Larvae Research Articles

In vitro assessment of assembly and sex pheromones in the control of larval stages of brown dog ticks

The increasing incidence of tick-borne diseases and the growing resistance to conventional acaricides underscore the urgent need for innovative and sustainable tick management strategies. This study investigates the potential of assembly and sex pheromones for the control of larval stages of Rhipicephalus sanguineus, the brown dog tick, which poses significant health risks to dogs. The ticks were collected from various canine hosts in Thrissur, Kerala and subjected to Petri-dish bioassays using vapour patches impregnated with synthetic assembly pheromone (AP) and sex pheromone (SP), both individually and in combination. The efficacy of these semiochemicals was assessed based on the attraction and clustering behaviour of unfed larvae and partially fed adult ticks over a 24-hour period. Results demonstrated a significantly higher attraction to AP-impregnated patches, with notable efficacy observed in causing tick aggregation, particularly beyond 30 min of exposure. While SP also showed potential in attracting ticks, the combination of SP and AP did not result in a synergistic effect. The study highlights the potential of using semiochemicals as an environmentally sustainable alternative to traditional acaricides, highlighting their role in integrated pest management strategies Keywords: Rhipicephalus sanguineus, adenine, guanine, xanthine, 2,6-DCP, Petri-dish bioassay

Identifying insect predators using camera traps reveal unexpected predator communities in oilseed rape fields

Cameras are common tools for ecologists studying species abundance, richness, and interactions. They are mainly used to study large animals but are increasingly used to study invertebrates. Camera traps could be a powerful tool to identify and quantify the natural enemies, such as predators, of specific agricultural pests. This knowledge can then be applied to adapt farming practices to favour the predators identified and improve conservation biocontrol.Here, cameras were used to identify predators of two insect pests of oilseed rape (Brassica napus): the pollen beetle (Brassicogethes aeneus) and the brassica pod midge (Dasineura brassicae). These insects are spring pests of oilseed rape and are both susceptible to predation by ground-dwelling arthropods during their development cycle. The objectives of this study were to 1) identify the predators of the pest species, and 2) investigate the hypothesis that the diel activity periods of the predators match the timing of pest larval presence on the ground. Field observations were conducted in two locations over two consecutive years in the UK.In contrast to previous reports, our data collected using cameras showed that larval stages of predatory beetles are responsible for most of the predation events and that adult stages were not frequently observed feeding on the prey. Our data also showed that the traditional method, using pitfall traps, failed to detect the presence of large numbers of beetle larvae. Interestingly, the pests dropped to the ground and were therefore most susceptible to predation at the time when predators were most active.These results give new insight on the predators of the two most important spring pests of oilseed rape in Europe. This knowledge can be used by practitioners to develop framing practices targeted on enhancing the abundance of the larval stages of these beneficial arthropods to better control the pests in a more sustainable way.

Imported arboviral disease cases and entomological surveillance of Aedes albopictus in Valencian Autonomous Region (Spain) along 2022

Abstract The Laboratory of Entomology and Pest Control (LEPC) of the University of Valencia, hired by the regional government of the Generalitat Valenciana, carried out the entomological surveillance of the Asian tiger mosquito along 2022 in the Valencian Autonomous Region, excepting the city of Valencia for which the surveillance was conducted by a private pest control company. The main purpose of this plan of action was both to continue the detection and monitoring of Aedes albopictus in the Valencian municipalities free of its presence until the end of 2021, and to carry out the entomological surveys related to imported arbovirus cases notified by the Regional Health System. The active search and detection of the mosquito breeding sites in both urban and peri-urban environments throughout the provinces of Alicante, Castellón and Valencia, has revealed the absence of new populations of this allochthonous mosquito in all those municipalities free of this species in December of 2021. Besides, the LEPC has conducted the mosquito surveys in 31 municipalities of the three provinces, in relation to the declaration of 33 cases of arthropod-borne diseases due to Chikungunya, dengue or Zika viruses, which were reported in travellers returning from countries of Latin America, Asia, or West Africa. The methods used to minimise the risk of local transmission of these viruses by Ae. albopictus were focused on detecting the presence of the tiger mosquito and other common native mosquito populations, at both adult and larval stages, in the surroundings of the residence of imported cases to guide the treatments of the detected resting and breeding sites. As result, tiger mosquito populations were recorded in 18 of the 33 municipalities where imported arbovirus cases were reported in 2022.

