Pupation Success Research Articles

Pathogenicity Efficacy of Entomopathogen Fungus Beauveria bassiana Against In Vitro Rice Stem Borer (Scirpophaga innotata)

White rice stem borer (Scirpophaga innotata) is one of the dangerous pests for rice (Oryzae sativa) because it attacks rice stems that can cause 50% loss of total production. The purpose of this study was to determine the pathogenicity of Entomopathogen Fungus Beauveria bassiana at different conidia dilutions (J), namely J1 (105), J2 (106), J3 (107), J4 (108), and J5 (109). The mortality of S. innotata larvae was tested with the density of B. bassiana of 2.08 × 106 conidia/ml. The effect of B. bassiana on the success transformation of S. innotata larvae to pupae for 14 observation day was also observed at the same B. bassiana density. This research resulted in the optimal pathogenicity of B. bassiana at dilution of 107 (treatment J3) which caused 100% death and 100% successful pupation. The lowest treatment was J5 (109 dilution) with a mortality rate of 50% and a successful pupation of 20%. The results of the probit analysis showed that the fastest average lethal time (LT50) value was found in treatment of 107 (J3) which reached 11.11 h. Keywords: Beauveria bassiana, Pathogenicity, Probit analysis, Scirpophaga innotata.

Galleria mellonella Invertebrate Model Mirrors the Pathogenic Potential of Mycoplasma alligatoris within the Natural Host

Most mycoplasmal infections result in chronic, clinically silent disease. In direct contrast, Mycoplasma alligatoris elicits a fulminant, multisystem disease in the natural host, Alligator mississippiensis (American alligator). The goals of the study were to better understand the disease in the natural host and to determine if the invertebrate model G. mellonella could serve as a surrogate alternate host. The survival of alligators infected intratracheally was dose dependent (p=0.0003), ranging from no mortality (102 CFU) to 100% mortality (108 CFU), with 60% mortality at the 104 and 105 CFU infectious dose. Microbial load in blood, joints, and brain was dose dependent, regardless of whether alligators were infected intratracheally or intravenously (p <0.002). Weight loss was similarly impacted (p <0.001). Experimental infection of the invertebrate Galleria mellonella mirrored the result in the natural host. In a dose response infection study, both larval survival curves and successful pupation curves were significantly different (p≤0.0001) and dose dependent. Infected insects did not emerge as moths (p <0.0001). Here, we describe the first study investigating G. mellonella as a surrogate model to assess the pathogenic potential of M. alligatoris. G. mellonella survival was dose dependent and impacted life stage outcome.

Struggling to survive: A comparison of Vanessa cardui larval survivorship on putative host plants

AbstractThe painted lady butterfly (Vanessa cardui) is a generalist herbivore with a global distribution. In North America, over 100 species have been identified as V. cardui larval host plants. This cosmopolitan species is commercially available in all life stages and can be reared on an artificial diet. As a result, V. cardui commonly serves as a model organism for education and outreach. However, gaps in our knowledge remain with regard to V. cardui ecology and larval host plant suitability for supporting complete development of larvae to reproductive adulthood. In a laboratory setting, we tested host plant suitability of five reported host plants commonly found in California sage scrub ecosystems by assessing survival of V. cardui caterpillars in its entirety. We also assessed how commercially sourced larvae survived on artificial diet: both raising the caterpillars exclusively on artificial diet as well as switching diets from artificial to thistle (a commonly reported host plant) and switching from thistle to artificial diet. We found that all commercially sourced caterpillars exhibited both high larval mortality and low pupation rates when reared on host plants, although larvae reared on Malacothamnus fasciculatus and Sphaeralcea ambigua survived longest. Moreover, M. fasciculatus and S. ambigua were the only host plants tested that supported successful pupation. This contrasted with 63% higher survival of wild‐collected larvae. Our findings suggest that commercially obtained V. cardui may struggle to utilize wild host plants and that future investigations into host plant suitability in wild populations are needed. Additionally, future research using commercial larvae should consider the implications for interpreting host plant suitability as larvae may exhibit adaptations to artificial diet or the loss of adaptation to consuming plant material.

Effects of larval diets on some biological parameters and morphometric and biochemical analysis of ovaries of Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae).

