A preliminary note on attraction and oviposition preferences of Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae).

Chrysomya rufifacies (Diptera: Calliphoridae) is a blow fly species of forensic importance, documented to have a strong preference for colonisation of substrate already inhabited by heterospecific blow fly larvae, thus exhibiting secondary colonisation behaviour. Larvae exhibit predatory behaviour that may be useful to support development where food substrate is limited or high competition exists, but they may alternately be drawn to pre-colonised substrate to capitalise on the advantages of collective exodigestion by previous/current colonisers. Previous authors have suggested female Ch. rufifacies may use visual orientation to detect substrate currently colonised by heterospecific larvae, rather than chemoreception of volatile organic compounds (VOCs), that signify condition of substrate, which would infer that active colonisation is likely a more important oviposition cue for Ch. rufifacies than substrate condition. This study addressed attraction as well as oviposition, examining whether the condition of substrate (either previously colonised or never colonised) or the presence of heterospecific larvae was more important in the initial choice of food source by female Ch. rufifacies where conspecifics were not present, and whether the condition of substrate and presence of heterospecific larvae affects the number of offspring deposited by a female. Attraction was studied using a Y-olfactometer system, and oviposition using a binary-choice assay, with females responding to pairwise choice between an array of meat conditions (fresh, larval aged or aged) and presence/absence of Lucilia sericata larvae. Females displayed a hierarchy of choice of larval aged substrate > aged substrate > fresh substrate, with the active presence of heterospecific larvae a secondary factor in choice. Females produced higher offspring numbers on meat that was either currently or previously colonised by heterospecific larvae, demonstrating the importance of heterospecific indicators of previous or current colonisation as an oviposition cue. This serves as an important consideration for entomologists working with Ch. rufifacies in any capacity where other blow fly species may be present, and most importantly for forensic entomologists where time of colonisation is utilised to estimate PMI.

Olfactory learning in Pieris brassicae butterflies is dependent on the intensity of a plant-derived oviposition cue.

Butterflies, like many insects, use gustatory and olfactory cues innately to assess the suitability of an oviposition site and are able to associate colours and leaf shapes with an oviposition reward. Studies on other insects have demonstrated that the quality of the reward is a crucial factor in forming associative memory. We set out to investigate whether the large cabbage white Pieris brassicae (Linnaeus) has the ability to associate an oviposition experience with a neutral olfactory cue. In addition, we tested whether the strength of this association is dependent on the gustatory response to the glucosinolate sinigrin, which is a known oviposition stimulus for P. brassicae. Female butterflies were able to associate a neutral odour with an oviposition experience after a single oviposition experience, both in a greenhouse and in a semi-natural outdoor setting. Moreover, butterflies performed best when trained with concentrations of sinigrin that showed the strongest response by specific gustatory neurons on the forelegs. Our study provides novel insight into the role of both gustatory and olfactory cues during oviposition learning in lepidopterans and contributes to a better understanding of how these insects might be able to adapt to a rapidly changing environment.

Heterogeneity in the projections and excitability of tyraminergic/octopaminergic neurons that innervate the Drosophila reproductive tract.

Aminergic nuclei in mammals are generally composed of relatively small numbers of cells with broad projection patterns. Despite the gross similarity of many individual neurons, recent transcriptomic, anatomic and behavioral studies suggest previously unsuspected diversity. Smaller clusters of aminergic neurons in the model organism Drosophila melanogaster provide an opportunity to explore the ramifications of neuronal diversity at the level of individual cells. A group of approximately 10 tyraminergic/octopaminergic neurons innervates the female reproductive tract in flies and has been proposed to regulate multiple activities required for fertility. The projection patterns of individual neurons within the cluster are not known and it remains unclear whether they are functionally heterogenous. Using a single cell labeling technique, we show that each region of the reproductive tract is innervated by a distinct subset of tyraminergic/octopaminergic cells. Optogenetic activation of one subset stimulates oviduct contractions, indicating that the cluster as a whole is not required for this activity, and underscoring the potential for functional diversity across individual cells. Using whole cell patch clamp, we show that two adjacent and morphologically similar cells are tonically inhibited, but each responds differently to injection of current or activation of the inhibitory GluCl receptor. GluCl appears to be expressed at relatively low levels in tyraminergic/octopaminergic neurons within the cluster, suggesting that it may regulate their excitability via indirect pathways. Together, our data indicate that specific tyraminergic/octopaminergic cells within a relatively homogenous cluster have heterogenous properties and provide a platform for further studies to determine the function of each cell.

