Monica A. M. Gruber

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.

Prevalence and genetic diversity of three bacterial endosymbionts (Wolbachia, Arsenophonus, and Rhizobiales) associated with the invasive yellow crazy ant (Anoplolepis gracilipes)

Invasive species carry pathogens, parasites and mutualistic microorganisms into their new range. We examined the prevalence and genetic diversity of three bacterial endosymbionts (Wolbachia, Arsenophonus, and Rhizobiales) in four Anoplolepis gracilipes (Smith) populations in the Pacific region, four populations from mainland Australia and one population from Christmas Island in the Indian Ocean. Wolbachia was found in eight of the nine sampling sites, with between 31.8 and 100% of ants infected. These infection rates are substantially higher than those previously observed for other invasive ants such as Linepithema humile and Solenopsis invicta. All sequences of Wolbachia were genetically identical. Arsenophonus was observed in five of the nine sampling sites, with infection rates ranging between 4.5 and 50.8%. Like Wolbachia, Arsenophonus can modify the sex-ratio of its hosts via male-killing. Arsenophonus was found to co-occur with Wolbachia in the same ants in five of the nine sampling sites. Rhizobiales is a clade of symbiotic bacteria mostly found in herbivorous ants. These bacteria help provide nitrogen to their hosts and were found in only three of the nine sampling sites with an infection rate of 1.6–11.8%. It also co-occurred with the other bacteria. There was no genetic variation in Arsenophonus and Rhizobiales samples, with the exception of a sequence from one Arsenophonus sample in Samoa that differed by a single base pair. These bacterial endosymbionts may contribute to the population variability in A. gracilipes and may be manipulated for the purpose of pest management.

Booms, busts and population collapses in invasive ants

The abundance of many invasive species can vary substantially over time, with dramatic population declines and local extinctions frequently observed in a wide range of taxa. We highlight population crashes of invasive ants, which are some of the most widespread and damaging invasive animals. Population collapse or substantial declines have been observed in nearly all of the major invasive ant species including the yellow crazy ant (Anoplolepis gracilipes), Argentine ants (Linepithema humile), big-headed or coastal brown ant (Pheidole megacephala), the tropical fire ant (Solenopsis geminata), red imported fire ants (Solenopsis invicta), and the little fire ant or electric ant (Wasmannia auropunctata). These declines frequently attract little attention, especially compared with their initial invasion phase. Suggested mechanisms for population collapse include pathogens or parasites, changes in the food availability, or even long-term effects of the reproductive biology of invasive ants. A critical component of the collapses may be a reduction in the densities of the invasive ant species, which are often competitively weak in low abundance. We propose that mechanisms causing a reduction in invasive ant abundance may initiate a local extinction vortex. Declines in abundance likely reduce the invasive ant’s competitive ability, resource acquisition and defense capability. These reductions could further reduce the abundance of an invasive ant species, and so on. Management of invasive ants through the use of pesticides is expensive, potentially ecologically harmful, and can be ineffective. We argue that pesticide use may even have the potential to forestall natural population declines and collapses. We propose that in order to better manage these invasive ants, we need to understand and capitalize on features of their population dynamics that promote population collapse.

No evidence of enemy release in pathogen and microbial communities of common wasps (Vespula vulgaris) in their native and introduced range.

