BACKGROUND: Lyme disease continues to expand in Canada and the USA and no single intervention is likely to curb the epidemic.
METHODS: We propose a platform to quantitatively assess the effectiveness of a subset of Ixodes scapularis tick management approaches. The platform allows us to assess the impact of different control treatments, conducted either individually (single interventions) or in combination (combined efforts), with varying timings and durations. Interventions include three low environmental toxicity measures in differing combinations, namely reductions in white-tailed deer (Odocoileus virginianus) populations, broadcast area-application of the entomopathogenic fungus Metarhizium anisopliae, and fipronil-based rodent-targeted bait boxes. To assess the impact of these control efforts, we calibrated a process-based mathematical model to data collected from residential properties in the town of Redding, southwestern Connecticut, where an integrated tick management program to reduce I.xodes scapularis nymphs was conducted from 2013 through 2016. We estimated parameters mechanistically for each of the three treatments, simulated multiple combinations and timings of interventions, and computed the resulting percent reduction of the nymphal peak and of the area under the phenology curve.
RESULTS: Simulation outputs suggest that the three-treatment combination and the bait boxes-deer reduction combination had the overall highest impacts on suppressing I. scapularis nymphs. All (single or combined) interventions were more efficacious when implemented for a higher number of years. When implemented for at least 4 years, most interventions (except the single application of the entomopathogenic fungus) were predicted to strongly reduce the nymphal peak compared with the no intervention scenario. Finally, we determined the optimal period to apply the entomopathogenic fungus in residential yards, depending on the number of applications.
CONCLUSIONS: Computer simulation is a powerful tool to identify the optimal deployment of individual and combined tick management approaches, which can synergistically contribute to short-to-long-term, costeffective, and sustainable control of tick-borne diseases in integrated tick management (ITM) interventions.

UNASSIGNED: Tick-borne pathogens, such as Borreliella spp., Rickettsia spp., and Anaplasma spp., are frequently detected in Germany. They circulate between animals and tick vectors and can cause mild to severe diseases in humans. Knowledge about distribution and prevalence of these pathogens over time is important for risk assessment of human and animal health.
UNASSIGNED: Ixodes ricinus nymphs were collected at different locations in 2009/2010 and 2019 in Germany and analyzed for tick-borne pathogens by real-time PCR and sequencing.
UNASSIGNED: Borreliella spp. were detected with a prevalence of 11.96% in 2009/2010 and 13.10% in 2019 with B. afzelii and B. garinii as dominant species. Borrelia miyamotoi was detected in seven ticks and in coinfection with B. afzelii or B. garinii. Rickettsia spp. showed a prevalence of 8.82% in 2009/2010 and 1.68% in 2019 with the exclusive detection of R. helvetica. The prevalence of Anaplasma spp. was 1.00% in 2009/2010 and 7.01% in 2019. A. phagocytophilum was detected in seven tick samples. None of the nymphs were positive for C. burnetii.
UNASSIGNED: Here, observed changes in prevalence were not significant after a decade but require longitudinal observations including parameters like host species and density, climatic factors to improve our understanding of tick-borne diseases.

Heavy metals are defined as an abiotic factor that affects the efficiency of biological pest control. This study constructed a cadmium (Cd)-polluted artificial diets-Hyphantria cunea-Arma chinensis food chain to analyze the effects of Cd exposure on the ability of A. chinensis to control H. cunea. The results revealed that Cd was transferred through the artificial diet to H. cunea larvae and A. chinensis nymphs via a biological amplification effect. After feeding on Cd-accumulated H. cunea larvae, the body weight of A. chinensis nymphs reduced, mortality increased, developmental duration prolonged, and the expression of growth regulatory genes (EX, cycE, and MER) decreased. Cd activated the antioxidant defense system of the nymphs, accompanied by a significant enhancement in the contents of H2O2 and MDA, marked damage to the midgut sub-microstructure, and a remarkable induction in the expression of genes crucial for the mitochondrial pathway/ER stress-apoptosis pathway. Cd significantly diminished the contents of total amino acids, glucose, free fatty acids, and expression of the genes (HK2, PFK, IDH1, and IDH2) essential for the TCA cycle and glycolysis in the nymphs. The preference of the A. chinensis nymphs to Cd-treated H. cunea larvae was evidently reduced. Cd diminished the search-ability, food intake, instantaneous attack rate, and maximum theoretical daily food intake but prolonged the feeding time of the nymphs. Taken together, Cd exposure reduces the ability of A. chinensis nymphs to control H. cunea and provides a new challenge for the efficiency of insect pest control using natural enemies. These findings have important reference value for optimizing pest control strategies in heavy metal polluted areas.