Temperature-Dependent Oviposition Models for Monochamus saltuarius (Coleoptera: Cerambycidae).

Monochamus saltuarius Gebler is a serious insect pest in Europe and East Asia regions, including Portugal, Spain, China, Japan, and Korea. It transfers the pine wood nematode Bursaphelenchus xylophilus to conifer trees, resulting in pine wilt disease (PWD). As temperature is a key factor influencing insect population dynamics, temperature-dependent models describing M. saltuarius oviposition could estimate population growth potential and evaluate outbreak risks. In this study, the longevity and fecundity of M. saltuarius females were measured under constant temperature conditions ranging from 20 to 32 °C, and temperature-dependent models were constructed. The longevity of M. saltuarius females ranged from 83.36 days to 22.92 days, with a total fecundity of 141 eggs and 52.77 eggs at 20 °C and 32 °C, respectively. To describe oviposition, we used a single-phase simulation describing oviposition as a single model and a two-phase simulation describing sexual maturation and oviposition as two separate models. These models effectively described M. saltuarius oviposition (r2 > 0.96) under constant temperature conditions, with the two-phase simulation demonstrating greater accuracy overall. Such models could facilitate assessments of PWD risks. The modeling framework of this study shows potential for predicting threats from various forestry and agricultural pests.

Male condition and seminal fluid affect female host-marking behavior in the Mexican fruit fly.

Mating and the transfer of seminal fluid components including male accessory glands (MAGs) proteins can affect oviposition behavior in insects. After oviposition, some species of fruit flies deposit a host-marking pheromone (HMP) on the fruit that discourages oviposition by other females of the same or different species or genus and reduces competition between larvae. However, we know very little about how mating, receiving seminal fluid, or male condition can affect female host marking behavior. Here, we tested how the physiological state of females (mated or unmated), the receipt of seminal fluid, and the condition of the male (wild or sterile) affect oviposition and host-marking behavior (HMB) in Anastrepha ludens (Diptera: Tephritidae). We also determined the efficiency of the host-marking pheromone from mated or unmated females in deterring oviposition. In a further examination of how seminal fluid may be affecting HMB we assessed if there were differences in the size of wild or sterile MAGs and the protein quantity transferred during mating. Our results indicate that receiving seminal fluid increased egg laying and increased time invested in host-marking (HM). Unmated females laid fewer eggs than mated females but invested the same amount of time in depositing host-marking pheromone, which had similar effectiveness in deterring oviposition as that of mated females. Females that mated with sterile males laid the same number of eggs as females that mated with wild males but spent less time depositing host-marking pheromone, which suggests that females detect the condition of the male and invest less in marking hosts. Finally, sterile males had larger accessory glands and transferred more MAGs proteins during mating compared to wild males. Seminal proteins could be manipulating HM behavior and female investment into their current reproductive effort. We are only beginning to understand how male condition and seminal fluid can affect female physiology and maternal investment in HMP.

Short-term responses of spider mites inform mechanisms of maize resistance to a generalist herbivore.

Plants are attacked by diverse herbivorous pests with different host specializations. While host plant resistance influences pest pressure, how resistance impacts the behaviors of generalist and specialist herbivores, and the relationship to resistance, is less well known. Here, we investigated the short-term (< 1 h) behavioral changes of a generalist herbivore, the two-spotted spider mite (TSM), and a specialist herbivore, the Banks grass mite (BGM), after introduction to no-choice Tanglefoot leaf-arenas (2 × 2 cm) of three maize inbred lines (B73, B75, and B96). The widely-used inbred line B73 is susceptible to spider mites, while B75 and B96 are known to be mite resistant, especially to TSM. Video tracking was used to record TSM and BGM walking, probing, feeding, resting, web-building and travel distance on arenas of each line. Mite oviposition was also recorded after 72 h. B75, a resistant line, decreased the feeding behavior (i.e., time) of both mite species compared to B73 (susceptible control) and B96. Moreover, TSM appeared to be sensitive to both resistant lines (B75 and B96) with reduced oviposition, and increased resting and web-building times compared to susceptible B73. In contrast, the specialist BGM showed no difference in oviposition, resting and web-building time across all maize inbred lines. Our findings of quite broad and short-term responses of TSM to B75 and B96 are consistent with a role for constitutive or rapidly induced plant defenses in maize in conferring TSM resistance. Other mechanisms of plant resistance may be needed, however, for defense against specialists like BGM.

