Genetic analysis reveals the putative native range and widespread double-clonal reproduction in the invasive longhorn crazy ant.

Clonal reproduction can provide an advantage for invasive species to establish as it can circumvent inbreeding depression which often plagues introduced populations. The world's most widespread invasive ant, Paratrechina longicornis, was previously found to display a double-clonal reproduction system, whereby both males and queens are produced clonally, resulting in separate male and queen lineages, while workers are produced sexually. Under this unusual reproduction mode, inbreeding is avoided in workers as they carry hybrid interlineage genomes. Despite the ubiquitous distribution of P. longicornis, the significance of this reproductive system for the ant's remarkable success remains unclear, as its prevalence is still unknown. Further investigation into the controversial native origin of P. longicornis is also required to reconstruct the evolutionary histories of double-clonal lineages. Here, we examine genetic variation and characterize the reproduction mode of P. longicornis populations sampled worldwide using microsatellites and mitochondrial DNA sequences to infer the ant's putative native range and the distribution of the double-clonal reproductive system. Analyses of global genetic variations indicate that the Indian subcontinent is a genetic diversity hotspot of this species, suggesting that P. longicornis probably originates from this geographical area. Our analyses revealed that both the inferred native and introduced populations exhibit double-clonal reproduction, with queens and males around the globe belonging to two separate, nonrecombining clonal lineages. By contrast, workers are highly heterozygous because they are first-generation interlineage hybrids. Overall, these data indicate a worldwide prevalence of double clonality in P. longicornis and support the prediction that the unusual genetic system may have pre-adapted this ant for global colonization by maintaining heterozygosity in the worker force and alleviating genetic bottlenecks.

Population genetic structure of the globally introduced big-headed ant in Taiwan.

Global commerce and transportation facilitate the spread of invasive species. The African big-headed ant, Pheidole megacephala (Fabricius), has achieved worldwide distribution through globalization. Since the late 19th century, Taiwan has served as a major seaport because of its strategic location. The population genetic structure of P. megacephala in Taiwan is likely to be shaped by international trade and migration between neighboring islands. In this study, we investigated the population genetics of P. megacephala colonies sampled from four geographical regions in Taiwan and elucidated the population genetic structures of P. megacephala sampled from Taiwan, Okinawa, and Hawaii. We observed a low genetic diversity of P. megacephala across regions in Taiwan. Moreover, we noted low regional genetic differentiation and did not observe isolation by distance, implying that long-distance jump dispersal might have played a crucial role in the spread of P. megacephala. We sequenced the partial cytochrome oxidase I gene and observed three mitochondrial haplotypes (TW1-TW3). TW1 and TW3 most likely originated from populations within the species' known invasive range, suggesting that secondary introduction is the predominant mode of introduction for this invasive ant. TW2 represents a novel haplotype that was previously unreported in other regions. P. megacephala populations from Taiwan, Okinawa, and Hawaii exhibited remarkable genetic similarity, which may reflect their relative geographic proximity and the historical connectedness of the Asia-Pacific region.

First Polycipivirus and Unmapped RNA Virus Diversity in the Yellow Crazy Ant, Anoplolepis gracilipes.

The yellow crazy ant, Anoplolepis gracilipes is a widespread invasive ant that poses significant threats to local biodiversity. Yet, compared to other global invasive ant species such as the red imported fire ant (Solenopsis invicta) or the Argentine ant (Linepithema humile), little is known about the diversity of RNA viruses in the yellow crazy ant. In the current study, we generated a transcriptomic database for A. gracilipes using a high throughput sequencing approach to identify new RNA viruses and characterize their genomes. Four virus species assigned to Dicistroviridae, two to Iflaviridae, one to Polycipiviridae, and two unclassified Riboviria viruses were identified. Detailed genomic characterization was carried out on the polycipivirus and revealed that this virus comprises 11,644 nucleotides with six open reading frames. Phylogenetic analysis and pairwise amino acid identity comparison classified this virus into the genus Sopolycivirus under Polycipiviridae, which is tentatively named "Anoplolepis gracilipes virus 3 (AgrV-3)". Evolutionary analysis showed that AgrV-3 possesses a high level of genetic diversity and elevated mutation rate, combined with the common presence of multiple viral strains within single worker individuals, suggesting AgrV-3 likely evolves following the quasispecies model. A subsequent field survey placed the viral pathogen "hotspot" of A. gracilipes in the Southeast Asian region, a pattern consistent with the region being recognized as part of the ant's native range. Lastly, infection of multiple virus species seems prevalent across field colonies and may have been linked to the ant's social organization.

