Suppressing tawny crazy ant (Nylanderia fulva) by RNAi technology

A bstract The tawny crazy ant(Nylanderia fulva)is a new invasive pest in the United States.At present,its management mainly relies on the use of synthetic insecticides,which are generally ineffective at producing lasting control of the pest,necessitating alternative environmentally friendly measures.In this study,we evaluated the feasibility of gene silencing to control this ant species.Six housekeeping genes encoding actin(NfActin),coatomer subunit β (NfCOPP),arginine kinase(NfArgK),and V-type proton ATPase subunits A(NfvATPaseA),B(NfvATPaseB)and E(NfvATPaseE)were cloned.Phylogenetic analysis revealed high sequence similarity to homologs from other ant species,particularly the Florida carpenter ant(Camponotus floridanus).To silence these genes,vector L4440 was used to generate six specific RNAi constructs for bacterial expression.Heat-inactivated,dsRNA-expressing Escherichia coli were incorporated into artificial diet.Worker ants exhibited reduced endogenous gene expression after feeding on such diet for 9 d.However,only ingestion of dsRNAs of NfCOPfi(a gene involved in protein trafficking)and NfArgK(a cellular energy reserve regulatory gene in invertebrates)caused modest but significantly higher ant mortality than the control.These results suggest that bacterially expressed dsRNA can be orally delivered to ant cells as a mean to target its vulnerabilities.Improved efficacy is necessary for the RNAi-based approach to be useful in tawny crazy ant management.     

Fecundity and longevity of Argentine ant (Hymenoptera: Formicidae) queens in response to irradiation

Irradiation is a post‐harvest quarantine treatment option to control ants and other hitchhiker pests on fresh horticultural products traded between countries. As little is known about irradiation effects on ants, radiotolerance of the Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae: Dolichoderinae), was studied to determine a dose sufficient for its control. Queens collected from Buenos Aires, Argentina, were irradiated with 30, 60, 90 Gy or left untreated as controls, and then followed for 8 weeks to evaluate their survival and fecundity. Overall queen survival and brood viability decreased with increasing irradiation dose. The number of eggs was reduced by 50%, 69% and 56% in the 30, 60 and 90 Gy doses, respectively, compared with untreated control queens. The percentage of eggs that developed into larvae decreased from 41.1% in the control to 22.5%, 1.4%, and 0% in the 30, 60, and 90 Gy treatments, respectively. Thus, the number of larvae was reduced by 69% in the 30 Gy treatment compared with the control, only one larva was observed in the 60 Gy treatment, and none in the 90 Gy treatment. Only one pupa was observed in the 30 Gy treatment and none in the 60 and 90 Gy treatments during the 8‐week experiment. Queens irradiated with 60 and 90 Gy had significantly reduced longevity compared with queens treated with lower doses or untreated queens. Radiation dose ≥90 Gy stopped brood development in Argentine ant queens and should be sufficient as a phytosanitary treatment. The radiotolerance of Argentine ant appears to be similar to that of two other important invasive ants.     

Immediate impacts of invasion by Wasmannia auropunctata (Hymenoptera: Formicidae) on native litter ant fauna in a New Caledonian rainforest

Abstract For the last 30 years, Wasmannia auropunctata (the little fire ant) has spread throughout the Pacific and represents a severe threat to fragile island habitats. This invader has often been described as a disturbance specialist. Here we present data on its spread in a dense native rainforest in New Caledonia. We monitored by pitfall trapping the litter ant fauna along an invasive gradient from the edge to the inner forest in July 1999 and March 2000. When W. auropunctata was present, the abundance and richness of native ants drops dramatically. In invaded plots, W. auropunctata represented more than 92% of all trapped ant fauna. Among the 23 native species described, only four cryptic species survived. Wasmannia auropunctata appears to be a highly competitive ant that dominates the litter by eliminating native ants. Mechanisms involved in this invasive success may include predation as well as competitive interactions (exploitation and interference). The invasive success of W. auropunctata is similar to that of other tramp ants and reinforces the idea of common evolutionary traits leading to higher competitiveness in a new environment.     

