Parasitoid Hymenoptera associated with necrophagous Diptera represent a potentially valuable but underexplored component of decomposition systems and forensic entomology. This study investigated parasitoid species associated with pupae of Chrysomya megacephala (Fabricius, 1794) exposed during pig cadaver decomposition in the Pampa biome of southern Brazil, and evaluated parasitoid occurrence across decomposition stages and sampling periods. Experiments were conducted in March, September, and November 2024, and January 2025 using 12 pig cadavers. Sentinel pupae were exposed daily throughout decomposition and recovered for parasitoid emergence and identification. Generalized linear mixed models were used to evaluate parasitoidism probability and species-specific occurrence across decomposition stages, while Weibull survival models were applied to analyze stage duration. A total of 5,280 pupae were exposed, of which 376 (7.12%) were parasitized, yielding 4,171 parasitoid adults. Four species were recorded: Nasonia vitripennis (Walker, 1836) (Hymenoptera: Pteromalidae), Pachycrepoideus vindemmiae (Rondani 1875) (Hymenoptera: Pteromalidae), Spalangia endius Walker, 1839 (Hymenoptera: Spalangiidae), and Tachinaephagus zealandicus Ashmead, 1904 (Hymenoptera: Encyrtidae). Parasitoidism probability was significantly lower during the fresh stage and increased from the bloated stage onward. Nasonia vitripennis showed greater contribution during the bloated stage, whereas P. vindemmiae was proportionally more frequent during the fresh stage. Decomposition progressed more rapidly during warmer months, particularly in November and January. Seasonal variation in parasitoidism was also observed, with higher rates during warmer sampling periods. These findings demonstrate that parasitoid assemblages vary across decomposition stages and environmental conditions, highlighting their ecological relevance in decomposition systems and their potential contribution to forensic entomology in subtropical environments.
In the last two decades, the field of molecular entomology has seen a shift toward next-generation sequencing techniques as a means of uncovering genetic and developmental processes. However, the standardization of methods is not well-established, and studies for insect-fungus consortia lack established protocols for advanced molecular techniques and downstream analysis compared to approaches applied in model systems involving insect-bacteria interactions. To investigate insect-microbe interactions, RNA sequencing and analysis is often used to identify genes involved in the symbiosis. But such protocols do not often consider insect-fungus systems, which vary significantly in community member abundance and/or fail to describe the details of the process from collection to data processing. This paper will introduce a comprehensive approach for RNA sequencing using two non-model insect-fungus consortia, which lack established, published protocols seen in model systems: the ambrosia beetle mutualism and cicada Massospora parasitism. The protocol includes a detailed TRIzol RNA extraction and quantification, RNA sequencing, and data processing using Nextflow pipeline software. Validation of a range of symbiotic interactions from mutualistic to parasitic is considered to justify this procedure to be utilized in a range of insect-fungus interactions with varied abundances and host interactions. Key features • Stepwise protocol for RNA extraction of samples containing insect and fungal tissue. • Novel dissection technique for beetle pupae. • Acquisition of transcriptomes of both host and symbiont with one protocol. • Direct comparisons of transcriptomes across life stages, stages of symbiosis, and/or by treatment.
