Insect Tissues Research Articles

Entomotoxicology as a tool for forensic investigations

Entomotoxicology is a rapidly developing field in forensic investigations that combines the disciplines of forensic entomology and toxicology. This paper reviews the application of entomotoxicology as a tool for forensic investigations and highlights its potential in providing valuable information on the presence and effects of drugs or toxins in insects found at a crime scene. The analysis of insect evidence in forensic investigations has long been used to estimate the time of death and other important factors. However, the incorporation of toxicology into this analysis can provide critical information on the presence and effects of drugs or toxins in the body. Entomotoxicology has the potential to be a valuable tool in forensic investigations, particularly in cases involving drug-related deaths or poisonings. By studying the presence and distribution of drugs or toxins in insect tissue, entomotoxicology can provide information on the timing and pattern of drug use. The use of entomotoxicology in forensic investigations is not without challenges. Factors such as post-mortem redistribution and contamination of the crime scene can impact the reliability of the results. There is also a need for standardized protocols and validation studies to ensure the accuracy and reliability of the results.

PHOTOSENSITIVE EFFECT OF THE BIOCHEMICAL AND ECONOMIC TRAITS OF Bombyx mori L.

The silkworm is by nature very delicate and sensitive to environmental conditions. The ecological factors, chiefly temperature, humidity and light during rearing show significant effect on growth larvae and finally cocoon crop quality. Silkworms are photosensitive and they have a tendency to crawl towards dim light. They do not like either strong light or complete darkness. In this way, based on various exposures of light on the effect of biochemical content and economic traits of silkworm were recorded. Biochemical content such as protein is highly essential for insects. The intracellular transport of the silk protein is dependent on hydrophobicity or hydrophilicity of the amino acid residues of the heavy and light chain of the silk protein and the haemolymph is the only extracellular fluid of insects with diverse functions and serves as reservoir for the products required for every physiological activity of the insect body, thus changes in the composition of haemolymph reflect the physiological and biochemical transformations taking place in the insect tissue. In this way of study, effect of light on biochemical content such as protein and carbohydrate of haemolyph and silk gland are significantly increased. The protein content was high in the haemolymph (17.48%) when the larvae exposed to 5 watt light and silk gland (66.67%) in 3 watt exposure group but fat body (44.33 %) when the larvae exposed to 8 watt. The carbohydrate content of haemolymph (35.54 %), silk gland (12.85%) and fat body (57.26 %) were significantly increased in 3 watt exposure group. In this findings, biochemical content had direct relation to economic traits of B. mori. The cocoon characters such as cocoon weight (28.42 per cent), pupal weight (20.9 per cent), shell weight (71 per cent) and shell ratio (33.48 per cent) were significantly increased in the 3 watt exposed group.

Studying Plant-Insect Interactions through the Analyses of the Diversity, Composition, and Functional Inference of Their Bacteriomes.

As with many other trophic interactions, the interchange of microorganisms between plants and their herbivorous insects is unavoidable. To test the hypothesis that the composition and diversity of the insect bacteriome are driven by the bacteriome of the plant, the bacteriomes of both the plant Datura inoxia and its specialist insect Lema daturaphila were characterised using 16S sRNA gene amplicon sequencing. Specifically, the bacteriomes associated with seeds, leaves, eggs, guts, and frass were described and compared. Then, the functions of the most abundant bacterial lineages found in the samples were inferred. Finally, the patterns of co-abundance among both bacteriomes were determined following a multilayer network approach. In accordance with our hypothesis, most genera were shared between plants and insects, but their abundances differed significantly within the samples collected. In the insect tissues, the most abundant genera were Pseudomonas (24.64%) in the eggs, Serratia (88.46%) in the gut, and Pseudomonas (36.27%) in the frass. In contrast, the most abundant ones in the plant were Serratia (40%) in seeds, Serratia (67%) in foliar endophytes, and Hymenobacter (12.85%) in foliar epiphytes. Indeed, PERMANOVA analysis showed that the composition of the bacteriomes was clustered by sample type (F = 9.36, p < 0.001). Functional inferences relevant to the interaction showed that in the plant samples, the category of Biosynthesis of secondary metabolites was significantly abundant (1.4%). In turn, the category of Xenobiotics degradation and metabolism was significantly present (2.5%) in the insect samples. Finally, the phyla Proteobacteria and Actinobacteriota showed a pattern of co-abundance in the insect but not in the plant, suggesting that the co-abundance and not the presence−absence patterns might be more important when studying ecological interactions.

