Oxygen extraction efficiency of the tidally-ventilated rectal gills of dragonfly nymphs.

Dragonfly nymphs breathe water using tidal ventilation, a highly unusual strategy in water-breathing animals owing to the high viscosity, density and low oxygen (O2) concentration of water. This study examines how well these insects extract O2 from the surrounding water during progressive hypoxia. Nymphs were attached to a custom-designed respiro-spirometer to simultaneously measure tidal volume, ventilation frequency and metabolic rate. Oxygen extraction efficiencies (OEE) were calculated across four partial pressure of oxygen (pO2) treatments, from normoxia to severe hypoxia. While there was no significant change in tidal volume, ventilation frequency increased significantly from 9.4 ± 1.2 breaths per minute (BPM) at 21.3 kPa to 35.6 ± 2.9 BPM at 5.3 kPa. Metabolic rate increased significantly from 1.4 ± 0.3 µl O2 min-1 at 21.3 kPa to 2.1 ± 0.4 µl O2 min-1 at 16.0 kPa, but then returned to normoxic levels as O2 levels declined further. OEE of nymphs was 40.1 ± 6.1% at 21.3 kPa, and did not change significantly during hypoxia. Comparison to literature shows that nymphs maintain their OEE during hypoxia unlike other aquatic tidal-breathers and some unidirectional breathers. This result, and numerical models simulating experimental conditions, indicate that nymphs maintain these extraction efficiencies by increasing gill conductance and/or lowering internal pO2 to maintain a sufficient diffusion gradient across their respiratory surface.

Wavelength-specific negatively phototactic responses of the burrowing mayfly larvae Ephoron virgo.

Mayflies are typically negatively phototactic during larval development, whereas the adults possess positive phototaxis. However, no extensive research has been done into the wavelength dependence of phototaxis in any mayfly larvae. We measured the repellency rate of Ephoron virgo larvae to light as a function of wavelength in the 368-743 nm spectral range. We established that the magnitude of repellence increased with decreasing wavelength and the maximal responses were elicited by 400 nm violet light. This wavelength dependence of phototaxis is similar to the recently reported spectral sensitivity of positive phototaxis of the twilight-swarming E. virgo adults. Negative phototaxis not only facilitates predation evasion: avoidance of the blue-violet spectral range could also promote the larvae to withdraw towards the river midline in the case of a drop in the water level, when the underwater light becomes enriched with shorter wavelengths as a result of the decreasing depth of overhead river water.

Predicting the habitat suitability of Schistosoma intermediate host Bulinus truncatus, its predatory aquatic insect Odonata nymph, and the associated aquatic plant Ceratophyllum demersum using MaxEnt.

Schistosomiasis is one of the most important parasitic diseases in tropical and subtropical areas. Its prevalence is associated with the distribution of freshwater snails, which are their intermediate hosts. Thus, control of freshwater snails is the solution to reduce the transmission of this disease. This will be achieved by understanding the relationship between the snails and their habitats including natural enemies and associated aquatic plants as well as the factors affecting their distribution. In this study, Maximum Entropy model (MaxEnt) was used for mapping and predicting the possible geographic distribution of Bulinus truncatus snail (the intermediate host of Schistosoma haematobium), Odonata nymph (predatory aquatic insect), and Ceratophyllum demersum (the associated aquatic plant) in Egypt based on topographic and climatic factors. The models of the investigated species were evaluated using the area under receiver operating characteristic curve. The results showed that the potential risk areas were along the banks of the Nile River and its irrigation canals. In addition, the MaxEnt models revealed some similarities in the distribution pattern of the vector, the predator, and the aquatic plant. It is obvious that the predictive distribution range of B. truncatus was affected by altitude, precipitation seasonality, isothermality, and mean temperature of warmest quarter. The presence of B. truncatus decreases with the increase of altitude and precipitation seasonality values. It could be concluded that the MaxEnt model could help introducing a predictive risk map for Schistosoma haematobium prevalence and performing better management strategies for schistosomiasis.

