Insects — Open Access Journal

Polyols are commonly used in food and medicines as sweeteners and preservatives but may also have insecticidal properties against some species of Diptera. Here we compared the insecticidal activity and feeding response of glycerol and propylene glycol (PG) on two tephritids: Anastrepha ludens and Anastrepha obliqua, and the drosophilid Drosophila suzukii. First, flies were exposed to solutions of 50% sucrose and the two polyols at concentrations of 1.67 M, 2.78 M and 4.18 M for 24 h and then observed at 24 h intervals for a period of three days. Both polyols elicited strong regurgitation behavior in the three flies and killed them. Regurgitation apparently also reduced flies’ body weight, and this was particularly apparent in insects that fed on 4.18 M PG solutions. A high percentage of individuals exposed to PG solutions perished after 72 h. The number of proboscis extensions, which is associated with feeding preference, was lower in the 4.18 M polyols + sucrose mixtures than in the 50% sucrose solution. Glycerol had a lower insecticidal effect in Anastrepha spp. and very little insecticidal effect in D. suzukii. Finally, elevated regurgitation and mortality was confirmed in A. ludens treated with 1.0–2.78 M of erythritol plus sucrose. Our results demonstrate that PG, and to a lower extent glycerol, have the potential for being used as a safer method of insect pest control. The hyper-regurgitation response may contribute to the insecticidal properties of these polyols in Diptera. Full article

Termites are social insects that live in colonies headed by reproductive castes. The breeding system is defined by the number of reproductive individuals in a colony and the castes to which they belong. There is tremendous variation in the breeding system of termites both within and among species. The current state of our understanding of termite breeding systems is reviewed. Most termite colonies are founded by a primary (alate-derived) king and queen who mate and produce the other colony members. In some species, colonies continue throughout their life span as simple families headed by the original king and queen. In others, the primary king and queen are replaced by numerous neotenic (nymph- or worker-derived) reproductives, or less commonly primary reproductives, that are descendants of the original founding pair leading to inbreeding in the colony. In still others, colonies can have multiple unrelated reproductives due to either founding the colonies as groups or through colony fusion. More recently, parthenogenetic reproduction has shown to be important in some termite species and may be widespread. A major challenge in termite biology is to understand the ecological and evolutionary factors driving the variation in termite breeding systems. Full article

Termites are social insects of economic importance that have a worldwide distribution. Identifying termite species has traditionally relied on morphometric characters. Recently, several mitochondrial genes have been used as genetic markers to determine the correlation between different species. Heavy metal accumulation causes serious health problems in humans and animals. Being involved in the food chain, insects are used as bioindicators of heavy metals. In the present study, 100 termite individuals of Anacanthotermesochraceus were collected from two Saudi Arabian localities with different geoclimatic conditions (Riyadh and Taif). These individuals were subjected to morphological identification followed by molecular analysis using mitochondrial 12S rRNA gene sequence, thus confirming the morphological identification of A. ochraceus. Furthermore, a phylogenetic analysis was conducted to determine the genetic relationship between the acquired species and other termite species with sequences previously submitted in the GenBank database. Several heavy metals including Ca, Al, Mg, Zn, Fe, Cu, Mn, Ba, Cr, Co, Be, Ni, V, Pb, Cd, and Mo were measured in both collected termites and soil samples from both study sites. All examined samples (termite and soil) showed high concentrations of metals with different concentrations and ratios. Generally, most measured metals had a significantly high concentration in soil and termites at Taif, except for Ca, Cd, Co, Cr, Cu, Mg, and Ni showing significantly high concentrations at Riyadh. Furthermore, termites accumulated higher amounts of heavy metals than the soil at both locations. The mean concentrations of the measured metals in soil samples were found to be in the descending order Ca ˃ Al ˃ Mg ˃ Zn ˃ Fe ˃ Cu ˃ Mn ˃ Ba ˃ Cr ˃ Co ˃ Be ˃ Ni ˃ V ˃ Pb ˃ Cd ˃ Mo, while it was Ca ˃ Mg ˃ Al ˃ Fe ˃ Zn ˃ Cu ˃ Mn ˃ Be ˃ Ba ˃ Pb ˃ Cr ˃ V ˃ Ni ˃ Cd ˃ Mo ˃ Co in termite specimens. The mean concentrations of the studied metals were determined in the soil and termite specimens at both locations. In addition, the contamination factor, pollution load index (PLI) and degree of contamination were calculated for all studied metals in different samples, indicating that both studied sites were polluted. However, Taif showed a significantly higher degree of pollution. Thus, the accurate identification of economically important insects, such as termites, is of crucial importance to plan for appropriate control strategies. In addition, termites are a good bioindicator to study land pollution. Full article