Background Cutaneous leishmaniasis (CL) is an emerging and rapidly escalating public health threat in Sri Lanka, where a complex interplay of environmental, vector-related, biological, and socio-economic factors governs transmission risk. Despite the rising case burden, no validated, country-specific framework exists to systematically quantify and prioritize these risk determinants. This study aimed to develop and validate a multi-level indicator framework for assessing CL transmission risk in Sri Lanka using a combined Delphi-entropy weight approach. Method A three-level hierarchical indicator framework was initially constructed through a systematic literature review of CL risk factors. A total of 65 domain experts engaged in leishmaniasis surveillance, clinical management, entomology, and epidemiological research in Sri Lanka were invited to participate. Seventeen experts provided informed consent and completed the survey. Expert consensus was elicited using two iterative Delphi rounds. An authority coefficient (Cr) was calculated for each expert by integrating familiarity (Cs) and judgment basis (Ca) scores to weight responses proportionally to expertise. Entropy weights were derived from the degree of dispersion in expert responses to provide objective, data-driven significance scores. Comprehensive indicator weights were obtained by integrating Delphi and entropy weights. Results The final framework comprised four primary indicators, 11 secondary indicators, and 46 tertiary indicators. Among the primary indicators, biological factors received the highest normalized Delphi weight (0.312), followed by environmental factors (0.229), interventions (0.200), and social factors (0.188). The top-ranked secondary indicators were climatic features (0.194), sand flies (0.152), geographic features (0.105), and dogs (0.095). Among tertiary indicators, the highest comprehensive weights were assigned to regular monitoring of sand fly density (0.118), annual average rainfall (0.113), indoor residual spraying (IRS) (0.096), sand fly density (0.067), and screening patients through mobile clinics (0.057). Notably, three of the five highest-weighted tertiary indicators were directly related to sand fly surveillance and control, highlighting the primacy of vector management in CL transmission risk reduction. A critical data gap was identified: the absence of systematic, national-level sand fly density monitoring, which constrains real-time risk assessment capacity. Patient screening through mobile clinics also emerged as a crucial measure, particularly for rural communities with limited healthcare access. Conclusion:The Delphi-entropy weight framework provides a validated, evidence-based tool for prioritizing CL transmission risk factors in Sri Lanka. The findings highlight the urgent need to establish a systematic national sand fly surveillance programme, analogous to existing dengue vector monitoring systems, and to reinstate targeted IRS in endemic areas. This framework can guide resource allocation and evidence-based policy decisions for CL prevention and control in Sri Lanka and may serve as a model for similar resource-limited, endemic settings.
Accurate minimum post-mortem interval estimation in forensic entomology relies on understanding necrophagous dipteran dispersal behavior. Post-feeding larval dispersal represents a critical phase impacting evidence recovery in forensic research. Currently, most studies suggest uniform radial dispersal patterns. Our study quantified spatial and temporal patterns of post-feeding larval dispersal in necrophagous Diptera under field conditions, examining how temperature, humidity, soil and season influence dispersal distance and directionality from a dead body. Twelve swine carcasses were deployed across wet and dry seasons in the Pedregal de San Ángel Ecological Reserve, Mexico City. Pitfall traps arranged in 9 concentric rings (1 to 7.4 m diameter) with 36 angular sectors monitored larval dispersal patterns. Among 2,545 individuals from 10 morphospecies were recorded, Lucilia sericata (Meigen) was the most dominant species (87.04%). Circular statistics rejected uniform distribution, revealing significant northwest directional preference (320.74°). Spatial distribution exhibited bimodal patterns with 50% of larvae within 130 cm and 95% within 290 cm. Dry season showed higher larvae concentration toward northwest compared to wet season. Predation by Solenopsis sp. Westwood ants during dry season forced larval aggregation in situ. Post-feeding larval dispersal exhibits consistent directional bias, complementing current uniform radial models. Our findings optimize entomological evidence collection by establishing search priorities: primary exhaustive searches within 3 m capture 95% of dispersing larvae, directional searches toward northwest up to 5 m increase recovery during wet seasons, and concentrated searches within 0.5 m maximize recovery during dry conditions when predation restricts dispersal. These quantified metrics provide operational guidelines for evidence recovery in Neotropical forensic investigations, improving search efficiency while reducing investigation time.
Sir Vincent Brian Wigglesworth (1899 -1994) is often cited as the leading figure most responsible for transforming entomology from a largely observational and descriptive natural science into a rigorous experimental biological discipline. His innovative investigations into the complexity of how insects work provided the foundations for modern insect physiology and endocrinology. By predicting three master hormones (prothoracicotropic hormone, ecdysone, and juvenile hormone), he managed to explain the processes of insect growth, development, and reproduction, but also established the fundamental mechanisms of neuroendocrine control across the animal kingdom. Wigglesworth's contributions were more than a collection of discoveries but constituted a unified, coherent theory of postembryonic development in insects governed by precisely timed neuroendocrine signals but extending into many other physiological processes.
