Ecology of Meimuna mongolica (Hemiptera: Cicadidae) Nymphs: Instars,Morphological Variation,Vertical Distribution and Population Density,Host-Plant Selection,and Emergence Phenology

The cicada Meimuna mongolica (Distant) (Hemiptera: Cicadidae) is one of the most important pests of economic forest in Guanzhong Plain of Shaanxi Province, China. Information about ecological characteristics and some sustainable control measures of this species is urgently required for its control. In this study, nymphal instars, morphological variation, vertical distribution, and population density in soil, and emergence phenology of nymphs of M. mongolica on three main host plants (Pinus tabuliformis Carr., Populus tomentosa Carr., and Pyrus xerophila Yü) were studied, based on combined morphological and molecular identification, investigation of the first-instar nymphs hatched from eggs and others excavated from soil, and investigation of exuviae in the adult emergence period. Five nymphal instars of M. mongolica were redetermined according to the distribution plots of the head capsule widths of the nymphs. Nymphs of third and fourth instars showed morphological variation, which is closely related to host-plant association. The mean densities of nymphs in soil under the three host plants were significantly different, indicating a distinct host preference. The nymphs could extend their distribution from the 0–10 cm soil layer to the 51–60 cm soil layer underground but not beyond 60 cm soil layer under all the three host plants. The 21–30 cm soil layer under all the three host plants has the highest nymphal population density. The sex ratio of the entire population was nearly 50:50, but males dominated in the early half of the duration of the emergence. These ecological characteristics of M. mongolica could provide important information for sustainable control measures.     

Link between Domoic Acid Production and Cell Physiology after Exchange of Bacterial Communities between Toxic Pseudo-nitzschia multiseries and Non-Toxic Pseudo-nitzschia delicatissima

Bacteria are known to influence domoic acid (DA) production by Pseudo-nitzschia spp., but the link between DA production and physiology of diatoms requires more investigation. We compared a toxic P. multiseries to a non-toxic P. delicatissima, investigating links between DA production, physiological parameters, and co-occurring bacteria. Bacterial communities in cultures of both species were reduced by antibiotic treatment, and each of the diatoms was inoculated with the bacterial community of the other species. The physiology of P. delicatissima was minimally affected by the absence of bacteria or the presence of alien bacteria, and no DA was detected. P. multiseries grew faster without bacteria, did not produce a significant amount of DA, and exhibited physiological characteristics of healthy cells. When grown with alien bacteria, P. multiseries did not grow and produced more DA; the physiology of these cells was affected, with decreases in chlorophyll content and photosynthetic efficiency, an increase in esterase activity, and almost 50% mortality of the cells. The alien bacterial community had morphological and cellular characteristics very different from the original bacteria, and the number of free-living bacteria per algal cell was much higher, suggesting the involvement of bacteria in DA production.     

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.     

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.     

Analysis of Differential Proteins in Two Wing-Type Females of Sogatella furcifera (Hemiptera: Delphacidae)

Sogatella furcifera (Horvath) is an important rice pest with the wing dimorphism, including macropterous and brachypterous morphs. The protein expression profiles in two wing-type adults and two wing-type disc fifth-instar nymphs were analyzed using two-dimensional gel protein electrophoresis and mass spectrometry. In adults and fifth-instar nymphs, 127 and 162 protein spots were detected, respectively. Fifty-five differentially expressed protein spots were identified between the long-winged adults and the short-winged adults, and 62 differentially expressed protein spots were found between the long-winged disc fifth-instar nymphs and short-winged disc fifth-instar nymphs. In long-winged and short-winged adults, six and seven specific protein spots were identified, respectively, with five and seven protein spots having more than threefold increased level, respectively. In long-winged and short-winged disc morph nymphs, 8 and 12 specific protein spots were identified, respectively, with 11 and 17 spots containing more than threefold increased level, respectively. Among the 16 identified proteins, five proteins are associated with muscle function, suggesting that muscle is a main tissue where the genes were differentially expressed between the two wing types. In addition, the content of a peptidase with an insulinase domain was higher (by 3.02 ± 0.59 fold) in the short-winged fifth-instar nymphs than in the long-winged fifth-instar nymphs, which suggests that this peptidase may be involved in wing differentiation by regulating insulin receptors. The results of this study provide some genetic clues for the wing differential development in S. furcifera and provide more references for future studies.     