Wheat, corn, and brown rice were treated with different combinations of a deltamethrin suspension concentrate (SC) formulation and a new emulsifiable concentrate (EC) formulation, at rates of either 0.5 or 1.0 ppm alone or in combination with 1.25 and 2.5 ppm methoprene (10 treatments in all, including an untreated control). Treated commodities were stored at ambient conditions on the floor of an empty grain bin in Manhattan, KS, USA, in 5-kg lots for individual replicates. The commodities were sampled and bio-assayed every three months for 15 months by exposing 10 mixed-sex parental adults of selected adult stored product insects on 70–80 g of the commodity. For all treatments, there was no regression of declining efficacy with respect to the month. Therefore, the data were combined for analysis. On wheat and brown rice, there was no reproduction of Rhyzopertha dominica (Fabricius) in any of the treatments, and there was no weight loss in either commodity that was caused by feeding of the parental adults or developing progeny. There was reproduction of Sitophilus oryzae (L.) on wheat but, for several combinations, the EC formulation gave better suppression of progeny compared to the standard SC. However, on brown rice, only the combination of 1.0 ppm deltamethrin EC and 2.5 ppm methoprene was different than other treatments with respect to progeny development, sample weight loss caused by feeding, and weight of the feeding damage itself. Progeny production was correlated with grain damage. No progeny of Tribolium castaneum (Herbst) developed on the treated corn, but there was some variation in insect damage, with less damage in those treatments involving the EC formulation. Progeny production of Sitophilus zeamais (Motschulsky) was at the lowest in the combination of 1.0 ppm deltamethrin EC and 2.5 ppm methoprene. The resulting insect damage was the lowest in this combination as well. Results of this study were used by the registrant (Central Life Sciences) in the United States (US) to modify the commercial formulation to replace the deltamethrin SC with the EC, at label rates of either 0.5 ppm EC+ 1.25 ppm methoprene, or 1.0 ppm EC + 2.5 ppm methoprene, on wheat, corn, and rice. Full article

The growing awareness that microbial symbionts residing in mosquito midguts can interrupt transmission of vector-borne diseases has stimulated interest in understanding their potential role in mosquito biology. Fluorescent proteins are powerful molecular markers that provide for detailed analysis of the function and behavior of specific midgut bacterial isolates without disturbing the normal gut microbiota. The aim of this study was to label bacterial isolates from the midgut of Ochlerotatus triseriatus, the primary vector of La Crosse virus, with green, yellow, and red fluorescent proteins (GFP, YFP, RFP) via electroporation. We also assessed the stability of GFP-, YFP-, and RFP-bearing plasmids and their effect on bacterial growth. Seven of eleven bacterial species could not be labeled despite several attempts. Labeling of Escherichia coli and Enterobacter cloacae was successfully achieved with all three fluorescent proteins. In contrast, labeling of Aerococcus viridans was achieved with GFP only and labeling of Aeromonas hydrophila was achieved with GFP and YFP only. The stability of GFP plasmid varied among bacterial species with A. hydrophila followed by E. cloacae having the most stable GFP label. In contrast, YFP and RFP plasmids were very stable in all bacterial species possessing these labels. GFP plasmid reduced the growth of labeled strains relative to wild type but this effect was not evident in YFP and RFP plasmids. These findings suggest that some mosquito midgut bacterial isolates can effectively be labeled with GFP, YFP and RFP plasmids allowing non-destructive studies on their functions within the vector. Full article