Reproductive parameters of a new biocontrol agent, Eupeodes americanus (Diptera: Syrphidae) and comparison with the commercialized Aphidoletes aphidimyza (Diptera: Cecidomyiidae).

The American hoverfly Eupeodes americanus (Wiedemann) (Diptera: Syrphidae) is an aphidophagous predator during its larval stage and is currently being evaluated for inclusion in biocontrol programs as a new biocontrol agent. However, little is known about its reproductive aptitudes. The objective of the present study was to determine the reproductive parameters of E. americanus and to compare them with those of a commercialized and widely used biological control agent for aphids, the aphid midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae). The preoviposition period, oviposition period, adult longevity, lifetime and daily fecundity, egg hatching rate, and fertility were determined for E. americanus females and compared to those of A. aphidimyza. Trials were conducted under laboratory conditions in rearing cages on the broad bean plant Vicia faba L. (Fabaceae), infested with pea aphids Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). The results revealed that the preoviposition period, oviposition period, and adult longevity were significantly longer in E. americanus than in A. aphidimyza. The daily fecundity and egg-hatching rate were similar in both species. However, lifetime fecundity and fertility were considerably higher in E. americanus than in A. aphidimyza. This study demonstrates that the reproductive capacity of E. americanus is clearly superior to that of A. aphidimyza and therefore supports its inclusion in the aphid pest management program as a new biocontrol agent.

Neem-Extract Formulation on Hypothenemus hampei Preference and Performance in Arabica Coffee Fruits and Artificial Diet.

Coffee berry borer (CBB) Hypothenemus hampei is a major biotic threat to coffee production worldwide. Studies have reported negative effects on CBB by oil-based formulations of neem (Azadirachta indica), but little information is available for other neem-extract formulations. This study evaluated CBB preference and performance in arabica coffee fruits and artificial diet treated with a neem-extract formulation (Openeem Plus®) in the field and laboratory conditions. Field experiments were performed using CBB females artificially infested in cherry or green coffee fruits confined in voile-fabric cages tied to branches of neem-treated and control plants, recording the adult mortality and offspring production. Dual-choice and no-choice bioassays assessed CBB preference and development in fruits and artificial diet treated with the neem extract compared to controls in the laboratory, respectively. As main results obtained in the field and laboratory experiments, the neem extract significantly reduced CBB oviposition in both cherry and green fruits, as well as in artificial diet compared to controls. However, the botanical product did not affect CBB adult survival and preference in the laboratory bioassays. The neem extract is promising for use in pest management strategies in sustainable arabica coffee crops by reducing CBB oviposition and offspring. These effects can contribute to lowering the pest population buildup along the crop cycle and damage potential to coffee production.

An overview of technologies available to monitor behaviours of mosquitoes.