The wheat stink bug, Aelia rostrata (Hemiptera: Pentatomidae), is a major pest of wheat in Central Anatolia. This species has an extended adult dormancy period lasting approximately 9 months. This period encompasses summer estivation and winter diapause in mountainous dormancy areas. Despite its economic importance and unusual dormancy biology, the molecular basis of seasonal dormancy in A. rostrata remains unexplored. Here, we combined RNA-seq, proteomics, and lipid content measurements to provide the first molecular insights into estivation and winter diapause responses in the fat body of the wheat stink bug. Transcriptomic enrichment analyses suggested that genes associated with protein synthesis, cytoplasmic translation, and ribosomal activity were enriched during estivation, whereas pathways related to catabolic processes, fatty acid metabolism, and pathogen response were associated with winter diapause. Complementary proteomic analyses identified mitochondrial and chitinase-like proteins with higher abundance during estivation, while proteins related to energy metabolism, including acyl-CoA dehydrogenases, were more abundant during winter diapause. Lipid measurements indicated significantly lower lipid reserves in post-diapause individuals compared to actively feeding adults, consistent with the utilization of lipid stores during dormancy and/or the transition to post-dormancy activity. Overall, this study provides the first transcriptomic and proteomic resource for A. rostrata and offers initial molecular insights into the physiological signatures associated with seasonal dormancy in this economically important pest.
The spotted lanternfly, Lycorma delicatula (White) (Hemiptera: Fulgoridae), is an invasive planthopper and significant agricultural pest in the United States. Current management relies heavily on insecticides, with limited options for organic or biological control. This study evaluated the overwintering survivability of Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae) spores on lanternfly egg masses to assess the feasibility of winter and spring, preemergence applications during vine dormancy. Laboratory bioassays suggested treatments of commercially available B. bassiana to spotted lanternfly egg masses resulted in infection of newly eclosed neonates. The optimal timing of spray applications on egg masses was approximately 2 wk before emergence. However, infection was also observed in newly emerged nymphs when treatments were applied even 20 wk before insect hatch. Treatments 20 wk before hatch significantly reduced the viability of conidial residues with wettable powder formulations exhibiting higher germination rates than oil-based formulations. These findings highlight the influence of environmental conditions on fungal spore persistence and underscore the potential for refining formulations and application timing to enhance biological control of spotted lanternfly. Two field trials demonstrated the applicability of mycoinsecticide treatments to overwintering spotted lanternfly egg masses. Both field trials demonstrated infection of hatching nymphs at levels greater than those observed in laboratory bioassays. This management approach would provide growers with a viable, low-toxicity, and Organic Materials Review Institute approved solution for managing spotted lanternfly months to weeks before the predicted hatch of spotted lanternfly.
Two grasshopper species, Dericorys albidula Serville, 1838 and Thrinchus arenosus Bey-Bienko, 1948, are reported as new records for China. Although they belong to different families, they share similar habitat preferences. Detailed morphological redescriptions and biological notes are provided for both species.
A new species of the genus Micreriodes Yoshimoto is described and illustrated in the fauna of the State of Paraná, Brazil: Micreriodes lautus sp. nov. The morphological differences with Micreriodes guamensis are also analyzed. This is the first time any specimen of the genus has ever been identified at the species level outside the Pacific region.
