Gene drives for invasive wasp control: Extinction is unlikely, with suppression dependent on dispersal and growth rates.

Gene drives offer a potentially revolutionary method for pest control over large spatial extents. These genetic modifications spread deleterious variants through a population and have been proposed as methods for pest suppression or even eradication. We examined the influence of local dispersal, long-distance and/or human-mediated dispersal, and variation in population growth on the success of a gene drive for the control of invasive social wasps (Vespula vulgaris). Our simulations incorporated a spatially realistic environment containing variable habitat quality in New Zealand. Pest eradication was not observed, except in extreme and unrealistic scenarios of constant, widespread, and spatially intense releases of genetically modified individuals every year for decades. Instead, the regional persistence of genetically modified and wild-type wasps was predicted. Simulations using spatially homogeneous versus realistic landscapes (incorporating uninhabitable areas and dispersal barriers) showed little difference in overall population dynamics. Overall, little impact on wasp abundance was observed in the first 15 years after introduction. After 25 years, populations were suppressed to levels <95% of starting populations. Populations exhibited "chase dynamics" with population cycles in space, with local extinction occurring in some areas while wasps became abundant in others. Increasing the wasps' local dispersal distance increased the spatial and temporal variability of the occupied area and population suppression. Varying levels of human-associated long-distance dispersal had little effect on population dynamics. Increasing intrinsic population growth rates interacted with local dispersal to cause higher mean populations and substantially higher levels of variation in population suppression and the total amount of landscape occupied. Gene drives appear unlikely to cause a rapid and widespread extinction of this and probably other pests but could offer long-term and cost-effective methods of pest suppression. The predicted level of <95% pest suppression would substantially reduce the predation pressure and competitive interactions of this invasive wasp on native species. However, the predicted long-term persistence of genetically modified pests will influence the ethics and likelihood of using gene drives for pest control, especially given concerns that modified wasps would eventually be transported back to their home range.

Can immune gene silencing via dsRNA feeding promote pathogenic viruses to control the globally invasive Argentine ant?

Pest control methods that can target pest species with limited environmental impacts are a conservation and economic priority. Species-specific pest control using RNA interference is a challenging but promising avenue in developing the next generation of pest management. We investigate the feasibility of manipulating a biological invader's immune system using double-stranded RNA (dsRNA) in order to increase susceptibility to naturally occurring pathogens. We used the invasive Argentine ant as a model, targeting the immunity-associated genes Spaetzle and Dicer-1 with dsRNA. We show that feeding with Spaetzle dsRNA can result in partial target gene silencing for up to 28 days in the laboratory and 5 days in the field. Dicer-1 dsRNA only resulted in partial gene knockdown after 2 days in the laboratory. Double-stranded RNA treatments were associated with significant gene expression disruptions across immune pathways in the laboratory and to a lower extent in the field. In total, 12 viruses and four bacteria were found in these ant populations. Some changes in viral loads in dsRNA-treated groups were observed. For example, Linepithema humile Polycipivirus 2 (LhuPCV2) loads increased after 2 days of treatment with Spaetzle and Dicer-1 dsRNA treatments in the laboratory. After treatment with the dsRNA in the field, after 5 days the virus Linepithema humile toti-like virus 1 (LhuTLV1) was significantly more abundant. However, immune pathway disruption did not result in a consistent increase in microbial infections, nor did it alter ant abundance in the field. Some viruses even declined in abundance after dsRNA treatment. Our study explored the feasibility of lowering a pest's immunity as a control tool. We demonstrate that it is possible to alter immune gene expression of pest species and pathogen loads, although in our specific system the affected pathogens did not appear to influence pest abundance. We provide suggestions on future directions for dsRNA-mediated immune disruption in pest species, including potential avenues to improve dsRNA delivery as well as the importance of pest and pathogen biology. Double-stranded RNA targeting immune function might be especially useful for pest control in systems in which viruses or other microorganisms are prevalent and have the potential to be pathogenic.

An invasive ant increases deformed wing virus loads in honey bees.