Abundance and taxonomic characterization of chigger mites (Acari: Trombiculidae and Walchiidae) associated with rodents in selected scrub typhus-prone areas in Southern and Western provinces of Sri Lanka

Larval trombiculid mites (chiggers) are the vectors and reservoirs of the potentially lethal infectious disease, scrub typhus (ST) caused by Orientia tsutsugamushi. Small rodents are natural hosts of parasitic larval stage of the chigger mites. This study focused on determining the abundance of chigger mites associated with rodents in Sri Lanka and the taxonomic characterization of field-caught chiggers. Field sampling was conducted in the districts of Galle, and Hambantota of the Southern Province, and Gampaha of the Western Province, in 2019 and 2020. Sampling sites were selected according to the patient distribution. A total of 58 small mammals (rodent species: Rattus rattus (Black rat), Rattus norvegicus (Brown rat), Tatera indica (Indian gerbil), Gollunda ellioti (Indian bush rat) and Suncus murinus (Asian house shrew)) were captured using baited traps. Chiggers from the captured rodents were speciated morphologically by visual inspection and morphometry using a camera-mounted light microscope (x100). A total of 394 chigger mites were collected. Three different genera were identified, including Leptotrombidium, Schoengastiella, and Microtrombicula. Leptotrombidium imphalum (72.59%; n=286) was the predominant species, followed by Schoengastiella punctata (8.12%; n=32). Some specimens were identifiable only up to genus level, Leptotrombidium sp. (3.55%; n=14) and Microtrombicula sp. (4.82%; n= 19). Some (7.11%; n=28) were not trombiculid mites, while 3.81%; n=15 was damaged beyond identification. Leptotrombidium imphalum was detected for the first time parasitizing the murids - Rattus novergicus and Tatera indica in the district of Galle, a new locality. In addition, S. punctata was recorded in a new locality in the Gampaha district, Western province with a new host association, Golunda ellioti. This study emphasizes the need for further entomological surveys in ST disease-endemic areas. Developing a morphological identification key for chigger mites in Sri Lanka is a top priority to facilitate field surveys.

Pink bollworm (Pectinophora gossypiella) movement with respect to rosette flowers in cotton: Novel insights for integrated pest management

Aim: This preliminary study aimed to investigate the movement patterns of Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) larvae in the rosette flowers. Methodology: Field experiments conducted on the migration behavior of pink bollworm larvae with respect to rosette flowers in cotton fields. Observations included monitoring infested flowers, larval stage duration within rosette flowers, and larval movement to soil for pupation, larval mortality post-drop, and larval migration to early bolls, all conducted without insecticide application to reflect natural conditions. Results: The movement of the larvae was tracked and recorded from the start of flowering to the boll formation stage. Nearly, 44.88% of the larvae present on the infested flowers had moved to the soil for pupation, while 14.29% had moved to newly formed bolls on the same rosette flower. Additionally, 30.90% of the larvae had moved to dry leaf debris on the soil, and the remaining 9.93% either escaped or died due to environmental factors such as damp soil. Notably, the study showed that dry soils or light debris accumulation were the most favorable environments for pink bollworm larvae. Interpretation: The present study has generated useful information regarding the movement of PBWs larvae from rosette flowers to bolls that contribute to infestation. Hence, by implementing integrated pest management strategies during the early crop periods between 60 and 100 DAS, green boll infestations can be avoided and economic yields improved. Key words: Larval movement, Pest carryover, Pest management, Rosette flower

Automated lepidopteran pest developmental stages classification via transfer learning framework.