Evidence of ecological niche shift in Rhododendron ponticum (L.) in Britain: Hybridization as a possible cause of rapid niche expansion

Biological invasions threaten global biodiversity and natural resources. Anticipating future invasions is central to strategies for combating the spread of invasive species. Ecological niche models are thus increasingly used to predict potential distribution of invasive species. In this study, we compare ecological niches of Rhododendron ponticum in its native (Iberian Peninsula) and invasive (Britain) ranges. Here, we test the conservation of ecological niche between invasive and native populations of R. ponticum using principal component analysis, niche dynamics analysis, and MaxEnt‐based reciprocal niche modeling. We show that niche overlap between native and invasive populations is very low, leading us to the conclusion that the two niches are not equivalent and are dissimilar. We conclude that R. ponticum occupies novel environmental conditions in Britain. However, the evidence of niche shift presented in this study should be treated with caution because of nonanalogue climatic conditions between native and invasive ranges and a small population size in the native range. We then frame our results in the context of contradicting genetic evidence on possible hybridization of this invasive species in Britain. We argue that the existing contradictory studies on whether hybridization caused niche shift in R. ponticum are not sufficient to prove or disprove this hypothesis. However, we present a series of theoretical arguments which indicate that hybridization is a likely cause of the observed niche expansion of R. ponticum in Britain.     

Transmission of Vairimorpha invictae (Microsporidia: Burenellidae) infections between red imported fire ant (Hymenoptera: Formicidae) colonies

Red imported fire ant, Solenopsis invicta, colonies were successfully infected with the microsporidium Vairimorpha invictae by introducing live larvae, pupae, or dead adults from V. invictae-infected field colonies collected in Argentina. Introductions with 4th instar larvae or non-melanized pupae obtained from infected field colonies, resulted in infection of 40% of the inoculated colonies. Introductions of 4th instars or melanized pupae produced from colonies that were initially infected in the laboratory, resulted in infections of 83% of the colonies, thus perpetuating the infection in other colonies. Infection was detected in 2 of 6 colonies after introducing adult worker caste ants that had died with V. invictae. The average number of adults and the volume of immature ants per colony were significantly lower in the infected than in the control colonies. Infected colonies had 86% fewer adults per colony and 82% less immature ants than the controls. A portion of the 16S rRNA gene of the V. invictae identified from these studies was amplified, cloned, and sequenced; the 1251 nucleotide amplicon was 100% identical to the 16S rRNA gene sequence recorded previously in the GenBank database, thus verifying the species as V. invictae. This is the first report of the artificial transmission of this pathogen to uninfected ant colonies, and demonstration of its ability to hinder growth in individual colonies.     

Molecular characterization,virulence and horizontal transmission of Beauveria pseudobassiana from Dendroctonus micans (Kug.) (Coleoptera: Curculionidae)

The great spruce bark beetle, Dendroctonus micans (Kugelann) (Coleoptera: Curculionidae), has been a potential threat for Turkey and the entire Eurasian spruce forests for many years. Control strategies which have been applied so far are still insufficient to prevent its damage. A previous study has shown that a Beauveria isolate (ARSEF 9271) proved to be an efficient microbial control agent against the great spruce bark beetle. In this study, this isolate was identified as B. pseudobassiana based on the partial sequence of EF1‐α and ITS sequence. A conidial suspension (1 × 10⁸/ml) of this fungus caused 100% mortality on both larvae and adults of D. micans within 5 and 6 days, respectively. Also, it caused 100% mycosis value on both larvae and adults. Mortality values of horizontal transmission experiments between larvae and adults which were contaminated with 1 × 10⁶/ml spore suspension at 25%, 50%, 75% and 100% rates were determined as 100% after 15 days at 20°C under the laboratory conditions. We also determined the decrease of the damage in spruce wood block (15 × 25 cm) when the contamination rate of the larvae was increased. Our results indicate that B. pseudobassiana ARSEF 9271 seems to be a very promising biocontrol agent against D. micans.     