The majority of invasive species are best known for their effects as predators. However, many introduced predators may also be substantial reservoirs for pathogens. Honey bee-associated viruses are found in various arthropod species including invasive ants. We examined how the globally invasive Argentine ant (Linepithema humile), which can reach high densities and infest beehives, is associated with pathogen dynamics in honey bees. Viral loads of deformed wing virus (DWV), which has been linked to millions of beehive deaths around the globe, and black queen cell virus significantly increased in bees when invasive ants were present. Microsporidian and trypanosomatid infections, which are more bee-specific, were not affected by ant invasion. The bee virome in autumn revealed that DWV was the predominant virus with the highest infection levels and that no ant-associated viruses were infecting bees. Viral spillback from ants could increase infections in bees. In addition, ant attacks could pose a significant stressor to bee colonies that may affect virus susceptibility. These viral dynamics are a hidden effect of ant pests, which could have a significant impact on disease emergence in this economically important pollinator. Our study highlights a perhaps overlooked effect of species invasions: changes in pathogen dynamics.

A global review of socioeconomic and environmental impacts of ants reveals new insights for risk assessment.

Risk assessments are fundamental to invasive species management and are underpinned by comprehensive characterization of invasive species impacts. Our understanding of the impacts of invasive species is growing constantly, and several recently developed frameworks offer the opportunity to systematically categorize environmental and socioeconomic impacts of invasive species. Invasive ants are among the most widespread and damaging invaders. Although a handful of species receives most of the policy attention, nearly 200 species have established outside their native range. Here, we provide a global, comprehensive assessment of the impacts of ants and propose a priority list of risk species. We used the Socioeconomic Impact Classification for Alien Taxa (SEICAT), Environmental Impact Classification for Alien Taxa (EICAT) and Generic Impact Scoring System (GISS) to analyze 642 unique sources for 100 named species. Different methodologies provided generally consistent results. The most frequently identified socioeconomic impacts were to human health. Environmental impacts were primarily on animal and plant populations, with the most common mechanisms being predation and competition. Species recognized as harmful nearly 20 years ago featured prominently, including Wasmannia auropunctata (little fire ant, electric ant), Solenopsis invicta (red imported fire ant), Anoplolepis gracilipes (yellow crazy ant), and Pheidole megacephala (African big-headed ant). All these species except W. auropunctata have been implicated in local extinctions of native species. Although our assessments affirmed that the most serious impacts have been driven by a small number of species, our results also highlighted a substantial number of less well publicized species that have had major environmental impacts and may currently be overlooked when prioritizing prevention efforts. Several of these species were ranked as high or higher than some of the previously recognized "usual suspects," most notably Nylanderia fulva (tawny crazy ant). We compared and combined our assessments with trait-based profiles and other lists to propose a consensus set of 31 priority species. Ever-increasing global trade contributes to growing rates of species introductions. The integrated approaches we used can contribute to robust, holistic risk assessments for many taxa entrained in these pathways.

Invasion Success and Management Strategies for Social Vespula Wasps.

Three species of Vespula have become invasive in Australia, Hawai'i, New Zealand, and North and South America and continue to spread. These social wasp species can achieve high nest densities, and their behavioral plasticity has led to substantial impacts on recipient communities. Ecologically, they affect all trophic levels, restructuring communities and altering resource flows. Economically, their main negative effect is associated with pollination and the apicultural industry. Climate change is likely to exacerbate their impacts in many regions. Introduced Vespula spp. likely experience some degree of enemy release from predators or parasites, although they are exposed to a wide range of microbial pathogens in both their native and introduced range. Toxic baits have been significantly improved over the last decade, enabling effective landscape-level control. Although investigated extensively, no effective biological control agents have yet been found. Emerging technologies such as gene drives are under consideration.

Pathogen shifts in a honeybee predator following the arrival of the Varroa mite.