The maize crop is highly susceptible to damage caused by its primary pests, which poses considerable challenges in manually identifying and controlling them at various larval developmental stages. To mitigate this issue, we propose an automated classification system aimed at identifying the different larval developmental stages of 23 instars of 4 major lepidopteran pests: the Asian corn borer, Ostrinia furnacalis (Guenée; Lepidoptera: Crambidae), the fall armyworm, Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae), the oriental armyworm, Mythimna separata (Walker; Lepidoptera: Noctuidae), and the tobacco cutworm, Spodoptera litura (Fabricius; Lepidoptera: Noctuidae). Employing 5 distinct Convolutional Neural Network architectures-Convnext, Densenet121, Efficientnetv2, Mobilenet, and Resnet-we aimed to automate the process of identifying these larval developmental stages. Each model underwent fine-tuning using 2 different optimizers: stochastic gradient descent with momentum and adaptive moment estimation (Adam). Among the array of models tested, Densenet121, coupled with the Adam optimizer, exhibited the highest classification accuracy, achieving an impressive 96.65%. The configuration performed well in identifying the larval development stages of all 4 pests, with precision, recall, and F1 score evaluation indicators reaching 98.71%, 98.66%, and 98.66%, respectively. Notably, the model was ultimately tested in a natural field environment, demonstrating that Adam_Densenet121 model achieved an accuracy of 90% in identifying the 23 instars of the 4 pests. The application of transfer learning methodology showcased its effectiveness in automating the identification of larval developmental stages, underscoring promising implications for precision-integrated pest management strategies in agriculture.

Effect of hypoxia in the post-hatching development of the salmon (Salmo salar L.) spinal cord

IntroductionHypoxia has a teratogenic effect on the fish during embryonic development. Nevertheless, the effects on the larval stage are not yet known. Therefore, the aim of this study was to assess the effects of hypoxia on the number of neurons and their apoptotic rate in the spinal cord of Salmo salar alevins after hatching.MethodsWe used a total of 400 alevins, establishing both hypoxia and control (normoxia) groups (n = 8), considering post-hatching days 1, 3, 5, and 7, each with 50 individuals. Transversal sections of 50 μm thickness were cut from the alevin body. We performed cresyl-violet staining and counted the spinal cord neurons. Also, immunohistochemistry for HIF-1α and caspase-3 were used. For statistical analysis ANOVA one-way and Tukey's Test were used.ResultsHIF-1α was expressed in spinal neurons in both the hypoxic and normoxic groups, with the former being significantly higher. Both the hypoxic and normoxic groups evidenced the process of neuronal apoptosis, with the hypoxic groups demonstrating a higher significance. The number of neurons in the spinal cord was significantly lower in the hypoxic group.DiscussionWe found that when oxygen levels in the aquatic environment were low in Salmo salar farming alevins post-hatch, the number of spinal neurons dropped by half. These results contribute to increasing our knowledge of the biological development of salmon, in particular the genesis of the spinal cord, and the effects of hypoxic conditions on the development of this structure of the nervous system.

Chironomid Pupal Exuviae Technique in Ecological Research of Man-Made Water Bodies

Reservoirs serve functional purposes such as irrigation and power generation. However, concerns are raised due to the alterations of the connected riverine ecosystems. Chironomidae (Diptera), a diverse aquatic macroinvertebrate group, are vital to the functioning of ecosystems and serve as water quality indicators. Their holometabolous development includes the pupal stage after four larval stages. The chironomid pupal skin (exuvia) is used in environmental assessments, where the Chironomid Pupal Exuvial Technique (CPET) is a recognized standard. The CPET method is adaptable to different freshwater environments and here was applied in the study of 28 man-made lakes in the Pannonian Lowlands and Dinaric Western Balkan Ecoregion in Croatia to obtain information on chironomid diversity and analyze the potential influence of environmental factors on the chironomid community. The lake surface was skimmed with an exuvial hand net (mesh size of 300 µm) along the lake edge with a transect length of 10 m in the area of accumulated debris of organic and inorganic matter. Individual exuviae were mounted in a Berlese mounting medium and identified by morphological characteristics to the lowest taxonomic level. During the study, 5698 chironomid pupal skins were collected, and 141 taxa (including 97 species) belonging to five subfamilies were identified. The tribe Tanytarsini comprised 40% of the identified taxa, with Paratanytarsus spp. being the most abundant. In the Dinaric ecoregion, Paratanytarsus bituberculatus dominated, while Microchironomus tener and the genus Cricotopus were the dominant taxa in the Pannonian ecoregion. Community structure in the Pannonian ecoregion was influenced by total organic carbon (TOC) and orthophosphates (PO43−), indicating higher anthropogenic pressure compared to the Dinaric ecoregion, where water conductivity influenced Chironomidae assemblages. The research has provided valuable and useful information on the chironomid diversity in man-made and highly altered water bodies, as some of the most vulnerable aquatic habitats to anthropogenic influence. The CPET method could be a useful tool for the ecological studies and bioassessment of water quality in Croatia.