The effects of three larval diets (beef meat, chicken meat, and beef liver) on development of Lucilia cuprina (Wiedemann) were evaluated. Egg hatching rates were higher on chicken meat and beef meat (99.5%) than on beef liver (96.5%). Pupation success was higher on chicken meat (99.0%) and beef meat (98.0%) than on beef liver (87.1%). Adult emergence rates were higher in flies reared on chicken meat (99.0%) and beef meat (98.5%) than on beef liver (93.5%). Proportions of female flies were somewhat higher in flies reared on chicken meat or beef meat (54-56% female) than on beef liver (52.5%). Flies that were reared on chicken meat and beef meat had shorter egg-adult development times (16.3 days) than flies reared on beef liver (18.5 days), and the generation time of flies reared on chicken meat or beef meat (51-52 days) was several days shorter than flies reared on beef liver (54 days). Flies reared on chicken meat had larger ovaries and more ovarioles per ovary when larvae were reared on chicken than flies reared on beef meat or beef liver. The ovaries of flies reared on chicken meat had significantly higher amounts of protein (21 mg/g body weight) than flies reared on beef liver (12 mg). Analysis of the larval diets showed that beef liver had the lowest amounts of protein and carbohydrate of the three tested diets, whereas the amounts of lipids did not differ among the diets.

TRNA-derived small RNAs are embedded in the gene regulatory network instructing Drosophila metamorphosis.

A class of noncoding RNAs, referred to as tsRNAs, is emerging with a potential to exert a new layer in gene regulation. These RNAs are breakdown products of tRNAs, either through active processing or passive cleavage or both. Since tRNAs are part of the general machinery for translation, their expression levels and activities are tightly controlled, raising the possibility that their breakdown products, tsRNAs, may provide a link between the overall translational status of a cell to specific changes in gene regulatory network. We hypothesize that Drosophila pupation, being a special developmental stage during which there is a global limitation of nutrients, represents a system in which such a link may readily reveal itself. We show that specific tsRNAs indeed show a dynamic accumulation upon entering the pupal stage. We describe experiments to characterize the mode of tsRNA action and, through the use of such gained knowledge, conduct a genome-wide analysis to assess the functions of dynamically expressed tsRNAs. Our results show that the predicted target genes are highly enriched in biological processes specific to this stage of development including metamorphosis. We further show that tsRNA action is required for successful pupation, providing direct support to the hypothesis that tsRNAs accumulated during this stage are critical to the gene expression program at this stage of development.

Development of a Diet Production System for Conopomorpha cramerella (Lepidoptera: Gracillariidae), a Major Cocoa Production Pest in Southeast Asia and the Pacific Islands.

The development of artificial diets for the cocoa pod borer Conopomorpha cramerella, a major pest of cocoa plants, has undergone significant advancements. In this study, we present the success rates of two diet formulations, MM1 and MM4, which have been progressively improved. Nutritional composition analysis revealed that the MM1 diet differed from the natural host, cocoa pods, in several aspects, including protein, carbohydrate, and vitamin C content. To address these differences, modifications were made to the diet compositions, leading to the MM4 diet version. These modifications resulted in improved diet quality and reduced contamination, leading to enhanced success rates in all stages of C. cramerella development. Larval development, pupation success rates, and adult emergence rates were significantly higher in the MM4 diet compared with the MM1 diet. Moreover, the duration of larval development and pupal stage decreased, while adult longevity increased with the MM4 diet. The overall development success of diet-reared insects from egg to adult was comparable with that of insects reared on cocoa pods. However, the cocoon formation, body length and fresh weight of the adults reared on the artificial diets were lower than those reared on cocoa pods. This diet formulation provides a promising approach for laboratory rearing of C. cramerella and opens avenues for further research and mass-rearing initiatives to mitigate the impact of this pest on cocoa production.

Biology, bionomics and life-table studies of Anopheles stephensi (Diptera: Culicidae) in Sri Lanka and estimating the vectorial potential using mathematical approximations.