Mosquito-borne diseases such as malaria, dengue, Zika, and chikungunya cause significant morbidity and mortality globally, resulting in over 600,000 deaths from malaria and around 36,000 deaths from dengue each year, with millions of people infected annually, leading to substantial economic losses. The existing mosquito control measures, such as long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS), helped to reduce the infections. However, mosquito-borne diseases are still among the deadliest diseases, forcing us to improve the existing control methods and look for alternative methods simultaneously. Advanced monitoring techniques, including remote sensing, and geographic information systems (GIS) have significantly enhanced the efficiency and effectiveness of mosquito control measures. Mosquitoes' behavioural traits, such as locomotion, blood-feeding, and fertility are the key determinants of disease transmission and epidemiology. Technological advancements, such as high-resolution cameras, infrared imaging, and artificial intelligence (AI) driven object detection models, including groundbreaking convolutional neural networks, have provided efficient and precise options to monitor various mosquito behaviours, including locomotion, oviposition, fertility, and host-seeking. However, they are not commonly employed in mosquito-based research. This review highlights the novel and significant advancements in behaviour-monitoring tools, mostly from the last decade, due to cutting-edge video monitoring technology and artificial intelligence. These advancements can offer enhanced accuracy, efficiency, and the ability to quickly process large volumes of data, enabling detailed behavioural analysis over extended periods and large sample sizes, unlike traditional manual methods prone to human error and labour-intensive. The use of behaviour-assaying techniques can support or replace existing monitoring techniques and directly contribute to improving control measures by providing more accurate and real-time data on mosquito activity patterns and responses to interventions. This enhanced understanding can help establish the role of behavioural changes in improving epidemiological models, making them more precise and dynamic. As a result, mosquito management strategies can become more adaptive and responsive, leading to more effective and targeted interventions. Ultimately, this will reduce disease transmission and significantly improve public health outcomes.

EVALUATION OF THE EFFICIENCY OF BEETROOT PEEL (BETA VULGARIS) IN OVITRAPS AS AN ATTRACTANT FOR SURVEILLANCE OF ARBOVIRUS VECTORS IN THE MUNICIPALITY OF AGRESTINA, STATE OF PERNAMBUCO, BRAZIL.

It is estimated that there are over 500 species of arboviruses worldwide, with more than 150 of them directly associated with human diseases, the majority of which are zoonotic. Among the main arboviruses circulating in Brazil, dengue, Zika, and chikungunya stand out, all transmitted through a common vector, Aedes aegypti. Given this scenario, the development and implementation of more efficient surveillance strategies become urgent. This study aims to compare and evaluate the efficiency of beetroot peel, Beta vulgaris, as an attractant for Ae. aegypti oviposition under field conditions in the municipality of Agrestina, State of Pernambuco, Brazil. Beetroot peel extract is a cheap and accessible source of geosmin, which holds significant potential as an attractant for mosquitoes due to its resemblance to microbial volatiles found in water bodies rich in organic material. During the study period (November 2023 to April 2024), 40 traps were set in each neighborhood, with 20 of each attractant. Two traps were used per property, one following the traditionally used model (beer yeast infusion) and another containing beetroot peel. Overall, in both analyzed neighborhoods (Campo Novo and Cohab), beetroot peel appeared to be a more efficient attractant for the oviposition of culicids. Beetroot peel showed the highest values in egg abundance and in the analyzed indices.

Taste cells expressing Ionotropic Receptor 94e reciprocally impact feeding and egg laying in Drosophila.

Chemosensory cells across the body of Drosophila melanogaster evaluate the environment to prioritize certain behaviors. Previous mapping of gustatory receptor neurons (GRNs) on the fly labellum identified a set of neurons in L-type sensilla that express Ionotropic Receptor 94e (IR94e), but the impact of IR94e GRNs on behavior remains unclear. We used optogenetics and chemogenetics to activate IR94e neurons and found that they drive mild feeding suppression but enhance egg laying. In vivo calcium imaging revealed that IR94e GRNs respond strongly to certain amino acids, including glutamate, and that IR94e plus co-receptors IR25a and IR76b are required for amino acid detection. Furthermore, IR94e mutants show behavioral changes to solutions containing amino acids, including increased consumption and decreased egg laying. Overall, our results suggest that IR94e GRNs on the fly labellum discourage feeding and encourage egg laying as part of an important behavioral switch in response to certain chemical cues.

Different genotypes and species of symbiotic fungi mediate the behavioral response of invasive Sirex noctilio fabricius (Hymenoptera: Siricidae).

