfalfa fields from June to September in 1999 and 2000. The relationship between each predatory taxa and E. fabae was examined using regression analysis. In 2000, all predators were found to be positively correlated with the presence of E. fabae during all periods sampled and most likely contributed to mortality. Orius insidiosus (Say) (Hemiptera: Anthoridae) was the most numerous insect predatory species; population numbers ranged from 0 to 1 and 0.1 to 3.7 adults per 0.25 m² in 1999 and 2000, respectively. Partial life tables were constructed for E. fabae nymphs for two alfalfa-growing periods. Nymphs were grouped into three age intervals: first and second, third and fourth, and fifth instars. For the first alfalfa growing period examined, E. fabae nymphal mortality was 70% in 1999 and 49% in 2000. During the last growing period of each season (August–September), total nymphal mortality was relatively low (<25%). Adult E. fabae density ranged from 5.4 to 25.6 and 1.4–9.2 per 0.25 m² in 1999 and 2000, respectively. E. fabae population peaks were similar for each age interval in all growing periods. This study provides further information on the population dynamics of E. fabae and its relationship with select predatory species in Iowa alfalfa fields.     

Variation in C:N:S Stoichiometry and Nutrient Storage Related to Body Size in a Holometabolous Insect (Curculio davidi) (Coleoptera: Curculionidae) Larva

Body size can be an important factor controlling consumer stoichiometry. In holometabolous insects, body size is typically associated with nutrient storage. Consumer stoichiometry is known to vary within species across a range of body sizes; however, the contribution of nutrient storage to this variation is not well understood. We used the fifth-instar larvae of the oak weevil (Coleoptera: Curculio davidi Fairmaire), which is characterized by a high capacity for nutrient storage, to investigate the effect of shifts in nutrient storage with body mass on variations in larva stoichiometry. Our results showed that weevil larvae with larger body mass had a lower carbon (C) content, reflecting decreases in the sequestration rate of C-rich lipids. Larger larvae had elevated concentrations of nitrogen (N), sulfur (S), and protein. The similar patterns of variation in elemental composition and macromolecule storage with body weight indicate that the shift in nutrient storage is the main factor causing the variation in larval stoichiometry with body weight. This finding was further supported by the low variation in residual larval biomass C, N, and S concentrations after lipid extraction. These results help decipher the physiological mechanism of stoichiometric regulation in growing organisms.     

Yield response of okra, Abelmoschus esculentus (L.) Moench to leaf damage by the flea beetle, Podagrica uniforma Jacoby (Coleoptera: Chrysomelidae)

Yield-related response of okra plants, Abelmoschus esculentus (L.) Moench, to artificial infestation of the flea beetle (Podagrica uniforma Jac.) at different densities (0, 5, 10, 20, 30, and 40 pairs per cage) was studied in screen house and field experiments. In both experiments, increase in beetle density resulted in a significant reduction (P < 0.05) in fruit production, fruit length, fruit width, fresh fruit weight, number of seeds per fruit, 100 seed weight and fresh fruit yield. In addition, higher densities caused more dry matter accumulation in the seeds than in the husk of okra fruits. Compensation was noticed at the 5- and 10-pair levels of infestation in some of the variables measured. Fresh fruit yield reduction was more than 50% when beetle density was increased beyond 20 pairs per cage in both experiments. The lowest density of P. uniforma at which significant reduction (P < 0.05) occurred in fresh fruit yield per cage, when compared with the control, was the 20-pair level, representing the damage threshold of the beetle at which initiation of control measures would be justified. Regression analysis indicated that flea beetle density was linearly associated with fruit damage and fresh fruit yield. Also, chi-square analysis showed that the models derived from the screen house and field experiments were not significantly different (P < 0.05) from each other, and either could be used for prediction purposes.     