Transgenerational phenotypic modification can alter organismal fitness, population demographics, and community interactions. For ectotherms, both dietary composition and temperature have important effects on organismal fitness, but they are rarely investigated together. Mormon crickets Anabrus simplex are capable of diapausing as eggs in the soil for multiple years with duration largely dependent on cumulative heat units or degree days. Because Mormon crickets can be abundant in the landscape in one year and disappear suddenly the next, I asked: does parental nutrition affect the duration of egg diapause? Beginning in the ultimate nymphal instar, Mormon crickets were fed a diet high in protein, one equal in protein to carbohydrate, or a diet high in carbohydrates and the time for eggs to develop after they were laid was measured. If parental nutrition affects temperature-sensitive egg diapause, then that change in sensitivity to temperature might also alter the relationship between embryonic development rate and temperature. I asked: does parental nutrition affect embryonic development rate as a function of temperature? To this end, I manipulated densities of Mormon cricket nymphs and protein-rich prey (grasshoppers) in field cages, collected eggs from the adult Mormon crickets, and measured the optimal temperature, maximum development rate, and thermal breadth for embryonic development of the offspring. I found that Mormon crickets fed a high protein diet laid eggs with shorter diapause. Consistent with this long-term result, those housed with the most grasshoppers to eat laid eggs that had the fastest maximum development rate, whereas those without grasshoppers laid eggs with slower maximum developmental rates but the broadest thermal breadth. Eggs from Mormon crickets housed with intermediate levels of grasshopper densities had a decline in peak development rate with an increase in density. In addition, Mormon crickets housed with more conspecifics laid eggs with faster development rates, whereas thermal breadth and the temperature optima were not affected by cricket density. As predicted, Mormon cricket diets significantly affected egg diapause and development rates. Contrary to expectations based on observed changes in diet preferences during a Mormon cricket outbreak, Mormon crickets fed high protein diets laid eggs with significantly shorter egg diapause and significantly faster egg development rates. Interestingly, doubling of Mormon cricket density caused eggs to develop in nearly half the time. This latter result indicates that Mormon cricket aggregations promote rapid development of progeny. Moreover, the tight, linear structure of migratory bands in which females intermittently stop to lay eggs assures that the progeny hatch and develop in dense cohorts. In this manner, the banding behavior might carry-over into subsequent generations as long as cohorts are dense and protein is available. With band thinning or protein restriction, females spread their bet-hedging and progeny remain longer as eggs in the soil.

Crickets have been extensively studied in recent insect nutritional research, but it remains largely unexplored how they balance the intake of multiple nutrients. Here, we used the nutritional geometry framework to examine the behavioural and physiological regulation of dietary protein and carbohydrate in nymphs of the two-spotted cricket, Gryllus bimaculatus (Orthoptera: Gryllidae). Growth, intake, utilization efficiencies, and body composition were measured from the eighth instar nymphs that received either food pairs or single foods with differing protein and carbohydrate content. When food choices were available, crickets preferentially selected a carbohydrate-biased protein:carbohydrate (P:C) ratio of 1:1.74. During this nutrient selection, carbohydrate intake was more tightly regulated than protein intake. When confined to nutritionally imbalanced foods, crickets adopted a nutrient balancing strategy that maximized the nutrient intake regardless of the nutrient imbalance, reflecting their omnivorous feeding habit. Intake was significantly reduced when crickets were confined to the most carbohydrate-biased food (P:C = 1:5). When nutrients were ingested in excess of the requirements, the post-ingestive utilization efficiencies of these nutrients were down-regulated, thereby buffering the impacts of nutrient imbalances on body nutrient composition. Crickets reared on the most carbohydrate-biased food (P:C = 1:5) suffered delayed development and reduced growth. Our data provide the most accurate description of nutrient regulation in G. bimaculatus and lay the foundation for further nutritional research in this omnivorous insect.