Biology, Ecology, and Management of the Invasive Longlegged Ant, Anoplolepis gracilipes.

The longlegged ant (Anoplolepis gracilipes) is one of the most damaging invasive tramp ants globally. It is generally found between latitudes 27°N and 27°S in Asia, although it has been introduced to other continents. Its native range remains debatable, but it is believed to be in Southeast Asia. Anoplolepis gracilipes invasion has many serious ecological consequences, especially for native invertebrate, vertebrate, and plant communities, altering ecosystem dynamics and functions. We examine and synthesize the literature about this species' origin and distribution, impacts on biodiversity and ecosystems, biology and ecology, chemical control, and potential biocontrol agents. We highlight emerging research needs on the origin and invasion history of this species, its reproductive mode, its relationship with myrmecophiles, and its host-microbial interactions, and we discuss future research directions.

The complete mitochondrial genome of a parthenogenetic ant Monomorium triviale (Hymenoptera: Formicidae).

Monomorium is one of the most species-rich yet taxonomically problematic ant genus. An East Asian species, M. triviale Wheeler, W.M., 1906, is reproduced by obligate thelytokous parthenogenesis and performs strict reproductive division of labor. We sequenced the M. triviale mitogenome using next-generation sequencing methods. The circular mitogenome of M. triviale was 16,290 bp in length, consisting of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNAs, and a single non-coding region of 568 bp. The base composition was AT-biased (82%). Gene order rearrangements were detected and likely to be unique to the genus Monomorium. We announce the M. triviale mitogenome as additional genomic resources for elucidating phylogenetic and taxonomic problems of Monomorium and comparative genomics of parthenogenetic ant species.

Improving invasive species management using predictive phenology models: an example from brown marmorated stink bug (Halyomorpha halys) in Japan.

BACKGROUND: In order to better understand the population dynamics of invasive species in their native range, we developed two predictive phenological models using the ubiquitous invasive insect pest, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), as the model organism. Our work establishes a zero-inflated negative binomial regression (ZINB) model, and a general additive mixed model (GAMM) based on 11 years of black light trap monitoring of H. halys at three locations in Japan. RESULTS: The ZINB model indicated that degree days (DD) have a significant effect on the trap catch of adult H. halys, and that precipitation has no effect. A dataset generated by 1000 simulations from the ZINB suggested that higher predicted trap catches equated to a lower probability of encountering a zero-count. The GAMM produced a cubic regression smooth curve which forecasts the seasonal phenology of H. halys as following a bell-shaped trend in Japan. Critical DD points during the field season in Japan included 261 DD for first H. halys adult detection and 1091 DD for peak activity. CONCLUSIONS: This study establishes the first models capable of forecasting native H. halys population dynamics based on DD. These robust models practically improve population forecasting of H. halys in the future and help fill gaps in knowledge pertaining to its native phenology, thus ultimately contributing to the progression of efficient management of this globally invasive species. © 2021 Society of Chemical Industry.

Comparison of Artificial Diets and Natural Prey for Mass Rearing of Orius strigicollis (Hemiptera: Anthocoridae) Using Demographic Characteristics to Optimize Cost-Efficiency.

