Evolution of horn length and lifting strength in the Japanese rhinoceros beetle Trypoxylus dichotomus.

What limits the size of nature's most extreme structures? For weapons like beetle horns, one possibility is a tradeoff associated with mechanical levers: as the output arm of the lever system-the beetle horn-gets longer, it also gets weaker. This "paradox of the weakening combatant" could offset reproductive advantages of additional increases in weapon size. However, in contemporary populations of most heavily weaponed species, males with the longest weapons also tend to be the strongest, presumably because selection drove the evolution of compensatory changes to these lever systems that ameliorated the force reductions of increased weapon size. Therefore, we test for biomechanical limits by reconstructing the stages of weapon evolution, exploring whether initial increases in weapon length first led to reductions in weapon force generation that were later ameliorated through the evolution of mechanisms of mechanical compensation. We describe phylogeographic relationships among populations of a rhinoceros beetle and show that the "pitchfork" shaped head horn likely increased in length independently in the northern and southern radiations of beetles. Both increases in horn length were associated with dramatic reductions to horn lifting strength-compelling evidence for the paradox of the weakening combatant-and these initial reductions to horn strength were later ameliorated in some populations through reductions to horn length or through increases in head height (the input arm for the horn lever system). Our results reveal an exciting geographic mosaic of weapon size, weapon force, and mechanical compensation, shedding light on larger questions pertaining to the evolution of extreme structures.

Egg development and viability in three species of Cyclocephala (Coleoptera: Scarabaeidae: Dynastinae).

Different species of Cyclocephala scarab beetles (Scarabaeidae, Dynastinae) perform key functional roles in both natural and agricultural systems, such as the cycling of organic matter and pollination, while also being known as destructive pests both as immatures and adults. Therefore, the identification of biological parameters is crucial for defining strategies for their conservation and efficient pest management. In a forest fragment within the Brazilian Atlantic Forest biodiversity hotspot, we field-captured adult individuals of Cyclocephala cearae, C. celata, and C. paraguayensis then reared and bred them under controlled temperature and humidity conditions. On a daily basis, we individually weighted eggs of all three species, from oviposition until hatching, and monitored egg development parameters (i.e., incubation duration, viability, and egg weight increase). Our findings provide novel empirical evidence showing (i) a positive correlation between egg weight and incubation duration, (ii) idiosyncratic characteristics on egg development, and (iii) a negative (involuntary) effect of manipulation on egg development and viability. Thus, the successful breeding and rearing of Cyclocephala spp. is correlated with egg integrity and the targeted species. Our analyses present a quantitative understanding of the egg phase and can assist in refining strategies for ovicidal activity and pest management of Cyclocephala spp. in agriculture systems. Moreover, they can provide a basis for new studies related to captivity breeding, pollinator management, and developmental biology for biodiversity conservation.

Coleoborers in Bambusa sp. in the Southern Amazon / Coleobrocas em Bambusa sp. na Amazonia Meridional

Bamboo (Bambusa sp.) is a grass species with high potential for exploitation, however, this raw material is easily attacked by xylophagous agents such as Coleoptera. The objective of this study is thus to analyse the presence of wood-boring beetles associated with Bambusa sp. in Alta Floresta, Mato Grosso, Brazil. The study was carried out in a bamboo plantation, from August 2016 to July 2017. Two models of ethanolic traps were used, PET Santa Maria and ESALQ-84. Three families of wood-boring beetles were observed in this study: Bostrichidae, Cerambycidae and Curculionidae. 2,144 individuals were identified, belonging to 19 species, in four subfamilies and 11 tribes, and the species Xyleborus affinis was the most frequently collected. Thus, this study constitutes an important contribution to the knowledge of coleoborers associated with bamboo plantations, with the registration of the occurrence of species, as well as contributing to the knowledge of the entomofauna associated with species cultivated in the Southern Amazon.

O bambu (Bambusa sp.) é uma espécie de gramínea com alto potencial de exploração de fibras, no entanto, essa matéria-prima é facilmente atacada por agentes xilófagos, como os coleópteros. O objetivo deste estudo foi analisar a presença de besouros associados a Bambusa sp. em Alta Floresta, Mato Grosso, Brasil. O estudo foi realizado em um plantio de bambu, no período de agosto de 2016 a julho de 2017. Foram utilizados dois modelos de armadilhas etanólicas, o PET Santa Maria e ESALQ-84. Três famílias de coleobrocas foram observadas neste estudo: Bostrichidae, Cerambycidae e Curculionidae. Foram identificados 2,144 indivíduos, pertencentes a 19 espécies, em quatro subfamílias e 11 tribos, e Xyleborus affinis foi a espécie com maior número de indivíduos coletados. Assim, este estudo constitui-se em uma importante contribuição para o conhecimento de coleobrocas associadas a plantios de bamboo, com o registro de ocorrência de espécies, bem como contribui para o conhecimento da entomofauna associada a espécies cultivadas na Amazônia Meridional.