When invasive species move to new environments they typically experience population bottlenecks that limit the probability that pathogens and parasites are also moved. The invasive species may thus be released from biotic interactions that can be a major source of density-dependent mortality, referred to as enemy release. We examined for evidence of enemy release in populations of the common wasp (Vespula vulgaris), which attains high densities and represents a major threat to biodiversity in its invaded range. Mass spectrometry proteomic methods were used to compare the microbial communities in wasp populations in the native (Belgium and England) and invaded range (Argentina and New Zealand). We found no evidence of enemy release, as the number of microbial taxa was similar in both the introduced and native range. However, some evidence of distinctiveness in the microbial communities was observed between countries. The pathogens observed were similar to a variety of taxa observed in honey bees. These taxa included Nosema, Paenibacillus, and Yersina spp. Genomic methods confirmed a diversity of Nosema spp., Actinobacteria, and the Deformed wing and Kashmir bee viruses. We also analysed published records of bacteria, viruses, nematodes and fungi from both V. vulgaris and the related invader V. germanica. Thirty-three different microorganism taxa have been associated with wasps including Kashmir bee virus and entomophagous fungi such as Aspergillus flavus. There was no evidence that the presence or absence of these microorganisms was dependent on region of wasp samples (i.e. their native or invaded range). Given the similarity of the wasp pathogen fauna to that from honey bees, the lack of enemy release in wasp populations is probably related to spill-over or spill-back from bees and other social insects. Social insects appear to form a reservoir of generalist parasites and pathogens, which makes the management of wasp and bee disease difficult.

The conundrum of the yellow crazy ant (Anoplolepis gracilipes) reproductive mode: no evidence for dependent lineage genetic caste determination

Asexual reproduction and hybridisation are often found among highly invasive plants and marine invertebrates. Recently, it has been suggested that clonality may enhance the success of invasive ants. In contrast, obligate hybridisation (dependent lineage genetic caste determination or DL GCD in ants) may decrease the chances of population persistence if one lineage is less prevalent than the other (asymmetry in lineage ratio). Genetic data available for the invasive yellow crazy ant (Anoplolepis gracilipes) suggest that it has an unconventional mode of reproduction that may involve asexual reproduction by workers or queens, or a form of genetic caste determination. Here, we investigated whether A. gracilipes reproduction involved DL GCD. The potential for worker reproduction was also assessed. We used microsatellite markers to assess the population structure of A. gracilipes workers, males, queens and sperm in queen spermathecae, from field collections in Arnhem Land. We found that a single queen lineage is present in Arnhem Land. The presence of a single lineage of queens discounts the possibility of DL GCD. Population structure separated queens and workers into different lineages, suggesting that these castes are determined genetically in A. gracilipes, or the mode of reproduction differs between workers and queens. Evidence for worker reproduction was weak. We conclude that the reproductive mode of A. gracilipes does not involve DL GCD. The resolution of the reproductive mode of A. gracilipes is complicated by a high prevalence of diploid males. The determination of the A. gracilipes reproductive mode remains a fascinating research question, and its resolution will improve our understanding of the contribution of the reproductive system to invasion success.

Genetic diversity is positively associated with fine‐scale momentary abundance of an invasive ant

Many introduced species become invasive despite genetic bottlenecks that should, in theory, decrease the chances of invasion success. By contrast, population genetic bottlenecks have been hypothesized to increase the invasion success of unicolonial ants by increasing the genetic similarity between descendent populations, thus promoting co-operation. We investigated these alternate hypotheses in the unicolonial yellow crazy ant, Anoplolepis gracilipes, which has invaded Arnhem Land in Australias Northern Territory. We used momentary abundance as a surrogate measure of invasion success, and investigated the relationship between A. gracilipes genetic diversity and its abundance, and the effect of its abundance on species diversity and community structure. We also investigated whether selected habitat characteristics contributed to differences in A. gracilipes abundance, for which we found no evidence. Our results revealed a significant positive association between A. gracilipes genetic diversity and abundance. Invaded communities were less diverse and differed in structure from uninvaded communities, and these effects were stronger as A. gracilipes abundance increased. These results contradict the hypothesis that genetic bottlenecks may promote unicoloniality. However, our A. gracilipes study population has diverged since its introduction, which may have obscured evidence of the bottleneck that would likely have occurred on arrival. The relative importance of genetic diversity to invasion success may be context dependent, and the role of genetic diversity may be more obvious in the absence of highly favorable novel ecological conditions.