Orius strigicollis (Poppius) has been commonly released to control minute pests due to its remarkable foraging and predation ability. Despite decades long history of mass-rearing using eggs of Cadra cautella Walker (Lepidoptera: Pyralidae) as food, the cost of this food impeded the predator's marketing potential. Finding cost-effective artificial diets for mass rearing programs is, therefore, the key to promote the use of this generalist predator in practice. The aim of this study was to explore suitability of alternative diets for mass rearing of this predator, using the age-stage, two-sex life table. Eight recipes of artificial diets were initially screened, and the candidate diets were subsequently evaluated using the population parameters of O. strigicollis. The highest intrinsic rate of increase was found in the O. strigicollis fed on the meridic combined Diet 1, where nymphs and adults were fed different diets. However, the intensive period of oviposition by females reared on oligidic Diet O3 contributed to the shorter rearing period (42 d) and lower rearing cost than that on combined Diet 1 (60 d). Besides, with the harvest rate of 0.919, the minimal population size of 10,774 individuals with a stable age-stage distribution was needed for daily harvesting 1,000 third instars of O. strigicollis reared on Diet O3 at the rearing cost of 0.295 NTD (new Taiwanese dollar) per nymph. Thus, Diet O3 was determined to be the most cost-effective recipe for the mass-rearing of O. strigicollis among those used in this study.

The role of anthropogenic disturbance and invasion of yellow crazy ant in a recent decline of land crab population.

The yellow crazy ant, Anoplolepis gracilipes was first reported in Kenting National Park, Taiwan, in the 2000s, raising the concern of this invasive ant's potential effects on the native land crab populations. We investigated the distribution and abundance of A. gracilipes and its adverse effect on the land crab populations at four land crab hotspots within the park. Our survey results indicated that A. gracilipes was widespread and abundant at three of the study sites (Hsiangchiaowan, Shadao, and Natural Spring), while the distribution was much more confined at the fourth site (Houwan). Land crab populations experienced a notable decline at all the study sites except Houwan, indicating an inverse relationship between the population of land crabs and distribution/abundance of A. gracilipes. Combining the results of visual observations, the decline of land crab populations can be attributed, at least partially, to direct attacks by A. gracilipes on land crabs in their natural habitat and during the breeding migration season. Generalized additive model showed that worker abundance of this ant is associated with human disturbance levels, suggesting that anthropogenic disturbance may have contributed to the decline in land crab populations via promoting the abundance of A. gracilipes.

Deformed Wing Virus in Two Widespread Invasive Ants: Geographical Distribution, Prevalence, and Phylogeny.

Spillover of honey bee viruses have posed a significant threat to pollination services, triggering substantial effort in determining the host range of the viruses as an attempt to understand the transmission dynamics. Previous studies have reported infection of honey bee viruses in ants, raising the concern of ants serving as a reservoir host. Most of these studies, however, are restricted to a single, local ant population. We assessed the status (geographical distribution/prevalence/viral replication) and phylogenetic relationships of honey bee viruses in ants across the Asia-Pacific region, using deformed wing virus (DWV) and two widespread invasive ants, Paratrechina longicornis and Anoplolepis gracilipes, as the study system. DWV was detected in both ant species, with differential geographical distribution patterns and prevenance levels between them. These metrics, however, are consistent across the geographical range of the same ant species. Active replication was only evident in P. longicornis. We also showed that ant-associated DWV is genetically similar to that isolated from Asian populations of honey bees, suggesting that local acquisition of DWV by the invasive ants may have been common at least in some of our sampled regions. Transmission efficiency of DWV to local arthropods mediated by ant, however, may vary across ant species.

Population Monitoring, Egg Parasitoids, and Genetic Structure of the Invasive Litchi Stink Bug, Tessaratoma papillosa in Taiwan.