BACKGROUND: Southeast Asia is regarded as a hotspot for the diversity of ixodid ticks. In this geographical region, Vietnam extends through both temperate and tropical climate zones and therefore has a broad range of tick habitats. However, molecular-phylogenetic studies on ixodid tick species have not been reported from this country.
METHODS: In this study, 1788 ixodid ticks were collected from cattle, buffalos and a dog at 10 locations in three provinces of northern Vietnam. Tick species were identified morphologically, and representative specimens were molecularly analyzed based on the cytochrome c oxidase subunit I (cox1) and 16S rRNA genes. Fifty-nine tick species that are indigenous in Vietnam were also reviewed in the context of their typical hosts in the region.
RESULTS: Most ticks removed from cattle and buffalos were identified as Rhipicephalus microplus, including all developmental stages. Larvae and nymphs were found between January and July but adults until December. Further species identified from cattle were Rhipicephalus linnaei, Rhipicephalus haemaphysaloides, Amblyomma integrum and Haemaphysalis cornigera. Interestingly, the latter three species were represented only by adults, collected in one province: Son La. The dog was infested with nymphs and adults of R. linnaei in July. Phylogenetically, R. microplus from Vietnam belonged to clade A of this species, and R. haemaphysaloides clustered separately from ticks identified under this name in China, Taiwan and Pakistan. Amblyomma integrum from Vietnam belonged to the phylogenetic group of haplotypes of an Amblyomma sp. reported from Myanmar. The separate clustering of H. cornigera from Haemaphysalis shimoga received moderate support.
CONCLUSIONS: Three tick species (R. linnaei, A. integrum and H. cornigera) are reported here for the first time in Vietnam, thus increasing the number of indigenous tick species to 62. Clade A of R. microplus and at least R. linnaei from the group of Rhipicephalus sanguineus sensu lato occur in the country. There is multiple phylogenetic evidence that different species might exist among the ticks that are reported under the name R. haemaphysaloides in South and East Asia. This is the first report of A. integrum in Southeastern Asia.

Gene duplication is a fundamental evolutionary process which provides opportunities to acquire new gene functions. In the case of the insulin receptors (InRs) in cockroaches and close-related insects, two successive duplications determined the occurrence of three InR genes: InR2, InR1 and InR3, the last two forming a sister cluster to InR2. The biological role of each of the gene duplicates and whether they resulted from neofunctionalization or subfunctionalization is still unclear. The analysis of the sequences from different lineages did not detect positive selection as driving the divergence of InR1 and InR3, discarding neofunctionalization, and suggesting that there is no functional divergence between both gene copies. Using the cockroach Blattella germanica as a model, we have determined that BgInR2 is the gene with the highest expression levels in all the tissues analyzed, both in adult females and males, as well as in nymphs and embryos. BgInR3 is second in expression levels while BgInR1 is expressed at lower levels and only in some tissues. The selective depletion by RNAi of each of the three InRs, analyzed in terms of phenotype and fat body transcriptomic profiles, resulted in essentially redundant effects, with a magnitude approximately proportional to the level of expression of the respective InR. Therefore, the results indicate that the InR duplicates likely experienced a subfunctionalization process, by which the three InRs maintained similar functions but contributing to those functions proportionally to their expression levels.