Ecological disturbances and abundance of anthropogenic pollutants in the aquatic ecosystem: Critical review of impact assessment on the aquatic animals

Anthropogenic toxins are discharged into the environment and distributed through a variety of environmental matrices. Trace contaminant detection and analysis has advanced dramatically in recent decades, necessitating further specialized technique development. These pollutants can be mobile and persistent in small amounts in the environment, and ecological receptors will interact with it. Despite the fact that few researches have been undertaken on invertebrate exposure, accumulation, and biological implications, it is apparent that a wide range of pollutants can accumulate in the tissues of aquatic insects, earthworms, amphipod crustaceans, and mollusks. Due to long-term stability during long-distance transit, a number of chemical and microbiological agents that were not previously deemed pollutants have been found in various environmental compartments. The uptake of such pollutants by the aquatic organism is done through the process of bioaccumulation when dangerous compounds accumulate in living beings while biomagnification is the process of a pollutant becoming more hazardous as it moves up the trophic chain. Organic and metal pollution harms animals of every species studied so far, from bacteria to phyla in between. The environmental protection agency says these poisons harm humans as well as a variety of aquatic organisms when the water quality is sacrificed in typical wastewater treatment systems. Contrary to popular belief, treated effluents discharged into aquatic bodies contain considerable levels of Anthropogenic contaminants. This evolution necessitates a more robust and recent advancement in the field of remediation and their techniques to completely discharge the various organic and inorganic contaminants.

The Dielectric Properties of Worker Bee Homogenate in a High Frequency Electric Field

Biological tissues, including insect tissues, are among lossy dielectric materials. The permittivity properties of these materials are described by loss factor ε″ and loss tangent tgδ. The dielectric properties of the worker honeybee body homogenate are tested in the range of high frequencies from 1 MHz to 6 GHz. The homogenate is produced by mixing whole worker honeybees and tested with an epsilometer from Compass Technology and a Copper Mountain Technologies vector circuit analyser VNA. Due to their consistency, the homogenate samples are placed inside polyurethane sachets. The measured permittivity relates to two components of a sample: homogenate and polyurethane. For five samples, two extremes were specified for the permittivity, loss factor ε″, and the loss tangent tgδ, for the frequency range 20 ÷ 80 MHz and 3 GHz. Four techniques of testing permittivity in biological tissues were used to determine the dielectric properties of the homogenate. A calculation model was developed featuring a minimum measurement error of the loss factor ε″ and the loss tangent tgδ. The power absorbed per unit volume is described for the whole frequency range.

Nucleic acid extraction and multiplex analysis for simultaneous detection of the corn stunt complex pathogens in plant and insect tissues

Nucleic acid extraction and multiplex analysis for simultaneous detection of the corn stunt complex pathogens in plant and insect tissues

Polerovirus N-terminal readthrough domain structures reveal molecular strategies for mitigating virus transmission by aphids

Poleroviruses, enamoviruses, and luteoviruses are icosahedral, positive sense RNA viruses that cause economically important diseases in food and fiber crops. They are transmitted by phloem-feeding aphids in a circulative manner that involves the movement across and within insect tissues. The N-terminal portion of the viral readthrough domain (NRTD) has been implicated as a key determinant of aphid transmission in each of these genera. Here, we report crystal structures of the NRTDs from the poleroviruses turnip yellow virus (TuYV) and potato leafroll virus (PLRV) at 1.53-Å and 2.22-Å resolution, respectively. These adopt a two-domain arrangement with a unique interdigitated topology and form highly conserved dimers that are stabilized by a C-terminal peptide that is critical for proper folding. We demonstrate that the PLRV NRTD can act as an inhibitor of virus transmission and identify NRTD mutant variants that are lethal to aphids. Sequence conservation argues that enamovirus and luteovirus NRTDs will follow the same structural blueprint, which affords a biological approach to block the spread of these agricultural pathogens in a generalizable manner.

The extended analogy of extraembryonic development in insects and amniotes.