Identification and characterization of microRNAs in the immature stage of the beneficial predatory bug Arma chinensis Fallou (Hemiptera: Pentatomidae).

MicroRNAs (miRNAs) are a type of small noncoding RNAs that regulate gene expression at the posttranscriptional level and can influence significant biological processes. Arma chinensis (Hemiptera: Pentatomidae) is a predaceous insect species that preys upon a wide variety of insect pests. It is important to explore and understand the molecular mechanisms involving miRNAs in regulating developmental and other gene expression for beneficial insects. However, examination of miRNAs associated with Hemiptera, especially predatory bugs, has been absent or scarce. This study represents the first comprehensive analysis of predatory bug A. chinensis encoded miRNAs through high throughput sequencing and predicts genes and biological processes regulated by the newly identified miRNAs through analyzing their differential expression in and across five nymphal instars. A total of 64 A. chinensis miRNAs, including 46 conserved miRNAs and 18 novel miRNAs, were identified by analysis of high throughput sequence reads mapped to the genome. A total of 2913 potential gene targets for these 64 miRNAs were predicted by comprehensive analyses utilizing miRanda, PITA, and RNAhybrid. Gene Ontology annotation of predicted target genes of A. chinensis suggested the key processes regulated by miRNAs involved biological processes, regulation of cellular processes, and transporter activity. Kyoto Encyclopedia of Genes and Genomes pathway predictions included the Toll and Imd signaling pathway, Valine, leucine and isoleucine degradation, Steroid biosynthesis, the AGE-RAGE signaling pathway in diabetic complications, and Alanine, aspartate and glutamate metabolism. This newly identified miRNAs through analyzing their differential expression, assessment of their predicted functions forms a foundation for further investigation of specific miRNAs.

Courtship, Mating Behavior, and Ovary Histology of the Nymph Parasitoid Psyllaephagus bliteus (Hymenoptera: Encyrtidae).

The potential of the parasitoid Psyllaephagus bliteus Riek for the biological control of the eucalyptus pest Glycaspis brimblecombei Moore (Hemiptera: Aphalaridae) nymphs is high. This research sought to analyze the courtship, male competition, and mating behavior of P. bliteus at the proportions of 1:1 and 2:1 males to female in a Petri dish (5 cm diameter), and to describe the ovary histology of virgin and mated females of this parasitoid. At 1:1, males touch the antennae and thorax-abdomen of females during courtship, but females avoid mate attempts before they are 48 h old. At 2:1, the competition between male parasitoids inhibits mating. The histology of ovaries of virgin and mated P. bliteus females is similar, with two well-defined germarium and vitellarium regions, with oocytes at different developmental stages, including mature ones rich in yolk and with eggshell. A clearer understanding of the reproductive behavior and histology of P. bliteus aids in the use of this parasitoid for the biological control of G. brimblecombei.

Verification of rabbits as intermediate hosts for Linguatula serrata (Pentastomida) in Australia.

We confirm the presence of nymphs of the introduced pentastomid, Linguatula serrata, in the introduced rabbit, Oryctolagus cuniculus, in Australia, based on morphological and molecular results. Two nymphs were collected from a single rabbit near the Kosciuszko National Park in New South Wales. Unlike reports of nymphs from domestic animals, these nymphs were not encapsulated, despite having the morphological features of infective nymphs. The possibility of different development pathways in lagomorphs is discussed. Examination of feral deer for L. serrata was unsuccessful and potential reasons for a lack of infection in these animals are postulated. Our results reiterate the need for a combined morphological and molecular approach to the identification of L. serrata. Further sampling of a range of feral and native animals is required to determine the true range of intermediate hosts and their relative importance in the transmission of L. serrata in Australia.

Apyrase with anti-platelet aggregation activity from the nymph of the camel tick Hyalomma dromedarii.