Chewing insects, pollinators, and predators on Acacia auriculiformis A. Cunn. ex Beth (Fabales: Fabaceae) plants fertilized with dehydrated sewage sludge / Insetos mastigadores, polinizadores e predadores em plantas de Acacia auriculiformis A. Cunn. ex Beth (Fabales: Fabaceae) fertilizada com lodo de esgoto desidratado

Fertilization with dehydrated sewage sludge can speed up the recovery process of degraded areas due to nutrients concentration, favoring the development of pioneer plants such as Acacia auriculiformis A. Cunn. ex Beth (Fabales: Fabaceae) and the emergence of insects. This study aimed the evaluation of chewing, pollinating insects, predators, their ecological indices and relationships on A. auriculiformis plants fertilized with dehydrated sewage sludge. The experimental design was completely randomized with two treatments (with and without dehydrated sewage sludge) and 24 repetitions. The prevalence of chewing insects Parasyphraea sp. (Coleoptera: Chrysomelidae), Nasutitermes sp. (Blattodea: Termitidae), and Tropidacris collaris (Stoll, 1813) (Orthoptera: Romaleidae), defoliation, and ecological indices of abundance of Coleoptera and Orthoptera were observed on fertilized A. auriculiformis. Acacia auriculiformis plants, with a superior number of branches/tree, revealed greater abundance of Coleoptera and Orthoptera, species richness of pollinating insects, defoliation, numbers of Parasyphraea sp. and T. collaris. The ones with larger leaves/branches displayed greater abundance of species richness of Coleoptera and Diabrotica speciosa (Germar, 1824) (Coleoptera: Chrysomelidae). Therefore, the use of A. auriculiformis plants, fertilized with dehydrated sewage sludge, is promising in the recovery of degraded areas due to the ecological indices increase of chewing and pollinators insects and spiders in the analyzed area.

A fertilização com lodo de esgoto desidratado pode acelerar o processo de recuperação de áreas degradadas devido à concentração de nutrientes, favorecendo o desenvolvimento de plantas pioneiras tais como Acacia auriculiformis A. Cunn. ex Beth (Fabales: Fabaceae) e de seus insetos. O objetivo deste trabalho foi avaliar os insetos mastigadores, polinizadores e predadores e seus índices e relações ecológicas em plantas de A. auriculiformis fertilizadas com lodo de esgoto desidratado, em área degradada, durante 24 meses. O delineamento foi inteiramente casualizados com dois tratamentos (com e sem adubação com lodo de esgoto desidratado) e 24 repetições (uma repetição = uma planta). O maior número de insetos mastigadores Parasyphraea sp. (Coleoptera: Chrysomelidae), Nasutitermes sp. (Blattodea: Termitidae) e Tropidacris collaris (Stoll, 1813) (Orthoptera: Romaleidae), de desfolha, e do índice ecológico abundância de Coleoptera e de Orthoptera foram maiores em plantas de A. auriculiformis fertilizadas do que nas não fertilizadas com lodo de esgoto desidratado. Plantas de A. auriculiformis, com maior quantidade de galhos/árvore, apresentaram maiores abundâncias de Coleoptera e Orthoptera, riqueza de espécies de insetos polinizadores, desfolha e números de Parasyphraea sp. e T. collaris, e as com maior folhas/galho os de riqueza de espécies de Coleoptera e Diabrotica speciosa (Germar, 1824) (Coleoptera: Chrysomelidae). Por tanto, a utilização de A. auriculiformis, adubada com lodo de esgoto desidratado, é promissora na recuperação de áreas degradadas devido ao aumento dos índices ecológicos de insetos mastigadores, polinizadores e aranhas na área.