Comparison of distinct gut bacterial communities in different stage of prediapause and nondiapause larvae in Loxostege sticticalis.

Symbiotic microorganisms in insects regulate multiple physiological functions, widely participating in nutrient metabolism, immune regulation, and crucial regulatory roles in development. However, little is known about how microbial factors might respond to the preparation of insect diapause. The gut bacterial communities of Loxostege sticticalis larvae induced at different photoperiod of long (LD16:8, nondiapause) and short (LD12:12, prediapause) daylength were compared, by 16S rRNA Illumina MiSeq. A total number of 42 phylum, 78 classes, 191 orders, 286 families, 495 genera, and 424 species were identified in the intestinal bacterial community of L. sticticalis larvae. Alpha diversity and beta diversity analyses revealed significant differences between nondiapause and prediapause larvae. In non-diapause larvae, the dominant intestinal bacteria were Firmicutes and Proteobacteria. In specific, in 3rd and 4th instar larvae, the main intestinal bacteria were Staphylococcus, while in 5th instar, it was JC017. For the prediapause larvae, the dominant phylum in 3rd instar larvae was Firmicutes, with the dominant genus of Staphylococcus, while in 4th and 5th instar larvae was Bacteroidota, with the dominant genus 4th instar was Staphylococcus, and in 5th instar was JC017. KEGG functional prediction analysis revealed that functional bacterial groups involved in metabolism had the highest abundance values. Specifically, the amino acid metabolism of metabolism-related functional genes in the 3rd instar prediapause larvae was significantly lower than that in the 4th and 5th instar prediapause larvae and the non-diapause treatment. However, the carbohydrate metabolism in 3rd instar prediapause larvae was significantly higher than that in 4th and 5th instar prediapause larvae and non-diapause treatments. The dominant bacterial phylum in the prediapause larvae at different stages of L. sticticalis was varied, and there were significant differences in community diversity and richness. These results suggest a complex interaction between the hosts' physiological state and its gut microbiota, indicating that bacterial communities may assist insects in adapting to diapause preparation by regulating their metabolic levels. This study lays the foundation for further understanding the physiological mechanisms by which intestinal microorganisms regulate overwintering dormancy in the L. sticticalis.

Genome-wide identification and expression profiling of the Wnt gene family in three abalone species.

The Wnt gene family plays pivotal roles in a variety of biological processes including cell proliferation and differentiation, apoptosis, and embryonic development. Identifying the Wnt signaling pathway in abalone could provide a basis for elucidating growth and development mechanisms and improving quality. To identify the number, protein physicochemical properties, gene structure, phylogenetic analysis, and expression profiles of the Wnt gene family in abalone. A comprehensive genome-wide analysis was performed to identify the Wnt gene family in the genomes of three abalone species (Haliotis discus hannai, H. rubra, and H. rufescens). Ten single-copy Wnt genes were identified in each abalone species, suggesting that the number of Wnt genes was relatively conserved in Haliotis. Eight Wnt gene subfamilies, including Wnt1, Wnt4, Wnt5, Wnt6, Wnt7, Wnt10, Wnt16, and WntA, are present in all three species. Each abalone species contains two species-specific subfamilies (Wnt9 and Wnt11 in H. discus hannai, Wnt2 and Wnt11 in H. rubra, and Wnt2 and Wnt9 in H. rufescens), reflecting polymorphisms of the Wnt genes in Haliotis. Interestingly, gastropods are characterised by the loss of Wnt8, suggesting a potential evolutionary specificity in gastropods. As expected, Wnt3 is absent in all protostomes, including the abalone. In addition, spatio-temporal expression profiling revealed differential expression levels of the Wnt genes at different developmental stages and in different tissues of H. discus hannai. HdWnt5 and HdWntA might participate in several processes during larval development stages, including germ layer formation and body axis elongation. HdWnt5 may be involved in eye and tentacle development. HdWnt10 may be related to muscle development, and HdWnt6 may be involved in shell formation in abalone. To our knowledge, the results of this study, which is the first genome-wide investigation of the Wnt gene family in abalone, lay the groundwork for future research on the evolution and function of the Wnt gene family in Gastropoda.