Emerging infectious diseases (EIDs) are a global threat to honeybees, and spillover from managed bees threaten wider insect populations. Deformed wing virus (DWV), a widespread virus that has become emergent in conjunction with the spread of the mite Varroa destructor, is thought to be partly responsible for global colony losses. The arrival of Varroa in honeybee populations causes a dramatic loss of viral genotypic diversity, favouring a few virulent strains. Here, we investigate DWV spillover in an invasive Hawaiian population of the wasp, Vespula pensylvanica, a honeybee predator and honey-raider. We show that Vespula underwent a parallel loss in DWV variant diversity upon the arrival of Varroa, despite the mite being a honeybee specialist. The observed shift in Vespula DWV and the variant-sharing between Vespula and Apis suggest that these wasps can acquire DWV directly or indirectly from honeybees. Apis prey items collected from Vespula foragers were positive for DWV, indicating predation is a possible route of transmission. We also sought cascading effects of DWV shifts in a broader Vespula pathogen community. We identified concurrent changes in a suite of additional pathogens, as well as shifts in the associations between these pathogens in Vespula. These findings reveal how hidden effects of the Varroa mite can, via spillover, transform the composition of pathogens in interacting species, with potential knock-on effects for entire pathogen communities.

The association between mitochondrial genetic variation and reduced colony fitness in an invasive wasp.

Despite the mitochondrion's long-recognized role in energy production, mitochondrial DNA (mtDNA) variation commonly found in natural populations was assumed to be effectively neutral. However, variation in mtDNA has now been increasingly linked to phenotypic variation in life history traits and fitness. We examined whether the relative fitness in native and invasive common wasp (Vespula vulgaris) populations in Belgium and New Zealand (NZ), respectively, can be linked to mtDNA variation. Social wasp colonies in NZ were smaller with comparatively fewer queen cells, indicating a reduced relative fitness in the invaded range. Interestingly, queen cells in this population were significantly larger leading to larger queen offspring. By sequencing 1,872 bp of the mitochondrial genome, we determined mitochondrial haplotypes and detected reduced genetic diversity in NZ. Three common haplotypes in NZ frequently produced many queens, whereas the four rare haplotypes produced significantly fewer or no queens. The entire mitochondrial genome for each of these haplotypes was sequenced to identify polymorphisms associated with fitness reduction. We found 16 variable sites; however, no nonsynonymous mutation that was clearly causing impaired mitochondrial function was detected. We discuss how detected variants may alter secondary structures, gene expression or mito-nuclear interactions, or could be associated with nuclear-encoded variation. Whatever the ultimate mechanism, we show reduced fitness and mtDNA variation in an invasive wasp population as well as specific mtDNA variants associated with fitness variation within this population. Ours is one of only a few studies that confirm fitness impacts of mtDNA variation in wild nonmodel populations.

Behavioural variation and plasticity along an invasive ant introduction pathway.

Once established in new areas, introduced species may exhibit changes in their biology due to phenotypic plasticity, novel selection pressures and genetic drift. Moreover, the introduction process itself has been hypothesised to act as a selective filter for traits that promote invasiveness. We tested the hypothesis that behaviours thought to promote invasiveness-such as increased foraging activity and aggression-are selected for during invasion by comparing traits among native and introduced populations of the widespread Argentine ant (Linepithema humile). We studied Argentine ant populations in the native range in Argentina and in three invaded regions along an introduction pathway: California, Australia and New Zealand. In each region, we set up 32 experimental colonies to measure foraging activity and interspecific aggression in a subset of the study regions. These colonies were subject to experimental manipulation of carbohydrate availability and octopamine, a biogenic amine known to modulate behaviour in insects, to measure variation in behavioural plasticity. We found variation in foraging activity among populations, but this variation was not consistent with selection on behaviour in relation to the invasion process. We found that colonies with limited access to carbohydrates exhibited unchanged exploratory behaviour, but higher exploitation activity and lower aggression. Colonies given octopamine consistently increased foraging behaviour (both exploration and exploitation), as well as aggression when also sugar-deprived. There was no difference in the degree of behavioural response to our experimental treatments along the introduction pathway. We did not find support for selection of behavioural traits associated with invasiveness along the Argentine ant's introduction pathway or clear evidence for an association between the introduction process and variation in behavioural plasticity. These results indicate that mechanisms promote behavioural variation in a similar fashion both in native and introduced ranges. Our results challenge the assumption that introduced populations always perform better in key behavioural traits hypothesised to be associated with invasion success.