Manure is an abundant waste source in dairy production with disposal creating significant challenges for producers. Recent work has shown that black soldier fly, Hermetia illucens (Linnaeus) (Diptera: Stratiomyidae), larvae can digest dairy manure; however, its high cellulose content and low nutrient value impact larval performance. Adding probiotics to black soldier fly feed can increase feed digestibility and weight gain. The goal of this study was to determine the influence of probiotic additions on black soldier fly larval performance when fed dairy manure. This study evaluated life-history traits of black soldier fly larvae fed dairy manure with single or combined probiotic treatments: Rhodococcus sp., Paenibacillus sp., and 1:1 combination, compared to a no-microbe control. Larvae were mass-reared on fresh dairy manure, and key metrics-bioconversion rate, survivorship, total biomass, and larval weight-were compared across treatments. Results showed a significant trial effect for all variables measured. Mean peak weights and the largest larval weights were 3.5-20.0% greater in at least one probiotic treatment compared to the control, while other life-history traits did not differ significantly. However, probiotic treatments reduced variation up to 17.0% in black soldier fly larval weight across trials, a trait not historically discussed in black soldier fly research. Developing processes that optimize larval weight in combination with reducing variation are critical for developing precise business models for predicting future production.
Climate change is shifting the distribution of insect vectors of disease, with consequences for human pathogen exposure. Predicting these range shifts requires understanding vectors' physiological limits. In North America, West Nile virus is transmitted primarily by two closely related and interbreeding Culex species-Cx. pipiens in the north and Cx. quinquefasciatus in the south-which, together with Cx. pipiens f. molestus comprise the North American Cx. pipiens Assemblage. Although the assemblage is widely distributed, little is known about how temperature shapes its species' ranges, particularly in the larval stage, where selection by temperature extremes is likely stronger than in adults. We compared larval thermal tolerance of first and fourth instars under high- and low-temperature extremes among five assemblage populations from the eastern United States. Southern Cx. quinquefasciatus populations were more tolerant of high temperatures than northern Cx. pipiens populations, whereas northern populations showed greater freezing tolerance in later stages. We also quantified larval macronutrient composition by colorimetric assay to characterize population-level differences in fitness-relevant energy stores. Populations differed in composition, but these differences did not align with thermal-tolerance patterns. Finally, we extended the first-instar high-temperature assay to two mid-latitude populations from within and near the hybrid zone in Maryland. Their thermal tolerance was intermediate to northern and southern populations', and LT50 was marginally correlated with collection-site climate after correcting for multiple comparisons. These findings provide empirical data on physiological variation within the Cx. pipiens Assemblage and inform future investigation of the genetic basis of thermal tolerance and range shifts.
Mosquitoes transmit a diverse range of deadly pathogens, including dengue and Plasmodium spp., when they consume the blood of vertebrate hosts in order to nutritionally provision their eggs. However, mosquito species from multiple genera have independently evolved the ability to produce eggs without blood. The molecular and physiological processes underlying the evolution of blood-independent reproduction are largely unresolved, in part because the two most well-studied models of mosquito reproductive physiology and genetics are obligate blood feeders. Herein, we integrate life-history, genomics, and gene expression to identify physiological processes and candidate genes associated with the evolution of blood-independent reproduction in a non-model vector mosquito, Aedes albopictus. Relative to a blood-dependent (control) population, females selected for blood-independent reproduction exhibit a longer larval development period and larger adult size, implying increased nutrient sequestration at the larval stage. Additionally, blood-independent larvae exhibit higher expression of amino acid and lipid storage genes (hex1.1 and lsd-2) relative to blood-dependent larvae. Linkage clusters and FST outliers identify a ca. 40 Mb genomic region that is highly differentiated between blood-independent and blood-dependent lines. Moreover, microRNAs and mRNAs related to reproductive physiology are differentially abundant between adults of the selected vs. control lines. Multiple lines of evidence indicate that larval nutrient sequestration and altered regulation of vitellogenesis have contributed to the rapid evolution of blood-independent reproduction in Ae. albopictus. Overlapping evidence also identifies proposed candidate genes affecting these processes. These results provide a foundation for developing novel paradigms to suppress disease transmission by vector mosquitoes.