Antennal Sensilla in the Parasitoid Sclerodermus sp. (Hymenoptera: Bethylidae)

Parasitoid wasps of the genus Sclerodermus (Hymenoptera: Bethylidae) are an important natural enemy of the Japanese pine sawyer beetle Monochamus alternatus Hope (Coleoptera: Cerambycidae). In this study, we used scanning electron microscopy to examine the external morphology of the antennal sensilla of Sclerodermus sp. Antennae of females and males comprised the scape, pedicel, and 11 flagellomere segments. Based on the morphology of the sensilla in each sex, seven types of sensillum were identified: sensilla trichodea (Tr.1, Tr.2 and Tr.3), sensilla basiconica (Ba.1, Ba.2, and Ba.3), sensilla styloconica (St.1 and St.2), sensilla placodea, sensilla coeloconica, sensilla squamiforma, and Bohm’s bristles. Tr.2, Ba.1, and St.1 were only found in females, whereas Ba.2, Ba.3, and St.2 were only observed in males. Sensilla placodea were the most common, given that they occur on the antennae of many parasitoid Hymenoptera, whereas sensilla Tr were the most abundant, being distributed over the entire antennal surface. These sensilla are likely to have roles in the host locating and habitat searching behavior of adult Sclerodermus wasps. Therefore, our findings provide a basis for further studies of the host location behavior of this and other species of parasitic wasp.     

Terpenoids from the octocorals Menella sp. (Plexauridae) and Lobophytum crassum (Alcyonacea)

A new germacrane-type sesquiterpenoid, menelloide E (1), and a new cembrane-type diterpenoid, lobocrassin F (2), were isolated from the octocorals Menella sp. and Lobophytum crassum, respectively. The structures of terpenoids 1 and 2 were determined by spectroscopic and chemical methods and compound 2 was found to display a significant inhibitory effect on the release of elastase by human neutrophils.     

Laboratory Evaluation of Acute Toxicity of the Essential Oil of Allium tuberosum Leaves and Its Selected Major Constituents Against Apolygus lucorum (Hemiptera: Miridae)

The aim of this research was to evaluate acute toxicity of the essential oil of leaves of Chinese chives, Allium tuberosum Rottler ex Spreng (Asparagales: Alliaceae) and its major constituents against Apolygus lucorum Meyer-Dür (Hemiptera: Miridae). The essential oil of A. tuberosum leaves was obtained by hydrodistillation and analyzed by gas chromatography and gas chromatography-mass spectrometry. The major constituents of the oil were sulfur-containing compounds, including allyl methyl trisulfide (36.24%), diallyl disulfide (27.26%), diallyl trisulfide (18.68%), and dimethyl trisulfide (9.23%). The essential oil of A. tuberosum leaves exhibited acute toxicity against Ap. lucorum with an LD₅₀ value of 20.03 μg per adult. Among the main compounds, diallyl trisulfide (LD₅₀ = 10.13 μg per adult) showed stronger acute toxicity than allyl methyl trisulfide (LD₅₀ = 21.10 μg per adult) and dimethyl trisulfide (LD₅₀ = 21.65 μg per adult). The LD₅₀ value of diallyl disulfide against Ap. lucorum was 28.10 μg per adult. The results indicated that the essential oil of A. tuberosum and its major constituents may have a potential to be developed as botanical insecticides against Ap. lucorum.     

Evaluation of augmentative release of Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae) for biological control of Parthenium hysterophorus L.

Augmentative release of biocontrol agents has been largely successful for the management of insect pests but it has not been a common approach for weed management. Augmentation methods need to be developed for weed management, especially for pernicious weeds like Parthenium hysterophorus L., commonly known as pathenium or carrot weed. The leaf beetle Zygogramma bicolorata is a potential biocontrol agent of P. hysterophorus. Initial release of biocontrol agents is subject to uncertainties as to whether timely population built-up will take place in sufficient numbers. Several augmentative releases may be required to ensure early establishment of the biocontrol agents, for successful biological control of noxious weeds including pathenium. We made augmentative releases of larvae or adults of Z. bicolorata each to three sites, severely infested with pathenium at Jabalpur, India consecutively for a period of three years. Initially 10 larvae or adults per sq m were released in each plot, followed by a second, third and fourth release of 3, 1.5 and 1.5 larvae or adults per sq m at an intervals of 3, 7 and 14 days after the first augmentation. The pathenium at augmented sites were completely defoliated in 45 and 60 days by larvae and adults respectively. There was also a reduction in the pathenium density and plant height in the augmented sites as compared to the non-augmented sites. Over a period of 3 years augmentation resulted in a noteworthy negative effect on the weed.     