Here we assessed population dynamics, natural enemy fauna (with emphasis on egg parasitoid), and population genetic structure (based on mitochondrial DNA) of the invasive litchi stink bug (LSB), Tessaratoma papillosa in Taiwan. Our major findings include: (1) fluctuations of LSB in numbers of adults, mating pairs, and egg masses over a 2-year period in Taiwan generally resemble those in the native populations; (2) Anastatus dexingensis and A. fulloi are among the most dominant LSB egg parasitoids, with the former consistently outnumbering the latter throughout Taiwan; (3) the presence of two genetically distinct clades suggests LSB in Taiwan most likely derived from multiple invasions. All these data practically improve our understanding of this invasive insect pest, particularly its ecological and genetic characteristics in the introduced area, which represents critical baseline information for the design of future integrated pest management strategies.

Ongoing Coevolution of Wolbachia and a Widespread Invasive Ant, Anoplolepis gracilipes.

While Wolbachia are commonly found among arthropods, intraspecific infection rates can vary substantially across the geographic populations. Here we report nearly 100% prevalence of Wolbachia in the global populations of the yellow crazy ant, Anoplolepis gracilipes. To understand coevolutionary history between Wolbachia and A. gracilipes, we identified single nucleotide polymorphisms (SNPs) in Wolbachia from the ant across 12 geographical regions and compared the phylogeny of SNP-based Wolbachia to patterns of the ant's mitochondrial DNA (mtDNA) variation. Our results revealed a strong concordance between phylogenies of Wolbachia and host mtDNA, providing immediate evidence of co-divergence. Among eight identified SNP loci separating the genetic clusters of Wolbachia, seven loci are located in potential protein-coding genes, three of which being non-synonymous SNPs that may influence gene functions. We found a Wolbachia hypothetical protein gene with signature of positive selection. These findings jointly allow us to characterize Wolbachia-ant coevolution and also raise a question about mechanism(s) underlying maintenance of high prevalence of Wolbachia during the colonization of this invasive ant.

Correction to: Complete genome sequences of two novel dicistroviruses detected in yellow crazy ants (Anoplolepis gracilipes).

Authors would like to correct the error in Fig. 1 which was incorrectly updated in the original publication.

Complete genome sequences of two novel dicistroviruses detected in yellow crazy ants (Anoplolepis gracilipes).

We report two novel RNA viruses from yellow crazy ants, (Anoplolepis gracilipes) detected using next-generation sequencing. The complete genome sequences of the two viruses were 10,662 and 8,238 nucleotides in length, respectively, with both possessing two open reading frames with three conserved protein domains. The genome organization is characteristic of members of the genus Triatovirus in the family Dicistroviridae. The two novel viruses were tentatively named "Anoplolepis gracilipes virus 1" and "Anoplolepis gracilipes virus 2" (AgrV-1 and AgrV-2). Phylogenetic analyses based on amino acid sequences of the non-structural polyprotein (ORF1) suggest that the two viruses are triatovirus-like viruses. This is the first report on the discovery of novel triatovirus-like viruses in yellow crazy ants with a description of their genome structure (two ORFs and conserved domains of RNA helicase, RNA-dependent RNA polymerase, and capsid protein), complete sequences, and viral prevalence across the Asia-Pacific region.

Analysis of Recent Interception Records Reveals Frequent Transport of Arboreal Ants and Potential Predictors for Ant Invasion in Taiwan.

We uncovered taxonomic diversity, country of origin and commodity type of intercepted ants at Taiwanese borders based on an 8 year database of 439 interception records. We found intercepted ants arrived predominantly via timber, a pattern likely reflecting the high domestic demand for foreign timber in Taiwan. The most frequently intercepted species were either arboreal or wood-dwelling ants, raising a concern of these ants constituting a next wave of ant invasion in Taiwan. Further analyses indicate that the taxonomic composition of intercepted ants does not match that of established non-native ant species, suggesting that interception data alone fails to provide adequate power to predict the establishment success of ants. Yet, interception frequency and selected life-history traits (i.e., flexible colony founding mode and general nesting habits) were shown to jointly serve as a practical predictor of the establishment risk of non-native ants. Consistent with other border interception databases, secondary introduction (i.e., species arriving from their introduced ranges instead of their native ranges) also represents a major pathway for transport of invasive ants into Taiwan, suggesting its role in shaping the global invasion of ants. Our findings offer baseline information for constructing a prediction framework for future ant invasions and assist in the decision-making process of quarantine authorities in Taiwan.