Knockout of a single Pax6 gene (toy but not ey) leads to compound eye deficiency and small head in honeybees

The compound eyes are crucial to honeybees, playing pivotal roles in color recognition, orientation, localization, and navigation processes. The development of compound eyes is primarily mastered by an evolutionarily conserved transcription factor Pax6. In honeybees, there are two Pax6 homologs: ey and toy. To gain a deeper understanding of their functions, we knock out both homologs using CRISPR/Cas9 technology. Intriguingly, we observe that toy knockout mutants have smaller heads without compound eyes and exhibit brain atrophy, while ey knockout mutants develop normal compound eyes, most of which die before/during their metamorphosis from pupa to adult. By comparing the head transcriptomes of four stages (larva, prepupa, pupa, and adult) in toy-knockout mutants versus normal controls, we identify significantly perturbed genes related to DNA binding transcription factors, neuron differentiation, and insect visual primordium development. Additionally, we find the interaction network of toy in honeybees differs obviously from that of D. melanogaster. Our findings suggest the two Pax6 genes serve distinct functions in honeybees and toy takes over the central function of ey in master-regulating the development of honeybee compound eyes. This adds new evidence for breaking the simplified view that some of conservative developmental toolkit genes function as all-or-nothing master regulators.

Distribution and genetic diversity of Bombyx mori nucleopolyhedrovirus in mass-reared silkworms in Thailand

Silk is an economically and culturally important product of Thailand that is threatened by the outbreaks of the Bombyx mori nucleopolyhedrovirus (BmNPV) in silkworm rearings. BmNPV infects the larval stages of the silkworm Bombyx mori, causing fatal viral infection and preventing the larvae from reaching the pupal stage. Numerous BmNPV isolates have been described from silk-producing countries from all over the world. In this study, the geographic distribution of BmNPV in Thailand was analyzed by collecting 18 samples from different regions. The BmNPV samples were analyzed by PCR and whole genome sequencing was performed for eight specimens. Their genome size ranged from 125,888 bp to 126,783 bp, comprising 138 open reading frames. Although the sequenced BmNPV genomes could be phylogenetically differentiated, no correlation between geographic and genetic distance was observed, indicating a close relationship between the BmNPV from Thailand.

Metazoan Parasites of Antimora rostrata (Günther, 1878) (Gadiformes: Moridae) from the Deep Sea in the Southeastern Pacific Ocean

A total of 127 specimens of the “Blue Antimora” Antimora rostrata (Günther, 1878) were obtained from 2015 to 2019 as bycatch from the artisanal fishery of the Patagonian toothfish (Dissostichus eleginoides (Smitt, 1898)) at depths between 1000 and 2200 m in Northern Chile (app. 22° S 70° W). All individuals were examined for parasites. A total of seventeen parasite taxa, two Copepoda, two Monogenea, seven Digenea, three Nematoda, and three Cestoda, were found, and twelve taxa were found as adults while five taxa were found at the larval stage. Anisakis sp. (Nematoda) and Trypanorhyncha gen. sp. (Cestoda) were the predominant species with a prevalence of 53.5% and 11.8%, respectively. The high prevalence of Anisakis sp. (>50%) suggests that A. rostrata may play a significant role in the life cycle of Anisakis sp. in the southeastern Pacific Ocean. The detected parasite community, consisting predominantly of parasites from pelagic environments rather than benthopelagic, suggests that A. rostrata may fulfill a crucial role as a predator of pelagic organism communities. Additionally, it may undertake vertical migrations in the southeastern Pacific Ocean.