Apyrase is one of the essential platelet aggregation inhibitors in hematophagous arthropods due to its ability to hydrolyze ATP and ADP molecules. Here, an apyrase (TNapyrase) with antiplatelet aggregation activity was purified and characterized from the nymphs of the camel tick Hyalomma dromedarii through anion exchange and gel filtration columns. The homogeneity of TNapyrase was confirmed by native-PAGE, SDS-PAGE as well as with isoelectric focusing. Purified TNapyrase had a molecular mass of 25 kDa and a monomer structure. TNapyrase hydrolyzed various nucleotides in the order of ATP > PPi > ADP > UDP > 6GP. The Km value was 1.25 mM ATP and its optimum activity reached at pH 8.4. The influence of various ions on TNapyrase activity showed that FeCl2, FeCl3 and ZnCl2 are activators of TNapyrase. EDTA inhibited TNapyrase activity competitively with a single binding site on the molecule and Ki value of 2 mM. Finally, TNapyrase caused 70% inhibition of ADP-stimulated platelets aggregation and is a possible target for antibodies in future tick vaccine studies.

Nymphal engorgement weight predicts sex of adult Amblyomma americanum, Amblyomma maculatum, Dermacentor andersoni, Dermacentor variabilis, and Rhipicephalus sanguineus ticks.

The engorgement weights of laboratory-raised nymphs of five common ticks in the USA, Amblyomma americanum, Amblyomma maculatum, Dermacentor andersoni, Dermacentor variabilis, and Rhipicephalus sanguineus, experimentally fed to repletion on an adult sheep (Ovis aries) were evaluated to determine the sex of molted adult ticks. Length of feeding period of nymphs, molting success and durations, and sex ratios between tick species were also compared. Individual replete nymphs were weighed and allowed to molt to adults in a humidity chamber. Length of feeding duration was different by species (F4,7785 = 1963.79; P < 0.0001); R. sanguineus nymphs became replete fastest, followed by A. americanum, D. variabilis, A. maculatum, and D. andersoni. Significant difference in molting success was not detected. The mean body weight of engorged nymphs of A. americanum (t = 32.3; df = 662), A. maculatum (t = - 9.70; df = 255), D. variabilis (t = 15.7; df = 751), and R. sanguineus (t = 5.17; df = 560; all P < 0.0001) molting into females was greater than that of nymphs molting into males, whereas heavier D. andersoni engorged nymphs became males (t = 8.71; df = 480; P < 0.0001). Amblyomma maculatum nymphs that molted to females fed to repletion faster (t = 3.33; df = 265; P ≤ 0.001) than nymphs that molted to males and a higher proportion (χ2 = 48.4; df = 1, P < 0.0001) of A. maculatum and D. andersoni (χ2 = 8.19; df = 1, P = 0.004) molted to females than males. Our study demonstrated biological and behavioral differences in and between engorging nymphs of five ixodid species. These findings may aid in studies evaluating the role of tick sex in transmission of tick-borne pathogens.

Redescription of Amblyomma integrum adults and immature stages.

Amblyomma integrum Karsch, 1879 (Acari: Ixodidae) is one of four Amblyomma Koch, 1844 species with eyes found in southern India and Sri Lanka. The immature stages of this species were poorly described. Therefore, accurate identification is difficult. Here we re-describe the male, female, nymph and larva of A. integrum and illustrate all the stages in greater detail for the first time. A set of diagnostic morphological characters is defined to distinguish this species from other sympatric species of eyed Amblyomma in any parasitic stage of development. Adults of A. integrum parasitize mostly various larger mammals whereas nymphs and larvae use mostly larger and medium mammals. Amblyomma integrum is recorded from India (Andhra Pradesh, Goa, Karnataka, Orissa and Tamil Nadu States) and throughout Sri Lanka.

Morphological and molecular description of immature stages of Ornithodoros savignyi (Acari: Argasidae).