It is fascinating that the amnion and serosa/chorion, two extraembryonic (EE) tissues that are characteristic of the amniote vertebrates (mammals, birds and reptiles), have also independently evolved in insects. In this review, we offer the first detailed, macroevolutionary comparison of EE development and tissue biology across these animal groups. Some commonalities represent independent solutions to shared challenges for protecting the embryo (environmental assaults, risk of pathogens) and supporting its development, including clear links between cellular properties (e.g. polyploidy) and physiological function. Further parallels encompass developmental features such as the early segregation of the serosa/chorion compared to later, progressive differentiation of the amnion and formation of the amniotic cavity from serosal–amniotic folds as a widespread morphogenetic mode across species. We also discuss common developmental roles for orthologous transcription factors and BMP signalling in EE tissues of amniotes and insects, and between EE and cardiac tissues, supported by our exploration of new resources for global and tissue-specific gene expression. This highlights the degree to which general developmental principles and protective tissue features can be deduced from each of these animal groups, emphasizing the value of broad comparative studies to reveal subtle developmental strategies and answer questions that are common across species.This article is part of the theme issue ‘Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom’.

Multilocus sequence typing of diverse phytoplasmas using hybridization probe-based sequence capture provides high resolution strain differentiation.

Phytoplasmas are insect-vectored, difficult-to-culture bacterial pathogens that infect a wide variety of crop and non-crop plants, and are associated with diseases that can lead to significant yield losses in agricultural production worldwide. Phytoplasmas are currently grouped in the provisional genus ‘Candidatus Phytoplasma’, which includes 49 ‘Candidatus’ species. Further differentiation of phytoplasmas into ribosomal groups is based on the restriction fragment length polymorphism (RFLP) pattern of the 16S rRNA-encoding operon, with more than 36 ribosomal groups (16Sr) and over 100 subgroups reported. Since disease symptoms on plants are not associated with phytoplasma identity, accurate diagnostics is of critical importance to manage disease associated with these microorganisms. Phytoplasmas are typically detected from plant and insect tissue using PCR-based methods targeting universal taxonomic markers. Although these methods are relatively sensitive, specific and are widely used, they have limitations, since they provide limited resolution of phytoplasma strains, thus necessitating further assessment of biological properties and delaying implementation of mitigation measures. Moreover, the design of PCR primers that can target multiple loci from phytoplasmas that differ at the sequence level can be a significant challenge. To overcome these limitations, a PCR-independent, multilocus sequence typing (MLST) assay to characterize an array of phytoplasmas was developed. Hybridization probe s targeting cpn60, tuf, secA, secY, and nusA genes, as well as 16S and rp operons, were designed and used to enrich DNA extracts from phytoplasma-infected samples for DNA fragments corresponding to these markers prior to Illumina sequencing. This method was tested using different phytoplasmas including ‘Ca. P. asteris’ (16SrI-B), ‘Ca. P. pruni’ (16SrIII-A),‘Ca. P. prunorum’ (16SrX-B), ‘Ca. P. pyri’ (16SrX-C), ‘Ca. P. mali’ (16SrX-A), and ‘Ca. P. solani’ (16SrXII-A). Thousands of reads were obtained for each gene with multiple overlapping fragments, which were assembled to generate full-length (typically >2 kb), high-quality sequences. Phytoplasma groups and subgroups were accurately determined based on 16S ribosomal RNA and cpn60 gene sequences. Hybridization-based MLST facilitates the enrichment of target genes of phytoplasmas and allows the simultaneous determination of sequences corresponding to seven different markers. In this proof-of-concept study, hybridization-based MLST was demonstrated to be an efficient way to generate data regarding ‘Ca. Phytoplasma’ species/strain differentiation.

Comparative Insecticide Uptake Characteristics of Chlorfenapyr Suggests an Additional Reason Why Bed Bugs (Hemiptera: Cimicidae) are so Difficult to Control.