Anopheles stephensi is an invasive mosquito in Sri Lanka that can potentially transmit malaria. The transmission intensity is linked with biology, bionomic and behavioral aspects of a vector that are associated with the Vectorial Capacity (VC). However, the influence of larval conditions eventually affects the vectorial potential of An. stephensi are not well understood. A colony of An. stephensi was established at the Regional Centre of the Open University of Sri Lanka, Jaffna District. The colony was maintained under confined conditions according to standard protocols. Biotypes of An. stephensi were characterized by referring to the number of egg ridges. Information on (a) biological aspects of eggs (duration for egg hatching, egg development and hatchability), (b) larval development time, larval survivorship pupation success, resting depth of larvae), (c) pupae (adult emergence rate, average time for adult emergence) and (d) adults (biting frequency, mating success gonotrophic cycle, fecundity, duration for egg-laying, percentage of sexes, adult survival/longevity) were evaluated under life-table analysis. Further, selected morphometric characters of each life cycle stage were recorded from the eggs (length and breadth), larvae (head length, width of head, length of thorax, width of thorax, length of abdomen, width of abdomen, and the total length of larvae), pupae (cephalothoracic length and width) and adults (length & width of wing, thorax and abdomen). The VC was calculated using a mathematical-based approach. Descriptive statistics, General Linear Model (GLM) and independent-sample t-test were used for the statistical analysis. All three biotypes were identified based on egg morphology. Mysorensis biotype (47%; n=470) was predominant followed by type (38.1%; n=381) and intermediate (14.9%; n=149). The mean egg length (F(2,997)=3.56; P=0.029) and breadth (F(2,997)=4.57; P=0.011) denoted significant differences among the three biotypes. The mating success of females observed was 80.7±4.45%. The mean hatching period was 1.9±0.03days, with a hatching rate of 86.2±0.77%. Overall, 8.0±0.14days were required for larval development and 30.3±0.14h were spent in the pupal stage. The pupation success was 94.5±0.37%, and the majority were males (53.1±0.73%). The mean fecundity was 106.5±6.38 eggs and a gonotrophic cycle of 3.4±0.06days. The female survival rate was 43.2±2.4%, with a mean biting frequency of 66.6±3.5%. The average VC of adult An. stephensi was estimated to be 18.7. The type biotype, which is an effective vector in the Indian subcontinent is present in Sri Lanka. According to the mathematical approximation, An. stephensi found locally has a vectorial capacity of over 18. Therefore, this study warrants the health authorities and vector control programmes to continue the entomological surveys, monitoring of vector densities and implementing appropriate vector control interventions based on biology and bionomic information of vectors.

Uric acid metabolism modulates diet-dependent responses to intraspecific competition in Drosophila larvae.

Intraspecific competition drives ecological specialization, and niche expansion. In holometabolous insects, larval intraspecific competition ("crowding") has lasting fitness consequences for individuals and shapes adaptive responses. However, our understanding of the molecular profile of larval crowding responses remains allusive. Here, I used a genetic construct in Drosophila melanogaster with disrupted uric acid metabolism (uro -) to demonstrate the role of the uric acid metabolism-and its interactive effects with larval crowding, diet, and urea concentration-on female oviposition and larval development. Uro - larvae developed faster in sugar-rich diets. However, uro - larvae pupated at lower heights in protein- and sugar-rich diets in crowded larval conditions and showed slower development in protein-rich mixed-genotype environments. uro - did not affect female oviposition nor larval pupation success. Overall, this study provides the first step toward an integrated understanding of the molecular pathways underpinning the responses to intraspecific competition in holometabolous insects.

Molecular characterization of insulin receptor (IR) in oriental fruit moth, Grapholita molesta (Lepidoptera: Tortricidae), and elucidation of its regulatory roles in glucolipid homeostasis and metamorphosis through interaction with miR-982490.

As an intermediate molecule in the Insulin/Insulin-like growth factor signalling pathway (IIS), the insulin receptor (IR) plays vital roles linking nutritional signals to the downstream regulation of metabolic homeostasis, development, metamorphosis, reproduction and stress responses. In the present study, we describe the molecular characteristics of IR in the cosmopolitan fruit boring pest, Grapholita molesta, and its predicted posttranscription regulator miR-982490, and elucidate its regulatory roles in glucolipid homeostasis and metamorphosis. Phylogenetic and domain analyses indicate that lepidopteran IRs normally cluster within families, and that four main domains are conserved in GmIR and those of other Lepidoptera. Bio-informatic prediction, synchronic expression profile evaluation and dual luciferase reporter assays indicated negative regulation of GmIR by miR-982490. Injection of miR-982490 agomir into fifth instar larvae yielded effects similar to dsGmIR injection, resulting in enhanced levels of trehalose and triglyceride in haemolymph, and reduced pupation success and pupal weight, both of which could be rescued by co-injection of dsGmIR and miR-982490 antagomir. We infer that GmIR regulates glucolipid homeostasis and affects G. molesta metamorphosis via interactions with its posttranscriptional regulator miR-982490. This study expands our understanding of the regulatory network of IIS in insect nutritional homeostasis and development.