Laboratory and field evaluation of entomopathogenic nematodes for control of Agriotes obscurus (L.) (Coleoptera: Elateridae)

The susceptibility of the dusky wireworm, Agriotes obscurus (L.) (Coleoptera: Elateridae), to different species and strains of entomopathogenic nematodes was tested in a virulence assay in the laboratory. Larvae were exposed to different nematode doses of 50 and 100 IJs/cm². At a dose of 50 IJs/cm², only a commercial strain Heterorhabditis bacteriophora Poinar and the native strain Steinernema carpocapsae (Weiser) B14 caused increased mortality compared with the control (11.1% and 13.3% mortality, respectively). At the higher dose tested, all strains (except Steinernema sp. D122) were virulent to A. obscurus larvae. Steinernema carpocapsae B14 caused higher mortality of wireworm (75.6%) and was used for the assay conducted in cages, with a dose of 100 IJs/cm², in field conditions. The results showed that S. carpocapsae B14 controlled 48.3% of A. obscurus larvae, demonstrating that some entomopathogenic nematodes have the potential to control larvae of A. obscurus. However, further work is needed to improve their efficacy.     

New records of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) predation by Brazilian Hemipteran predatory bugs

The tomato borer Tuta absoluta, native to western South America, is an extremely devastating pest in tomato crops in most of South America, Europe and Africa North of the Sahel, causes yield losses up to 100% and decreases fruit quality in open field and greenhouse crops if control methods are not applied. In Brazil two other important lepidopteran pests – Neoleucinodes elegantalis and Helicoverpa zea – occur in tomato, as well as thrips, whiteflies and aphids. For control of these pests, frequent applications of pesticides of up to 5 times per week are needed, and these resulted in the appearance of resistant populations to a number of active ingredients and decimation of natural enemies. Biological control may offer a better, safer and more sustainable opportunity for pest management. Mirid predatory bugs are currently used with success in southern Europe to control T. absoluta and other pests. In Brazil, four Hemipteran predatory bugs, not yet known to attack T. absoluta, were found to successfully prey on eggs and larvae of this pest. The first results on their predation capacity, development, survival and reproduction on T. absoluta on tomato plants are presented.     

Evidence for a recent horizontal transmission and spatial spread of Wolbachia from endemic Rhagoletis cerasi (Diptera: Tephritidae) to invasive Rhagoletis cingulata in Europe

The widespread occurrence of Wolbachia in arthropods and nematodes suggests that this intracellular, maternally inherited endosymbiont has the ability to cross species boundaries. However, direct evidence for such a horizontal transmission of Wolbachia in nature is scarce. Here, we compare the well‐characterized Wolbachia infection of the European cherry fruit fly, Rhagoletis cerasi, with that of the North American eastern cherry fruit fly, Rhagoletis cingulata, recently introduced to Europe. Molecular genetic analysis of Wolbachia based on multilocus sequence typing and the Wolbachia surface protein wsp showed that all R. cingulata individuals are infected with wCin2 identical to wCer2 in R. cerasi. In contrast, wCin1, a strain identical to wCer1 in R. cerasi, was present in several European populations of R. cingulata, but not in any individual from the United States. Surveys of R. cingulata from Germany and Hungary indicated that in some populations, the frequency of wCin1 increased significantly in just a few years with at least two independent horizontal transmission events. This is corroborated by the analysis of the mitochondrial cytochrome oxidase II gene that showed association of wCin1 with two distinct haplotypes in Germany, one of which is also infected with wCin1 in Hungary. In summary, our study provides strong evidence for a very recent inter‐specific Wolbachia transmission with a subsequent spatial spread in field populations.     

First Philippine record of the parasitoid,Comperiella sp. (Hymenoptera: Encyrtidae): a potential biocontrol agent against Aspidiotus rigidus (Hemiptera: Diaspididae)

The encyrtid genus Comperiella Howard has so far not been reported in the Philippines, where there is currently an outbreak of the coconut scale insect Aspidiotus rigidus Reyne particularly in the southern parts of the island of Luzon and in some areas in Mindanao. Among Comperiella species, only C. unifasciata Ishii has been reported as a parasitoid of A. rigidus. We report not only new sightings of this parasitoid genus in the Philippines from surveys conducted in parts of the provinces of Laguna and Batangas, but also the discovery of a possibly new species that, like C. unifasciata, has been found to parasitize A. rigidus at a high rate. These findings have presented a potential of biological control against the coconut scale insect problem that has threatened the coconut industry in the country.     