The Toxicity and Detoxifying Mechanism of Cycloxaprid and Buprofezin in Controlling Sogatella furcifera (Homoptera: Delphacidae)

The effects of cycloxaprid (a modified neonicotinoid insecticide) and buprofezin (a thiadiazine insecticide) on mortality of the white-backed planthopper (WBPH), Sogatella furcifera, were determined in laboratory assays. Cycloxaprid killed WBPH nymphs and adults but buprofezin killed only nymphs, and cycloxaprid acted faster than buprofezin. One day after infestation, mortality of third-instar nymphs was >65% with cycloxaprid at 125 mg liter⁻¹ but was <38% with buprofezin at 148 mg liter⁻¹. By the 4th day after infestation, however, control of nymphs by the two insecticides was similar, and cycloxaprid at 125 mg liter⁻¹ caused ≥80% mortality of adults but buprofezin at 148 mg liter⁻¹ (the highest rate tested) caused almost no adult mortality. LC₅₀ values for cycloxaprid were lowest with nymphs, intermediate with adult males, and highest with adult females. Although buprofezin was slower acting than cycloxaprid, its LC₅₀ for nymphs 5 d after infestation was 3.79-fold lower than that of cycloxaprid. Mean carboxylesterase (CarE) specific activity of nymphal WBPH treated with cycloxaprid and buprofezin was higher than that of control, but there was no significant difference between cycloxaprid and control (no insecticide), and it was significantly higher for buprofezin than those of cycloxaprid and control. For glutathione S-transferase and mixed function oxygenase, the specific activity of nymphal WBPH treated with buprofezin was significantly higher than those of cycloxaprid and control, too.     

Inventory of Fatty Acid Desaturases in the Pennate Diatom Phaeodactylum tricornutum

The diatom Phaeodactylum is rich in very long chain polyunsaturated fatty acids (PUFAs). Fatty acid (FA) synthesis, elongation, and desaturation have been studied in depth in plants including Arabidopsis, but for secondary endosymbionts the full picture remains unclear. FAs are synthesized up to a chain length of 18 carbons inside chloroplasts, where they can be incorporated into glycerolipids. They are also exported to the ER for phospho- and betaine lipid syntheses. Elongation of FAs up to 22 carbons occurs in the ER. PUFAs can be reimported into plastids to serve as precursors for glycerolipids. In both organelles, FA desaturases are present, introducing double bonds between carbon atoms and giving rise to a variety of molecular species. In addition to the four desaturases characterized in Phaeodactylum (FAD2, FAD6, PtD5, PtD6), we identified eight putative desaturase genes. Combining subcellular localization predictions and comparisons with desaturases from other organisms like Arabidopsis, we propose a scheme at the whole cell level, including features that are likely specific to secondary endosymbionts.     

Characterization of Solanum chomatophilum resistance to 2 aphid potato pests, Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer)

The aphids Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer) (Hemiptera: Aphididae) are responsible for yield reduction in potato (Solanum tuberosum) production by direct phloem feeding and by spreading viruses. Breeding resistant traits from Solanum chomatophilum into the potato germplasm provides alternative means to control aphid infestations. Integrated pest management strategy, using plant resistance, benefits from the characterization of the resistance and of its impact on aphid biology. Our objective was to characterize the resistance of S. chomatophilum by assessing the effects of accessions, plant parts on aphid performance, and by assessing the impact of the resistance factors on different aphid developmental stages and on alate morph production. Detailed aphid performance was obtained by measuring fecundity, survival, percentage of nymphs that reached adult moult, and population growth using whole plant and clip cage experimental designs. Accession and plant physiological age, but not aphid developmental stage, influenced all life-history parameters, except for alate morph production which was not induced on the resistant accessions. Plant part influence was independent of plant species and accession. Both experimental designs resulted in congruent resistance levels at the accession level for each of the two aphid species, supporting the use of any of them in S. chomatophilum resistance screening. PI243340 was resistant to both aphid species, while PI365324 and PI310990 were also resistant to M. euphorbiae and M. persicae, respectively.     

