Fast-tracking bespoke DNA reference database generation from museum collections for biomonitoring and conservation.

Despite recent advances in high-throughput DNA sequencing technologies, a lack of locally relevant DNA reference databases limits the potential for DNA-based monitoring of biodiversity for conservation and biosecurity applications. Museums and national collections represent a compelling source of authoritatively identified genetic material for DNA database development, yet obtaining DNA barcodes from long-stored specimens may be difficult due to sample degradation. Here we demonstrate a sensitive and efficient laboratory and bioinformatic process for generating DNA barcodes from hundreds of invertebrate specimens simultaneously via the Illumina MiSeq system. Using this process, we recovered full-length (334) or partial (105) COI barcodes from 439 of 450 (98%) national collection-held invertebrate specimens. This included full-length barcodes from 146 specimens which produced low-yield DNA and no visible PCR bands, and which produced as little as a single sequence per specimen, demonstrating high sensitivity of the process. In many cases, the identity of the most abundant sequences per specimen were not the correct barcodes, necessitating the development of a taxonomy-informed process for identifying correct sequences among the sequencing output. The recovery of only partial barcodes for some taxa indicates a need to refine certain PCR primers. Nonetheless, our approach represents a highly sensitive, accurate and efficient method for targeted reference database generation, providing a foundation for DNA-based assessments and monitoring of biodiversity.

High Dietary Niche Overlap Between Non-native and Native Ant Species in Natural Ecosystems.

Ants represent a highly diverse and ecologically important group of insects found in almost all terrestrial ecosystems. A subset of ant species have been widely transported around the globe and invade many natural ecosystems, often out-competing native counterparts and causing varying impacts on recipient ecosystems. Decisions to control non-native ant populations require an understanding of their interactions and related impacts on native communities. We employed stable isotope analysis and metabarcoding techniques to identify potential dietary niche overlap and identify gut contents of 10 ant species found in natural ecosystems in Aotearoa New Zealand. Additionally, we looked at co-occurrence to identify potential competitive interactions among native and non-native ant species. Ants fed mainly across two trophic levels, with high dietary overlap. Relative to other ant species sampled, two non-native ant species, Linepithema humile and Technomyrmex jocosus, were found to feed at the lowest trophic level. The largest isotopic niche overlap was observed between the native Monomorium antarcticum and the invasive Ochetellus glaber, with analyses revealing a negative co-occurrence pattern. Sequence data of ant gut content identified 51 molecular operational taxonomic units, representing 22 orders and 34 families, and primarily consisting of arthropod DNA. Although we generally found high dietary overlap among species, negative occurrence between a dominant, non-native species and a ubiquitous native species indicates that species-specific interactions could be negatively impacting native ecosystems. Our research progresses and informs the currently limited knowledge around establishing protocols for metabarcoding to investigate ant diet and interactions between native and non-native ant species.

Dataset of host records for introduced parasitoid wasp species (Hymenoptera) in New Zealand.

BACKGROUND: The introduction of species to new regions is occurring at an increasing rate. These introductions typically consist of species that are deliberately introduced for the purposes of biological control of pests or of species that are accidentally introduced through human-mediated transport networks.Understanding the potential and actual impacts of these introduced species requires comprehensive information on their geographic distributions and biological associations.However, apart from a few well-known case studies, such information is lacking for many introduced species which severely hinders further assessment of risks and impact. NEW INFORMATION: A dataset is provided on host associations, geographic distributions and dates of collection for both deliberately and accidentally-introduced parasitoid wasp species (Hymenoptera) in New Zealand. Information was obtained by digitising specimens from the New Zealand Arthropod Collection. Dates of records range from 1921 to 2017.The dataset includes 1265 specimen records, representing 127 parasitoid species from 12 families, with host records for 177 host species from 61 families and eight insect orders.These data provide baseline information to help evaluate the risk from introduced parasitoids to non-target and native species.

Notogaster, a new genus of Microgastrinae (Hymenoptera: Braconidae) from New Zealand.

A new genus of Microgastrinae parasitoid wasp endemic to New Zealand, Notogaster gen. nov. Fernández-Triana and Ward, is described, with ten new species: Notogaster avilai sp. nov., N. charlesi sp. nov., N. macdonaldae sp. nov., N. martini sp. nov., N. poultonae sp. nov., N. sucklingi sp. nov., N. toddae sp. nov., N. walkeri sp. nov., N. withersae sp. nov. and N. wornerae sp. nov. Based on some features, Notogaster resembles the genus Pholetesor Mason, although morphological and molecular data reveal they are not closely related. Notogaster is found throughout New Zealand, although many species are predominantly in the South Island. Species have been collected from a range of habitats, elevations, and collecting techniques. No host information is currently available.

Tersilochinae (Hymenoptera: Ichneumonidae) from New Zealand. Part 2. Review of genera Kiwi gen. nov. and Zealochus Khalaim.