Cold treatment at 0.0 °C with different exposure durations (0–12 days) was applied to the Mediterranean fruit fly Ceratitis capitata (Wiedemann) fed on a lab diet. The examined developmental stages were early eggs (<6 h), late eggs (>42 h), first instar, second instar and third instar larvae. Pupation, adult emergence and sex ratios of survived flies were investigated to study the C. capitata responses to this low temperature treatment. Our results showed that exposure time at low temperature has a clear effect on pupation and adult emergence. Based on pupation ratios, the first and third instar are the most cold tolerant stages, with LT99 = 7.3 for both of them. Cold tolerance at both stages are very close and no significant differences were detected. There were no significant differences on C. capitata sex ratios among different stages after treatment. This study improves our understanding of C. capitata responses to cold treatment, which may assist in the improvement of the current treatment strategies to control this destructive horticulture pest species. Full article

Mosquito-borne infections are a constant problem in Vietnam, and mosquito vector control is a primary approach to control these infections. Essential oils represent environmentally friendly alternatives to synthetic pesticides for mosquito control. The essential oils of two weedy species in Vietnam, Erechtites hieraciifolius and E. valerianifolius, have been obtained by hydrodistillation and analyzed by gas chromatography–mass spectrometry. The essential oils have been screened for mosquito larvicidal activity against Aedes albopictus, Ae. aegypti, and Culex quinquefasciatus. The essential oil from the aerial parts of E. hieraciifolius was rich in α-pinene (14.5%), limonene (21.4%), and caryophyllene oxide (15.1%), while E. valerianifolius essential oil was dominated by myrcene (47.8%) and α-pinene (30.2%). Both essential oils showed good larvicidal activity against Ae. albopictus (24-h LC₅₀ 10.5 and 5.8 μg/mL, respectively) and Ae. aegypti (24-h LC₅₀ 10.6 and 12.5 μg/mL, respectively). The essential oil of E. valerianifolius also showed good activity against Cx. quinquefasciatus larvae (24-h LC₅₀ = 40.7 μg/mL). Thus, Erechtites essential oils may serve as low-cost vector control agents for mosquito-borne infections. Full article

Dengue virus (DENV) is transmitted by mosquitoes and is a major public health concern. The study of innate mosquito defense mechanisms against DENV have revealed crucial roles for the Toll, Imd, JAK-STAT, and RNAi pathways in mediating DENV in the mosquito. Often overlooked in such studies is the role of intrinsic cellular defense mechanisms that we hypothesize to work in concert with the classical immune pathways to affect organismal defense. Our understanding of the molecular interaction of DENV with mosquito host cells is limited, and we propose to expand upon the recent results from a genome-scale, small interfering RNA (siRNA)-based study that identified mammalian host proteins associated with resistance to dengue/West Nile virus (DENV/WNV) infection. The study identified 22 human DENV/WNV resistance genes (DVR), and we hypothesized that a subset would be functionally conserved in Aedes aegypti mosquitoes, imparting cellular defense against flaviviruses in this species. We identified 12 homologs of 22 human DVR genes in the Ae. aegypti genome. To evaluate their possible role in cellular resistance/antiviral defense against DENV, we used siRNA silencing targeted against each of the 12 homologs in an Ae. aegypti cell line (Aag2) infected with DENV2 and identified that silencing of the two candidates, AeFKBP1 and AeATCAY, homologs of human FKBP1B and ATCAY, were associated with a viral increase. We then used dsRNA to silence each of the two genes in adult mosquitoes to validate the observed antiviral functions in vivo. Depletion of AeFKBP1 or AeATCAY increased viral dissemination through the mosquito at 14 days post-infection. Our results demonstrated that AeFKBP1 and AeATCAY mediate resistance to DENV akin to what has been described for their homologs in humans. AeFKBP1 and AeATCAY provide a rare opportunity to elucidate a DENV-resistance mechanism that may be evolutionarily conserved between humans and Ae. aegypti. Full article