Insecticides interact with biochemical sites, disrupting homeostasis, leading to moribundity and death. Considering this process of intoxication, investigating insecticide uptake ensures an understanding of exposure profile. This study evaluated insecticide uptake by bed bugs, Cimex lectularius L., as affected by time and formulation, and a comparative uptake by German cockroaches, Blattella germanica (L.). Insects exposed to chlorfenapyr residues from two Phantom insecticide formulations (i.e., Aerosol and Suspended Concentrate (SC)) were solvent extracted and analyzed by GC-ECD. Chlorfenapyr uptake by bed bugs significantly increased with time, but the rate of external chlorfenapyr adsorption between the two formulations was not significantly different. However, initial exposure and uptake to the aerosol-treated versus SC-treated surfaces were significantly greater by 2.7x. Absorption-rates of chlorfenapyr residues inside the insect, associated with internal insect tissues or exoskeleton-bound, were significantly lower compared with external adsorption, with aerosol and SC formulations accumulating at 4.8x and 14.4x less, respectively. An unexpected formulation effect occurred with aerosol-treated papers and aging of residues. A reformulation of the aerosol provided a greater adsorption of chlorfenapyr at 19.1x that of the original formula after 2 hr. External versus internal uptake of chlorfenapyr by cockroaches was inverse to that seen in bed bugs, with greater chlorfenapyr absorption. Obstructing cockroach mouthparts prevented an internal uptake of chlorfenapyr and provided a similar exposure profile to bed bugs. This work demonstrates a need to evaluate insecticide uptake with formulation and insect morphology to maximize dose, especially in such a morphologically and environmentally isolated insect as the bed bug.

In vitro Insect Fat Cultivation for Cellular Agriculture Applications.

Cell-cultured fat could provide important elements of flavor, nutrition, and texture to enhance the quality and therefore expand consumer adoption of alternative meat products. In contrast to cells from livestock animals, insect cells have been proposed as a relatively low-cost and scalable platform for tissue engineering and muscle cell-derived cultured meat production. Furthermore, insect fat cells have long been cultured and characterized for basic biology and recombinant protein production but not for food production. To develop a food-relevant approach to insect fat cell cultivation and tissue engineering, Manduca sexta cells were cultured and induced to accumulate lipids in 2D and 3D formats within decellularized mycelium scaffolding. The resultant in vitro fat tissues were characterized and compared to in vivo fat tissue data by imaging, lipidomics, and texture analyses. The cells exhibited robust lipid accumulation when treated with a 0.1 mM soybean oil emulsion and had "healthier" fat profiles, as measured by the ratio of unsaturated to saturated fatty acids. Mycelium scaffolding provided a simple, food-grade approach to support the 3D cell cultures and lipid accumulation. This approach provides a low-cost, scalable, and nutritious method for cultured fat production.

What Goes in Must Come Out? The Metabolic Profile of Plants and Caterpillars, Frass, And Adults of Asota (Erebidae: Aganainae) Feeding on Ficus (Moraceae) in New Guinea.

Insect herbivores have evolved a broad spectrum of adaptations in response to the diversity of chemical defences employed by plants. Here we focus on two species of New Guinean Asota and determine how these specialist moths deal with the leaf alkaloids of their fig (Ficus)hosts. As each focal Asota species is restricted to one of three chemically distinct species of Ficus, we also test whether these specialized interactions lead to similar alkaloid profiles in both Asota species. We reared Asota caterpillars on their respective Ficus hosts in natural conditions and analyzed the alkaloid profiles of leaf, frass, caterpillar, and adult moth samples using UHPLC-MS/MS analyses. We identified 43 alkaloids in our samples. Leaf alkaloids showed various fates. Some were excreted in frass or found in caterpillars and adult moths. We also found two apparently novel indole alkaloids-likely synthesized de novo by the moths or their microbiota-in both caterpillar and adult tissue but not in leaves or frass. Overall, alkaloids unique or largely restricted to insect tissue were shared across moth species despite feeding on different hosts. This indicates that a limited number of plant compounds have a direct ecological function that is conserved among the studied species. Our results provide evidence for the importance of phytochemistry and metabolic strategies in the formation of plant-insect interactions and food webs in general. Furthermore, we provide a new potential example of insects acquiring chemicals for their benefit in an ecologically relevant insect genus.