Rab GTPase is critical for autophagy processes and is implicated in insect immunity against viruses. In this study, we aimed to investigate the role of FoRabs in the autophagic regulation of antiviral defense against tomato spotted wilt orthotospovirus (TSWV) in Frankliniella occidentalis. Transcriptome analysis revealed the downregulation of FoRabs in viruliferous nymph and adults of F. occidentalis in response to TSWV infection. Manipulation of autophagy levels with 3-MA and Rapa treatments resulted in a 5- to 15-fold increase and a 38-64% decrease in viral titers, respectively. Additionally, interference with FoRab10 in nymphs and FoRab29 in adults led to a 20-90% downregulation of autophagy-related genes, a decrease in ATG8-II (an autophagy marker protein), and an increase in the TSWV titers by 1.5- to 2.5-fold and 1.3- to 2.0-fold, respectively. In addition, the leaf disk and the living plant methods revealed increased transmission rates of 20.8-41.6 and 68.3-88.3%, respectively. In conclusion, FoRab10 and FoRab29 play a role in the autophagic regulation of the antiviral defense in F. occidentalis nymphs and adults against TSWV, respectively. These findings offer insights into the intricate immune mechanisms functional in F. occidentalis against TSWV, suggesting potential targeted strategies for F. occidentalis and TSWV management.

Usutu virus (USUV) is an emerging flavivirus that is maintained in an enzootic cycle with mosquitoes as vectors and birds as amplifying hosts. In Europe, the virus has caused mass mortality of wild birds, mainly among Common Blackbird (Turdus merula) populations. While mosquitoes are the primary vectors for USUV, Common Blackbirds and other avian species are exposed to other arthropod ectoparasites, such as ticks. It is unknown, however, if ticks can maintain and transmit USUV. We addressed this question using in vitro and in vivo experiments and field collected data. USUV replicated in IRE/CTVM19 Ixodes ricinus tick cells and in injected ticks. Moreover, I. ricinus nymphs acquired the virus via artificial membrane blood-feeding and maintained the virus for at least 70 days. Transstadial transmission of USUV from nymphs to adults was confirmed in 4.9% of the ticks. USUV disseminated from the midgut to the haemocoel, and was transmitted via the saliva of the tick during artificial membrane blood-feeding. We further explored the role of ticks by monitoring USUV in questing ticks and in ticks feeding on wild birds in the Netherlands between 2016 and 2019. In total, 622 wild birds and the Ixodes ticks they carried were tested for USUV RNA. Of these birds, 48 (7.7%) carried USUV-positive ticks. The presence of negative-sense USUV RNA in ticks, as confirmed via small RNA-sequencing, showed active virus replication. In contrast, we did not detect USUV in 15,381 questing ticks collected in 2017 and 2019. We conclude that I. ricinus can be infected with USUV and can transstadially and horizontally transmit USUV. However, in comparison to mosquito-borne transmission, the role of I. ricinus ticks in the epidemiology of USUV is expected to be minor.

Chagas disease affects around 8 million people globally, with Latin America bearing approximately 10,000 deaths each year. Combatting the disease relies heavily on vector control methods, necessitating the identification of new targets. Within insect genomes, genes harboring small open reading frames (smORFs - < 100 amino acids) present numerous potential candidates. In our investigation, we elucidate the pivotal role of the archetypal smORF-containing gene, mille-pattes/polished-rice/tarsalless (mlpt/pri/tal), in the post-embryonic development of the kissing bug Rhodnius prolixus. Injection of double-stranded RNA targeting mlpt (dsmlpt) during nymphal stages yields a spectrum of phenotypes hindering post-embryonic growth. Notably, fourth or fifth stage nymphs subjected to dsmlpt do not undergo molting. These dsmlpt nymphs display heightened mRNA levels of JHAMT-like and EPOX-like, enzymes putatively involved in the juvenile hormone (JH) pathway, alongside increased expression of the transcription factor Kr-h1, indicating changes in the hormonal control. Histological examination reveals structural alterations in the hindgut and external cuticle of dsmlpt nymphs compared to control (dsGFP) counterparts. Furthermore, significant changes in the vector\'s digestive physiology were observed, with elevated hemozoin and glucose levels in the posterior midgut of dsmlpt nymphs. Importantly, dsmlpt nymphs exhibit impaired metacyclogenesis of Trypanosoma cruzi, the causative agent of Chagas disease, underscoring the crucial role of proper gut organization in parasite differentiation. Thus, our findings constitute the first evidence of a smORF-containing gene\'s regulatory influence on vector physiology, parasitic cycle, and disease transmission.