Fall armyworm (FAW) was reported for the first time in Africa in 2016. FAW is widely distributed in Ethiopia, causing significant damage to maize. Nine synthetic insecticides belonging to different chemical groups and 11 pesticidal plants (botanicals) were tested for their efficacy against FAW under laboratory, greenhouse, and field conditions. In the laboratory, Radiant, Tracer, Karate, and Ampligo caused over 90% larval mortality 72 h after application. Malathion had moderate activity, causing 51.7% mortality 72 h after application, while Carbaryl was less effective, causing 28% mortality 72 h after application. In the greenhouse experiment, all synthetic insecticides reduced foliar damage to maize compared to the untreated control. Chemical sprays did not affect plant height, stem thickness, or leaf number. The highest fresh weight (471 g) was obtained from plants treated with Radiant. Among the botanicals tested, Azadirachta indica, Schinnus molle, and Phytolacca dodecandra resulted in the highest percentage larval mortality (>95%) 72 h after application. In the field, non-treated control plants showed extensive leaf injury compared to the synthetic insecticide- and botanical-treated plants. The synthetic insecticides and botanicals that showed high efficacy against FAW larvae can be used as components for integrated pest management (IPM) plans for FAW under smallholder farmer conditions in Ethiopia and elsewhere in Africa. Full article

The leafhopper Erasmoneura vulnerata (Fitch) (Hemiptera: Cicadellidae) is native of Northern and Central America where it occurs on wild and cultivated grapes as well as on a number of secondary hosts. This species was recorded for the first time in Europe (Italy, Veneto region) in 2004. Since then it has spread over Northern Italy and Slovenia. Studies on the biology of E. vulnerata in America are limited and thus its phenology was investigated on Vitis labrusca L. and Vitis vinifera L. plants under field and semi-field conditions. These observations suggest that E. vulnerata can complete 2–3 generations per year. The development of E. vulnerata from first instar nymphs to adults was studied under controlled conditions (about 23 °C). Developmental times lasted from 16.1 days in July–August to 19.5 days in September, and this variability was probably due to grape cultivar and plant susceptibility. Data were consistent with the number of generations previously reported. Erasmoneura vulnerata was more abundant on vines close to overwintering sites than on those located 100–250 m from these sites and contiguous to commercial vineyards. Full article

Plant sugar is an essential dietary constituent for mosquitoes, and hemipteran honeydew is one of the many forms of plant sugar that is important to mosquitoes. Many insects rely on volatile honeydew semiochemicals to locate aphids or honeydew itself. Mosquitoes exploit volatile semiochemicals to locate sources of plant sugar but their attraction to honeydew has not previously been investigated. Here, we report the attraction of female yellow fever mosquitoes, Aedes aegypti, to honeydew odorants from the green peach aphid, Myzus persicae, and the pea aphid, Acyrthosiphon pisum, feeding on fava bean, Vicia faba. We used solid phase micro-extraction and gas chromatography-mass spectrometry to collect and analyze headspace odorants from the honeydew of A. pisum feeding on V. faba. An eight-component synthetic blend of these odorants and synthetic odorant blends of crude and sterile honeydew that we prepared according to literature data all attracted female A. aegypti. The synthetic blend containing microbial odor constituents proved more effective than the blend without these constituents. Our study provides the first evidence for anemotactic attraction of mosquitoes to honeydew and demonstrates a role for microbe-derived odorants in the attraction of mosquitoes to essential plant sugar resources. Full article

Ants are a biodiverse group of insects that have evolved toxic venom containing many undiscovered bioactive molecules. In this study, we found that the venom of the ruby ant Myrmica rubra is a rich source of peptides. LC-MS analysis revealed the presence of 142 different peptides varying in molecular weight, sequence length, and hydrophobicity. One of the most abundant peaks was selected for further biochemical and functional characterization. Combined Edman degradation and de novo peptide sequencing revealed the presence of a novel decapeptide (myrmicitoxin) with the amino acid sequence NH₂-IDPKLLESLA-CONH₂. The decapeptide was named U-MYRTX-MRArub1 and verified against a synthetic standard. The amidated peptide was tested in a synthetic form to determine the antimicrobial activity towards the bacterial pathogens and insecticidal potential against pea aphids (Acyrthosiphon pisum). This peptide did not show antimicrobial activity but it significantly reduced the survival of aphids. It also increased the sensitivity of the aphids to two commonly used chemical insecticides (imidacloprid and methomyl). Since ant venom research is still in its infancy, the findings of this first study on venom peptides derived from M. rubra highlight these insects as an important and rich source for discovery of novel lead structures with potential application in pest control. Full article