This study was designed to provide more details about larva, first nymph, and second nymph of Ornithodoros savignyi using a combination of light microscopy (LM), scanning electron microscopy (SEM), and partial sequence of mitochondrial 16S ribosomal ribonucleic acid (rRNA). The main characteristics of larva are wrinkled integument with many grooves, gnathosoma without camerostome cheeks, hypostome with a pair of large teeth apically, and tarsus without humps. The comparisons between the first and the second nymphs are different shape and distribution of dorsal grooves; a few spots without mammilla on the dorsal surface of the second nymph; 27 and 63-65 pairs of setae on the dorsal surface of the first and second nymphs, respectively; small holes on mammillae that are more dense in the second nymph; basis capitulum with two pairs of small setae in the second nymph; and one pair of sate in the first nymph, hypostome with dental formula 2/2 in the first nymph, and 3/3 in the second nymph. The partial 16S rRNA sequence of the second nymph that was determined as O. savignyi (450 bp) was deposited in GenBank under the accession number KU163242.

Successful Feeding of Amblyomma coelebs (Acari: Ixodidae) Nymphs on Humans in Brazil: Skin Reactions to Parasitism.

Identifying the tick species that successfully feed on humans would increase knowledge of the epidemiology of several tick-borne diseases. These species salivate into the host, increasing the risk of pathogen transmission. However, there is a lack of data in the literature regarding the ticks that prefer to feed on humans. Herein, we describe the successful feeding of Amblyomma coelebs Neumann nymphs on two of the authors after accidental tick bites occurred during field surveys in two preserved areas of Mato Grosso do Sul, Brazil. One of the host-parasite interactions was closely monitored, and the tick development, gross host skin alterations, and related sensations are presented.

Seasonal Changes in the Caste Distribution of Foraging Populations of Formosan Subterranean Termite in New Orleans, Louisiana.

This study examined the relationship between temperature, precipitation, soil composition, levels of feeding damage, and the caste distribution (workers, soldiers, nymphs) of the Formosan subterranean termite, Coptotermes formosanus Shiraki, collected in underground monitoring stations over a 12 mo period. Because nymphs are the caste that develops into alates, the seasonal abundance of nymphs was examined over a 5 yr period. Numbers of workers, soldiers, and soldier/worker ratio were significantly affected by month. Recruitment and retention of foraging termites in stations was significantly affected by the level of feeding damage. The number of nymphs collected in monitoring stations was highly variable. In the 12 mo test, there was a significant correlation between numbers of nymphs and level of feeding damage, temperature, precipitation, and soil composition. Over a 5 yr period, significantly more nymphs were collected in 2011 than in 2007 and 2008. Peak nymph collections varied from year to year. Overall, peak nymph collections were more likely to occur in Mar., Sept., and Oct. Increasing our knowledge of the environmental factors that influence recruitment and retention of foraging termites in monitoring stations could influence termite bait placement and improve baiting strategies for termite control. Identifying the key factors that cause aggregations of nymphs in underground stations could increase our ability to predict the intensity and location of alate swarms.

Description of the immature stages of Sigara (Aphelosigara) tucma.

Descriptions of the last three nymphal instars of the water boatman Sigara (Aphelosigara) tucma Bachmann (Hemiptera: Heteroptera: Corixidae) are provided, for the first time, for specimens collected in northern Argentina; the egg is redescribed. Adults and nymphs were collected in the field. The eggs were obtained by dissection from females. The immature stages were fixed for microscopic examination and illustration and were described with an emphasis on morphometry and chaetotaxy of selected structures. The last three nymphal instars of S. tucma can be easily recognized by the body and head lengths and widths; the number of transverse sulcations of the rostrum; the chaetotaxy of trochanters, protibiotarsus, midlegs, metatibia, and metatarsus; and the grade of development of the wing pads. The eggs of the genus Sigara can be distinguished by the size and the chorionic surface. The chaetotaxy of the mesonotum, metafemur, and metatibia distinguish groups of species belonging to the last nymphal instar of the genus Sigara. A key to the last three nymphal instars of the species of Sigara from Argentina is provided.