Reversible mitophagy drives metabolic suppression in diapausing beetles

Many insects enter a state of dormancy (diapause) during winter in which they lower their metabolism to save energy. Metabolic suppression is a hallmark of diapause, yet we know little about the mechanisms underpinning metabolic suppression in winter or how it is reversed in the spring. Here, we show that metabolic suppression in dormant Colorado potato beetles results from the breakdown of flight muscle mitochondria via mitophagy. Diapausing Colorado potato beetles suppress their metabolism by 90%, and this lowered metabolic rate coincides with a similar reduction in flight muscle mitochondrial function and density. During early diapause, beetles increase the expression of mitophagy-related transcripts (Parkin and ATG5) in their flight muscle coincident with an increase in mitophagy-related structures in the flight muscle. Knocking down Parkin expression with RNA interference in diapausing beetles prevented some mitochondrial breakdown and partially restored the whole animal metabolic rate, suggesting that metabolic suppression in diapausing beetles is driven by mitophagy. In other animals and in models of disease, such large-scale mitochondrial degradation is irreversible. However, we show that as diapause ends, beetles reverse mitophagy and increase the expression of PGC1α and NRF1 to replenish flight muscle mitochondrial pools. This mitochondrial biogenesis is activated in anticipation of diapause termination and in the absence of external stimuli. Our study provides a mechanistic link between mitochondrial degradation in insect tissues over the winter and whole-animal metabolic suppression.

In-Field LAMP Detection of Flavescence Dorée Phytoplasma in Crude Extracts of the Scaphoideus titanus Vector

One of the most destructive diseases affecting grapevine in Europe is caused by Flavescence Dorée phytoplasma (FDp), which belongs to the 16Sr-V group and is a European Union quarantine pathogen. Although many molecular techniques such as loop-mediated isothermal amplification (LAMP) are widely used for the rapid detection of FDp in infected grapevine plants, there is no developed isothermal amplification assay for FDp detection in the insect vectors that are fundamental for the spread of the disease. For this reason, a simple in-field real-time LAMP protocol was optimized and developed for the specific detection of FDp in the insect vector Scaphoideus titanus. The LAMP assay was optimized to work with crude insect extracts obtained by manually shaking a single insect in a buffer for 5 min. Such a simple, sensitive, specific, economic, and user-friendly LAMP assay allowed the detection of FDp in S. titanus in less than half an hour, directly in the field. The developed insect tissue preparation procedure, combined with the LAMP protocol, promptly revealed the presence of FDp in infected S. titanus directly in the vineyards, allowing for monitoring of the spread of the pathogen in the field and to apply timely strategies required for the mandatory control of this pathogen.

Bioaccumulation and bioamplification of pharmaceuticals and endocrine disruptors in aquatic insects

Environmental fate of emerging contaminants such as pharmaceuticals and endocrine disrupting compounds at the aquatic terrestrial boundary are largely unexplored. Aquatic insects connect aquatic and terrestrial food webs as their life cycle includes aquatic and terrestrial life stages, thus they represent an important inter-habitat linkage not only for energy and nutrient flow, but also for contaminant transfer to terrestrial environments. We measured the concentrations of pharmaceuticals and endocrine disrupting compounds in the larval and adult tissues (last larval stages and teneral adults) of five Odonata species sampled in a wastewater-impacted river, in order to examine their bioaccumulation and bioamplification at different taxonomic levels. Twenty different compounds were bioaccumulated in insect tissues, with majority having higher concentrations (up to 90% higher) in aquatic larvae compared to terrestrial adults (reaching 88 ng/g for 1H-benzotriazole). However, increased concentration in adults was observed for seven compounds in at least one suborder (41% of the accumulated), confirming contaminants bioamplification across the metamorphosis. Both, bioaccumulation and bioamplification differed at various taxa levels; the order (Odonata), suborder (Anisoptera and Zygoptera) and species level. Highest variability was observed between Anisoptera and Zygoptera, due to the underlying differences in their ecology. Generally, Zygoptera had higher concentrations of contaminants in both larvae and adults. Additionally, we aimed at predicting effects of contaminant properties on bioaccumulation and bioamplification patterns using the commonly used physicochemical and pharmacokinetic descriptors on both order and suborder levels, however, neither of the two processes could be consistently predicted with simple linear models. Our study highlights the importance of taxonomy in studies aiming at advancing the understanding of contaminant exchange between aquatic and terrestrial food webs, as higher taxonomic categories include ecologically diverse groups, whose contribution to “the dark side of subsidies” could substantially differ.