In the second paper on New Zealand Tersilochinae (Ichneumonidae) we review two endemic New Zealand genera, Kiwi gen. nov. (eight species) and Zealochus Khalaim (three species). Nine species are described as new to science: Kiwi barrattae sp. nov., K. canterberus sp. nov., K. earlyi sp. nov., K. gronous sp. nov., K. oreteus sp. nov., K. ruzelus sp. nov., K. waitakerus sp. nov., Zealochus abominosus sp. nov. and Z. stepheni sp. nov. Zealochus gauldi Khalaim is transferred to Kiwi (comb. nov.). Identification keys to species of Kiwi and Zealochus occurring in New Zealand are provided.

Tersilochinae (Hymenoptera: Ichneumonidae) from New Zealand. Part 1. Generic key and three new genera.

The Tersilochinae (Ichneumonidae) from New Zealand are revised in part, with three new endemic genera and seventeen new species described: Aotearoazeus gen. nov., A. bullivantus sp. nov., A. coronetus sp. nov., A. probles sp. nov., Barycnellus gen. nov., B. aucklandellus sp. nov., B. conlisus sp. nov., B. cuvierensis sp. nov., B. globosus sp. nov., B. robustus sp. nov., Diaparsis zealandica sp. nov., Gauldiana gen. nov., G. arantia sp. nov., G. aspiringa sp. nov., G. dubia sp. nov., G. kaweka sp. nov., G. minuta sp. nov., G. nigra sp. nov., G. rotoitia sp. nov., and G. triangulata sp. nov. Zealochus postfurcalis is transferred to the genus Gauldiana (comb. nov.). The genus Diaparsis is recorded from New Zealand for the first time. Keys to genera and species of Tersilochinae occurring in New Zealand are provided.

Recurrent bridgehead effects accelerate global alien ant spread.

Biological invasions are a major threat to biological diversity, agriculture, and human health. To predict and prevent new invasions, it is crucial to develop a better understanding of the drivers of the invasion process. The analysis of 4,533 border interception events revealed that at least 51 different alien ant species were intercepted at US ports over a period of 70 years (1914-1984), and 45 alien species were intercepted entering New Zealand over a period of 68 years (1955-2013). Most of the interceptions did not originate from species' native ranges but instead came from invaded areas. In the United States, 75.7% of the interceptions came from a country where the intercepted ant species had been previously introduced. In New Zealand, this value was even higher, at 87.8%. There was an overrepresentation of interceptions from nearby locations (Latin America for species intercepted in the United States and Oceania for species intercepted in New Zealand). The probability of a species' successful establishment in both the United States and New Zealand was positively related to the number of interceptions of the species in these countries. Moreover, species that have spread to more continents are also more likely to be intercepted and to make secondary introductions. This creates a positive feedback loop between the introduction and establishment stages of the invasion process, in which initial establishments promote secondary introductions. Overall, these results reveal that secondary introductions act as a critical driver of increasing global rates of invasions.

Variation in the diversity and richness of parasitoid wasps based on sampling effort.

Parasitoid wasps are a mega-diverse, ecologically dominant, but poorly studied component of global biodiversity. In order to maximise the efficiency and reduce the cost of their collection, the application of optimal sampling techniques is necessary. Two sites in Auckland, New Zealand were sampled intensively to determine the relationship between sampling effort and observed species richness of parasitoid wasps from the family Ichneumonidae. Twenty traps were deployed at each site at three different times over the austral summer period, resulting in a total sampling effort of 840 Malaise-trap-days. Rarefaction techniques and non-parametric estimators were used to predict species richness and to evaluate the variation and completeness of sampling. Despite an intensive Malaise-trapping regime over the summer period, no asymptote of species richness was reached. At best, sampling captured two-thirds of parasitoid wasp species present. The estimated total number of species present depended on the month of sampling and the statistical estimator used. Consequently, the use of fewer traps would have caught only a small proportion of all species (one trap 7-21%; two traps 13-32%), and many traps contributed little to the overall number of individuals caught. However, variation in the catch of individual Malaise traps was not explained by seasonal turnover of species, vegetation or environmental conditions surrounding the trap, or distance of traps to one another. Overall the results demonstrate that even with an intense sampling effort the community is incompletely sampled. The use of only a few traps and/or for very short periods severely limits the estimates of richness because (i) fewer individuals are caught leading to a greater number of singletons; and (ii) the considerable variation of individual traps means some traps will contribute few or no individuals. Understanding how sampling effort affects the richness and diversity of parasitoid wasps is a useful foundation for future studies.

Global rise in emerging alien species results from increased accessibility of new source pools.