An improved artificial diet for larvae of the seven-spotted ladybird beetle Coccinella septempunctata L.

The seven-spotted ladybird beetle, Coccinella septempunctata, is an important predator of soft-bodied insect pests that can be reared on both natural and artificial diets. In this study, an artificial diet for larvae was optimized by using a four-factor and three-level orthogonal design. Shrimp, pollen, lard, and honey were selected as the four key factors, and their effects on larval survival, development, and pupation were assessed. The results showed that an increased shrimp content of 21.85% in the diet could shorten developmental time and increase larval survival. Increased pollen content of 4.86% improved both pupation and survival, and a lard content of 3.03% shortened developmental time and increased survival and pupation success. The biological indices of ladybird larvae were optimal when honey content was 10.02%.

Effects of elevated temperatures on the development of immature stages of Anopheles gambiae (s.l.) mosquitoes.

This study investigated the effects of temperature on the development of the immature stages of Anopheles gambiae (s.l.) mosquitoes. Mosquito eggs were obtained from laboratory established colonies and reared under eight temperature regimes (25, 28, 30, 32, 34, 36, 38 and 40°C), and 80 ± 10% relative humidity. Larvae were checked daily for development to the next stage and for mortality. Pupation success, number of adults produced and sex ratio of the newly emerged adults were recorded. Larval survival was monitored every 24h, and data were analysed using Kaplan-Meier survival analysis. Analysis of variance was used where data followed normal distribution, and a Kruskal-Wallis test where data were not normally distributed. Larval and pupal measurements were log-transformed and analysed using ordinary least square regression with robust standard errors. Increasing the temperature from 25 to 36°C decreased the development time by 10.57days. Larval survival (X2 (6) = 5353.12, p < 0.001) and the number of adults produced (X2 (5) = 28.16, p < 0.001) decreased with increasing temperature. Increasing temperatures also resulted in significantly smaller larvae and pupae (p < 0.001). At higher temperatures, disproportionately more male than female mosquitoes were produced. Increased temperature affected different developmental stages in the life cycle of An. gambiae (s.l.) mosquitoes, from larval to adult emergence. This study contributes to the knowledge on the relationship between temperature and Anopheles mosquitoes and provides useful information for modelling vector population dynamics in the light of climate change.

Altered capsaicin levels in domesticated chili pepper varieties affect the interaction between a generalist herbivore and its ectoparasitoid

Plant domestication has commonly reduced levels of secondary metabolites known to confer resistance against insects. Chili pepper is a special case because the fruits of different varieties have been selected for lower and higher levels of capsaicin, the main compound associated with defence. This may have important consequences for insect herbivores and their natural enemies. Despite the widespread consumption of chili peppers worldwide, the effects of capsaicin on insects are poorly understood. Here, we investigated the effect of capsaicin on a generalist herbivore, Spodoptera latifascia (Lepidoptera: Noctuidae) and its ectoparasitoid, Euplectrus platyhypenae (Hymenoptera: Eulophidae). Using chili varieties with three pungency levels: non-pungent (Padron), mild (Cayenne) and highly pungent (Habanero), as well as artificial diets spiked with three different levels of synthetic capsaicin, we determined whether higher capsaicin levels negatively affect the performance of these insects. Overall, capsaicin had a negative effect on both herbivore and parasitoid performance, particularly at high concentrations. Caterpillars reared on highly pungent fruits and high-capsaicin diet had longer development time, reduced pupation success, lower adult emergence, but also lower parasitism rates than caterpillars reared on mild or non-capsaicin treatments. In addition, we found that the caterpillars were capable of sequestering capsaicinoids in their haemolymph when fed on the high pungent variety with consequences for parasitoids’ performance and oviposition decisions. These results increase our understanding of the role of capsaicin as a chemical defence against insects and its potential implications for pest management.