Symbiotic bacterial communities in ants are modified by invasion pathway bottlenecks and alter host behavior.

Biological invasions are a threat to global biodiversity and provide unique opportunities to study ecological processes. Population bottlenecks are a common feature of biological invasions and the severity of these bottlenecks is likely to be compounded as an invasive species spreads from initial invasion sites to additional locations. Despite extensive work on the genetic consequences of bottlenecks, we know little about how they influence microbial communities of the invaders themselves. Due to serial bottlenecks, invasive species may lose microbial symbionts including pathogenic taxa (the enemy release hypothesis) and/or may accumulate natural enemies with increasing time after invasion (the pathogen accumulation and invasive decline hypothesis). We tested these alternate hypotheses by surveying bacterial communities of Argentine ants (Linepithema humile). We found evidence for serial symbiont bottlenecks: the bacterial community richness declined over the invasion pathway from Argentina to New Zealand. The abundance of some genera, such as Lactobacillus, also significantly declined over the invasion pathway. Argentine ants from populations in the United States shared the most genera with ants from their native range in Argentina, while New Zealand shared the least (120 vs. 57, respectively). Nine genera were present in all sites around the globe possibly indicating a core group of obligate microbes. In accordance with the pathogen accumulation and invasive decline hypothesis, Argentine ants acquired genera unique to each specific invaded country. The United States had the most unique genera, though even within New Zealand these ants acquired symbionts. In addition to our biogeographic sampling, we administered antibiotics to Argentine ants to determine if changes in the micro-symbiont community could influence behavior and survival in interspecific interactions. Treatment with the antibiotics spectinomycin and kanamycin only slightly increased Argentine ant interspecific aggression, but this increase significantly decreased survival in interspecific interactions. The survival of the native ant species also decreased when the symbiotic microbial community within Argentine ants was modified by antibiotics. Our work offers support for both the enemy release hypothesis and that invasive species accumulate novel microbial taxa within their invaded range. These changes appear likely to influence invader behavior and survival.

The long-term population dynamics of common wasps in their native and invaded range.

Populations of introduced species are often thought to perform differently, or experience different population dynamics, in their introduced range compared to their native habitat. Differences between habitats in climate, competition or natural enemies may result in populations with varying density dependence and population dynamics. We examined the long-term population dynamics of the invasive common wasp, Vespula vulgaris, in its native range in England and its invaded range in New Zealand. We used 39 years of wasp density data from four sites in England, and 23 years of data from six sites in New Zealand. Wasp population time series was examined using partial rate correlation functions. Gompertz population models and multivariate autoregressive state-space (MARSS) models were fitted, incorporating climatic variation. Gompertz models successfully explained 59-66% of the variation in wasp abundance between years. Density dependence in wasp populations appeared to act similarly in both the native and invaded range, with wasp abundance in the previous year as the most important variable in predicting intrinsic rate of increase (r). No evidence of cyclic population dynamics was observed. Both the Gompertz and MARSS models highlighted the role of weather conditions in each country as significant predictors of annual wasp abundance. The temporal evolution of wasp populations at all sites was best modelled jointly using a single latent dynamic factor for local trends, with the inclusion of a latent spring weather covariate. That same parsimonious multivariate model structure was optimal in both the native and invaded range. Density dependence is overwhelmingly important in predicting wasp densities and 'wasp years' in both the native and invaded range. Spring weather conditions in both countries have a major influence, probably through their impact on wasp colony initiation and early development. The population dynamics in the native range and invaded range show no evidence of cyclic boom-and-bust dynamics. Invasive species may not exhibit different population dynamics despite considerable variation in abundances throughout their distribution.

Changes in the Bacteriome of Honey Bees Associated with the Parasite Varroa destructor, and Pathogens Nosema and Lotmaria passim.