Bioprospecting from Marine Sediments of New Brunswick,Canada: Exploring the Relationship between Total Bacterial Diversity and Actinobacteria Diversity

Actinomycetes are an important resource for the discovery of natural products with therapeutic properties. Bioprospecting for actinomycetes typically proceeds without a priori knowledge of the bacterial diversity present in sampled habitats. In this study, we endeavored to determine if overall bacterial diversity in marine sediments, as determined by 16S rDNA amplicon pyrosequencing, could be correlated with culturable actinomycete diversity, and thus serve as a powerful tool in guiding future bioprospecting efforts. Overall bacterial diversity was investigated in eight marine sediments from four sites in New Brunswick, Canada, resulting in over 44,000 high quality sequences (x = 5610 per sample). Analysis revealed all sites exhibited significant diversity (H’ = 5.4 to 6.7). Furthermore, statistical analysis of species level bacterial communities (D = 0.03) indicated community composition varied according to site and was strongly influenced by sediment physiochemical composition. In contrast, cultured actinomycetes (n = 466, 98.3% Streptomyces) were ubiquitously distributed among all sites and distribution was not influenced by sediment composition, suggesting that the biogeography of culturable actinomycetes does not correlate with overall bacterial diversity in the samples examined. These actinomycetes provide a resource for future secondary metabolite discovery, as exemplified by the antimicrobial activity observed from preliminary investigation.     

Developmental Biology and Effects of Adult Diet on Consumption,Longevity, and Fecundity of Colaspis crinicornis (Coleoptera: Chrysomelidae)

The chrysomelid beetle Colaspis crinicornis Schaeffer (Coleoptera: Chrysomelidae) occurs primarily in the Great Plains region of the United States. Little is known about the biology and ecology of this species, but over the last decade, it has become increasingly common in the corn, Zea mays L., and soybean, Glycine max (L.) Merrill, agroecosystem of southeastern Nebraska. As part of a larger comprehensive project to understand the natural history and pest potential of this species, laboratory experiments were conducted to study the developmental biology, morphological characters of immature stages, and the effect of adult diet on consumption, longevity, and fecundity. Females readily deposited egg clusters in the soil, and percentage egg hatch was high under laboratory conditions. Larvae and pupae were confirmed to be soil-dwelling stages. C. crinicornis has relatively short egg, pupal, and adult stages with the majority of its life cycle spent in the larval stage. Results of choice and no-choice adult feeding experiments indicate that diets of corn or soybean leaves did not significantly affect consumption, longevity, or fecundity of adult C. crinicornis, suggesting that corn and soybean leaves are similarly suitable food sources for adults. The ability to effectively utilize tissues from very different plant families as adult food sources suggests that C. crinicornis is polyphagous in the field.     

Diverse bacterial PKS sequences derived from okadaic acid-producing dinoflagellates

Okadaic acid (OA) and the related dinophysistoxins are isolated from dinoflagellates of the genus Prorocentrum and Dinophysis. Bacteria of the Roseobacter group have been associated with okadaic acid producing dinoflagellates and have been previously implicated in OA production. Analysis of 16S rRNA libraries reveals that Roseobacter are the most abundant bacteria associated with OA producing dinoflagellates of the genus Prorocentrum and are not found in association with non-toxic dinoflagellates. While some polyketide synthase (PKS) genes form a highly supported Prorocentrum clade, most appear to be bacterial, but unrelated to Roseobacter or Alpha-Proteobacterial PKSs or those derived from other Alveolates Karenia brevis or Crytosporidium parvum.     

Sequential sampling plans for estimating density of glassy-winged sharpshooter, Homalodisca vitripennis (Hemiptera: Cicadellidae) on citrus

The glassy-winged sharpshooter, Homalodisca vitripennis (Germar), is a serious pest of grapes and other crop and ornamental plants mainly through its role as a vector of the bacterium Xylella fastidiosa Wells. Citrus harbors large populations of this insect throughout much of the year in areas where the pest is problematic and improved understanding of the population dynamics and management of H. vitripennis on citrus may be key to its management in the broader agricultural landscape. In turn, the study of population dynamics and the development of management strategies require effective and efficient sampling methods. Within-tree sampling distribution studies revealed that adults and nymphs were more abundant and less variable in the upper strata of citrus trees (>1.5 m). They occurred in greater numbers on the southern quadrants of trees but relative variability did not differ due to cardinal direction. We developed and validated several fixed-precision sequential sampling plans for estimating the density of nymphs and adults of H. vitripennis using a pole bucket sampling method. Based on validation from resampling of independent data sets, Green’s sequential sampling model, based on the Taylor’s power law, provided the best overall performance in terms of providing mean density estimates with levels of precision equal to or better than the desired precision over a range of possible insect densities. Average sampling costs varied from about 21 to 189 min for a desired precision of 0.25 depending on insect density and whether the goal is to sample nymphs, adults or both stages combined. Further, the sampling plans developed on orange trees were robust, being equally effective on orange and lemon trees and on trees treated or not with insecticides.     