Evaluation of inhibitory, immunomodulatory, survival, and growth effects of host-derived Weissella confusa on Macrobrachium rosenbergii challenged with Vibrio parahaemolyticus

Macrobrachium rosenbergii is a highly valuable prawn species in aquaculture due to its current growing demand in the market. However, various bacterial diseases caused by Vibrio parahaemolyticus have been observed to induce mortality in larval, juvenile, and adult stages of M. rosenbergii. While antibiotics are commonly used to manage these diseases, they contribute to antimicrobial resistance, a global concern with serious environmental and health implications. Consequently, the scientific community is exploring host-derived beneficial microorganisms as functional feed additives, offering eco-friendly alternatives that boost shrimp and prawn immunity. The purpose of this study was to examine the effects of host-derived beneficial microorganisms from the gut of M. rosenbergii, on the inhibition, immunomodulation, survival, and growth of M. rosenbergii when exposed to V. parahaemolyticus. The result of this study showed that Weissella confusa C6, a lactic acid bacterium isolated from the gut of M. rosenbergii, significantly inhibits the growth of V. parahaemolyticus R1 as demonstrated in agar well diffusion assay and co-culture assay. Additionally, it showed immunomodulatory effects, indicated by elevated total hemocyte count (THC) and phenoloxidase (PO) activity during the bacterial challenge test. Furthermore, supplementing the diet with W. confusa led to significant improvements in M. rosenbergii, including increased survival rates, weight gain, specific growth rate, better feed conversion ratio, and reduced density of V. parahaemolyticus in the prawn's gut. Therefore, it can be considered a viable option for use as a feed supplement in prawn farming, enhancing M. rosenbergii's immune system, disease resistance, and overall health.

Proteomic analysis of extracellular vesicles and extracellular vesicle-depleted excretory-secretory products of Toxocara canis and Toxocara cati larval cultures

Toxocara canis and Toxocara cati are parasitic nematodes in the order Ascaridida, which inhabit the small intestines of dogs and cats, respectively, as adults. Although often nonpathogenic as adults, nematodes within this genus are capable of causing widespread disease throughout the host while in a larval stage, during which time larvae migrate throughout the body in a process termed larva migrans. Larvae are also capable of surviving within host tissues in an encysted arrested stage, without immune clearance by the host. The ability of larvae to survive within host tissues during migration and encystment may be attributed to immunomodulatory molecules released by the excretory cells of larvae in excretory-secretory (ES) products. ES products of parasites contain a variety of molecules, including proteins, lipids, and extracellular vesicles (EVs). Toxocara excretory-secretory (TES) products have been studied to some degree, with proteomic analysis of TES proteins described previously; however, investigation of the EVs within TES is lacking, despite the suggested role for these molecules in host interaction and potential immunomodulation. To further characterize the protein cargo within EVs in TES, EVs were isolated from larval cultures of T. canis and T. cati via ultrafiltration, with concurrent collection of EV-depleted TES filtrate for additional study. Isolated EVs and EV-depleted TES from both T. canis and T. cati were submitted for proteomic analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS). Proteomic identification results revealed 140 proteins across all samples, with 16 shared by all samples, and 76 total proteins shared between T. canis and T. cati, present within EVs and EV-depleted TES. There were 17 proteins shared exclusively by EV samples, and 15 were shared exclusively between EV-depleted TES samples. Many shared proteins were associated with the host immune response. Several proteins were specific to either T. canis or T. cati, highlighting the potential use of these proteins as diagnostic tools in the differentiation of etiologic agents in cases of toxocariasis. The results of this study build upon previously reported proteomic evaluations of TES, contributing new information in regards to newly identified proteins, EV protein cargo within TES, and potential immunomodulatory functions of these proteins.