The honey bee, Apis mellifera, is a globally important species that suffers from a variety of pathogens and parasites. These parasites and pathogens may have sublethal effects on their bee hosts via an array of mechanisms, including through a change in symbiotic bacterial taxa. Our aim was to assess the influence of four globally widespread parasites and pathogens on the honey bee bacteriome. We examined the effects of the ectoparasitic mite Varroa destructor, the fungal pathogens Nosema apis and Nosema ceranae, and the trypanosome Lotmaria passim. Varroa was detected by acaricidal treatment, Nosema and L. passim by PCR, and the bacteriome using MiSeq 16S rRNA gene sequencing. Overall, the 1,858,850 obtained sequences formed 86 operational taxonomic units (OTUs) at 3 % dissimilarity. Location, time of year, and degree of infestation by Varroa had significant effects on the composition of the bacteriome of honey bee workers. Based on statistical correlations, we found varroosis more important factor than N. ceranae, N. apis, and L. passim infestation influencing the honey bee bacteriome and contributing to the changes in the composition of the bacterial community in adult bees. At the population level, Varroa appeared to modify 20 OTUs. In the colonies with high Varroa infestation levels (varroosis), the relative abundance of the bacteria Bartonella apis and Lactobacillus apis decreased. In contrast, an increase in relative abundance was observed for several taxa including Lactobacillus helsingborgensis, Lactobacillus mellis, Commensalibacter intestini, and Snodgrassella alvi. The results showed that the "normal" bacterial community is altered by eukaryotic parasites as well as displaying temporal changes and changes associated with the geographical origin of the beehive.

Corruption, development and governance indicators predict invasive species risk from trade.

Invasive species have an enormous global impact, with international trade being the leading pathway for their introduction. Current multinational trade deals under negotiation will dramatically change trading partnerships and pathways. These changes have considerable potential to influence biological invasions and global biodiversity. Using a database of 47 328 interceptions spanning 10 years, we demonstrate how development and governance socio-economic indicators of trading partners can predict exotic species interceptions. For import pathways associated with vegetable material, a significantly higher risk of exotic species interceptions was associated with countries that are poorly regulated, have more forest cover and have surprisingly low corruption. Corruption and indicators such as political stability or adherence to rule of law were important in vehicle or timber import pathways. These results will be of considerable value to policy makers, primarily by shifting quarantine procedures to focus on countries of high risk based on their socio-economic status. Further, using New Zealand as an example, we demonstrate how a ninefold reduction in incursions could be achieved if socio-economic indicators were used to select trade partners. International trade deals that ignore governance and development indicators may facilitate introductions and biodiversity loss. Development and governance within countries clearly have biodiversity implications beyond borders.

Invasive ants carry novel viruses in their new range and form reservoirs for a honeybee pathogen.

When exotic animal species invade new environments they also bring an often unknown microbial diversity, including pathogens. We describe a novel and widely distributed virus in one of the most globally widespread, abundant and damaging invasive ants (Argentine ants, Linepithema humile). The Linepithema humile virus 1 is a dicistrovirus, a viral family including species known to cause widespread arthropod disease. It was detected in samples from Argentina, Australia and New Zealand. Argentine ants in New Zealand were also infected with a strain of Deformed wing virus common to local hymenopteran species, which is a major pathogen widely associated with honeybee mortality. Evidence for active replication of viral RNA was apparent for both viruses. Our results suggest co-introduction and exchange of pathogens within local hymenopteran communities. These viral species may contribute to the collapse of Argentine ant populations and offer new options for the control of a globally widespread invader.

Relative roles of climatic suitability and anthropogenic influence in determining the pattern of spread in a global invader.