Eleganolone,a Diterpene from the French Marine Alga Bifurcaria bifurcata Inhibits Growth of the Human Pathogens Trypanosoma brucei and Plasmodium falciparum

Organic extracts of 20 species of French seaweed have been screened against Trypanosoma brucei rhodesiense trypomastigotes, the parasite responsible for sleeping sickness. These extracts have previously shown potent antiprotozoal activities in vitro against Plasmodium falciparum and Leishmania donovani. The selectivity of the extracts was also evaluated by testing cytotoxicity on a mammalian L6 cell line. The ethyl acetate extract of the brown seaweed, Bifurcaria bifurcata, showed strong trypanocidal activity with a mild selectivity index (IC₅₀ = 0.53 µg/mL; selectivity index (SI) = 11.6). Bio-guided fractionation led to the isolation of eleganolone, the main diterpenoid isolated from this species. Eleganolone contributes only mildly to the trypanocidal activity of the ethyl acetate extract (IC₅₀ = 45.0 µM, SI = 4.0). However, a selective activity against P. falciparum erythrocytic stages in vitro has been highlighted (IC₅₀ = 7.9 µM, SI = 21.6).     

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.     

Intercolony Comparisons of Gut Microbiome Composition From Lab Reared Eastern Subterranean Termites (Blattodea: Rhinotermitidae)

Termites are social insects living in colonies composed of worker, soldier, and reproductive castes. Termite hindguts are inhabited by all three domains of life- Eukarya (protists), Bacteria, and Archaea. These gut microorganisms are horizontally and vertically transferred by nestmates and reproductives, respectively. Prior evidence suggests that every colony potentially has a different gut microbiome that was transferred vertically and horizontally over time. However, we do not know if different colonies reared in the laboratory on the same diet will ultimately demonstrate similar microbial composition and structure. Therefore, we looked at gut bacteria in Eastern subterranean termite (Reticulitermes flavipes) colonies that were reared in the laboratory with identical diets and rearing conditions. Based on16S rRNA gene sequencing, the observed features, and Shannon’s diversity were significantly different between the colonies while differences in Pielou evenness and Faith phylogenetic diversity were not statistically significant. In addition, the microbial community structures were significantly different between colonies. Based on ANCOM (Analysis of Composition of Microbiomes), the taxa Elizabethkingia (Bacteroidetes: Flavobacteriales) and Chryseobacterium (Bacteroidetes: Flavobacteriales) were differentially abundant between the colonies. These results suggest that providing the exact same diet and rearing environment for >2 yr cannot result in identical gut microbiomes between termite colonies.     

Insecticidal efficacy of phosphine fumigation at low pressure against major stored-product insect species in a commercial dried fig processing facility

We investigated the application of phosphine at low pressure for various exposure durations against major stored-product insects in a commercial dried fig processing facility in Central Greece. Trials were carried out inside a chamber, in which phosphine, in the form of aluminium phosphide pellets, was introduced with the use of a phosphine generator. The generator unit was also equipped with a vacuum pump to achieve low pressure inside the chamber. The chamber was filled with pallets with boxes containing figs. The insects tested were Tribolium confusum (all life stages), Ephestia elutella (eggs and larvae), Sitophilus oryzae (adults), Sitophilus granarius (adults), Rhyzopertha dominica (adults), Oryzaephilus surinamensis (adults) and Prostephanus truncatus (adults). Moreover, wheat grains containing immature stages of S. oryzae were also used. All insect-life stage combinations were exposed to phosphine at low pressure for 18, 48 and 72 h. In most cases, significant differences in mortality of insects treated with phosphine at low pressure compared to the control treatments were recorded. However, complete control (100%) was recorded only in the case of O. surinamensis adults and T. confusum larvae after exposure for 48 and 72 h, respectively. We conclude that the combined application of phosphine and low pressure at short exposure durations (up to 72 h) cannot provide sufficient control at least against the stored-product insect species and life stages tested in the present study.     

