Acute toxicity effects of pesticides on beneficial organisms - Dispelling myths for a more sustainable use of chemicals in agricultural environments.

Agricultural practitioners, researchers and policymakers are increasingly advocating for integrated pest management (IPM) to reduce pesticide use while preserving crop productivity and profitability. Using selective pesticides, putatively designed to act on pests while minimising impacts on off-target organisms, is one such option - yet evidence of whether these chemicals control pests without adversely affecting natural enemies and other beneficial species (henceforth beneficials) remains scarce. At present, the selection of pesticides compatible with IPM often considers a single (or a limited number of) widely distributed beneficial species, without considering undesired effects on co-occurring beneficials. In this study, we conducted standardised laboratory bioassays to assess the acute toxicity effects of 20 chemicals on 15 beneficial species at multiple exposure timepoints, with the specific aims to: (1) identify common and diverging patterns in acute toxicity responses of tested beneficials; (2) determine if the effect of pesticides on beetles, wasps and mites is consistent across species within these groups; and (3) assess the impact of mortality assessment timepoints on International Organisation for Biological Control (IOBC) toxicity classifications. Our work demonstrates that in most cases, chemical toxicities cannot be generalised across a range of beneficial insects and mites providing biological control, a finding that was found even when comparing impacts among closely related species of beetles, wasps and mites. Additionally, we show that toxicity impacts increase with exposure length, pointing to limitations of IOBC protocols. This work challenges the notion that chemical toxicities can be adequately tested on a limited number of 'representative' species; instead, it highlights the need for careful consideration and testing on a range of regionally and seasonally relevant beneficial species.

Wolbachia infection status and molecular diversity in the species of tribe Tagiadini Mabille, 1878 (Lepidoptera: Hesperiidae) collected in China.

Wolbachia, one of the most ubiquitous heritable symbionts in lepidopteran insects, can cause mitochondrial introgression in related host species. We recently found mito-nuclear discordance in the Lepidopteran tribe Tagiadini Mabille 1878 from which Wolbachia has not been reported. In this study, we found that 13 of the 46 species of Tagiadini species tested were positive for Wolbachia. Overall, 14% (15/110) of Tagiadini specimens were infected with Wolbachia and nine new STs were found from 15 isolates. A co-phylogenetic comparison, divergence time estimation and Wolbachia recombination analysis revealed that mito-nuclear discordance in Tagiadini species is not mediated by Wolbachia, but Wolbachia acquisition in Tagiadini appears to have occurred mainly through horizontal transmission rather than codivergence.

Defence and nutrition synergistically contribute to the distinct tolerance of rice subspecies to the stem borer, Chilo suppressalis.

Damage caused by the rice striped stem borer (SSB), Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), is much more severe on indica/xian rice than on japonica/geng rice (Oryza sativa) which matches pest outbreak data in cropping regions of China. The mechanistic basis of this difference among rice subspecies remains unclear. Using transcriptomic, metabolomic and genetic analyses in combination with insect bioassay experiments, we showed that japonica and indica rice utilise different defence responses to repel SSB, and that SSB exploited plant nutrition deficiencies in different ways in the subspecies. The more resistant japonica rice induced patterns of accumulation of methyl jasmonate (MeJA-part of a defensive pathway) and vitamin B1 (VB1 -a nutrition pathway) distinct from indica cultivars. Using gene-edited rice plants and SSB bioassays, we found that MeJA and VB1 jointly affected the performance of SSB by disrupting juvenile hormone levels. In addition, genetic variants of key biosynthesis genes in the MeJA and VB1 pathways (OsJMT and OsTH1, respectively) differed between japonica and indica rice and contributed to performance differences; in indica rice, SSB avoided the MeJA defence pathway and hijacked the VB1 nutrition-related pathway to promote development. The findings highlight important genetic and mechanistic differences between rice subspecies affecting SSB damage which could be exploited in plant breeding for resistance.

Introduction of Aedes aegypti mosquitoes carrying wAlbB Wolbachia sharply decreases dengue incidence in disease hotspots.