Abundance and Frequency of the Asiatic Oak Weevil (Coleoptera: Curculionidae) and Defoliation on American,Chinese, and Hybrid Chestnut (Castanea)

The Asiatic oak weevil, Cyrtepistomus castaneus Roelofs (Coleoptera: Curculionidae), is a nonnative defoliator of trees in the Fagaceae family in the United States but has not been studied on Castanea species in the southern Appalachian Mountains. Planted trees of Castanea dentata (Marsh.) Borkh. (Fagales: Fagaceae), Castanea mollissima Blume (Fagales: Fagaceae), and four hybrid breeding generations were evaluated in 2012 for insect defoliation and C. castaneus abundance and frequency. Defoliation was visually assessed throughout the growing season at two sites in the southern Appalachian Mountains (western North Carolina and eastern Tennessee). C. castaneus abundance and frequency were monitored on trees using beat sheets and emergence was recorded from ground traps. Asiatic oak weevils were more abundant and more frequently collected on American chestnut (Ca. dentata) and its most closely related BC₃F₃ hybrid generation than on the Asian species Ca. mollissima. In most months, C. castaneus colonization of hybrid generations was not significantly different than colonization of parental species. Frequency data for C. castaneus suggested that adults were distributed relatively evenly throughout the study sites rather than in dense clusters. Emergence of C. castaneus was significantly higher under a canopy dominated by Quercus species than under non-Quercus species or open sky. C. castaneus emergence began in May and peaked in late June and early July. These results may be useful for resource managers trying to restore blight-resistant chestnut to the Southern Appalachians while minimizing herbivory by insect pests.     

Within- and Between-Species Variation of Wing Venation in Genus Monochamus (Coleoptera: Cerambycidae)

We have described the morphological variation of five Western Palaearctic species of Monochamus Dejean, 1821. The variation was assessed using wing measurements. Special emphasis was placed on the differences between Monochamus sartor (F., 1787) and Monochamus urussovii (Fischer-Waldheim, 1805). There was an interesting pattern of variation between the two species. Individuals of M. sartor from the Carpathians differed markedly from individuals of M. urussovii from Siberia, but individuals from north-eastern Poland were intermediate between those two populations. The intermediate individuals were more similar to the Siberian M. urussovii than to the Carpathian M. sartor despite the relatively large geographic distance between north-eastern Poland and Siberia. The occurrence of the intermediate individuals in north-eastern Poland may be the effect of hybridization between M. urussovii and M. sartor, which might have occurred after secondary contact between the two species in the Holocene.     

Attack Pattern and Reproductive Ecology of Tomicus brevipilosus (Coleoptera: Curculionidae) on Pinus yunnanensis in Southwestern China

Tomicus brevipilosus (Eggers) (Coleoptera: Curculionidae, Scolytinae) was recently discovered as a new pest of Yunnan pine (Pinus yunnanensis Franchet) in Yunnan Province in southwestern China. However, little was known on its reproductive biology and pattern of trunk attack on Yunnan pine. The objectives of this study were to better understand the reproductive biology of T. brevipilosus by investigating the seasonality of trunk attacks by parent adults for the purpose of reproduction (i.e., breeding attacks) and the within-tree pattern of these attacks. Our results showed that T. brevipilosus breeding attacks in P. yunnanensis generally started in early March and ended in early June in Anning County, Yunnan. T. brevipilosus exhibited two general patterns of infestation. From early March to mid-April, T. brevipilosus bred preferentially in the trunks of Yunnan pine trees that were already infested by Tomicus yunnanensis Kirkendall and Faccoli and Tomicus minor (Hartig), colonizing spaces along the trunk (mostly in the mid- and lower trunk) that were not already occupied by the other two Tomicus species. Later, from about mid-April to early June, when there were no Yunnan pine trees newly infested by T. yunnanensis and T. minor, T. brevipilosus attacked Yunnan pine by itself, infesting the lower parts of the trunk first and then infesting progressively upward along the trunk into the crown. Infestation by T. brevipilosus extends the total period that P. yunnanensis trees are under attack by Tomicus beetles in southwestern China, which helps explain why Yunnan pine has suffered high levels of tree mortality in recent decades.     