Future semiochemical control of codling moth, Cydia pomonella

Codling moth (CM), Cydia pomonella, is a significant pest of apple (Malus domestica) and other orchard crops worldwide, posing challenges due to the decrease in registered insecticides, rising resistance, and a changing climate. The pest exhibits a strong resistance capacity to both synthetic and natural insecticides, while shifting seasonal temperatures disrupt the reliability of phenology-temperature models for predicting targeted control strategies. Alternative control strategies are necessary to future-proof control of this pest. Current control methods primarily rely on chemical insecticide sprays or granulosis virus applications during egg hatching. This review focuses explicitly on semiochemical-based manipulation of CM adults and larvae for control in orchards. Topics covered include the role of semiochemicals in integrated pest management, area-wide control, mating disruption, female attractants, larval kairomones, and incorporation into monitoring and control strategies. The potential of CM repellents in a push–pull strategy is also discussed. Primary sources for identifying relevant literature included GoogleScholar and ResearchGate, with a focus on papers published since 2013 but also include relevant papers from 2003. Nine review papers and 119 papers were reviewed. The review emphasizes that effective control necessitates an area-wide approach targeting all life stages (eggs, larvae, pupae, and adults). Comprehensive monitoring is crucial for identifying CM “hot-spots” and enhancing targeted interventions. Growers must consider landscape context when designing control programs. Lastly, recommendations are provided for future research and CM management strategies. There are opportunities to explore and exploit female kairomone attractants and repellents in control strategies and modify monitoring traps to be more attractive and autonomous.

Impact of Benzodiazepine Delorazepam on Growth and Behaviour of Artemia salina Nauplii.

Benzodiazepines, a significant group of newly recognised water contaminants, are psychotropic medications prescribed for common anxiety symptoms and sleep disorders. They resist efficient degradation during sewage treatment and endure in aquatic environments. Their presence in aquatic matrices is increasing, particularly after the recent pandemic period, which has led many people to systematically use benzodiazepines to manage anxiety. In previous studies, an important interference of this class of drugs on both the larval and adult stages of some aquatic species has been demonstrated, with effects on behaviour and embryonic development. This study examined the influence of delorazepam, a diazepam metabolite, on Artemia salina development to gain insight into responses in naupliar larvae. Results demonstrated that treatments (1, 5, and 10 µg/L) increase the hatching percentage and induce a desynchronisation in growth. Mortality was only slightly increased (close to 10% at six days post-hatching), but lipid reserve consumption was modified, with the persistence of lipid globules at the advanced naupliar stages. Locomotory activity significantly decreased only at 10 µg/L treatment. No teratogenic effects were observed, though modest damages were noticed in the posterior trunk and eyes, two targets of environmental toxicity. The negative impact of delorazepam on Artemia salina adds to those already reported in other species of invertebrates and vertebrates, which are not yet considered targets of these drugs. This study underscores the need for further research and immediate attention to this class of contaminants and the importance of monitoring their presence during environmental risk assessments.

Chromosome-level genome assembly of Scathophaga stercoraria provides new insights into the evolutionary adaptations of dung flies

The yellow dung fly Scathophaga stercoraria is a widely distributed species in high-altitude regions of the Northern Hemisphere. It plays important roles as a decomposer, predator, and pollinator in the ecosystem. As a staple model organism, S. stercoraria serves as a standard test species for assessing the toxicity of drug residues in livestock dung and has been the focus of numerous studies. The genetic mechanisms underlying the ecological adaptability of S. stercoraria remain poorly understood. To fill the gap, we first assembled a high-quality chromosome-level genome of S. stercoraria, resulting in a final assembly size of 549.64 Mb, with a contig N50 of 4.06 Mb, and 92.53 % of the sequence anchored to six chromosomes. Gene family analysis revealed an expansion of Toll (Toll1), GNBP3, Cyp303a1, Cyp4d14, Cyp6g1, OR67d, and yolk protein genes in the S. stercoraria genome. Transcriptome analysis indicated that most genes in the trypsin and carboxypeptidase gene families are predominantly expressed during the larval stage, whereas the α-Amylase gene family is mainly expressed during the adult stage. Additionally, PGRP-SC is highly expressed during the larval stage, OBPs are primarily expressed during the adult stage, and yolk protein genes exhibit female-biased expression. Our study not only provides a new resource for the dung flies genomic pool, but also identifies the expression patterns of key ecologically adaptative genes and gene families at the developmental stages, which provides new insights into the ecological adaptive evolution of dung flies.