Evidence for Common Horizontal Transmission of Wolbachia among Ants and Ant Crickets: Kleptoparasitism Added to the List.

While Wolbachia, an intracellular bacterial symbiont, is primarily transmitted maternally in arthropods, horizontal transmission between species has been commonly documented. We examined kleptoparasitism as a potential mechanism for Wolbachia horizontal transmission, using ant crickets and their host ants as the model system. We compared prevalence and diversity of Wolbachia across multiple ant cricket species with different degrees of host specificity/integration level. Our analyses revealed at least three cases of inter-ordinal Wolbachia transfer among ant and ant crickets, and also showed that ant cricket species with high host-integration and host-specificity tend to harbor a higher Wolbachia prevalence and diversity than other types of ant crickets. This study provides empirical evidence that distribution of Wolbachia across ant crickets is largely attributable to horizontal transmission, but also elucidates the role of intimate ecological association in successful Wolbachia horizontal transmission.

Vibrational communication between a myrmecophilous butterfly Spindasis lohita (Lepidoptera: Lycaenidae) and its host ant Crematogaster rogenhoferi (Hymenoptera: Formicidae).

Ants are a dominant insect group in terrestrial ecosystems and many myrmecophilous species evolve to associate with ants to gain benefits. One iconic example is myrmecophilous butterflies that often produce ant-mimicking vibrational calls to modulate ant behaviors. Despite its popularity, empirical exploration of how butterflies utilize vibrational signals to communicate with ants is scarce. In this study, we reported that the myrmecophilous butterfly Spindasis lohita produce three types of larval calls and one type of pupal call, while its tending ant, Crematogaster rogenhoferi emit a single type of call. The results of discriminant analysis revealed that calls of the two species are quantitatively similar in their signal attributes; the potential role of butterfly calls are further confirmed by the playback experiments in which certain ant behaviors including antennation, aggregation, and guarding were induced when one of the butterfly calls was played to C. rogenhoferi workers. The findings in the current study represent the very first evidence on vibrational communication between Spindasis and Crematogaster and also imply that S. lohita may have been benefited from ant attendance due to the ability to produce similar calls of the ant C. rogenhoferi.

The Association between Virus Prevalence and Intercolonial Aggression Levels in the Yellow Crazy Ant, Anoplolepis Gracilipes (Jerdon).

The recent discovery of multiple viruses in ants, along with the widespread infection of their hosts across geographic ranges, provides an excellent opportunity to test whether viral prevalence in the field is associated with the complexity of social interactions in the ant population. In this study, we examined whether the association exists between the field prevalence of a virus and the intercolonial aggression of its ant host, using the yellow crazy ant (Anoplolepis gracilipes) and its natural viral pathogen (TR44839 virus) as a model system. We delimitated the colony boundary and composition of A. gracilipes in a total of 12 study sites in Japan (Okinawa), Taiwan, and Malaysia (Penang), through intercolonial aggression assay. The spatial distribution and prevalence level of the virus was then mapped for each site. The virus occurred at a high prevalence in the surveyed colonies of Okinawa and Taiwan (100% infection rate across all sites), whereas virus prevalence was variable (30%-100%) or none (0%) at the sites in Penang. Coincidentally, colonies in Okinawa and Taiwan displayed a weak intercolonial boundary, as aggression between colonies is generally low or moderate. Contrastingly, sites in Penang were found to harbor a high proportion of mutually aggressive colonies, a pattern potentially indicative of complex colony composition. Our statistical analyses further confirmed the observed correlation, implying that intercolonial interactions likely contribute as one of the effective facilitators of/barriers to virus prevalence in the field population of this ant species.