The Influence of Gut Microbiota on the Fecundity of Henosepilachna vigintioctopunctata (Coleoptera: Coccinellidae)

The gut microbiota of insects usually plays an important role in the development and reproduction of their hosts. The fecundity of Henosepilachna vigintioctopunctata (Fabricius) varies greatly when they develop on different host plants. Whether and how the gut microbiota regulates the fecundity of H. vigintioctopunctata was unknown. To address this question, we used 16S rRNA sequencing to analyze the gut microbiomes of H. vigintioctopunctata adults fed on two host plant species (Solanum nigrum and Solanum melongena) and one artificial diet. The development of the ovaries and testes was also examined. Our results revealed that the diversity and abundance of gut microorganisms varied significantly in insects reared on different diets. The gut microbiota of H. vigintioctopunctata raised on the two host plants was similar, with Proteobacteria being the dominant phylum in both groups, whereas Firmicutes was the dominant phylum in the group reared on the artificial diet. The predominant microbiota in the S. nigrum group were Acinetobacter soli and Acinetobacter ursingii (Acinetobacter, Moraxellaceae); Moraxella osloensis (Enhydrobacter, Moraxellaceae); and Empedobacter brevis (Empedobacter, Weeksellaceae). The microbiota in this group are associated with high lipid metabolism. In addition, the beetles’ ovaries and testes were more highly developed in the S. nigrum group than in the other two groups. These findings provide valuable information for elucidating the complex roles the gut microbiota play in the fecundity of H. vigintioctopunctata, and may also contribute to developing future novel control strategies involving this economically important pest.     

Biological Activity of trans-2-Hexenal Against Bradysia odoriphaga (Diptera: Sciaridae) at Different Developmental Stages

trans-2-Hexenal, one of the C6 green leaf volatiles, is potentially useful for the control of Bradysia odoriphaga Yang et Zhang. In this study, the biological activity of trans-2-hexenal on B. odoriphaga was assessed in the laboratory. trans-2-Hexenal was observed to kill B. odoriphaga in different developmental stages at a relatively low concentration under fumigation. The respiration rate in the male treatment group decreased from 131.44 to 4.07 nmol/g·min with a prolonged fumigation time, while the respiration rate in females decreased from 128.82 to 24.20 nmol/g·min. Male adults exhibited a more sensitive electroantennogram response at 0.05–500 μl/ml at the dose of 10.0 μl than female adults. Moreover, trans-2-hexenal had a repellent effect on adults based on the results with a Y-tube olfactometer at 10.0 μl, as shown by the deterrent rate of male and female adults with 96.67% and 98.33%, respectively. The results showed that trans-2-hexenal had good biological activity in different developmental stages of B. odoriphaga, which could reduce the need for, and risks associated with, the use of traditional insecticides and enable nonharmful management.     

Evaluation of entomopathogenic fungi against Cataenococcus ensete [Williams and Matile-Ferrero, (Homoptera: Pseudococcidae)] on enset

Enset root mealybug, Cataenococcus ensete Williams and Matile-Ferrero, (Homoptera: Pseudococcidae) is an economically important pest on enset. Adults and nymphs of overlapping generations feed on the corms and roots of enset and are difficult to control once established. Isolates of Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnickoff) Sorokin that originated from Ethiopia were evaluated for their efficacies against enset root mealybugs under laboratory, pots and field conditions. Of the tested isolates, two strains (FF and PPRC-56) of B. bassiana were found to be highly pathogenic to adults, causing 97 and 100% mortality, respectively 20 days after inoculation under laboratory conditions. On the other hand, the isolates PPRC-56, FF, PPRC-6 and Mm induced mean mortalities of 97, 95, 96 and 83%, respectively, to the adult enset root mealybugs on the pot experiment, and 51.33, 38.67, 29.33 and 19.33% mortality at the first site and 54.00, 42.67, 32.00 and 25.33% at the second site, respectively, in field experiments. The study showed B. bassiana and M. anisopliae can be used in the integrated management of enset root mealybug.