Because invasive species threaten the integrity of natural ecosystems, a major goal in ecology is to develop predictive models to determine which species may become widespread and where they may invade. Indeed, considerable progress has been made in understanding the factors that influence the local pattern of spread for specific invaders and the factors that are correlated with the number of introduced species that have become established in a given region. However, few studies have examined the relative importance of multiple drivers of invasion success for widespread species at global scales. Here, we use a dataset of >5,000 presence/absence records to examine the interplay between climatic suitability, biotic resistance by native taxa, human-aided dispersal, and human modification of habitats, in shaping the distribution of one of the world's most notorious invasive species, the Argentine ant (Linepithema humile). Climatic suitability and the extent of human modification of habitats are primarily responsible for the distribution of this global invader. However, we also found some evidence for biotic resistance by native communities. Somewhat surprisingly, and despite the often cited importance of propagule pressure as a crucial driver of invasions, metrics of the magnitude of international traded commodities among countries were not related to global distribution patterns. Together, our analyses on the global-scale distribution of this invasive species provide strong evidence for the interplay of biotic and abiotic determinants of spread and also highlight the challenges of limiting the spread and subsequent impact of highly invasive species.

Beekeepers Support the Use of RNA Interference (RNAi) to Control Varroa destructor.

Current Varroa mite management strategies rely heavily on the use of pesticides, adversely affecting honey bee health and leaving toxic residues in hive products. To explore the likelihood of RNAi technology being utilised as an alternative control method for pests like Varroa, the opinions of beekeepers on the use of this new biotechnology were obtained using a mixed-methodology approach. In-person surveys and focus groups using the Q method were conducted to discover the willingness of beekeepers to utilise Varroa-targeting RNAi treatments in their hives, and to gain feedback to inform decisions before the implementation of this new technology. Overall, the beekeepers saw potential in RNAi being used to control Varroa in their hives and were eager to have access to an alternative to pesticide treatments. Participants raised concerns about unknown long-term effects on bees and other non-target species, and the potential of an uninformed public preventing them from accessing a new Varroa treatment. While further research and discussion is needed before RNAi treatments for Varroa become commercially available, RNAi technology presents a promising, species-specific and non-toxic solution for Varroa management.

RNA interference as a next-generation control method for suppressing Varroa destructor reproduction in honey bee (Apis mellifera) hives.

BACKGROUND: The Varroa mite (Varroa destructor) is considered to be the greatest threat to apiculture worldwide. RNA interference (RNAi) using double-stranded RNA (dsRNA) as a gene silencing mechanism has emerged as a next-generation strategy for mite control. RESULTS: We explored the impact of a dsRNA biopesticide, named vadescana, designed to silence the calmodulin gene in Varroa, on mite fitness in mini-hives housed in a laboratory. Two dosages were tested: 2 g/L dsRNA and 8 g/L dsRNA. Vadescana appeared to have no effect on mite survival, however, mite fertility was substantially reduced. The majority of foundress mites exposed to vadescana failed to produce any offspring. No dose-dependent effect of vadescana was observed, as both the low and high doses inhibited mite reproduction equally well in the mini-hives and neither dose impacted pupal survival of the honey bee. Approximately 95% of bee pupae were alive at uncapping across all treatment groups. CONCLUSION: These findings suggest that vadescana has significant potential as an effective alternative to conventional methods for Varroa control, with broader implications for the utilization of RNAi as a next-generation tool in the management of pest species. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A neurotoxic pesticide changes the outcome of aggressive interactions between native and invasive ants.

Neurotoxic pesticides, such as neonicotinoids, negatively affect the cognitive capacity and fitness of non-target species, and could also modify interspecific interactions. We tested whether sublethal contamination with neonicotinoid could affect foraging, colony fitness and the outcome of behavioural interactions between a native (Monomorium antarcticum) and an invasive ant species (Linepithema humile). The foraging behaviour of both ants was not affected by neonicotinoid exposure. Colonies of the invasive species exposed to the neonicotinoid produced significantly fewer brood. In interspecific confrontations, individuals of the native species exposed to the neonicotinoid lowered their aggression towards the invasive species, although their survival probability was not affected. Exposed individuals of the invasive species interacting with non-exposed native ants displayed increased aggression and had their survival probability reduced. Non-exposed individuals of the invasive species were less aggressive but more likely to survive when interacting with exposed native ants. These results suggest that non-target exposure of invaders to neonicotinoids could either increase or decrease the probability of survival according to the exposure status of the native species. Given that, in any community, different species have different food preferences, and thus different exposure to pesticides, non-target exposure could potentially change the dynamics of communities and influence invasion success.