Genetic Diversity and Wolbachia Infection Patterns in a Globally Distributed Invasive Ant.

Understanding the phylogeographic history of an invasive species may facilitate reconstructing the history and routes of its invasion. The longhorn crazy ant, Paratrechina longicornis, is a ubiquitous agricultural and household pest throughout much of the tropics and subtropics, but little is known about the history of its spread. Here, we examine worldwide genetic variation in P. longicornis and its associated Wolbachia bacterial symbionts. Analyses of mtDNA sequences of 248 P. longicornis workers (one per colony) from 13 geographic regions reveal two highly diverged mtDNA clades that co-occur in most of the geographic regions. These two mtDNA clades are associated with different Wolbachia infection patterns, but are not congruent with patterns of nDNA (microsatellite) variation. Multilocus sequence typing reveals two distinct Wolbachia strains in P. longicornis, namely, wLonA and wLonF. The evolutionary histories of these two strains differ; wLonA appears to be primarily transmitted maternally, and patterns of mtDNA and nDNA variation and wLonA infection status are consistent with a relatively recent Wolbachia-induced selective sweep. In contrast, the observed patterns of mtDNA variation and wLonF infections suggest frequent horizontal transfer and losses of wLonF infections. The lack of nDNA structure among sampled geographic regions coupled with the finding that numerous mtDNA haplotypes are shared among regions implies that inadvertent long-distance movement through human commerce is common in P. longicornis and has shaped the genetic structure of this invasive ant worldwide.

Fitness advantages of the biased use of paired laterally symmetrical penises in an insect.

The evolution of laterality, that is the biased use of laterally paired, morphologically symmetrical organs, has attracted the interest of researchers from a variety of disciplines. It is, however, difficult to quantify the fitness benefits of laterality because many organs, such as human hands, possess multimodal functions. Males of the earwig Labidura riparia (Insecta: Dermaptera: Labiduridae) have morphologically similar laterally paired penises, only one of which is used for inseminating the female during a single copulation bout, and thus provide a rare opportunity to address how selection pressure may shape the evolution of population-level laterality. Our population studies revealed that in 10 populations, located at 2.23-43.3° north, the right penis is predominantly used for copulating (88.6%). A damaged penis was found in 23% of rare left-handers, suggesting that the left penis can function as a spare when the right one is damaged. By pairing L. riparia females with surgically manipulated males, we found that males forced to use the right penis outperformed left-handed males in copulation (the probability of establishing genital coupling during the 1-hr observation period: odds ratio [OR] of 3.50) and insemination (probability of transferring a detectable amount of sperm: OR of 2.94). This right-handed advantage may be due to the coiled morphology of the sperm storage organ with a right-facing opening. Thus, female genital morphology may play a significant role in the evolution of handedness and may have acted as a driving force to reduce penis number in related taxa.

Apoptosis as a primary defense mechanism in response to viral infection in invasive fire ant Solenopsis invicta.

Apoptosis is considered an innate defense mechanism of insect hosts at the early stage of pathogen infection. The present study attempts to determine whether apoptosis is involved in defending the fire ant, Solenopsis invicta from a natural viral pathogen Solenopsis invicta virus 1 (SINV-1). Results of TEM examination and TUNEL assay both revealed the signature of apoptosis in the midgut epithelium of SINV-1-infected fire ant larvae. A time-course study was conducted to monitor changes in the dynamics of SINV-1 viral titers and apoptosis levels in the midgut epithelium of SINV-1-infected larvae. We found that the viral titer significantly decreases as apoptosis level increases, suggesting that the apoptotic epithelium constitutes a barrier against dissemination of SINV-1. These findings serve as the very first empirical evidence for the virus-induced apoptosis in ants and also help explain some previously observed mortality patterns and behavioral alterations associated with SINV-1 in fire ants.