Depredación, reproducción y desarrollo de la catarina Hippodamia convergens (Coleoptera: Coccinellidae) / Predation, reproduction, and development of the lady beetle Hippodamia convergens (Coleoptera: Coccinellidae)

Introducción: Mantener poblaciones de insectos depredadores en el agroecosistema ofreciéndoles presas y recursos alimenticios provenientes de las plantas es importante para mejorar el control biológico y contribuir con la conservación. Objetivo: Determinar si el consumo de presas y recursos alimenticios florales potencia los atributos biológicos del depredado Hippodamia convergens. Métodos: Las larvas de Hippodamia convergens fueron alimentadas con dos especies de áfidos, mientras que los adultos fueron alimentados con áfidos, espigas de maíz y dos especies de plantas no cultivadas. La presencia de azúcar en el intestino del depredador se comprobó mediante la prueba de antronas frías, y la presencia de polen mediante el método de acetólisis. Los experimentos demográficos se realizaron durante los primeros 40 días después de la emergencia de la hembra bajo condiciones controladas (25 °C ± 0.5; 75 % ± 1.75 HR; L12:D12). Resultados: H. convergens no consumió fructosa de Sorghum halepense, Parthenium hysterophorus o Zea mays, sino únicamente polen. En condiciones controladas (25 °C ± 0.5, 75 % ± 1.75 HR), el tiempo de desarrollo (huevo-adulto) del depredador fue más corto (21.36 días) cuando consumió Rhopalosiphum maidis en lugar de Uroleucon nigrotibium (24.6 días), mientras que la supervivencia (L1- adulto) y la proporción sexual no cambió. La fecundidad promedio fue mayor (55.5 huevos / 40 días) al consumir U. nigrotibium con polen de P. hysterophorus, que solo U. nigrotibium (22.5 huevos / 40 días), o R. maidis con polen de Zea mays (11 huevos / 40 días). La tasa intrínseca de crecimiento natural fue mayor al consumir U. nigrotibium con polen de P. hysterophorus (0.055), que U. nigrotibium (0.034) o R. maidis con polen de Z. mays (0.019). La tasa de depredación (L1-L5) fue mayor al consumir R. maidis (0.65) que U. nigrotibium (0.51). Conclusiones: Las especies de áfidos y su combinación con polen de plantas no cultivadas afectan de manera diferente el tiempo de desarrollo, reproducción y tasa de depredación de H. convergens.

Introduction: Keeping populations of predatory insects in the agroecosystem by offering them prey, as well as food resources from plants, is important for enhancing conservation biological control. Objective: To determine if the consumption of prey and floral food resources enhances the biological attributes of the predator Hippodamia convergens. Methods: We fed the beetle larvae two species of aphids; and the adults were fed aphids, maize tassels, and two non-cultivated plant species. We checked gut sugar in the predator by the cold anthrone test and pollen presence by the acetolysis method. Demographic experiments were done in the first 40 days after female emergence, under controlled conditions (25 °C ± 0.5; 75 % ± 1.75 Relative Humidity; Light 12 h: Darkness 12 h). Results: H. convergens did not consume fructose, but only pollen from Sorghum halepense, Parthenium hysterophorus or Zea mays. Developmental time (egg-adult) of the predator was shorter (21.4 days) when it consumed Rhopalosiphum maidis than Uroleucon nigrotibium (24.6 days); survival (L1-adult) and sex ratio did not change. Average fecundity was higher (55.5 eggs / 40 days) when consuming U. nigrotibium with pollen from P. hysterophorus, than U. nigrotibium (22.5 eggs / 40 days) or R. maidis with pollen from Zea mays (11 eggs / 40 days). The intrinsic rate of natural growth was higher when consuming U. nigrotibium with pollen from P. hysterophorus (0.055), than U. nigrotibium (0.034) or R. maidis with pollen from Z. mays (0.019). Predation rate (L1-L5) was higher when consuming R. maidis (0.65) than U. nigrotibium (0.51). Conclusions: Aphid prey species and its combination with pollen from maize and non-cultivated plants affected development time, reproduction, and predation rate of H. convergens.

Larval morphology of the Neotropical diving beetle genus Hemibidessus Zimmermann, 1921 (Coleoptera: Dytiscidae: Bidessini).

We describe for the first time the larva of the diving-beetle genus Hemibidessus Zimmermann, 1921, based on instars II and III of H. conicus (Zimmermann, 1921). We include detailed morphometric and chaetotaxic analyses of the cephalic capsule, head appendages, legs, last abdominal segment and urogomphi in order to discover useful characters for distinguishing Hemibidessus larvae from those of other known Bidessini. Hemibidessus larvae characteristically have elongate legs armed with natatory setae and elongate urogomphi that bear numerous secondary setae, features commonly associated with an increased swimming ability. These characters are shared with Brachyvatus Zimmermann, 1921, suggesting a close phylogenetic relationship between these two genera. The primary seta UR8 is inserted apically on urogomphomere 2 in Hemibidessus larvae, a feature so far only shared with the mature larva of Yola Gozis, 1886. Other characters that combined with those previously mentioned are useful to distinguish Hemibidessus from other bidessines are the absence of a ventroapical spinula on antennomere 3 and the presence of secondary dorsal setae on protarsus.