De Novo Transcriptome Profiling of Mustard Aphid (Lipaphis erysimi) and Differential Expression of Transcripts Associated with Feeding and Non-Feeding Conditions and Developmental Stages.

Lipaphis erysimi is a specialist aphid of the Indian subcontinent that causes significant yield losses in oilseed Brassicas. Several aphid genes have been used as preferred targets in RNAi-based transgenic plants for aphid resistance. In order to enhance the repertoire of potential target genes for aphid control and to identify the genes associated with aphid feeding and development, we performed a two-way comparative study of differential gene expression profiles between (i) feeding and non-feeding adults and (ii) adult and nymph developmental stages of L. erysimi. De novo RNA-seq of aphids using Illumina technology generated a final transcriptome comprising 52,652 transcripts. Potential transcripts for host selection, detoxification, salivary proteins and effectors, molecular chaperones and developmental genes were identified. Differential gene expression studies identified variations in the expression of 1502 transcripts between feeding and non-feeding adults and 906 transcripts between nymphs and adults. These data were used to identify novel target genes for RNAi-based aphid control and facilitate further studies on the molecular basis of aphid feeding and development.

Wax "tails" enable planthopper nymphs to self-right midair and land on their feet.

The striking appearance of wax 'tails' - posterior wax projections on planthopper nymphs - has captivated entomologists and naturalists alike. Despite their intriguing presence, the functional roles of these structures remain largely unexplored. This study leverages high-speed imaging to uncover the biomechanical implications of these wax formations in the aerial dynamics of planthopper nymphs (Ricania sp.). We quantitatively demonstrate that removing wax tails significantly increases body rotations during jumps. Specifically, nymphs without wax projections undergo continuous rotations, averaging 4.3 ± 1.9 per jump, in contrast to wax-intact nymphs, who narrowly complete a full rotation, averaging only 0.7 ± 0.2 per jump. This suggests that wax structures effectively counteract rotation through aerodynamic drag forces. These stark differences in body rotation correlate with landing success: nymphs with wax intact achieve a near perfect landing rate of 98.5%, while those without wax manage only a 35.5% success rate. Jump trajectory analysis reveals transitions from parabolic to Tartaglia shapes at higher take-off velocities for wax-intact nymphs, illustrating how wax structures assist nymphs in achieving stable, controlled descents. Our findings confirm the aerodynamic self-righting functionality of wax tails in stabilizing planthopper landings, advancing our understanding of the complex interplay between wax morphology and aerial maneuverability, with broader implications for the evolution of flight in wingless insects and bioinspired robotics.

Ixodes ricinus tick bacteriome alterations based on a climatically representative survey in Hungary.

IMPORTANCE: Climate-sensitive disease vectors, such as ticks, respond to the environment with changes in their microbiome. These changes can affect the emergence or re-emergence of various vector-borne pathogens, such as the causative agent of Lyme borreliosis (LB) or tick-borne encephalitis. This aspect is particularly emphasized in light of climate change. The climatically representative assessment of microbiome differences in various developmental stages of the most common Central European tick species, Ixodes ricinus, deepens our understanding of the potential climatic factors behind microbial relative abundance and interaction changes. This knowledge can support the development of novel disease vector control strategies.

Multiple physiological cohorts comprise seasonal activity of wild Amblyomma americanum (Acari: Ixodidae) nymphs.

Nymphs of the hard tick Amblyomma americanum (L.) are an important life stage in the maintenance and transmission of tick-borne pathogens. As pathogen composition can vary across developmental cohorts, it is essential to understand the demographic structure of the questing population. Amblyomma americanum nymphs often display a second peak in activity during late summer, but it is unknown whether this peak represents older overwintered ticks or younger newly molted ticks. The objective of this study was to examine the heme concentration in field-captured A. americanum nymphs to determine if the questing population consists of one physiological cohort or multiple cohorts in a season. Ticks were collected from March to August in an old field of primarily non-native grasses, and heme concentration was used to assess physiological age. LOESS modeling depicted that heme concentration in the population declined from March to early July but increased in later sampling sessions. As ticks cannot replenish declining heme stores without a blood meal, a late-summer spike in heme concentration demonstrates that newly molted nymphs are entering the active population. The vector potential of these newly emerging nymphs may differ from those collected earlier in the year as pathogen diversity depends on reservoir host dynamics and timing of larval feeding.