Sample preparation optimization of insects and zebrafish for whole-body mass spectrometry imaging.

Appropriate sample preparation is one of the most critical steps in mass spectrometry imaging (MSI), which is closely associated with reproducible and reliable images. Despite that model insects and organisms have been widely used in various research fields, including toxicology, drug discovery, disease models, and neurobiology, a systematic investigation on sample preparation optimization for MSI analysis has been relatively rare. Unlike mammalian tissues with satisfactory homogeneity, freezing sectioning of the whole body of insects is still challenging because some insect tissues are hard on the outside and soft on the inside, especially for some small and fragile insects. Herein, we systematically investigated the sample preparation conditions of various insects and model organisms, including honeybees (Apis cerana), oriental fruit flies (Bactrocera dorsalis), zebrafish (Danio rerio), fall armyworms (Spodoptera frugiperda), and diamondback moths (Plutella xylostella), for MSI. Three cutting temperatures, four embedding agents, and seven thicknesses were comprehensively investigated to achieve optimal sample preparation protocols for MSI analysis. The results presented herein indicated that the optimal cutting temperature and embedding agent were -20°C and gelatin, respectively, providing better tissue integrity and less mass spectral interference. However, the optimal thickness for different organisms can vary with each individual. Using this optimized protocol, we exploited the potential of MSI for visualizing the tissue-specific distribution of endogenous lipids in four insects and zebrafish. Taken together, this work provides guidelines for the optimized sample preparation of insects and model organisms, facilitating the expansion of the potential of MSI in the life sciences and environmental sciences.

Aquatic midges take an acid trip

Many years ago in Copenhagen, future Nobel Prize winner August Krogh did what he did best: study a peculiar animal for a very particular reason. He focused on the gas-filled sacs of Chaoborus midges, which he found curiously reminiscent of fish swim bladders. Through a series of experiments, Krogh demonstrated that the midges used the sacs as ballasts, expanding them to float and contracting them to sink, but he couldn't figure out how they accomplished this feat and eventually moved on to other interests. Now published in Current Biology, graduate student Evan McKenzie of the University of British Columbia and colleagues solve Krogh's century old mystery, revealing that it all came down to a common insect protein used in a unique way.Chaoborus air sacs are unusual: they are completely cut off from the respiratory system and enclosed by a wall made of alternating bands of chitin, a hard polymer found in insect shells, and an unidentified protein. Curious about these mysterious bands and their potential role in regulating buoyancy, the authors examined the sacs under a microscope. Surprisingly, the sacs glowed bright blue under UV light. As zoologists well-versed in the idiosyncrasies of insects, they knew that this could only mean one thing: the unknown protein was resilin; a stretchy, biological rubber found in the elastic tendons of locusts, fleas and other arthropods. An unusual quirk of resilin is that it swells under certain conditions such as high pH. The scientists reasoned that reversible swelling or shrivelling of the resilin bands within the air sac wall could alter the sac's volume and, therefore, adjust the midge's buoyancy. They tested this idea by exposing freshly dissected air sacs to acidic or basic conditions, finding that the air sacs shrunk up to 20% in acid and expanded by 45% in base, confirming their hunch.The midges only have to change the pH around their air sacs to float or to sink. Like many organisms, insects control the pH of their guts, mitochondria and other compartments with specialized proteins called proton pumps that move hydrogen ions across cell membranes. The researchers tested if these ubiquitous proteins also controlled the pH of the air sac by exposing them to drugs that directly or indirectly interfered with proton pumps. When the team did so, the sacs expanded like they did in basic conditions, confirming that pH controlled the air sac volume and that a specific type of proton pump called vacuolar-type H+ ATPase controlled the pH.Other insect tissues turn this proton pumping ATPase on or off using cellular signalling pathways involving compounds like cyclic AMP or cyclic GMP. To find out whether the midges use either of these pathways to regulate their buoyancy, the researchers exposed dissected air sacs to synthetic versions of AMP or GMP, each attuned to different branches of the signalling pathway, and observed which compounds caused the sacs to expand. This experiment revealed not only that cyclic AMP co-ordinated the expansion of the air sac, but also identified two more classes of proteins, protein kinase A and cyclic AMP-activated exchange proteins, which are involved in the pH-control signalling pathway, setting the foundation for deeper dives into the unusual physiology of these insects.Chaoborus midges are the only planktonic insects – in fact the only non-fish – that regulate their buoyancy using a gas bladder and they are the only organism to use resilin as a pH-activated means of motion. Unique on three counts, it is perhaps not surprising that they caught the attention of August Krogh or that it took scientists over 100 years to understand how their unique organ works.