Modelling the emergence dynamics of the western corn rootworm beetle (Diabrotica virgifera virgifera).

The western corn rootworm is an invasive species to Europe and is a major agricultural pest that causes widespread economic and yield losses to maize producers. The Gompertz curve was originally used to model human population mortality. It is a sigmoidal curve where the beginning and end of a period shows the slowest time for growth, and adequately describes observed dynamics of many phenomena. We propose the use of the Gompertz function in a Bayesian Hierarchical framework to model the emergence dynamics of the western corn rootworm beetle. The proposed model includes the use of climatic variables to assess how weather can influence the observed dynamics. We apply the model to Austrian monitoring data collected in 2004-2015.

Oncideres saga Dalman: first record damaging Stryphnodendron adstringens Mart. Coville (Fabaceae) in Brazil / Oncideres saga Dalman: primeiro registro em Stryphnodendron adstringens Mart. Coville (Fabaceae) no Brasil

Oncideres females girdle tree branches of the Fabaceae family, interrupting the sap flow and turning the wood conditions ideal for their larvae development. The bark of Stryphnodendron adstringens (Mart.) Coville, a species native to the Brazilian Cerrado, is widely used in the traditional medicine. The objectives were to report, for the first time, Oncideres saga (Dalman), using S. adstringens as a host and to describe the pattern of branch girdling and oviposition distribution by this insect on these branches. The diameter at the base and the length of the girdled branches were measured and the number of incisions made by the O. saga females to oviposit, per branch section (basal, median and apical), counted. The emerged specimens were counted and the diameter of the exit holes measured. The average diameter at the base of the girdled branches was 2.5 ± 0.16 cm and the length was 90.6 ± 4.6 cm. The average number of incisions per branch was 37.7 ± 2.7. Damage by O. saga can reduce the growth and cause losses on S. adstringens, a tree with great extractivism potential.

As fêmeas de Oncideres anelam galhos de árvores da família Fabaceae, interrompendo o fluxo de seiva, tornando a madeira ideal para o desenvolvimento de suas larvas. A casca de Stryphnodendron adstringens (Mart.) Coville uma espécie nativa do cerrado brasileiro, é amplamente utilizada na medicina tradicional. Os objetivos foram relatar, pela primeira vez, Oncideres saga (Dalman), usando S. adstringens como hospedeiro e descrever o padrão dos galhos anelados e a distribuição das posturas desse inseto. O diâmetro na base e o comprimento dos galhos anelados foram medidos e o número de incisões feitas pelas fêmeas por seção do galho (basal, mediana e apical) quantificados. Os espécimes emergidos foram contados e o diâmetro dos orifícios de emergência medidos. O diâmetro médio, dos galhos anelados, na base foi de 2,58 ± 0,16 cm e o comprimento de 90,6 ± 4,6 cm. O número médio de posturas por galho foi 37,7 ± 2,7. Danos por O. saga podem reduzir o crescimento e causar perdas em S. adstringens, árvore com grande potencial extrativista.

Anadenanthera colubrina (Fabaceae) logs in the Atlantic Forest biome: first host plant for Thoracibidion lineatocolle (Col: Cerambycidae) and a new host for Temnopis megacephala (Col: Cerambycidae) / Troncos de Anadenanthera colubrina (Fabaceae) no bioma da Mata Atlântica: primeira planta hospedeira de Thoracibidion lineatocolle (Col: Cerambycidae) e uma nova planta hospedeira para Temnopis megacephala (Col: Cerambycidae)

Wood-boring beetles develop in live trees and dead wood, performing ecological services such as decomposition and regulation of forest resources. Species of the Cerambycidae family, widely distributed in the world, bore into the trunks of trees and dead wood in native and cultivated areas. The objective is to report the first host plant for Thoracibidion lineatocolle (Thomson, 1865) (Coleoptera: Cerambycidae) and a new host plant for Temnopis megacephala (Germar, 1824) (Coleoptera: Cerambycidae) in the Brazilian Atlantic Forest biome. Three logs, with one-meter-long by 20 cm in diameter, were cut from the trunk of a healthy Anadenanthera colubrina (Fabaceae) tree in October 2013 and tied in the understory at 1.5m high in the Rio Doce State Park, Minas Gerais State, Brazil. The logs, exposed in the forest, were each removed after 40, 80 and 120 days and stored individually in a cardboard box in the "Laboratório de Campo do Projeto de Ecologia de Longa Duração (PELD-CNPq)" in the Rio Doce State Park. A total of 94 individuals of T. lineatocolle and 228 of T. megacephala emerged from the A. colubrina logs. This is the first report of a host plant for T. lineatocolle and a new host plant for T. megacephala.