In northeast China, the invasive woodwasp., Sirex noctilio, attacks Pinus sylvestris var. mongolica Litv and often shares habitat with native Sirex nitobei. Previous research showed that S. noctilio can utilize the volatiles from its symbiotic fungus (A. areolatum IGS-BD) to locate host trees. Consequently, symbiotic fungi (A. areolatum IGS-D and A. chailletii) carried by S. nitobei may influence the behavioral selection of S. noctilio. This study aimed to investigate the impact of fungal odor sources on S. noctilio's behavior in laboratory and field experiments. Our observations revealed that female woodwasps exhibited greater attraction toward the fungal volatiles of 14-day-old Amylostereum IGS-D in a "Y"-tube olfactometer and wind tunnel. When woodwasps were released into bolts inoculated separately with three strains in the field, females of S. noctilio exhibited a preference for those bolts pre-inoculated with A. areolatum IGS-BD. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that the volatiles emitted by the two genotypes of A. areolatum were similar yet significantly distinct from those of Ampelopsis chailletii. Hence, we postulate that the existence of native A. areolatum IGS-D could potentially facilitate the colonization of S. noctilio in scenarios with minimal or no A. areolatum IGS-BD present in the host.

Oviposition strategies in Pieridae butterflies and the role of an egg-killing plant trait therein.

Most herbivorous insects are host-plant specialists that evolved detoxification mechanisms to overcome their host plant's toxins. In the evolutionary arms-races between Pieridae butterflies and Brassicaceae plants, some plant species have evolved another defence against the pierids: egg-killing. Underneath the eggs, leaves develop a so-called hypersensitive response (HR)-like cell death. Whether some butterflies have evolved oviposition strategies to counter-adapt against egg-killing remains to be studied. In this study, we assessed the oviposition site location of Pieridae butterflies on their natural host plants. We described the plant tissue on which we located the eggs of the most common Pieridae in the Netherlands: Gonepteryx rhamni, Anthocharis cardamines, Pieris rapae, P. napi, P. brassicae and P. mannii. Additionally, we assessed expression of HR-like cell death in response to the deposited butterfly eggs. We found that both A. cardamines and G. rhamni mainly oviposited on the floral stem and the branch, respectively, and oviposited on host plants from lineages not expected to kill pierid eggs. Accordingly, no HR-like cell death was seen. All Pieris eggs found were located on leaves of their host, the only tissue found to express HR-like cell death. Furthermore, each Pieris species was found to at least occasionally oviposit on Brassica nigra. This was the only plant species in this survey that expressed HR-like cell death in response to the eggs of P. rapae, P. napi and P. brassicae. Our observations demonstrate that HR-like cell death remains an effective defence strategy against these Pieris species and as such did not find evidence for the hypothesized counterstrategies. Surveying certain key species and disentangling the micro-evolution of oviposition strategies within a species would allow us to further investigate potential counter-adaptations that evolved against HR-like cell death. This study provides the basis for further investigation of potential counter-adaptations to egg-killing defences.

Laboratory studies of movement and microenvironment choices of engorged adult female Amblyomma maculatum (Acari: Ixodidae).

Microenvironmental factors affect ovipositional choices and behavior in ticks. In this study, engorged female Amblyomma maculatum Koch were released in an observation arena covered with garden soil. The arena was evenly split into wet and dry sides, each containing 5 different types of structures (totaling 10). Upon release, observations at particular time points were made over 2 days as to speed, distance, actual paths traveled, and ultimate site selection, presumably for oviposition. In addition, time-lapse videography was utilized to record the track of each individual tick. This scenario was replicated 3 times with different cohorts of ticks (n = 5 per replicate). Of the total 15 ticks released in the arena, all attained a final site selection by 24 h. These final sites were 7/15 (47%) edge of arena; 2/15 (13%) under bark; 2/15 (13%) open soil; 2/15 (13%) at or near release point; 1/15 (7%) tunnel with animal fur; and 1/15 (7%) tunnel with chicken feathers. At final site selection, 11/15 (73%) subsequently burrowed into the soil, 3 of which were completely buried. Time-lapse videography revealed that most ticks (80%) explored the arena in a "looping back" fashion. Overall, engorged Gulf Coast ticks moved at an average speed of 0.09 mm/s, and the total distance traversed by the ticks averaged 2.37 m.

Host Plants and Fertilization Mediated Life History of American Serpentine Leaf Miner, Liriomyza trifolii (Burgess) (Diptera: Agromyzidae).