Fumigant Toxicity and Repellence Activity of Camphor Essential Oil from Cinnamonum camphora Siebold Against Solenopsis invicta Workers (Hymenoptera:Formicidae)

The red imported fire ant (RIFA) Solenopsis invicta Buren causes severe damage to humans and animals as well as the environment. Chemical treatment is the main strategy of RIFA management, which also is potentially toxic to the environment. Plant essential oils (EOs) are considered as potential substance that can be used to control insects. This study aimed to identify the chemical composition of camphor EO and investigate the insecticidal activity on RIFAs. The chemical composition of the EO was analyzed by gas chromatography/mass spectrometry and gas chromatography with flame ionization detection. Results revealed that 36.61% camphor and 30.05% cineole were the major components. The insecticidal activity of camphor EO was assessed against RIFA workers by conducting two different bioassays: fumigant toxicity and repellence. Fumigant toxicity assay results showed that the lethal dose (LC₅₀) of the EO at 24 h was 1.67 and 4.28 μg/ml for minor and major workers, respectively; knockdown time (KT₅₀) was 10.82 and 14.73 h. At 2.55 μg/ml, the highest average mortality of the ants was 84.89% after 72 h. Camphor EO exhibited fumigant toxicity against minor and major workers as indicated by the effects on attacking, feeding, and climbing behaviors. This EO was also strongly repellent to the two size workers of the colony as observed in their behavior against Tenebrio molitor treated with 5 µl EO. The fumigant toxicity and repellence of camphor EO against RIFA indicated that this substance could be a potential alternative for the development of eco-friendly products used to control pests.     

Evidence for Bombus occidentalis (Hymenoptera: Apidae) Populations in the Olympic Peninsula,the Palouse Prairie,and Forests of Northern Idaho

Since the mid-1990s, Bombus occidentalis (Green) has declined from being one of the most common to one of the rarest bumble bee species in the Pacific Northwest of the United States. Although its conservation status is unresolved, a petition to list this species as endangered or threatened was recently submitted to the U.S. Fish and Wildlife Service. To shed light on the conservation situation and inform the U.S. Fish and Wildlife Service decision, we report on the detection and abundance of B. occidentalis following bumble bee collection between 2012 and 2014 across the Pacific Northwest. Collection occurred from the San Juan Islands and Olympic peninsula east to northern Idaho and northeastern Oregon, excluding the arid region in central Washington. B. occidentalis was observed at 23 collection sites out of a total of 234. With the exception of three sites on the Olympic peninsula, all of these were in the southeastern portion of the collection range.     

Intracellular Immunohistochemical Detection of Tetrodotoxin in Pleurobranchaea maculata (Gastropoda) and Stylochoplana sp. (Turbellaria)

Tetrodotoxin (TTX), is a potent neurotoxin targeting sodium channels that has been identified in multiple marine and terrestrial organisms. It was recently detected in the Opisthobranch Pleurobranchaea maculata and a Platyhelminthes Stylochoplana sp. from New Zealand. Knowledge on the distribution of TTX within these organisms is important to assist in elucidating the origin and ecological role of this toxin. Intracellular micro-distribution of TTX was investigated using a monoclonal antibody-based immunoenzymatic technique. Tetrodotoxin was strongly localized in neutral mucin cells and the basement membrane of the mantle, the oocytes and follicles of the gonad tissue, and in the digestive tissue of P. maculata. The ova and pharynx were the only two structures to contain TTX in Stylochoplana sp. Using liquid chromatography-mass spectrometry, TTX was identified in the larvae and eggs, but not the gelatinous egg cases of P. maculata. Tetrodotoxin was present in egg masses of Stylochoplana sp. These data suggest that TTX has a defensive function in adult P. maculata, who then invest this in their progeny for protection. Localization in the digestive tissue of P. maculata potentially indicates a dietary source of TTX. Stylochoplana sp. may use TTX in prey capture and for the protection of offspring.     