Evolutionary history of the little fire ant Wasmannia auropunctata before global invasion: inferring dispersal patterns,niche requirements and past and present distribution within its native range

The evolutionary history of invasive species within their native range may involve key processes that allow them to colonize new habitats. Therefore, phylogeographic studies of invasive species within their native ranges are useful to understand invasion biology in an evolutionary context. Here we integrated classical and Bayesian phylogeographic methods using mitochondrial and nuclear DNA markers with a palaeodistribution modelling approach, to infer the phylogeographic history of the invasive ant Wasmannia auropunctata across its native distribution in South America. We discuss our results in the context of the recent establishment of this mostly tropical species in the Mediterranean region. Our Bayesian phylogeographic analysis suggests that the common ancestor of the two main clades of W. auropunctata occurred in central Brazil during the Pliocene. Clade A would have differentiated northward and clade B southward, followed by a secondary contact beginning about 380 000 years ago in central South America. There were differences in the most suitable habitats among clades when considering three distinct climatic periods, suggesting that genetic differentiation was accompanied by changes in niche requirements, clade A being a tropical lineage and clade B a subtropical and temperate lineage. Only clade B reached more southern latitudes, with a colder climate than that of northern South America. This is concordant with the adaptation of this originally tropical ant species to temperate climates prior to its successful establishment in the Mediterranean region. This study highlights the usefulness of exploring the evolutionary history of invasive species within their native ranges to better understand biological invasions.     

Exotic wildland weeds serve as reservoirs for a newly introduced cole crop pest,Bagrada hilaris (Hemiptera: Pentatomidae)

The role of wildland weeds that serve as alternate hosts for insect pests has not been directly examined for the potential to sustain pest populations or facilitate pest outbreaks. The development of weed biological control programmes is also complicated by weed species that are closely related to economically important or native plants, especially rare or special status species. In recent surveys in southern California, USA, we found a newly introduced insect pest of cole crops, Bagrada hilaris Burmeister (Bagrada bug), feeding on Brassicaceae weeds in riparian areas adjacent to agricultural fields where cole crops are routinely grown. Insect populations grew to levels well over action thresholds and caused severe damage to populations of the invasive weed, Lepidium latifolium (perennial pepperweed). The numerical response of B. hilaris on L. latifolium and other Brassicaceae weeds in natural areas may pose a significant challenge to effectively managing pest populations in crops. However, the accidental introduction of this insect provides the opportunity to examine plant–insect interactions with important implications for development of biologically based control methods for weeds.     

The impact of Broussonetia papyrifera (L.) vent. on community characteristics in the forest and forest–savannah transition ecosystems of Ghana

Following its introduction in the late 1960s, Broussonetia papyrifera L. Vent. Moraceae (paper mulberry) has emerged as a major exotic invasive species in Ghana's forest ecosystems. This study was carried out to assess the effects of B. papyrifera invasion on community composition in forest and forest–savannah transition ecosystems. Comparative and removal experiments were conducted in paired B. papyrifera invaded versus uninvaded plots. In the comparative assessment, species composition was found to be similar in both invaded and uninvaded plots. However, relative per cent cover of resident species and guilds including Chromolaena odorata, indigenous broadleaves and indigenous grasses were significantly lower in invaded plots. Seven months after B. papyrifera was experimentally removed from invaded stands, cover by indigenous broadleaves increased by 35%, as against only 5% in control plots at the forest site. However, at the transition site, the increase in per cent cover of indigenous broadleaves (18%) was not significantly different from control (2.5%) plots. We conclude that B. papyrifera has the capacity to reduce the abundance of indigenous broadleaf species, although its removal is more likely to favour regeneration in a forest than a forest–savannah transition ecosystem.     