Herbivorous insects depend on the host plant to optimize their overall reproductive success, and balanced fertilization may alter the plant's quality against herbivory. Life history traits of the Liriomyza trifolii (Burgess) were determined under laboratory conditions using either unfertilized and fertilized plants of bean [Phaseolus vulgaris L. (Fabaceae)], chrysanthemum [Chrysanthemum × morifolium (Asteraceae)], potato [Solanum tuberosum (Solanaceae)], bell pepper [Capsicum annuum (Solanaceae)], and tomato [Solanum lycopersicum (Solanaceae)]. Results indicated that L. trifolii completed development on all studied unfertilized and fertilized plants. Nevertheless, a higher performance of the leaf miner was observed on bean and bell pepper plants compared to the other plants. Furthermore, there was an interaction of the host plant and fertilization with Calcium Aria or Sitam negatively affecting the fitness-related traits of the leaf miner. Application of these fertilizers resulted in delayed immature development of L. trifolii, decreased survival rate, and reduced adult longevity and fecundity. The activity of cinnamyl alcohol dehydrogenase (CAD), peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) enzymes, as well as phenolic, flavonoid, and lignin content were higher in Calcium Aria + Sitam fertilized plants, intermediate in Calcium Aria and Sitam treated plants, and the lower in unfertilized plants. The development and survival of L. trifolii on different host plants, considering fertilization options, become important for deploying cultural control practices against this important pest species.

Effects of alloferon and its analogues on reproduction and development of the Tenebrio molitor beetle.

As the most numerous group of animals on Earth, insects are found in almost every ecosystem. Their useful role in the environment is priceless; however, for humans, their presence may be considered negative or even harmful. For years, people have been trying to control the number of pests by using synthetic insecticides, which eventually causes an increased level of resistance to applied compounds. The effects of synthetic insecticides have encouraged researchers to search for alternatives and thus develop safe compounds with high specificity. Using knowledge about the physiology of insects and the functionality of compounds of insect origin, a new class of bioinsecticides called peptidomimetics, which are appropriately modified insect analogues, was created. One promising compound that might be successfully modified is the thirteen amino acid peptide alloferon (HGVSGHGQHGVHG), which is obtained from the hemolymph of the blue blowfly Calliphora vicinia. Our research aimed to understand the physiological properties of alloferon and the activity of its peptidomimetics, which will provide the possibility of using alloferon or its analogues in the pharmaceutical industry, as a drug or adjuvant, or in agriculture as a bioinsecticide. We used alloferon and its three peptidomimetics, which are conjugates of the native peptide with three unsaturated fatty acids with various chain lengths: caprylic, myristic, and palmitic. We tested their effects on the morphology and activity of the reproductive system and the embryogenesis of the Tenebrio molitor beetle. We found that the tested compounds influenced the growth and maturation of ovaries and the expression level of the vitellogenin gene. The tested compounds also influenced the process of egg laying, embryogenesis, and offspring hatching, showing that alloferon might be a good peptide for the synthesis of effective bioinsecticides or biopharmaceuticals.

Evaluation of the effect of agroclimatic variables on the probability and timing of olive fruit fly attack.

Agroclimatic variables may affect insect and plant phenology, with unpredictable effects on pest populations and crop losses. Bactrocera oleae Rossi (Diptera: Tephritidae) is a specific pest of Olea europaea plants that can cause annual economic losses of more than one billion US dollars in the Mediterranean region. In this study, we aimed at understanding the effect of olive tree phenology and other agroclimatic variables on B. oleae infestation dynamics in the Umbria region (Central Italy). Analyses were carried out on B. oleae infestation data collected in 79 olive groves during a 7-year period (from 2015 to 2021). In July-August, B. oleae infestation (1% attack) was negatively affected by altitude and spring mean daily temperatures and positively by higher winter mean daily temperatures and olive tree cumulative degree days. In September-October, infestation was negatively affected by a positive soil water balance and high spring temperatures. High altitude and cumulative plant degree days were related to delayed attacks. In contrast, high winter and spring temperatures accelerated them. Our results could be helpful for the development of predictive models and for increasing the reliability of decision support systems currently used in olive orchards.

Molecular Basis of CO2 Sensing in Hyphantria cunea.