Besouros broqueadores se desenvolvem em árvores vivas e madeira morta, realizando serviços ecológicos como decomposição e regulação de recursos da floresta. Espécies da família Cerambycidae, amplamente distribuídas no mundo, perfuram o caule de árvores e madeira morta em áreas nativas e cultivadas. O objetivo é relatar a primeira planta hospedeira de Thoracibidion lineatocolle (Thomson, 1865) (Coleoptera: Cerambycidae) e uma nova planta hospedeira para Temnopis megacephala (Germar, 1824) (Coleoptera: Cerambycidae) no bioma da Mata Atlântica brasileira. Três toras, com um metro de comprimento por 20 cm de diâmetro, foram cortadas de uma árvore sadia de Anadenanthera colubrina (Fabaceae) em outubro de 2013 e amarradas no sub-bosque a 1,5m de altura no Parque Estadual do Rio Doce, estado de Minas Gerais, Brasil. As toras, expostas na floresta, foram removidas, cada uma, após 40, 80 e 120 dias e armazenadas, individualmente, em caixas de papelão no “Laboratório de Campo do Projeto de Ecologia de Longa Duração (PELD-CNPq)” no Parque Estadual do Rio Doce. Um total de 94 indivíduos de T. lineatocolle e 228 de T. megacephala emergiu das toras de A. colubrina. Esse é o primeiro registro de uma planta hospedeira para T. lineatocolle e o de uma nova planta hospedeira para T. megacephala.

Notes on the biological development of the darkling beetle Blaps nefrauensis nefrauensis Seidlitz, 1893 (Coleoptera: Tenebrionidae) / Notas sobre o desenvolvimento biológico do besouro escuro Blaps nefrauensis nefrauensis Seidlitz, 1893 (Coleoptera: Tenebrionidae)

Several endemic species of Blaps occur in Tunisia, and the species Blaps nefrauensis nefrauensis has been reported in Moulares (urban zone in west-central Tunisia), where it lives and reproduces in home gardens and old buildings. The aim of this work is to study the life cycle of the darkling beetle, considering both field and laboratory rearing conditions. As a result, the beetle species has different developmental stages (egg, larva, prepupa, pupa, and adult) that last about 15 months. Each year during the same period, adults emerge (early summer) and expire (late autumn), larvae hatch (late summer) and pupate (early summer). There is only one generation per year. Females began laying eggs in late July. The eggs were ovoid, white, and about 2.7 mm in length and 1.5 mm in width. Embryogenesis took an average of nine days. The first instar larvae were at initially only 4.5 mm long and ivory white in color. A brief description of the newly egg hatched larva was provided; thus, the nerve fibers innervating the apical setae in the antennae and ligula were detected. Further light microscopic examination of the embryo before hatching from the egg pointed out that the antennal sensilla are protected during the embryogenesis stage.

Várias espécies endêmicas de Blaps ocorrem na Tunísia, e a espécie Blaps nefrauensis nefrauensis foi relatada em Moulares (zona urbana no centro-oeste da Tunísia), onde vive e se reproduz em jardins domésticos e prédios antigos. O objetivo deste trabalho é estudar o ciclo de vida do besouro escuro, considerando as condições de criação em campo e em laboratório. Como resultado, a espécie de besouro tem diferentes estágios de desenvolvimento (ovo, larva, prepupa, pupa e adulto) que duram cerca de 15 meses. Todos os anos, durante o mesmo período, os adultos emergem (início do verão) e expiram (final do outono), as larvas eclodem (final do verão) e se tornam pupas (início do verão). Existe apenas uma geração por ano. As fêmeas começaram a botar ovos no final de julho. Os ovos eram ovóides, brancos, com cerca de 2,7 mm de comprimento e 1,5 mm de largura. A embriogênese demorou em média nove dias. As larvas de primeiro instar tinham inicialmente apenas 4,5 mm de comprimento e uma cor branca marfim. Foi fornecida uma breve descrição da larva recém-eclodida; assim, as fibras nervosas que inervam as cerdas apicais nas antenas e ligulas foram detectadas. Um exame microscópico de luz posterior do embrião antes da eclosão do ovo mostrou que as sensilas antenais são protegidas durante o estágio de embriogênese.