Genetic and behavioural evidence for a city-wide supercolony of the invasive Argentine ant Linepithema humile (Mayr) (Hymenoptera: Formicidae) in southeastern Australia

The success of invasive ants is frequently attributed to genetic and behavioural shifts in colony structure during or after introduction. The Argentine ant ( Linepithema humile ), a global invader, differs in colony genetic structure and behaviour between native populations in South America and introduced populations in Europe, Japan, New Zealand and North America. However, little is known about its colony structure in Australia. We investigated the genetic structure and behaviour of L. humile across Melbourne, Victoria by quantifying variation at four microsatellite loci and assaying intraspecific aggression at neighbourhood (30–200 m), fine (1–3.3 km) and regional (5–82 km) spatial scales. Hierarchical analyses across these scales revealed that most genetic variation occurred among workers within nests (∼98%). However, although low genetic differentiation occurred among workers between nests at the fine and regional scales (∼2%), negligible differentiation was detected among workers from neighbouring nests. Spatial genetic autocorrelation analysis confirmed that neighbouring nests were genetically more similar to each other. Lack of aggression within and across these scales supported the view that L. humile is unicolonial and forms a large supercolony across Melbourne. Comparisons of genetic structure of L. humile among single nests sampled from Adelaide, Brisbane, Hobart and Perth with Melbourne showed no greater levels of genetic differentiation or dissimilar spatial structure, suggesting an Australia-wide supercolony.     

互花米草黄酮含量分析及其生态学意义

互花米草原产于北美洲大西洋沿岸和墨西哥海湾,经引种和其他原因,已扩散到包括中国在内的多个国家和地区,并对被入侵地生态系统的结构和功能均造成了显著影响。有关该物种的入侵机制和控制策略是当前入侵生态学研究领域的热点。互花米草生物质材料中富含黄酮类物质,与该物种向海扩张的能力可能存在一定联系。鉴于黄酮广泛的应用价值,能否借助黄酮成分的开发带动生物质的收割,以达到有效控制该物种的目的,也是当前研究者所关注的问题。以芦丁为标准参照物,NaNO2-Al（ NO3）3法比较温室互花米草不同器官中总黄酮含量的差异,分析盐城滩涂互花米草越冬芽黄酮含量与所在地理位置之间的关系,采用L9（34）正交试验对干储互花米草叶总黄酮的提取工艺进行优化。互花米草叶片中黄酮类物质远高于根、根状茎、茎、叶鞘、颖稃及种子。盐城滩涂上互花米草越冬芽的黄酮含量随着由陆向海方向基本呈升高趋势。互花米草总黄酮最佳提取工艺为料液比1∶30,乙醇浓度70％,水浴温度80℃,水浴时间2h。互花米草植物材料中,尤其在叶部,黄酮含量十分丰富。该黄酮物质与互花米草在海岸带盐沼生态系统中的适应和向海扩张也存在正相关关系,可作为探索互花米草入侵机制的一个方向。利用本研究中所得优化提取工艺可从互花米草干储生物质中获得稳定的黄酮来源。配合黄酮经济成分的开发而推动生物质材料的收割利用,将成为互花米草生态控制的一项潜在策略。     

Genomic footprints of a biological invasion: Introduction from Asia and dispersal in Europe of the topmouth gudgeon (Pseudorasbora parva)

Facilitated by the intensification of global trading, the introduction and dispersal of species to areas in which they are historically non‐native is nowadays common. From an evolutionary standpoint, invasions are paradoxical: not only non‐native environments could be different from native ones for which introduced individuals would be ill‐adapted, but also small founding population size should be associated with reduced adaptive potential. As such, biological invasions are considered valuable real‐time evolutionary experiments. Here, we investigated the population structure and adaptive potential of the highly invasive topmouth gudgeon (Pseudorasbora parva) across Europe and East Asia. We RAD‐sequenced 301 specimens from sixteen populations and three distinct within‐catchment invaded regions as well as two locations in the native range. With 13,785 single nucleotide polymorphisms, we provide conclusive evidence for a genome‐wide signature of two distinct invasion events, in Slovakia and Turkey, each originating from a specific area in the native range. A third invaded area, in France, appears to be the result of dispersal within the invasive range. Few loci showed signs of selection, the vast majority of which being identified in the Slovakian region. Functional annotation suggests that faster early stage development, resistance to pollution and immunocompetence contribute to the invasion success of the local habitats. By showing that populations in the invasive range have different evolutionary histories, our study reinforces the idea that populations, rather than species, are the units to consider in invasion biology.     