Carbon dioxide (CO2) released by plants can serve as a cue for regulating insect behaviors. Hyphantria cunea is a widely distributed forestry pest that may use CO2 as a cue for foraging and oviposition. However, the molecular mechanism underlying its ability to sense CO2 has not been elucidated. Our initial study showed that CO2 is significantly attractive to H. cunea adults. Subsequently, 44 H. cunea gustatory receptors (GRs) were identified using transcriptome data, and 3 candidate CO2 receptors that are specifically expressed in the labial palps were identified. In vivo electrophysiological assays revealed that the labial palp is the primary organ for CO2 perception in H. cunea, which is similar to findings in other lepidopteran species. By using the Xenopus oocyte expression system, we showed that the HcunGR1 and HcunGR3 co-expressions produced a robust response to CO2, but HcunGR2 had an inhibitory effect on CO2 perception. Finally, immunohistochemical staining revealed sexual dimorphism in the CO2-sensitive labial pit organ glomerulus (LPOG). Taken together, our results clarified the mechanism by which H. cunea sense CO2, laying the foundation for further investigations into the role of CO2 in the rapid spread of H. cunea.

Taking it with a grain of salt: tolerance to increasing salinization in Culex pipiens (Diptera: Culicidae) across a low-lying delta.

BACKGROUND: Salinity, exacerbated by rising sea levels, is a critical environmental cue affecting freshwater ecosystems. Predicting ecosystem structure in response to such changes and their implications for the geographical distribution of arthropod disease vectors requires further insights into the plasticity and adaptability of lower trophic level species in freshwater systems. Our study investigated whether populations of the mosquito Culex pipiens, typically considered sensitive to salt, have adapted due to gradual exposure. METHODS: Mesocosm experiments were conducted to evaluate responses in life history traits to increasing levels of salinity in three populations along a gradient perpendicular to the North Sea coast. Salt concentrations up to the brackish-marine transition zone (8 g/l chloride) were used, upon which no survival was expected. To determine how this process affects oviposition, a colonization experiment was performed by exposing the coastal population to the same concentrations. RESULTS: While concentrations up to the currently described median lethal dose (LD50) (4 g/l) were surprisingly favored during egg laying, even the treatment with the highest salt concentration was incidentally colonized. Differences in development rates among populations were observed, but the influence of salinity was evident only at 4 g/l and higher, resulting in only a 1-day delay. Mortality rates were lower than expected, reaching only 20% for coastal and inland populations and 41% for the intermediate population at the highest salinity. Sex ratios remained unaffected across the tested range. CONCLUSIONS: The high tolerance to salinity for all key life history parameters across populations suggests that Cx. pipiens is unlikely to shift its distribution in the foreseeable future, with potential implications for the disease risk of associated pathogens.

Spotted Lanternflies Respond to Natural Pheromone Lures for Mate-Finding and Oviposition.

Using semiochemicals collected from spotted lanternflies Lycorma delicatula (Hemiptera: Fulgoridae) (SLF) and deployed in the field with circle traps, we demonstrated that SLF responded to SLF pheromones: in particular, this was the case for males while seeking mates and for females while ovipositing. The attractants consisted of SLF body extract emitted from diffuser lures and SLF honeydew on burlap ribbons, collected from heavily infested locations. Traps with attractants were deployed in field sites with very light SLF infestations to avoid competing signals of pre-existing aggregations. The number of SLF equivalents emitted by each diffuser per trapping period was used in a dose-response analysis. Three trees per block received either (1) a control hexane lure and a clean ribbon, (2) a lure containing SLF extract and a clean ribbon, or (3) a lure containing SLF extract and a honeydew-laden ribbon. Ten blocks were sampled three times per week for twelve weeks. We found a significant positive dose-response by males to SLF body extract only in the presence of SLF honeydew, indicating a synergistic effect between honeydew volatiles and body volatiles. This dose-response occurred for five weeks after mating started, after which males no longer responded. Subsequently, females had a significant positive dose-response to SLF extract only in the presence of honeydew when oviposition was their primary activity, continuing for two weeks, suggesting that females may use pheromones to aggregate for oviposition. The extract in the absence of honeydew did not result in a positive dose-response, nor did the hexane control. These findings suggest that SLF respond synergistically to the combination of pheromones present in both SLF honeydew and SLF bodies. Thus, combining key components from both sources may aid the development of semiochemical lures for SLF.