The bamboo mosquito, Tripteroides bambusa (Yamada) (Diptera: Culicidae), is a common insect across East Asia. Several studies have looked at the ecology of Tr. bambusa developmental stages separately, but little is known about the factors associated with the persistence (how often) and abundance (how many individuals) of Tr. bambusa stages simultaneously studied across a heterogeneous landscape. Here, we ask what environmental and landscape factors are associated with the persistence and abundance of Tr. bambusa stages across the altitudinal gradient of Mt. Konpira, Nagasaki City, Japan. During a season-long study we counted 8065 (7297 4th instar larvae, 670 pupae and 98 adults) Tr. bambusa mosquitoes. We found that persistence and abundance patterns were not associated among stages, with the exception of large (4th instar) and small (1st to 3rd instars) larvae persistence, which were positively correlated. We also found that relative humidity was associated with the persistence of Tr. bambusa aquatic stages, being positively associated with large and small larvae, but negatively with pupae. Similarly, landscape aspect changed from positive to negative the sign of its association with Tr. bambusa pupae and adults, highlighting that environmental associations change with life stage. Meanwhile, Tr. bambusa abundance patterns were negatively impacted by more variable microenvironments, as measured by the negative impacts of kurtosis and standard deviation (SD) of environmental variables, indicating Tr. bambusa thrives in stable environments, suggesting this mosquito species has a finely grained response to environmental changes. Full article

To monitor and quantify the changes in pollinator communities over time, it is important to have robust survey techniques of insect populations. Pan traps allow for the assessment of the relative insect abundance in an environment and have been promoted by the Food and Agricultural Organization (FAO) as an efficient data collection methodology. It has been proposed that fluorescent pan traps are particularly useful, as it has been suggested that they capture high numbers of insects in an unbiased fashion. We use a simultaneous presentation of fluorescent and non-fluorescent pan trap colours to assess how flower-visiting insects of different orders respond to visual stimuli and reveal a significant interaction between trap fluorescence and captured insect type. In particular, Coleoptera (beetles) and Lepidoptera (butterflies and moths) were captured significantly more frequently by fluorescent traps, whilst Dipterans (flies) were captured significantly less frequently by this type of pan trap. Hymenopterans (bees and wasps) showed no significant difference in their preference for fluorescent or non-fluorescent traps. Our results reveal that the use of fluorescent pan traps may differently bias insect capture rates when compared to the typical experience of colour flower-visiting insects in natural environments. Correction factors may, therefore, be required for interpreting insect pan trap data collected with different methodologies. Full article

Chikungunya virus (CHIKV) is a medically important mosquito-borne virus transmitted to humans by infected Aedes (Stegomyia) species. In 2013–2014, Ae. aegypti transmitted CHIKV to humans in the Caribbean and in 2005–2006, Ae. albopictus transmitted CHIKV on La Réunion Island (Indian Ocean basin). CHIKV LR2006 OPY1 from the La Réunion epidemic was associated with a mutation (E1:A226V) in the viral E1 glycoprotein that enhanced CHIKV transmission by Ae. albopictus. CHIKV R99659 from the Caribbean outbreak did not have the E1:A226V mutation. Here, we analyzed the salivary glands and saliva of Ae. albopictus strains from New Jersey, Florida, Louisiana and La Réunion after infection with each virus to determine their transmission potential. We infected the Ae. albopictus strains with blood meals containing 3–7 × 10⁷ PFU/mL of each virus and analyzed the mosquitoes nine days later to maximize infection of their salivary glands. All four Ae. albopictus strains were highly susceptible to LR2006 OPY1 and R99659 viruses and their CHIKV disseminated infection rates (DIR) were statistically similar (p = 0.3916). The transmission efficiency rate (TER) was significantly lower for R99659 virus compared to LR2006 OPY1 virus in all Ae. albopictus strains and Ae. aegypti (Poza Rica) (p = 0.012) suggesting a salivary gland exit barrier to R99659 virus not seen with LR2006 OPY1 infections. If introduced, LR2006 OPY1 virus poses an increased risk of transmission by both Aedes species in the western hemisphere. Full article