Diploscapter formicidae sp n. (Rhabditida: Diploscapteridae), from the ant Prolasius advenus (Hymenoptera: Formicidae) in New Zealand

Diploscapter formicidae sp. n. was collected from the ant Prolasius advenus and its nests in native beech forests of the South Island, New Zealand. This is a new host record for the nematode genus and the first report of an ant associate from the southern hemisphere. Diploscapter formicidae sp. n. appears to be native to New Zealand. No males were found from collections from 16 nests, in agreement with previously published data on the other members of this genus, suggesting that males are absent or very rare. The adult females have bilateral symmetry of the head, characteristic dorsal and ventral projections of the putative cheilostom with paired hook-like structures or hamuli, expansive membranous lateral lip flaps or laciniae, gymnostom and stegostom with parallel walls, a swollen procorpus, large terminal bulb with a strong valve, paired ovaries with medial vulva, and a short conoid tail with slender pointed or spicate tip. Scanning electron micrographs of the structure of the head confirmed that the lateral laciniae with finger-like tines or filopodia are moveable (alternately covering and exposing the mouth). These lateral lip flaps are located posterior to the stoma, but anterior to the pore-like amphidial openings. The anterior margin of the cheilostom possesses apomorphic lateral bell-shaped projections and the hamuli are broader and less pointed than other species that have been examined. Molecular phylogeny of near full-length small subunit, D2/D3 expansion segments of the large subunit rRNA gene and heat shock protein 90 (Hsp90) gene showed that D. formicidae sp. n. is monophyletic with the Diploscapter species and isolates available in GenBank, but is on an independent trajectory supporting separate species status.

A metatranscriptomic survey of the invasive yellow crazy ant, Anoplolepis gracilipes , identifies several potential viral and bacterial pathogens and mutualists

Thriving populations of invasive species often decline, but the mechanisms behind such declines are rarely elucidated. The microbial community living within insects can have significant impacts on host health and population dynamics and may be one such mechanism. Here, we use Illumina RNAseq and 16S amplicon sequencing to compare the bacterial communities, and investigate the presence of viruses, in queens of the invasive yellow crazy ant, Anoplolepis gracilipes, from populations in various stages of decline or expansion in Australia. Black queen cell virus and sequences with homology to the Dicistroviridae family of viruses were detected in queens from declining ant populations. This is the first instance of sequences with homology to a virus being found in A. gracilipes and may indicate a new virus. The overall bacterial communities were not statistically different between the population types and a number of potential mutualists were discovered. Several putative pathogens were also detected, including Candidatus Rhabdochlamydia, the entomopathogen Serratia marcescens and the reproductive parasite Candidatus Cardinium. These taxa are known to significantly affect host biology in other organisms. Our survey has identified several candidates that may be responsible for population declines of this invasive ant, and therefore may have potential as biological control agents.

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 association between invasive Lantana camara and seedlings/saplings of a plant community in Mudumalai Tiger Reserve, India.

We examined changes in a community of seedlings/saplings 10–150 cm tall associated with the presence of a widely invasive plant, Lantana camara and environmental covariates along 67 randomly located transects, in Mudumalai, India. We compared plant species assemblage and grass cover in L. camara- invaded and uninvaded plots in three habitats. Multivariate analyses revealed a significant association of all environmental covariates with plant species assemblage. Pairwise tests indicated that L. camara was significantly associated with changes in plant species assemblage and grass cover within the moist and dry deciduous forest, but not in the thorn forest. The relationship between L. camara and that of elephant browse plants varied with species. A linear regression analysis indicated that L. camara invasion was the only significant predictor of grass occupancy. Our results indicate that in addition to other factors, L. camara was associated with altering plant species assemblage, some elephant browse plants and grass cover in the moist and dry deciduous forest. It appears that L. camara can have a major effect on diversity within this reserve, but whether this effect is by L. camara driving the change or being associated with other habitat change requires further experimental evidence.