Application timing affects the efficacy of Bacillus thuringiensis subsp. kurstaki against Tortrix viridana in deciduous oak forests

The green oak leaf roller moth, Tortrix viridana, is one of the main defoliators of deciduous oaks in the Mediterranean region. Although aerial applications of Btk-based insecticides represent the most effective method to control the larval populations of this pest at a large spatial scale, the optimal time window for sprayings has not yet been defined. Accordingly, experimental trials were conducted in a Quercus pubescens forest district in Sardinia (Italy) involving both large-scale aerial treatments and small-scale applications from the ground. The aim of our work was to study how different larval development stages and/or sprouting phenological phases of the host trees affect Btk efficacy. Aerial sprayings were conducted in 2003 and 2004 in two different areas of about 300 ha, selected according to altitude and exposure to simulate three different application timings against different larval instars and sprouting phenological phases. From-the-ground applications were carried out in 2021 in a Q. pubescens forest stand by applying Btk-based insecticides at different timings, and the overall Q. pubescens phenological stage and the distribution of T. viridana larval development stages were assessed accordingly. The effectiveness of Btk applications was evaluated by estimating larval mortality 7, and 14 days after aerial applications, and 7, 14 and 21 days after ground applications. Defoliation due to T. viridana was also evaluated at the end of larval development in all years. In both 2003 and 2004, the average larval mortality due to Btk exceeded 80 % two weeks after aerial applications across all application timings. Damage caused by T. viridana varied with Btk application timing, with the later applications showing a higher defoliation than other timings. In ground application experiments, larval mortality significantly increased when Btk was sprayed against a larval population primarily composed of IV instars, which were feeding on sprouts with leaves that were spread out and had not fully developed internodes, thus resulting in a significative reduction of insect damage. Our findings indicate that, in accordance with the strict synchrony between the larval development and the oak flushing, the optimal timing for Btk application against T. viridana can be defined based on the bud burst phenological phase. In particular, the prevalence of sprout with expanding leaf surface allowing insecticidal droplet interception and the simultaneous presence of susceptible larval instars (within the IV), resulted in an increased treatment efficacy.

Cu(II)-Dependent Spine Development Injury in Zebrafish (Danio rerio) with Organ Heterogeneous Cu Imbalance.

Growing evidence suggests that the imbalance of Cu leads to multiorgan diseases or other adverse effects, but the underlying mechanisms remain largely unknown. Herein, we used zebrafish to uncover the mystery of organ heterogeneous responses to Cu stress and Cu(II)-dependent spine developmental injury in the early organogenesis stage. We first demonstrated that Cu(I) was distributed in the entire body, but high contents of Cu(II) were accumulated in the yolk sac and eye in normal zebrafish larvae. Cu exposure from birth to 144 hpf caused no obvious damage to Cu-metabolizing organs (liver and intestine), despite the elevated Cu(I) and Cu(II) levels. However, the spine was more sensitive to the Cu exposure. In the spine region, the Cu(I) level remained stable, whereas the level of Cu(II) significantly increased, which was highly associated with spine development injury. A significant negative correlation between Cu(II) and the spine-related parameters was identified. Moreover, cuproptosis caused spine development deformation during the early embryogenesis stage. Spine-related pathways such as somitegenesis significantly changed in the early embryogenesis period, and 5 spine-related pathways were significantly altered in the larval stage at 96 hpf. Our study suggested that Cu stress induced organ heterogeneous Cu imbalance and Cu(II)-dependent spine development injury in zebrafish.