Modeling the impact of temperature on the population abundance of the ambrosia beetle Xyleborus affinis (Curculionidae: Scolytinae) under laboratory-reared conditions.

Modeling the impact of temperature on each life stage of a beetle population represents a continuing challenge. This study evaluates the effects of five temperature treatments (20, 23, 26, 29 and 32 °C) on population abundance and timing of a colony of ambrosia beetles Xyleborus affinis reared under laboratory conditions and use this data to develop demographic and phenological models. Abundances at each life stage (eggs, larvae, pupae and adult) were examined through periodic destructive sampling; given that it was not possible to track individuals. To assess the effects of temperature on oviposition, development and survival rates we developed a novel estimation strategy based on cohorts, which does not require individual developmental data. Since oviposition was entirely unwitnessed, we assessed competing empirical ovipositional models. Rates of development were computed using a modal rate curve for each life stage, and rates were projected to cohorts in life stages assuming log-normal developmental variance. Temperature-driven survival rates were assumed to be logistic with a quadratic exponent to capture modal temperature dependence. Parameters were estimated simultaneously using minimum negative log posterior likelihood, assuming Poisson distribution of observations and using priors to inform unobserved developmental rates and enforce mechanistic constraints on oviposition models. A parabolic function best described oviposition rate. Optimal developmental temperatures were 30.5 °C, 29 °C and 27.5 °C, with maximum developmental rates of 0.26/day, 0.12/day and 0.23/day for eggs, larvae and pupae, respectively. The survival rates in the range 20-29 °C were equal to 1 in the eggs-to-larvae transition, from 0.72 to 0.35 in larvae-to-pupae transition, and from 0.2 to 0.89 in pupae-to-adults transition. This procedure effectively characterized the direct thermal effects on development and survival of each life stage in the X. affinis under laboratory conditions and would be suitable for estimating temperature dependence for other species in which individual observations are not possible.

Structural and functional insights into the first Bacillus thuringiensis vegetative insecticidal protein of the Vpb4 fold, active against western corn rootworm.

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major maize pest in the United States causing significant economic loss. The emergence of field-evolved resistant WCR to Bacillus thuringiensis (Bt) traits has prompted the need to discover and deploy new insecticidal proteins in transgenic maize. In the current study we determined the crystal structure and mode of action (MOA) of the Vpb4Da2 protein (formerly known as Vip4Da2) from Bt, the first identified insecticidal Vpb4 protein with commercial level control against WCR. The Vpb4Da2 structure exhibits a six-domain architecture mainly comprised of antiparallel ß-sheets organized into ß-sandwich layers. The amino-terminal domains 1-3 of the protein share structural homology with the protective antigen (PA) PA14 domain and encompass a long ß-pore forming loop as in the clostridial binary-toxB module. Domains 5 and 6 at the carboxyl-terminal half of Vpb4Da2 are unique as this extension is not observed in PA or any other structurally-related protein other than Vpb4 homologs. These unique Vpb4 domains adopt the topologies of carbohydrate-binding modules known to participate in receptor-recognition. Functional assessment of Vpb4Da2 suggests that domains 4-6 comprise the WCR receptor binding region and are key in conferring the observed insecticidal activity against WCR. The current structural analysis was complemented by in vitro and in vivo characterizations, including immuno-histochemistry, demonstrating that Vpb4Da2 follows a MOA that is consistent with well-characterized 3-domain Bt insecticidal proteins despite significant structural differences.

Bacillus thuringiensis Spores and Cry3A Toxins Act Synergistically to Expedite Colorado Potato Beetle Mortality.

The insect integument (exoskeleton) is an effective physiochemical barrier that limits disease-causing agents to a few portals of entry, including the gastrointestinal and reproductive tracts. The bacterial biopesticide Bacillus thuringiensis (Bt) enters the insect host via the mouth and must thwart gut-based defences to make its way into the body cavity (haemocoel) and establish infection. We sought to uncover the main antibacterial defences of the midgut and the pathophysiological features of Bt in a notable insect pest, the Colorado potato beetle Leptinotarsa decemlineata (CPB). Exposing the beetles to both Bt spores and their Cry3A toxins (crystalline δ-endotoxins) via oral inoculation led to higher mortality levels when compared to either spores or Cry3A toxins alone. Within 12 h post-exposure, Cry3A toxins caused a 1.5-fold increase in the levels of reactive oxygen species (ROS) and malondialdehyde (lipid peroxidation) within the midgut - key indicators of tissue damage. When Cry3A toxins are combined with spores, gross redox imbalance and 'oxidation stress' is apparent in beetle larvae. The insect detoxification system is activated when Bt spores and Cry3A toxins are administered alone or in combination to mitigate toxicosis, in addition to elevated mRNA levels of candidate defence genes (pattern-recognition receptor, stress-regulation, serine proteases, and prosaposin-like protein). The presence of bacterial spores and/or Cry3A toxins coincides with subtle changes in microbial community composition of the midgut, such as decreased Pseudomonas abundance at 48 h post inoculation. Both Bt spores and Cry3A toxins have negative impacts on larval health, and when combined, likely cause metabolic derangement, due to multiple tissue targets being compromised.