Feeling the heat? Substantial variation in temperatures does not affect the proportion of males born in Australia.

The global proportion of male births has been shown to vary with climate, with a higher proportion of male births documented in colder climates. Here we examined the hypothesis that ambient temperature predicts fluctuations in the proportion of male births in Australia and within seven Australian states using historical annual data spanning 1910-2009. We predicted that within states with tropical ambient temperatures the proportion of male births would decrease when ambient temperatures are higher. First, considering the national composite births for the whole of Australia, the proportion of males born ranged only from 0.510 to 0.517. We observed no relationship between the proportion of male births and the mean annual temperature in the concurrent or previous years for Australia as a whole. Second, the analyses for the seven Australian regions provided a total of 14 estimated relationships. We found no association between ambient temperature in the previous or concurrent year and the proportion of males born for all 14 comparisons. Thus, we found no evidence of a negative relationship between ambient temperatures in the concurrent or previous year and the proportion of males born in the tropical regions of Australia, and no evidence of any relationship in the more temperate Australian regions or in Australia as a whole.

Effects of Deformed Wing Virus-Targeting dsRNA on Viral Loads in Bees Parasitised and Non-Parasitised by Varroa destructor.

The Varroa destructor mite is a devastating parasite of honey bees; however the negative effects of varroa parasitism are exacerbated by its role as an efficient vector of the honey bee pathogen, Deformed wing virus (DWV). While no direct treatment for DWV infection is available for beekeepers to use on their hives, RNA interference (RNAi) has been widely explored as a possible biopesticide approach for a range of pests and pathogens. This study tested the effectiveness of three DWV-specific dsRNA sequences to lower DWV loads and symptoms in honey bees reared from larvae in laboratory mini-hives containing bees and varroa. The effects of DWV-dsRNA treatment on bees parasitised and non-parasitised by varroa mites during development were investigated. Additionally, the impact of DWV-dsRNA on viral loads and gene expression in brood-parasitising mites was assessed using RNA-sequencing. Bees parasitised during development had significantly higher DWV levels compared to non-parasitised bees. However, DWV-dsRNA did not significantly reduce DWV loads or symptoms in mini-hive reared bees, possibly due to sequence divergence between the DWV variants present in bees and varroa and the specific DWV-dsRNA sequences used. Varroa mites from DWV-dsRNA treated mini-hives did not show evidence of an elevated RNAi response or significant difference in DWV levels. Overall, our findings show that RNAi is not always successful, and multiple factors including pathogen diversity and transmission route may impact its efficiency.

Gene silencing for invasive paper wasp management: Synthesized dsRNA can modify gene expression but did not affect mortality.

Invasive paper wasps such as Polistes dominula are a major pest and problem for biodiversity around the globe. Safe and highly targeted methods for the control of these and other social wasp populations are needed. We attempted to identify potentially-lethal gene targets that could be used on adult paper wasps in a gene silencing or RNA interference (RNAi) approach. Double-stranded RNA (dsRNA) was designed to target genes for which silencing has proven lethal in other insects. dsRNA was provided either orally to foragers or directly injected into the wasps. We also provided the dsRNA unprotected or protected from degradation by gut nucleases in two different forms (lipofectamine and carbon quantum dots). The effects of oral delivery of 22 different gene targets to forager wasps was evaluated. The expression of five different genes was successfully reduced following dsRNA ingestion or injection. These gene targets included the FACT complex subunit spt16 (DRE4) and RNA-binding protein fusilli (FUSILLI), both of which have been previously shown to have potential as lethal targets for pest control in other insects. However, we found no evidence of significant increases in adult wasp mortality following ingestion or injection of dsRNA for these genes when compared with control treatments in our experiments. The methods we used to protect the dsRNA from digestive degradation altered gene expression but similarly did not influence wasp mortality. Our results indicate that while many of the same gene targets can be silenced and induce mortality in other insects, dsRNA and RNAi approaches may not be useful for paper wasp control.