Aedes aegypti is the most important mosquito vector of arboviruses worldwide, originating on islands of the southwest Indian Ocean before spreading to Africa and, more recently, the Americas and Asia. Migration out of Africa was enabled by the evolution of behaviors that increased human-vector contact, and as Ae. aegypti became more domesticated, arboviral competence may have also increased. Here we investigate variation in vector competence between present-day populations of Ae. aegypti from Florida (USA) and Tanzania (East Africa) and use a genomics approach to examine genetic differentiation potentially underlying traits important for arbovirus transmission. Using vector-competence experiments with dengue virus serotype 2, we demonstrate greater susceptibility to infection in American mosquitoes, along with higher viral loads and a shorter time to virus dissemination. We then sought to identify highly divergent genes potentially underlying these differences and discovered more than 1.6 million single nucleotide polymorphisms via exome sequencing. Of these, 1058 were highly differentiated between populations (FST > 0.9) with more than half mapping to 17 outlier genes influencing chromatin remodeling, lipid metabolism, glycosylation, circadian rhythms, olfaction, and saliva bioactivity. Eight of these outlier genes formed a highly divergent genomic island on chromosome 2. Allele-frequency patterns of outlier genes across Ae. aegypti's range highlight genetic differentiation within sub-Saharan Africa, as well as evolutionary changes accompanying the shift towards domestication in the Americas and Asia. This work offers insights into phenotypic variation and evolutionary divergence within Ae. aegypti, including naturally occurring genetic variants that may influence transmission-related characters.
A guilty pleasure of ours is having coffee on a Saturday morning with a friend at a neighborhood cafe where the chocolate croissants are divine and the pace of the day is unhurried. We linger longer than intended, cups refilled, as our conversations wander across art, history, science, travel, wine, and the linguine marinara we had for dinner last night. On one such occasion, the conversa-tion shifted to the recognition that many of the plants producing our favorite foods and beverages face growing threats from viral pathogens about which we know surprisingly little.An adapted version of this editorial was published in The Hill on May 28, 2026 (https://thehill.com/opinion/energy-environment/5896747-coffee-chocolate-wine-plant-viruses/).
Removal of senescent and damaged cells is fundamental for tissue homeostasis. While macrophage recognition and clearance of apoptotic cells are well characterized, the ultimate fate of the digested material remains poorly understood. Here, we explore current knowledge on the fate of engulfed material and examine how the metabolic nature of engulfed cargo shapes downstream signaling and phagocyte polarization. We also discuss emerging evidence that macrophages act as metabolic hubs, recycling and supplying nutrients to surrounding tissues. Drawing on studies in invertebrate phagocytes, we explore the evolutionary origins of this "nurturing" function and highlight its conservation in mammals, emphasizing its physiological relevance and potential contributions to metabolic disease.
Parasitoid wasp venoms contain diverse bioactive molecules that manipulate host physiology, most notably by inducing rapid paralysis, yet the paralytic components of many agriculturally important species remain uncharacterized. Bracon brevicornis, an ectoparasitoid of lepidopteran pests, induces rapid and sustained paralysis in its insect hosts, but the molecular basis of this effect remains unknown. Using LC-MS/MS analysis of venom gland extracts, we identified five peptides derived from the C-terminal region of a putative paralytic protein. Two representative peptides-Peptide 1 (946.48 Da; GLFDFIVH) and an N-acetylated Peptide 2 (1,791.93 Da; QAAAQAAAAGIQLPSKPAS)-were validated by LC-ESI-QToF MS/MS analysis, synthesized, and subjected to functional assays. Bioassays revealed dose-dependent paralysis induced by both synthetic peptides, confirming that they retain intrinsic paralytic activity. However, the onset of paralysis was consistently slower than that caused by the crude venom extract, suggesting that additional venom constituents act synergistically to enhance or accelerate the paralytic response. Overall, our findings identify Peptides 1 and 2 as active fragments of the B. brevicornis paralytic protein and indicate that venom activity arises from multiple interacting components. This work provides the first molecular insight into B. brevicornis venom-mediated paralysis and establishes a foundation for mechanistic and applied studies.