Entomopathogenic nematode management of small hive beetles (Aethina tumida) in three native Alabama soils under low moisture conditions.

The goal was to determine the efficacy of entomopathogenic nematodes (EPNs) on Aethina tumida small hive beetle (SHB) in Alabama soils. The objectives were to (i) determine the pupation success of SHB wandering larvae; (ii) determine the efficacy of EPNs on SHB wandering larvae in natural and autoclaved soil; and (iii) determine the efficacy of EPNs on SHB wandering larvae in three Alabama soil types at typical low moisture levels. The Alabama soils were Kalmia loamy sand (KLS), Benndale fine sandy loam (BFSL), and Decatur silt loam (DSL). Heterorhabditis bacteriophora, H. indica, Steinernema carpocapsae, S. feltiae, S. kraussei, and S. riobrave were tested at population densities of 5, 10, 20, 40, and 80 third-stage infective EPN juveniles (IJ3) per 130 cm3 soil. Pupation success in SHB population densities of 5, 10, and 20 wandering larvae per Petri dish were similar. Of the six EPN species, S. carpocapsae achieved the highest efficacy across all EPN population densities in both natural and autoclaved soil. Steinernema riobrave and H. indica achieved the next highest efficacies; however, they were significantly less effective than S. carpocapsae. Steinernema carpocapsae parasitized 87% SHB wandering larvae across all population densities tested. Steinernema carpocapsae achieved the best efficacy colonizing 94% of the SHB in the KLS soil, 80% in the BFSL soil, and 47% in the DSL soil. In conclusions, S. carpocapsae is be a promising biological control EPN to implement into a management system on SHB.

Development of an Alternative Low-Cost Larval Diet for Mass Rearing of Aedes aegypti Mosquitoes.

Background Aedes aegypti is a major vector of arboviruses that may be controlled on an area-wide basis, using novel approaches such as Sterile Insect Technique (SIT) and Incompatible Insect Technique (IIT). Larval diet is a critical factor to be considered in mass rearing of Aedes mosquitoes for SIT and IIT programs. Therefore, the current study is aimed at evaluating the effects of two novel diets developed from dry fish powder on the growth and development of immature stages and adult fitness-related characteristics of Ae. aegypti in Sri Lanka. Method Three batches of the first instar Ae. aegypti larva, each containing 250 larvae, were exposed to three different larval diets as standard dry fish powder (D1), dry fish powder meal and brewer's yeast (D2), and International Atomic Energy Agency- (IAEA-) recommended diet (D3), separately. Morphometric and developmental parameters of the 4th instar larvae, pupae, and adult mosquitoes reared under different dietary treatments were measured. The entire experimental setup was replicated thrice. A General Linear Model (GLM) in the form of two-way ANOVA was used for the statistical analysis. Results Significant diet-based variations were observed in the head length, head width, thoracic length, thoracic width, abdominal length, abdominal width, and total length (F2,87 > 4.811; P < 0.05) of Ae. aegypti larvae. The highest pupation success and the larval size were observed from the larvae fed the D2 diet, while the lowest was reported from D1. All adult morphometric parameters of adult male and female Ae. aegypti mosquitoes also denoted significant dietary variations, reporting the best-sized adults from the D2 diet (F2,87 > 3.54; P < 0.05). Further, significantly higher fecundity and male longevity were also shown by the adult Ae. aegypti (F2,6 > 7.897; P < 0.01) mosquitoes reared under diet D2. Conclusion Based on all the growth and developmental parameters, the D2 diet tends to perform similar to the IAEA-recommended diet in mass rearing of Ae. aegypti mosquitoes, while being more inexpensive. Therefore, larval diet D2 could be suggested as the ideal diet for mass rearing of Ae. aegypti for IIT and SIT-based vector control in Sri Lanka.

Development and lifespan of Culicoides obsoletus s.s. (Meigen) and other livestock-associated species reared at different temperatures under laboratory conditions.