Many African termite species are true soil-feeders: how can they coexist, sometimes with high densities? How do they separate their trophic niches? Preliminary results suggest that two coexisting Cubitermes species forage in different soil layers, and stable C and N isotopes show that they feed on different organic material. Cubitermes aff. ugandensis forages near the soil surface whereas C. aff. sankurensis forages in deeper layers; however, unexpectedly, the former shows a higher δ¹⁵N than the latter, highlighting, for the first time, a trophic niche differentiation between two sympatric true soil feeders bearing different enteric valve patterns. Full article

Behavioral symptoms and mortality associated with intoxication with insecticides fipronil and indoxacarb were determined in field-collected eastern subterranean termites, Reticulitermes flavipes (Kollar), and Formosan subterranean termites, Coptotermes formosanus Shiraki. Behaviors and mortality were evaluated at three temperatures (16, 22, and 28 °C) and three concentrations of fipronil (0.5, 1, and 5 ppm) and indoxacarb (50, 75, and 100 ppm). LT₅₀ (median lethal time to kill 50% of the termites) values declined with increasing concentrations and temperatures for both fipronil-exposed eastern and Formosan subterranean termites, whereas these values were not always the highest at 16 °C for indoxacarb-treated termites. The greatest change (reduction) in LT₅₀ values occurred for both species between 16 and 22 °C at the lowest concentration of each insecticide. Intoxication and moribundity were the most frequently observed behaviors for fipronil-exposed termites, whereas intoxication, ataxia, and moribundity were observed for most concentration and temperature combinations for indoxacarb-exposed termites. The inherent toxicity of fipronil was higher than that of indoxacarb. The higher presence and duration of intoxication behaviors may positively affect the performance of indoxacarb against subterranean termite colonies. Full article

The global trade of honeybee hive products imposes the risk of the introduction of exotic pests. However, data on the potential of specific products enabling pest survival are often lacking. This holds especially true for ectoparasitic mites Tropilaelaps spp., which are mandatory pests of honeybees in many countries. Here, we evaluated the longevity of Tropilaelaps mercedesae mites associated with empty honeycomb and dry pollen as two possible global import routes. Mites were able to survive up to three days in dry pollen and up to six days in empty honeycomb, thereby suggesting a sufficient time window for the potential introduction of T. mercedesae into mite-free countries via import of these hive products. Full article

Elicitors are biofactors that induce resistance in plants against different insect pests. This in vitro study evaluated the impact of a novel elicitor protein PeBC1, extracted from a necrotrophic fungus Botrytis cinerea, on the development and fecundity parameters of green peach aphid (Myzus persicae) on common beans (Phaseolus vulgaris L.). Three different concentrations of PeBC1 elicitor (i.e., 33.56, 25.43, 19.33 µg mL⁻¹) were applied at three different temperature regimes (i.e., 18, 21, and 25 °C). Elicitor treatments were applied topically on the bean plants at 3-leaf stage and newly emerged (0–6 h old) apterous adult aphids were exposed to these treated leaves. In addition to the biological parameters of aphids, the relative expression levels of key genes associated with jasmonic acid (JA) and salicylic acid (SA) plant defense pathways were also determined through RT-qPCR. Results of bioassays revealed that the application of PeBC1 elicitor protein exhibited pronounced and significant (p < 0.05) sub-lethal effects on green peach aphids. The fecundity was reduced and the nymphal development time was prolonged by different concentrations of PeBC1 elicitor and temperature regimes. Gene expression studies showed that the exogenous application of PeBC1 induced a significant upregulation of the expression levels of JA and SA pathway-associated genes in bean plants. As compared to control, elicitor-treated plants exhibited an induced resistance against aphids. Our findings suggest the potential use of PeBC1 elicitor protein in future bio-intensive management strategies against sap-sucking insect pests such as green peach aphids. Full article