Madagascar's malaria burden has been increasing for several years despite continued investments in vector control. Supplemental measures to reduce vector populations and outdoor biting, such as larval source management (LSM), may be required in some settings. Madagascar piloted drone-delivered microbial larviciding using Bacillus thuringiensis israelensis (Bti) in rice fields in two districts with populations unfamiliar with drones during Feb-Jul 2022. We assessed knowledge and perceptions among purposefully selected rice farmers, rice field workers, and community members during focus group discussions (FGDs) in the two districts three to four months after the intervention. In total, 293 people participated in 39 FGDs, which were analyzed using a combined deductive-inductive approach. Despite initial concerns, both Bti-based larviciding and using drones to apply larvicides were ultimately well accepted by rice field owners and workers and community members in rice-growing areas of Madagascar. Key concerns spanned safety of Bti for people, livestock, and the environment, and harms related to drones crashing or being used for nefarious purposes. Participants perceived decreases in mosquitoes and malaria and increased rice yield related to the intervention. Receipt of project messaging appeared to differ by demographic group, with rice field workers frequently reporting having missed community meetings while working in the fields; many participants also reported that text-heavy communications were difficult to understand. Participants appreciated the involvement of personnel from health, agriculture, and environmental sectors, and they recommended expanding such intersectoral collaboration, involving trusted local officials, hiring local actors to help plan and implement the activities, and combining larvicide and fertilizer application in subsequent projects. Future LSM activities could benefit from social behavior change (SBC) efforts addressing safety and privacy concerns linked to the larvicide and the delivery method and highlight potential health and financial benefits related to decreased malaria burdens in the community, improved quality of life associated with fewer mosquito bites, the efficiency of drones in applying larvicide products, and the need to continue using ITNs. SBC will be most effective if provided early and repeatedly as needed, via channels appropriate to the community's language and literacy levels and in venues that are geographically and temporally convenient to those potentially impacted by the intervention.
Zea mays (maize) is a globally important cereal crop and a key system for studying plant development and stress responses. Proteome profiling and phosphoproteome profiling provide direct, quantitative readouts of protein abundance and phosphorylation states, which offer insights into aspects of regulation and cellular function that transcript-level measurements alone cannot provide. Robust and reproducible methods are essential for generating accurate and biologically relevant data in proteomics studies. The complexity of plant tissues, however, poses challenges for developing reliable sample preparation workflows. Here, we describe a detailed sample preparation protocol for quantitative proteome and phosphoproteome profiling in maize. The protocol encompasses protein extraction, filter-aided sample preparation (FASP), peptide desalting, tandem mass tag (TMT)-based labeling for quantitative multiplexing, and complementary TiO2 and Fe-NTA enrichment steps, yielding peptides suitable for analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). This approach enables the quantitative profiling of protein abundance and phosphorylation dynamics in maize tissues.
The gut microbiota orchestrates host health by influencing nutrition, immunity, and behavior. Bifidobacterium species are early colonizers maintaining gut homeostasis in honeybees. Despite these critical roles, strain-level dynamics and driving forces behind microbial competition during social transmission remain poorly understood. Here, we established six field-mimicking colonies to track Bifidobacterium communities across successive generations of newly emerged workers. Social transmission markedly reshaped community structure, generating distinct trajectories among phylotypes. OTU10 consistently dominated, reaching a median relative abundance of 77%, whereas OTU61 and OTU43146 were progressively outcompeted or lost. Comparative genomics of five representative species identified 69 carbohydrate-active enzyme families, with GH43 glycosyl hydrolases driving genomic divergence. Bifidobacterium polysaccharolyticum encoded an expanded CAZyme repertoire, supporting rapid growth on glucose and arabinan and indicating an r-selected strategy. In vitro and in vivo competition assays demonstrated that nutrient availability and priority effects act as key ecological filters. Pollen favored Bifidobacterium apousia, whereas sucrose-only diets promoted B. polysaccharolyticum. Moreover, early colonizers excluded later arrivals to some extent, highlighting priority effects. Together, these findings reveal how pollen polysaccharides and transmission bottlenecks interact to structure the strain-level landscape of the social microbiome.