Partial replacement of resident Aedes aegypti mosquitoes with introduced mosquitoes carrying certain strains of inherited Wolbachia symbionts can result in transmission blocking of dengue and other viruses of public health importance. Wolbachia strain wAlbB is an effective transmission blocker and stable at high temperatures, making it particularly suitable for hot tropical climates. Following trial field releases in Malaysia, releases using wAlbB Ae. aegypti have become operationalized by the Malaysian health authorities. We report here on an average reduction in dengue fever of 62.4% (confidence intervals 50-71%) in 20 releases sites when compared to 76 control sites in high-rise residential areas. Importantly the level of dengue reduction increased with Wolbachia frequency, with 75.8% reduction (61-87%) estimated at 100% Wolbachia frequency. These findings indicate large impacts of wAlbB Wolbachia invasions on dengue fever incidence in an operational setting, with incidence expected to further decrease as wider areas are invaded.

Plasticity of cold and heat stress tolerance induced by hardening and acclimation in the melon thrips.

Extreme temperatures threaten species under climate change and can limit range expansions. Many species cope with changing environments through plastic changes. This study tested phenotypic changes in heat and cold tolerance under hardening and acclimation in the melon thrips, Thrips palmi Karny (Thysanoptera: Thripidae), an agricultural pest of many vegetables. We first measured the critical thermal maximum (CTmax) of the species by the knockdown time under static temperatures and found support for an injury accumulation model of heat stress. The inferred knockdown time at 39 °C was 82.22 min. Rapid heat hardening for 1 h at 35 °C slightly increased CTmax by 1.04 min but decreased it following exposure to 31 °C by 3.46 min and 39 °C by 6.78 min. Heat acclimation for 2 and 4 days significantly increased CTmax at 35 °C by 1.83, and 6.83 min, respectively. Rapid cold hardening at 0 °C and 4 °C for 2 h, and cold acclimation at 10 °C for 3 days also significantly increased cold tolerance by 6.09, 5.82, and 2.00 min, respectively, while cold hardening at 8 °C for 2 h and acclimation at 4 °C and 10 °C for 5 days did not change cold stress tolerance. Mortality at 4 °C for 3 and 5 days reached 24.07 % and 43.22 % respectively. Our study showed plasticity for heat and cold stress tolerance in T. palmi, but the thermal and temporal space for heat stress induction is narrower than for cold stress induction.

Diversity and distribution of sodium channel mutations in Aedes albopictus (Diptera: Culicidae).

There is growing interest in insecticide resistance in the mosquito, Aedes albopictus (Skuse), as its potential for spreading diseases is increasing as urbanization and control efforts intensify. Here we review the presence and diversity of mutations in the voltage-sensitive sodium channel (Vssc) gene associated with pyrethroid resistance and report on additional surveys of these mutations in new populations with an analysis of their spread. The known diversity of these mutations has increased in recent years including the identification of 26 non-synonymous mutations, although phenotypic data associating mutations with resistance remain limited. We provide data on mutations in several new locations including those in Timor Leste, Indonesia, and Vanuatu. We use population genomic data from ddRAD analyses of target populations with the 1534C mutation to identify single nucleotide polymorphisms (SNPs) associated with the mutant to test for clustering of SNPs based on the presence of the 1534C mutation rather than population origin. Our findings suggest spread of resistance alleles via genetic invasion, which is further supported by patterns from a genome-wide principal components analysis. These data point to movement of resistance alleles across wide areas with likely impacts on local control options.

Mosquitoes provide a transmission route between possums and humans for Buruli ulcer in southeastern Australia.

Buruli ulcer, a chronic subcutaneous infection caused by Mycobacterium ulcerans, is increasing in prevalence in southeastern Australia. Possums are a local wildlife reservoir for M. ulcerans and, although mosquitoes have been implicated in transmission, it remains unclear how humans acquire infection. We conducted extensive field survey analyses of M. ulcerans prevalence among mosquitoes in the Mornington Peninsula region of southeastern Australia. PCR screening of trapped mosquitoes revealed a significant association between M. ulcerans and Aedes notoscriptus. Spatial scanning statistics revealed overlap between clusters of M. ulcerans-positive Ae. notoscriptus, M. ulcerans-positive possum excreta and Buruli ulcer cases, and metabarcoding analyses showed individual mosquitoes had fed on humans and possums. Bacterial genomic analysis confirmed shared single-nucleotide-polymorphism profiles for M. ulcerans detected in mosquitoes, possum excreta and humans. These findings indicate Ae. notoscriptus probably transmit M. ulcerans in southeastern Australia and highlight mosquito control as a Buruli ulcer prevention measure.