Culicoides Latreille (Diptera: Ceratopogonidae) transmit arboviruses affecting wild and domestic ruminants such as bluetongue (BTV) and Schmallenberg virus (SBV). The sub-adult development and lifespan of Culicoides obsoletus s.s. (Meigen), Culicoides circumscriptus Kieffer and Culicoides paolae Boorman were examined at three different temperatures under laboratory conditions. Insects were collected from field between spring and autumn 2015 in two livestock farms located in Majorca (Spain). Gravid females were held individually at 18, 25 or 30 °C. Low temperatures increased the adult lifespan, time to oviposit and rate of development, whereas high temperatures increased the number of eggs, successful pupation and adult emergence as well as the larvae growth rate. The results showed that C. obsoletus s.s. have optimum development at 18 °C, whereas the optimal rearing temperature for C. circumscriptus and C. paolae was under warmer conditions of 25-30 °C. Variations in temperature/humidity and assays with different materials and substrates for oviposition should be considered in future studies. Understanding the requirements of the different species of Culicoides optimizing the results should be of special interest for predicting environmental change effects on these species, in addition to determining the rearing conditions for candidate Culicoides vectors.

Successful Pupation of Small Hive Beetle, Aethina tumida (Coleoptera: Nitidulidae), in Greenhouse Substrates.

The small hive beetle, Aethina tumida Murray, is an invasive pest that has spread globally. Western honey bees, Apis mellifera Linnaeus (Hymenoptera: Apidae), are considered the most important host and infestations can lead to collapse of colonies. Larvae feed on honey, pollen, and brood inside the hive and leave the hive as postfeeding wandering larvae to pupate in the surrounding soil. Other host species include bumble bees, stingless bees, and solitary bees, all of which can facilitate small hive beetle reproduction and are used for greenhouse crop pollination worldwide. Here, we investigated if small hive beetles can complete their life cycle when soil is absent by pupating in plant root-supporting substrates commonly used in greenhouses. Wandering small hive beetle larvae were introduced into containers with coconut fiber, perlite, a mixture of both and stone wool substrates to investigate pupation success and development time. Sand was used as control substrate. In all but one substrate (perlite), small hive beetles developed into adults equally well as they did in the sand. Development time ranged between 23 and 37 d and was not different from that of the control. We showed that small hive beetles can pupate in greenhouse substrates. This could constitute a problem for greenhouse pollination as well as it could facilitate small hive beetle survival in areas which otherwise would be deemed unsuitable or marginal environments for small hive beetles to become established. Our study highlights the opportunistic nature of the small hive beetle as an invasive species.

Adverse responses of Cabera pusaria caterpillars to high dietary manganese concentration

AbstractThe adaptation of insects to environmental changes can constitute a crucial factor in their development and activity. The response of Cabera pusaria L. (Lepidoptera: Geometridae) caterpillars to high manganese (Mn) concentrations in the diet was studied. Birch leaves were treated by dipping in MnCl2·4H2O solutions, thereby achieving Mn contents of 370 (T0), 695 (T1), 3 198 (T2), and 6 302 mg kg−1 (T3). The reactions were determined by observing caterpillar mortality, development time, food consumption, and pupal weight. Manganese concentrations in larval excrement, pupae, and food were determined. Manganese in the diet at unnaturally increased concentrations caused great stress for caterpillars. All individuals in the treatment with the highest Mn concentration (T3) died during rearing and successful pupation occurred in only four individuals in T2. Even in the case of caterpillars from T1 (twofold higher than T0) a negative reaction (increased food consumption and prolonged development) was recorded. We also determined significantly increased Mn concentration in pupae from T1 (T2 and T3 were not included in this evaluation due to mortality) and excrement (T1‒T3) compared with T0 having a natural Mn concentration. Caterpillars were seen to eliminate negatively acting dietary Mn by its translocation to excrement. However, the highest mortality rate in T2 and T3 and negative reactions of individuals in T1 very likely demonstrate energy insufficiency and the high energy requirements of Mn elimination mechanisms.

Uptake and toxicity of clothianidin to monarch butterflies from milkweed consumption.