Virtual Screening of Plant Volatile Compounds Reveals a High Affinity of Hylamorpha elegans (Coleoptera: Scarabaeidae) Odorant-Binding Proteins for Sesquiterpenes From Its Native Host

Hylamorpha elegans (Burmeister) is a native Chilean scarab beetle considered to be a relevant agricultural pest to pasture and cereal and small fruit crops. Because of their cryptic habits, control with conventional methods is difficult; therefore, alternative and environmentally friendly control strategies are highly desirable. The study of proteins that participate in the recognition of odorants, such as odorant-binding proteins (OBPs), offers interesting opportunities to identify new compounds with the potential to modify pest behavior and computational screening of compounds, which is commonly used in drug discovery, may help to accelerate the discovery of new semiochemicals. Here, we report the discovery of four OBPs in H. elegans as well as six new volatiles released by its native host Nothofagus obliqua (Mirbel). Molecular docking performed between OBPs and new and previously reported volatiles from N. obliqua revealed the best binding energy values for sesquiterpenic compounds. Despite remarkable divergence at the amino acid level, three of the four OBPs evaluated exhibited the best interaction energy for the same ligands. Molecular dynamics investigation reinforced the importance of sesquiterpenes, showing that hydrophobic residues of the OBPs interacted most frequently with the tested ligands, and binding free energy calculations demonstrated van der Waals and hydrophobic interactions to be the most important. Altogether, the results suggest that sesquiterpenes are interesting candidates for in vitro and in vivo assays to assess their potential application in pest management strategies.     

Portable acoustic device for detection of coconut palms infested by Rynchophorus ferrugineus (Coleoptera: Curculionidae)

Acoustic methods have been developed and used to detect insects in concealed habitats. The larvae of red palm weevil, Rynchophorus ferrugineus (Olivier), a serious pest of the coconut palm, Cocos nucifera L. feed on the soft tissues inside the stem and bud region. Detection of infested coconut palms in the early stages by the conventional method of checking for external symptoms is time consuming, labour intensive and costly. This paper describes the development of a portable and efficient acoustic device and its potential in detection of infested palms in the field. The device comprises a sensor to mount on the palm and to acquire the sounds of red palm weevil larvae, an electronic unit that processes the acquired sounds and a set of headphones to receive the output sound by the listener. It is light weight, user-friendly and powered by batteries. The highest accuracy of identifying infested palms was obtained when the palm was checked at four positions; either side of palm base and bases of the two lowermost leaves. The infested palms were detected with over 97% accuracy, while the probability of not detecting uninfested palms was over 92%. A second check increased the rate of accuracy. Many difficulties encountered with conventional methods could be overcome by the use of this device.     

Transovarial Effect of Novaluron on Tribolium castaneum (Coleoptera: Tenebrionidae) After Termination of Direct Contact

The insect growth regulator novaluron (Rimon 10 EC, Makhteshim-Agan Ltd, Israel) is used against many field pests on corn, vegetables, orchards, forests, and cotton plantations. Previously, we studied various effects of novaluron on stored grain pests. Termination in Tribolium castaneum (Herbst) eggs hatching after treating adults with novaluron and following restoration after adult transfer to untreated media was observed. The objective of this study was to investigate the restoration of T. castaneum egg hatch following transfer of adults from treated media to untreated favorable and unfavorable media. The time needed for hatching restoration of 50% of eggs laid by adults transferred from novaluron (1 ppm) treated flour to untreated flour (RT₅₀) was 2.7 d. RT₅₀ for those transferred to untreated wheat grain was 4.1 d. RT₉₀ in flour was 3.6 d, in grain—6.1 d. Varieties of RTs in grain and in flour with nonoverlapping confidence intervals indicate that RTs were significantly different. Delay of eggs hatching restoration for adults transferred from treated flour to unfavorable media (Petri dishes with limited amount of flour, lying of eggs not detected) was observed. RT₅₀ in flour was 2.1 d and RT₉₀—3.1 d, while RT₅₀ in the unfavorable media was 3.4 d and RT₉₀ 6.5 d. Delayed effect of egg hatching restoration after adult transfer to unfavorable media provides evidence of the significant role of insect physiological state in novaluron excretion and (or) degradation by T. castaneum females.