Identification and Characterization of CYP6 Family Genes from the Oriental Fruit Moth (Grapholita molesta) and Their Responses to Insecticides.

Simple SummaryEight CYP6 family genes were identified in Grapholita molesta (Busck). The expression of individual gene members differed between the developmental stages and insect tissues. High expression was found in third/fourth-instar larvae and in the midgut and Malpighian tubules. The response patterns of the genes exhibited diverse response patterns to the three representative insecticides were diverse.Cytochrome P450 (CYP) monooxygenases comprise a superfamily of proteins that detoxify xenobiotics and plant secondary metabolites in insects. The CYP6 family is unique to the class Insecta, and its members participate in the metabolism of exogenous substances. In this study, we sequenced and characterized the full-length cDNAs of eight CYP6 family genes from Grapholita molesta (Busck), a global pest of pome fruits. P450 genes with the exception of CYP6AN35, which was most highly expressed in adults, consistently showed high expression in third- or fourth-instar larvae. The analysis of different tissues of adults showed that most of these genes were predominantly expressed in the midgut, Malpighian tubules, and/or fat body. The expression of these eight CYP6 genes was differentially affected by three representative insecticides: malathion (organophosphate), deltamethrin (pyrethroid), and chlorantraniliprole (carbamate). All eight CYP6 genes responded to malathion treatment. Only three CYP6 genes were highly expressed in deltamethrin-treated individuals. Chlorantraniliprole treatment exerted weak effects on gene expression. Interestingly, CYP6AN35 was a highly expression level in the adult head and its expression was induced by all three insecticides. CYP6AN35 may be a key gene in the metabolism of insecticides. This study provides a fundamental understanding of the functions of the CYP6 gene family in insecticide metabolism in G. molesta.

Engineering insects from the endosymbiont out.

Insects are an incredibly diverse group of animals with species that benefit and harm natural ecosystems, agriculture, and human health. Many insects have consequential associations with microbes: bacterial symbionts may be embedded in different insect tissues and cell types, inherited across insect generations, and required for insect survival and reproduction. Genetically engineering insect symbionts is key to understanding and harnessing these associations. We summarize different types of insect-bacteria relationships and review methods used to genetically modify endosymbiont and gut symbiont species. Finally, we discuss recent studies that use this approach to study symbioses, manipulate insect-microbe interactions, and influence insect biology. Further progress in insect symbiont engineering promises to solve societal challenges, ranging from controlling pests to protecting pollinator health.

Molecular identification of seed‐feeding flies dissected from herbarium specimens clarifies the 100‐year history of parasitism by Japanagromyza tokunagai in Japan

AbstractInsect pests, particularly seed‐feeding flies, can cause serious damage to the fruits of many wild orchid species. Evidence of fly damage has been observed in 55 orchid species, belonging to 25 genera, distributed throughout Japan. Although some authors suggested that the observed increase in fly damage to orchids in recent years is related to the introduction of exotic species, the timing and route of spread remain unclear. We examined herbarium specimens of seven orchid species, collected between 1923 and 2016, to collect coarctate pupae and empty puparia, with minimal damage to the specimens. Genomic DNA was then extracted from the insect tissue and species were identified using mitochondrial cytochrome oxidase subunit 1 barcode regions. Long amplicons (660 base pairs [bp]) were obtained from 45.2% of samples and short amplicons (162 bp) were obtained from 90.3% of samples. In total, 51 of 62 samples had nucleotide sequences homologous to Japanagromyza tokunagai, with specimens distributed across collection years and geographic locations. Therefore, the severe damage to Japanese orchids observed in recent years is not related to the recent introduction of an exotic species; it may be related to a rapid increase in a native species. This work highlights the value of herbaria as important resources for elucidating the history and distribution of parasitic insects.