Recent concern for the adverse effects from neonicotinoid insecticides has centered on risk for insect pollinators in general and bees specifically. However, natural resource managers are also concerned about the risk of neonicotinoids to conservation efforts for the monarch butterfly (Danaus plexippus) and need additional data to help estimate risk for wild monarch butterflies exposed to those insecticides. In the present study, monarch butterfly larvae were exposed in the laboratory to clothianidin via contaminated milkweed plants from hatch until pupation, and the effects upon larval survival, larval growth, pupation success, and adult size were measured. Soils dosed with a granular insecticide product led to mean clothianidin concentrations of 10.8–2,193 ng/g in milkweed leaves and 5.8–58.0 ng/g in larvae. Treatment of soils also led to clothianidin concentrations of 2.6–5.1 ng/g in adult butterflies indicating potential for transfer of systemic insecticides from the soil through plants and larvae to adult butterflies. Estimated LC50s for total mortality (combined mortality of larvae and pupae) and EC50 for larval growth were variable but higher than the majority of concentrations reported in the literature for clothianidin contamination of leaves.

Effect of Larval Nutritional Regimes on Morphometry and Vectorial Capacity of Aedes aegypti for Dengue Transmission.

Background Nutritional level in larval diet of mosquito vectors influence on life history traits and vectorial capacity (VC). Therefore, the present study was carried out to assess the effect of larval diet concentration on vector bionomic and VC of Aedes aegypti in Sri Lanka. Method Three batches of 400 Ae. aegypti larvae (first instar) were reared under different concentrations of larval diet (6%, 8%, and 10%; Volume/Volume), which was prepared by mixing 12.5 g of tuna meal, 9.0 g of bovine liver powder, and 3.5 g of Brewer's yeast, in 100 ml of distilled water. The effect of larval diet concentration on different morphometric and functional parameters of larvae (length and width of head, abdomen, survival rate, and pupation success), pupae (length and width of cephalothorax, survival rate, and adult emersion), adult (length and width of thorax, abdomen, survival rate, longevity, biting frequency and fecundity of adults) were examined. In addition, VC of Ae. aegypti was evaluated. The General Linear Model (GLM) was used for the statistical analysis. Results Larval head length, head width, thoracic width, abdominal length, abdominal width, total length, and survival rate significantly increased with higher doses of larval diet (P < 0.05). In case of pupae, length, and width of cephalothorax, survival rate and adult emergence rate denoted an increasing trend with the elevated larval diets. However, the variations of survival rate and adult emergence rate were statistically significant (P < 0.05). In adults, all morphometric parameters (thoracic length, abdominal length, abdominal width, and wing length) significantly increased with elevating larval diets levels (except for thoracic width) along with the biting frequency, fecundity, and survival rate (P < 0.05) of adult females. The VC also denoted significant variations (F4,14 = 24.048; P < 0.05) with the larval diet concentration, whereby the highest VC of 196.37 was observed at 10% treatment. Conclusion Larval food availability has a significant influence on the adult fitness and thus may affect the incidence of dengue due to variations in the VC of Ae.

Environment and host plant quality interact to determine the pupation success of a biological control agent, Agasicles hygrophila, on a nontarget host, Alternanthera sessilis

Understanding insect performance on nontarget native plants is critical in the risk assessment of weed biological control agents. A combination of laboratory and field host tests could help us better understand how abiotic and biotic environments interact to determine insect feeding and development on potential nontarget plants. In this study, we examined the performance of Agasicles hygrophila Selman and Vogt, a beetle used for biological control of the invasive plant alligator weed, Alternanthera philoxeroides (Mart.) Griseb., on a nontarget native plant, Alternanthera sessilis (L.) R.Br. ex DC. Our laboratory tests with various sized A. sessilis stems showed that larvae could pupate and successfully develop into adults on A. sessilis regardless of the stem size, but with larger stems improving the pupation rate. The insect was also found to pupate, and the pupae survive, when there was no stem available under laboratory conditions. When rearing pupae in a Petri dish, increased moisture improved pupal and adult survival, while most pupae were unable to develop to adults under dry conditions. In our field surveys, however, pupae were only found in stems, and no pupae appeared to survive without stems. Our predation tests further showed that pupae exposed to ambient environments (with no stems) had the lowest emergence rate, relative to pupae developing in A. sessilis stems. Together, these results indicate that A. sessilis stems facilitate pupal survival by providing moisture and protecting pupae from attack by natural enemies, explaining the difference in pupation rates between laboratory tests and field observations. Our study provides strong evidence that successful pupation of A. hygrophila on A. sessilis in nature is jointly determined by the environment and the host plant quality. Thus, considering abiotic and biotic factors that influence plant quality, insect survival and development could help to improve the assessment of the ecological host range of the insect biological control agent.