Constitutively expressed small heat shock protein LsHsp21.5 not only enhances heat tolerance but also helps to maintain reproduction in female Laodelphax striatellus.

Coping with stressful conditions and maintaining reproduction are two key biological processes that affect insect population dynamics. Small heat shock proteins (sHSPs) are involved in the stress response and the development of insects. The sHsp gene Laodelphax striatellus (Hemiptera: Delphacidae) sHsp 21.5 (LsHsp21.5) showed constitutive, stage- and organ-specific expression in L. striatellus, a pest that damages cultivated rice in east Asia. The expression of LsHsp21.5 was highest in the ovary, with 43.60, 12.99 and 1.45 time higher expression here than in the head, gut and female fat bodies, respectively. The expression of this gene was weakly affected by heat or cold shock. The gene provided in vitro protection against heat damage to malate dehydrogenase and in vivo protection against heat stress in Escherichia coli (Enterobacteriales: Enterobacteriaceae) BL21(DE3) and L. striatellus. Moreover, L. striatellus reproduction decreased by 1.85 times when the expression of LsHsp21.5 was inhibited by RNA interference. The expression of some genes related to reproduction, such as the homologous gene of chorion protein, also declined. These results suggest that LsHsp21.5 expression not only protects other proteins against stress but also helps maintain the stable expression of some reproduction-related genes under non-stressful conditions, with impacts on L. striatellus fecundity.

A new spider mite elicitor triggers plant defence and promotes resistance to herbivores.

Herbivore-associated elicitors (HAEs) are active molecules produced by herbivorous insects. Recognition of HAEs by plants induces defence that resist herbivore attacks. We previously demonstrated that the tomato red spider mite Tetranychus evansi triggered defence in Nicotiana benthamiana. However, our knowledge of HAEs from T. evansi remains limited. Here, we characterize a novel HAE, Te16, from T. evansi and dissect its function in mite-plant interactions. We investigate the effects of Te16 on spider mites and plants by heterologous expression, virus-induced gene silencing assay, and RNA interference. Te16 induces cell death, reactive oxygen species (ROS) accumulation, callose deposition, and jasmonate (JA)-related responses in N. benthamiana leaves. Te16-mediated cell death requires a calcium signalling pathway, cytoplasmic localization, the plant co-receptor BAK1, and the signalling components SGT1 and HSP90. The active region of Te16-induced cell death is located at amino acids 114-293. Moreover, silencing Te16 gene in T. evansi reduces spider mite survival and hatchability, but expressing Te16 in N. benthamiana leaves enhances plant resistance to herbivores. Finally, Te16 gene is specific to Tetranychidae species and is highly conserved in activating plant immunity. Our findings reveal a novel salivary protein produced by spider mites that elicits plant defence and resistance to insects, providing valuable clues for pest management.

Mitochondrial DNA variation in Aedes aegypti (Diptera: Culicidae) mosquitoes from Jeddah, Saudi Arabia.

Wolbachia (Hertig 1936) (Rickettsiales: Ehrlichiaceae) has emerged as a valuable biocontrol tool in the fight against dengue by suppressing the transmission of the virus through mosquitoes. Monitoring the dynamics of Wolbachia is crucial for evaluating the effectiveness of release programs. Mitochondrial (mtDNA) markers serve as important tools for molecular tracking of infected mitochondrial backgrounds over time but require an understanding of the variation in release sites. In this study, we investigated the mitochondrial lineages of Aedes aegypti (Linnaeus 1762) in Jeddah, Saudi Arabia, which is a prospective release site for the "wAlbBQ" Wolbachia-infected strain of this mosquito species. We employed a combination of comprehensive mitogenomic analysis (including all protein-coding genes) and mtDNA marker analysis (cox1 and nad5) using data collected from Jeddah. We combined our mitogenome and mtDNA marker data with those from previous studies to place mitochondrial variation in Saudi Arabia into a broader global context. Our findings revealed the presence of 4 subclades that can be broadly categorized into 2 major mitochondrial lineages. Ae. aegypti mosquitoes from Jeddah belonged to both major lineages. Whilst mitogenomic data offered a higher resolution for distinguishing Jeddah mosquitoes from the wAlbBQ strain, the combination of cox1 and nad5 mtDNA markers alone proved to be sufficient. This study provides the first important characterization of Ae. aegypti mitochondrial lineages in Saudi Arabia and offers essential baseline information for planning future molecular monitoring efforts during the release of Wolbachia-infected mosquitoes.