Establishment,spread, and impact of an invasive planthopper on its invasive host plant: Prokelisia marginata (Homoptera: Delphacidae) exploiting Spartina anglica (Poales: Poaceae) in Britain

1. Since its recent arrival in Britain, the planthopper Prokelisia marginata has spread widely around saltmarshes on the east and south coast of England and south Wales, feeding on Common Cordgrass, Spartina anglica, itself an invasive non-native species. 2. Results suggest that P. marginata populations in Britain benefit from a degree of natural enemy release. No evidence of parasitism was found in over 71 000 eggs, nymphs, and adults inspected. The only potential natural enemy control was suggested by a positive correlation between the densities of planthoppers and generalist spiders. 3. Experimental exposure under both glasshouse and field conditions to typical field densities of planthoppers resulted in significant negative effects on a number of host plant performance metrics. 4. Spartina anglica is important for stabilising estuarine sediments and has been deliberately planted for this purpose in the past. Its weakening as a result of heavy planthopper herbivory could have serious consequences for the long-term stability of Britain's vulnerable saltmarsh habitats.     

Effects of Metarhizium robertsii on the bloodsucking mosquito Aedes flavescens and non‐target predatory insects (Odonata)

The effects of different concentrations and methods of treatment with Metarhizium robertsii Bisch., Rehner & Humber conidia on the non‐target aquatic dragonfly larvae Lestes sponsa Hansemann, Lestes dryas Kirby and Aeshna affinis Vander Linden and on the target bloodsucking mosquito larvae Aedes (O.) flavescens (Muller) were analysed. We found that dragonflies are significantly less susceptible than mosquitoes to the fungus. Larvae of L. sponsa larvae were more susceptible to wet conidia than dry conidia. However, the mortality of the air‐breathing larvae of A. affinis was significantly higher after treatment with dry conidia relative to aqueous suspension. The results help to minimize the negative effects of entomopathogenic fungi on non‐target predator insects under the control of mosquito larvae.     

First report on Wolbachia endosymbiosis in freshwater Aphelocheirus aestivalis (Heteroptera: Aphelocheiridae) and its potential impact on genetic diversity of host

Alphaproteobacteria Wolbachia have been described as endosymbionts of approximately half of all aquatic insect species. These bacteria might affect not only reproduction but also the genetic diversity of its hosts. In the present study we identified Wolbachia endosymbiosis in freshwater true bug Aphelocheirus aestivalis F., 1794 (Heteroptera: Aphelocheiridae). Despite the fact that A. aestivalis is widely distributed in Europe, it occurs rather locally, often in isolated populations. Taking into account that Wolbachia, close relationships and past demographic phenomena could affect the genetic diversity of its host, we analyzed mitochondrial (COI and 16S) and nuclear (internal transcribed spacer 2) markers determined for A. aestivalis individuals collected from five populations. Moreover, we compared obtained COI sequences with those deposited in GenBank. Analyses revealed low genetic differentiation among samples tested, as well as low variation among determined COI sequences and those downloaded from the database. Although Wolbachia infection could correlate with decreasing mitochondrial diversity of its host, we suggest that low genetic variation observed in tested A. aestivalis samples (at both mitochondrial and nuclear levels) is a result of populations’ close relationships, past demographic phenomena or is characteristic for this species. Detailed analysis of the wsp gene fragment revealed two distinct strains of Wolbachia infecting A. aestivalis. Both of them belong to supergroup A, also found in other arthropods.