Nondetrimental impact of two concomitant entomopathogenic fungi on life history parameters of a generalist predator, Coccinella septempunctata (Coleoptera: Coccinellidae).

The non-persistent impact of biocontrol agents can be revealed for pest control when associated entomopathogenic fungi (EPFs) negatively affect the natural enemies. In this assay, impacts of Beauvaria bassiana (Balsamo) Vuillemin, and Metarhizium anisopliae (Metschnikoff) Sorokin were studied for their compatibility or side effects on life table parameters of an important generalist predator, Coccinella septempunctata L. The results indicated non-significant impacts of both EPFs on life table parameters of C. septempunctata. The development time (egg-adult) was not significantly different in control (69.79 days) and EPFs treated C. septempunctata (69.35-80.07 days). Both fungi did not induce any significant changes in the fecundity, adult pre-oviposition period (APOP), total preoviposition period (TPOP), and mean generation time (T) as compared to control treatment. Similarly, no difference in fecundity rate of C. septempunctata was observed after EPFs treatment (287.7-288.5) compared to control (290.0). The highest net reproductive rate (R0) occurred in control (87.05 offspring individual-1) and M. anisopliae (86.31 offspring individual-1) as compared to B. bassiana treated beetles (76.97 offspring individual-1). The age-specific fecundity curves indicated that the C. septempunctata had a similar fecundity rate in both EPFs treatments and control. This study demonstrates no significant side effects of B. bassiana and M. anispoliae on the performance and biology of C. septempunctata. Considering the compatibility of both EPFs with C. septempunctata, their combinations can be recommended in various integrated pest management programs.

The structure of the cereal leaf beetle (Oulema melanopus) microbiome depends on the insect's developmental stage, host plant, and origin.

Cereal leaf beetle (CLB, Oulema melanopus, Coleoptera, Chrysomelidae) is a serious agricultural pest that causes considerable damages to agricultural production. The aim of this study was to characterize the bacterial communities associated with larvae and imagoes of CLB collected from various cereal host species and locations. The bacterial profile was characterized by 16S rRNA gene sequencing at the V3-V4 hypervariable region. Using taxonomy-based analysis, the bacterial community of CLB containing 16 phyla, 26 classes, 49 orders, 78 families, 94 genera, and 63 species of bacteria was identified. The abundance of Wolbachia, Rickettsia, and Lactococcus genus was significantly higher in CLB imagoes than in larvae. Statistical analysis confirmed that the bacterial community of the larvae is more diverse in comparison to imagoes and that insects collected from spring barley and wheat are characterized by a much higher biodiversity level of bacterial genera and species than insects collected from other cereals. Obtained results indicated that the developmental stage, the host plant, and the insect's sampling location affected the CLB's microbiome. Additionally, the CLB core microbiome was determined. It consists of 2 genera (Wolbachia and Rickettsia) shared by at least 90% tested CLB insects, regardless of the variables analysed.

Is developmental rate polymorphism constant? Influence of temperature on the occurrence and constancy of slow and fast development in Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae).

Temperature plays an important role in different biological activities of organisms. The relationship between temperature and insect development has long been recognized as an important environmental parameter in modeling insect population dynamics. Although few studies have investigated the existence of developmental rate polymorphism within a cohort, the role of abiotic and biotic factors on such developmental variation has so far been meagerly investigated. The present study was designed to investigate the effect of thermal extremes on the developmental rate polymorphism and its influence on reproductive potential of Parthenium beetle, Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae). The study will also be helpful in solving hitherto whether the existence of slow and fast developers within a cohort has a purely genetic basis or this developmental polymorphism is presided by environmental factors. Our result reveals a clear bimodal pattern of distribution with two peaks at each temperature (20, 25, 27, 30 and 35 °C) where the first peak represents the fast developers and second peak represents the slow developers. Both developmental variants took the longest duration for development at 20 °C followed by 25, 27, 30 °C and minimum at 35 °C. More fast developers were found at higher temperatures. Slow developing individuals were heavier than the fast developing individuals regardless of rearing temperature. Slow developers have higher reproductive success in terms of fecundity and egg viability than the fast developers. The results of this study denote the constancy of the developmental rate polymorphism within a cohort and the possibility that this polymorphism was owing to the exogenous cues inclined differential rates of mortality.