Lanternflies (Hemiptera: Fulgoridae) of Taiwan.

The family Fulgoridae belongs to the order Hemiptera, superfamily Fulgoridea, with approximately 770 described species worldwide. Their extraordinary appearance attracts the attention of both entomologists and the public. In addition to the evolutionary issue of their special appearance, certain species are also considered notorious pests (e.g., Lycorma delicatula). Several issues widely exist in previous taxonomic studies of lanternflies: (1) application of uncertain morphological characters leads to synonymy or misidentification; (2) descriptions of male genitalia are incomplete; (3) information of nymphal morphology is insufficient. Therefore, this study aims to provide a comprehensive taxonomic study of Fulgoridae from Taiwan. In this study, eight species in six genera from Taiwan were reported, of which Limois westwoodii was recorded for the first time from Taiwan. Lycorma olivacea was proposed as a new junior synonym of L. meliae. The fifth-instar nymph of Saiva formosana was described for the first time. Detailed descriptions of these lanternflies and an identification key to adults of Fulgoridae from Taiwan were also included.

A remarkable new species of the flat bug genus Nesoproxius (Hemiptera, Aradidae), the first Oceanian representative with brachyptery.

A new flat bug species, Nesoproxiuskishimotoisp. nov., from the Oceanian region (Ogasawara Islands, Japan) is described. It is the first brachypterous representative in the genus Nesoproxius. The sexual dimorphism, nymph, and habitat are also described for the first time in this genus. A key to the species of Nesoproxius is also provided.

20E biosynthesis gene CYP306A1 confers resistance to imidacloprid in the nymph stage of Bemisia tabaci by detoxification metabolism.

BACKGROUND: Difference in physiology level between the immature and mature stages of insects likely contribute to different mechanisms of insecticide resistance. It is well acknowledged that insect 20-hydroxyecdysone (20E) plays an important role in many biological processes in the immature stage, whether 20E confers insecticide resistance at this specific stage is still poorly understood. By gene cloning, reverse transcription quantitative real-time PCR, RNA interference (RNAi) and in vitro metabolism experiments, this study aimed to investigate the potential role of 20E-related genes in conferring imidacloprid (IMD) resistance in the immature stage of the whitefly Bemisia tabaci Mediterranean. RESULTS: After identification of low to moderate IMD resistance in the whitefly, we found CYP306A1 of the six 20E-related genes was overexpressed in the nymph stage of the three resistant strains compared to a laboratory reference susceptible strain, but not in the adult stage. Further exposure to IMD resulted in an increase in CYP306A1 expression in the nymph stage. These results together imply that CYP306A1 may be implicated in IMD resistance in the nymph stage of the whitefly. RNAi knockdown of CYP306A1 increased the mortality of nymphs after treatment with IMD in bioassay, suggesting a pivotal role of CYP306A1 in conferring IMD resistance in the nymph stage. Additionally, our metabolism experiments in vivo showed that the content of IMD reduced by 20% along with cytochrome P450 reductase and heterologously expressed CYP306A1, which provides additional evidence for the important function of CYP306A1 in metabolizing IMD that leads to the resistance. CONCLUSION: This study uncovers a novel function of the 20E biosynthesis gene CYP306A1 in metabolizing imidacloprid, thus contributing to such resistance in the immature stage of the insect. These findings not only advance our understanding of 20E-mediated insecticide resistance, but also provide a new target for sustainable pest control of global insect pests such as whitefly. © 2023 Society of Chemical Industry.