How Can Genomics Help or Hinder Wildlife Conservation?

Genomic data are becoming increasingly affordable and easy to collect, and new tools for their analysis are appearing rapidly. Conservation biologists are interested in using this information to assist in management and planning but are typically limited financially and by the lack of genomic resources available for non-model taxa. It is therefore important to be aware of the pitfalls as well as the benefits of applying genomic approaches. Here, we highlight recent methods aimed at standardizing population assessments of genetic variation, inbreeding, and forms of genetic load and methods that help identify past and ongoing patterns of genetic interchange between populations, including those subjected to recent disturbance. We emphasize challenges in applying some of these methods and the need for adequate bioinformatic support. We also consider the promises and challenges of applying genomic approaches to understand adaptive changes in natural populations to predict their future adaptive capacity.

The effects of insecticide seed treatments on the parasitism and predation of Myzus persicae (Homoptera: Aphididae) in canola.

The green peach aphid, Myzus persicae (Sulzer) (Homoptera: Aphididae), is a major pest of brassica plants, with the ability to transmit > 100 viruses. Although the adoption of Integrated Pest Management is increasing, chemical treatment remains the predominant method used to control M. persicae globally. Insecticide seed treatments, typically with neonicotinoid active ingredients, have become commonplace in canola crops, and are viewed as a "softer" alternative to foliar sprays but may nevertheless impact natural enemies of M. persicae. In this study, the effects of canola seed treatments, containing imidacloprid, thiamethoxam, and a mixture of thiamethoxam + lambda-cyhalothrin, were investigated on the parasitoid wasp, Aphidius colemani Viereck (Hymenoptera: Braconidae) and the green lacewing, Mallada signatus (Schneider) (Neuroptera: Chrysopidae), both important natural enemies of M. persicae. Laboratory trials were undertaken using whole plants, with lethal and sublethal effects assessed by measuring several traits. Compared with untreated plants, more aphid mummies were produced and more A. colemani were reared on plants treated with thiamethoxam + lambda-cyhalothrin and more aphid mummies were produced on imidacloprid plants. Imidacloprid reduced the time A. colemani spent searching for M. persicae and thiamethoxam reduced its cleaning time. However, after A. colemani were removed from treated plants, there were no such effects observed, suggesting these impacts were relatively short-lived. We found no significant effects of seed treatments on M. signatus. These results point to the complexity of ecotoxicology studies involving multiple trophic levels and indicate that seed treatments may have variable impacts on key fitness traits of natural enemies.

Rapid turnover of pathogen-blocking Wolbachia and their incompatibility loci.

At least half of all insect species carry maternally inherited Wolbachia alphaproteobacteria, making Wolbachia the most common endosymbionts in nature. Wolbachia spread to high frequencies is often due to cytoplasmic incompatibility (CI), a Wolbachia-induced sperm modification that kills embryos without Wolbachia. Several CI-causing Wolbachia variants, including wMel from Drosophila melanogaster, also block viruses. Establishing pathogen-blocking wMel in natural Aedes aegypti mosquito populations has reduced dengue disease incidence, with one study reporting about 85% reduction when wMel frequency is high. However, wMel transinfection establishment is challenging in many environments, highlighting the importance of identifying CI-causing Wolbachia variants that stably persist in diverse hosts and habitats. We demonstrate that wMel-like variants have naturally established in widely distributed holometabolous dipteran and hymenopteran insects that diverged approximately 350 million years ago, with wMel variants spreading rapidly among these hosts over only the last 100,000 years. Wolbachia genomes contain prophages that encode CI-causing operons (cifs). These cifs move among Wolbachia genomes - with and without prophages - even more rapidly than Wolbachia move among insect hosts. Our results shed light on how rapid host switching and horizontal gene transfer contribute to Wolbachia and cif diversity in nature. The diverse wMel variants we report here from hosts present in different climates offer many new options for broadening Wolbachia-based biocontrol of diseases and pests.

