Social insects host a diversity of viruses. We examined New Zealand populations of the globally widely distributed invasive Argentine ant (Linepithema humile) for RNA viruses. We used metatranscriptomic analysis, which identified six potential novel viruses in the Dicistroviridae family. Of these, three contigs were confirmed by Sanger sequencing as Linepithema humile virus-1 (LHUV-1), a novel strain of Kashmir bee virus (KBV) and Black queen cell virus (BQCV), while the others were chimeric or misassembled sequences. We extended the known sequence of LHUV-1 to confirm its placement in the Dicistroviridae and categorised its relationship to closest relatives, which were all viruses infecting Hymenoptera. We examined further for known viruses by mapping our metatranscriptomic sequences to all viral genomes, and confirmed KBV, BQCV, LHUV-1 and Deformed wing virus (DWV) presence using qRT-PCR. Viral replication was confirmed for DWV, KBV and LHUV-1. Viral titers in ants were higher in the presence of honey bee hives. Argentine ants appear to host a range of' honey bee' pathogens in addition to a virus currently described only from this invasive ant. The role of these viruses in the population dynamics of the ant remain to be determined, but offer potential targets for biocontrol approaches.Social insects carry a range of viruses that can have a major effect on host population dynamics. Perhaps the best known viral community is from honey bees, which has been the focus of considerable study due to their economic importance. A range of different factors are likely to contribute to colony collapse and bee declines in general, with viruses frequently considered key players 1, 2 . A recent review noted honey bees host 24 viruses, primarily in the Dicistroviridae and Iflaviridae families 3 . Of these, the Deformed wing virus (DWV) has been suggested as a likely candidate for the majority of global honey bee colony losses during the past 50 years 4 . Such viruses, however, are not restricted to honey bees. There is increasing evidence that these 'honey bee' viruses are common in a wide range of insect hosts [5][6][7] . Other social insects have been found to carry their own unique suites of viral pathogens. For example, over the last decade four viruses have been described from the red imported fire ant (Solenopsis invicta) 8 . These were the first viruses fully described from ants. Three of these viruses are positive-sense, single-stranded RNA (ssRNA) viruses, with one (Solenopsis invicta virus-1, SINV-1) assigned taxonomically to the Dicistroviridae family, one (Solenopsis invicta virus-3, SINV-3) in a proposed new family, Solinviviridae 9 and the third currently unclassified (Solenopsis invicta virus-2, SINV-2) 8, 9 ; The fourth virus is a DNA virus, and has been placed in the family Parvoviridae 10 . One of the three ssRNA viruses, SINV-3, shows promise as a biocontrol agent as it can cause significant mortality in laboratory fire ant colonies 11 . Metatranscriptomic and pyrosequencing techniques have proven particula...
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 malekilling. 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.
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.
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