Developmental models of the carrion beetle Thanatophilus rugosus (Linnaeus, 1758) (Coleoptera: Silphidae).

Coleoptera are currently considered a fundamental tool to help solve criminal investigations, allowing forensic entomologists to estimate post-mortem intervals and obtain other ecology-related information. Thanatophilus rugosus (Linnaeus, 1758) is an important necrophagous beetle distributed through most of the Palaearctic region, where it is readily found on human bodies and animal carcasses. In this study, the new thermal summation models for all the developmental stages of Thanatophilus rugosus are provided. Beetles were reared at six different constant and ecologically relevant temperatures (12, 14, 16, 18, 20, and 22 °C), and their developmental times were measured. Thermal summation constants were calculated for each developmental stage (egg, three larval instars, post-feeding stage, and pupa).

Carbon Dioxide-Enriched Atmosphere to Control Oryzaephilus surinamensis L. On Stored Saqie Date Fruits.

<b>Background and Objective:</b> Control of the stored date insects by an eco-friendly method is essential to maintain the fruit for prolonged periods, especially that dates are eaten fresh. The search for a safe method, as an alternative to commonly used chemical pesticides, is needed. The objective is to investigate CO₂ enriched atmosphere on the Mortality Percentage (MP) of the Saw-toothed grain beetle, <i>O. surinamensis</i>, life stages with special emphasis on reducing damage to stored dates. <b>Materials and Methods:</b> The effect of high levels of CO₂ as an alternative control method against the Saw-toothed grain beetle, <i>Oryzaephilus surinamensis</i> L., at different life stages was studied on infested 'Saqie' dates. Four CO₂ pressures (25, 50, 75 and 90 kPa, balance is nitrogen) were tested for 6, 12, 18, 24, 48, 72 and 96 hrs intervals. The response of different life stages of <i>O. surinamensis</i> to the different treatments varied according to CO₂ level, developmental stage and exposure period. <b>Results:</b> Mortality (%) was higher during the larval stage, followed by adults, pupae and eggs, in descending order. The larvae and adult stages were more sensitive to CO₂ treatment than the pupal and egg. Exposure time was more effective on eggs, larval and adult MP than the CO₂ atmosphere level. Mortality% at 96 hrs exposure time was almost 100% with CO₂ atmospheres of 50, 75 and 90 kPa. Mathematical equations were developed to model the relationship between mortality% and CO₂ treatments using multiple regression analyses for each life stage. <b>Conclusion:</b> The results confirmed that CO₂ could be applied to final food products during packaging to control the residual occurrence of insect pests after storage and before the packaging process to prevent further infestation in the final packages.

Effect of food source availability in the salivary gland transcriptome of the unique burying beetle Nicrophorus pustulatus (Coleoptera: Silphidae).

Nicrophorus is a genus of beetles that bury and transform small vertebrate carcasses into a brood ball coated with their oral and anal secretions to prevent decay and that will serve as a food source for their young. Nicrophorus pustulatus is an unusual species with the ability to overtake brood of other burying beetles and whose secretions, unlike other Nicrophorus species, has been reported not to exhibit antimicrobial properties. This work aims to better understand how the presence or absence of a food source influences the expression of genes involved in the feeding process of N. pustulatus. To achieve that, total RNA was extracted from pooled samples of salivary gland tissue from N. pustulatus and sequenced using an Illumina platform. The resulting reads were used to assemble a de novo transcriptome using Trinity. Duplicates with more than 95% similarity were removed to obtain a "unigene" set. Annotation of the unigene set was done using the Trinotate pipeline. Transcript abundance was determined using Kallisto and differential gene expression analysis was performed using edgeR. A total of 651 genes were found to be differentially expressed, including 390 upregulated and 261 downregulated genes in fed insects compared to starved. Several genes upregulated in fed beetles are associated with the insect immune response and detoxification processes with only one transcript encoding for the antimicrobial peptide (AMP) defensin. These results confirm that N. pustulatus does not upregulate the production of genes encoding AMPs during feeding. This study provides a snapshot of the changes in gene expression in the salivary glands of N. pustulatus following feeding while providing a well described transcriptome for the further analysis of this unique burying beetle.