Evolution of Wolbachia reproductive and nutritional mutualism: insights from the genomes of two novel strains that double infect the pollinator of dioecious Ficus hirta.

Wolbachia is a genus of maternally inherited endosymbionts that can affect reproduction of their hosts and influence metabolic processes. The pollinator, Valisia javana, is common in the male syconium of the dioecious fig Ficus hirta. Based on a high-quality chromosome-level V. javana genome with PacBio long-read and Illumina short-read sequencing, we discovered a sizeable proportion of Wolbachia sequences and used these to assemble two novel Wolbachia strains belonging to supergroup A. We explored its phylogenetic relationship with described Wolbachia strains based on MLST sequences and the possibility of induction of CI (cytoplasmic incompatibility) in this strain by examining the presence of cif genes known to be responsible for CI in other insects. We also identified mobile genetic elements including prophages and insertion sequences, genes related to biotin synthesis and metabolism. A total of two prophages and 256 insertion sequences were found. The prophage WOjav1 is cryptic (structure incomplete) and WOjav2 is relatively intact. IS5 is the dominant transposon family. At least three pairs of type I cif genes with three copies were found which may cause strong CI although this needs experimental verification; we also considered possible nutritional effects of the Wolbachia by identifying genes related to biotin production, absorption and metabolism. This study provides a resource for further studies of Wolbachia-pollinator-host plant interactions.

Egg-associated secretions from the brown planthopper (Nilaparvata lugens) activate rice immune responses.

The brown planthopper (BPH, Nilaparvata lugens) is a notorious sap-sucking insect pest that damages rice (Oryza sativa) plants throughout Asia. During BPH feeding, saliva enters rice plant tissues, whereas during oviposition egg-associated secretions (EAS) are deposited in damaged plant tissue. Dynamic changes in rice to planthopper salivary effectors have been widely reported. However, the effects of EAS from planthopper on rice immunity remains largely unexplored. In this study, we found that both infestation of rice by gravid BPH female adults and treatment with the EAS elicited a strong and rapid accumulation of jasmonic acid (JA), JA-isoleucine, and hydrogen peroxide in rice. EAS enhanced plant defenses not only in rice but also in tobacco, and these impaired the performance of BPH on rice, as well as the performance of aphids and whiteflies on tobacco. High-throughput proteome sequencing of EAS led to 110 proteins being identified and 53 proteins with 2 or more unique peptides being detected. Some proteins from BPH EAS were also found in the salivary proteome from herbivores, suggesting potential evolutionary conservation of effector functions across feeding and oviposition; however, others were only identified in EAS, and these are likely specifically related to oviposition. These findings point to novel proteins affecting interactions between planthoppers and rice during oviposition, providing an additional source of information for effector studies.

Heterogeneous patterns of heterozygosity loss in isolated populations of the threatened eastern barred bandicoot (Perameles gunnii).

Identifying and analysing isolated populations is critical for conservation. Isolation can make populations vulnerable to local extinction due to increased genetic drift and inbreeding, both of which should leave imprints of decreased genome-wide heterozygosity. While decreases in heterozygosity among populations are frequently investigated, fewer studies have analysed how heterozygosity varies among individuals, including whether heterozygosity varies geographically along lines of discrete population structure or with continuous patterns analogous to isolation by distance. Here we explore geographical patterns of differentiation and individual heterozygosity in the threatened eastern barred bandicoot (Perameles gunnii) in Tasmania, Australia, using genomic data from 85 samples collected between 2008 and 2011. Our analyses identified two isolated demes undergoing significant genetic drift, and several areas of fine-scale differentiation across Tasmania. We observed discrete genetic structures across geographical barriers and continuous patterns of isolation by distance, with little evidence of recent or historical migration. Using a recently developed analytical pipeline for estimating autosomal heterozygosity, we found individual heterozygosities varied within demes by up to a factor of two, and demes with low-heterozygosity individuals also still contained those with high heterozygosity. Spatial interpolation of heterozygosity scores clarified these patterns and identified the isolated Tasman Peninsula as a location where low-heterozygosity individuals were more common than elsewhere. Our results provide novel insights into the relationship between isolation-driven genetic structure and local heterozygosity patterns. These may help improve translocation efforts, by identifying populations in need of assistance, and by providing an individualised metric for identifying source animals for translocation.

How often are male mosquitoes attracted to humans?

Many mosquito species live close to humans where females feed on human blood. While male mosquitoes do not feed on blood, it has long been recognized that males of some species can be attracted to human hosts. To investigate the frequency of male mosquito attraction to humans, we conducted a literature review and human-baited field trials, as well as laboratory experiments involving males and females of three common Aedes species. Our literature review indicated that male attraction to humans is limited to a small number of species, including Ae. aegypti and Ae. albopictus. In our human-baited field collections, only 4 out of 13 species captured included males. In laboratory experiments, we found that male Ae. notoscriptus and Ae. vigilax showed no attraction to humans, while male Ae. aegypti exhibited persistent attraction for up to 30 min. Both male and female Ae. aegypti displayed similar preferences for different human subjects, suggesting that male Ae. aegypti respond to similar cues as females. Additionally, we found that mosquito repellents applied to human skin effectively repelled male mosquitoes. These findings shed light on mosquito behaviour and have implications for mosquito control programmes, particularly those involving the release or monitoring of the male mosquito population.

Evaluation of In2Care mosquito stations for suppression of the Australian backyard mosquito, Aedes notoscriptus (Diptera: Culicidae).

Aedes notoscriptus (Skuse) is a container-inhabiting mosquito endemic to Australia that vectors arboviruses and is suspected to transmit Mycobacterium ulcerans, the cause of Buruli ulcer. We evaluated the effectiveness of the In2Care station, which suppresses mosquito populations via the entomopathogenic fungus, Beauveria bassiana, and the insect growth regulator pyriproxyfen, the latter of which is autodisseminated among larval habitats by contaminated mosquitoes. A field trial was conducted using 110 In2Care stations in a 50,000 m2 area and results were compared to 4 control areas that did not receive the treatment. Efficacy was evaluated by comparing egg counts and measuring larvicidal impact in surrounding breeding sites. Laboratory experiments validated the effect of B. bassiana on adult survival. Results of this field trial indicate that, 6 wk after the In2Care stations were deployed, treatment site ovitraps contained 43% fewer eggs than control site ovitraps, and 33% fewer eggs after 10 wk, suggesting that the In2Care station was able to reduce the egg density of Ae. notoscriptus. Population reduction remained evident for up to 3 wk after In2Care stations were removed. Treatment site ovitraps had significantly fewer Ae. notoscriptus eclosing than control site ovitraps, confirming the pyriproxyfen autodissemination feature of the stations. An average reduction of 50% in adult eclosion was achieved. Exposure to B. bassiana resulted in four-times higher mortality among adult mosquitoes. Additionally, using fresh In2Care nettings led to an 88% decrease in average survival compared to 4-wk-old nettings. The use of In2Care stations has potential for suppressing Ae. notoscriptus egg density.

Testing evolutionary adaptation potential under climate change in invertebrates (mostly Drosophila): findings, limitations and directions.

A (quite) large set of experiments has been undertaken to assess the potential for evolutionary changes in invertebrates under current and future climate change conditions. These experimental studies have established some key principles that could affect climate change adaptation, yet there remain substantial obstacles in reaching a meaningful predictive framework. This Review starts with exploring some of the traits considered in individuals and approaches used in assessing evolutionary adaptation relevant to climate, and some of the core findings and their substantial limitations, with a focus on Drosophila. We interpret results in terms of adaptive limits based on population processes versus fundamental mechanistic limits of organisms. We then consider the challenges in moving towards a predictive framework and implications of the findings obtained to date, while also emphasizing the current limited context and the need to broaden it if links to changes in natural populations are to be realized.