Our ability to predict the identity of future invasive alien species is largely based upon knowledge of prior invasion history. Emerging alien species-those never encountered as aliens before-therefore pose a significant challenge to biosecurity interventions worldwide. Understanding their temporal trends, origins, and the drivers of their spread is pivotal to improving prevention and risk assessment tools. Here, we use a database of 45,984 first records of 16,019 established alien species to investigate the temporal dynamics of occurrences of emerging alien species worldwide. Even after many centuries of invasions the rate of emergence of new alien species is still high: One-quarter of first records during 2000-2005 were of species that had not been previously recorded anywhere as alien, though with large variation across taxa. Model results show that the high proportion of emerging alien species cannot be solely explained by increases in well-known drivers such as the amount of imported commodities from historically important source regions. Instead, these dynamics reflect the incorporation of new regions into the pool of potential alien species, likely as a consequence of expanding trade networks and environmental change. This process compensates for the depletion of the historically important source species pool through successive invasions. We estimate that 1-16% of all species on Earth, depending on the taxonomic group, qualify as potential alien species. These results suggest that there remains a high proportion of emerging alien species we have yet to encounter, with future impacts that are difficult to predict.

Using spatially explicit surveillance models to provide confidence in the eradication of an invasive ant.

Effective detection plays an important role in the surveillance and management of invasive species. Invasive ants are very difficult to eradicate and are prone to imperfect detection because of their small size and cryptic nature. Here we demonstrate the use of spatially explicit surveillance models to estimate the probability that Argentine ants (Linepithema humile) have been eradicated from an offshore island site, given their absence across four surveys and three surveillance methods, conducted since ant control was applied. The probability of eradication increased sharply as each survey was conducted. Using all surveys and surveillance methods combined, the overall median probability of eradication of Argentine ants was 0.96. There was a high level of confidence in this result, with a high Credible Interval Value of 0.87. Our results demonstrate the value of spatially explicit surveillance models for the likelihood of eradication of Argentine ants. We argue that such models are vital to give confidence in eradication programs, especially from highly valued conservation areas such as offshore islands.

Metal Bioaccumulation by Estuarine Food Webs in New England, USA.

Evaluating the degree of metal exposure and bioaccumulation in estuarine organisms is important for understanding the fate of metals in estuarine food webs. We investigated the bioaccumulation of Hg, methylmercury (MeHg), Cd, Se, Pb, and As in common intertidal organisms across a watershed urbanization gradient of coastal marsh sites in New England to relate metal exposure and bioaccumulation in fauna to both chemical and ecological factors. In sediments, we measured metal and metalloid concentrations, total organic carbon (TOC) and SEM-AVS (Simultaneously extracted metal-acid volatile sulfides). In five different functional feeding groups of biota, we measured metal concentrations and delta 15N and delta 13C signatures. Concentrations of Hg and Se in biota for all sites were always greater than sediment concentrations whereas Pb in biota was always lower. There were positive relationships between biota Hg concentrations and sediment concentrations, and between biota MeHg concentrations and both pelagic feeding mode and trophic level. Bioavailability of all metals measured as SEM-AVS or Benthic-Sediment Accumulation Factor was lower in more contaminated sites, likely due to biogeochemical factors related to higher levels of sulfides and organic carbon in the sediments. Our study demonstrates that for most metals and metalloids, bioaccumulation is metal specific and not directly related to sediment concentrations or measures of bioavailability such as AVS-SEM.

Poles Apart: Comparing Trends of Alien Hymenoptera in New Zealand with Europe (DAISIE).

Developing generalisations of invasive species is an important part of invasion biology. However, trends and generalisations from one part of the world may not necessarily hold elsewhere. We present the first inventory and analysis of all Hymenoptera alien to New Zealand, and compare patterns from New Zealand with those previously published from Europe (DAISIE). Between the two regions there was broad correlation between families with the highest number of alien species (Braconidae, Encyrtidae, Pteromalidae, Eulophidae, Formicidae, Aphelinidae). However, major differences also existed. The number of species alien to New Zealand is higher than for Europe (334 vs 286), and major differences include: i) the much lower proportion of intentionally released species in New Zealand (21% vs 63% in Europe); and ii) the greater proportion of unintentionally introduced parasitoids in New Zealand (71.2% vs 22.6%). The disharmonic 'island' nature of New Zealand is shown, as a high proportion of families (36%) have no native representatives, and alien species also represent >10% of the native fauna for many other families. A much larger proportion of alien species are found in urban areas in New Zealand (60%) compared to Europe (~30%), and higher numbers of alien species were present earlier in New Zealand (especially <1950). Differences in the origins of alien species were also apparent. Unlike Europe, the New Zealand data reveals a change in the origins of alien species over time, with an increasing dominance of alien species from Australasia (a regional neighbour) during the past 25 years. We recommend that further effort be made towards the formation, and analysis, of regional inventories of alien species. This will allow a wider range of taxa and regions to be examined for generalisations, and help assess and prioritise the risk posed by certain taxa towards the economy or environment.

Microbial community structure in the gut of the New Zealand insect Auckland tree weta (Hemideina thoracica).

The endemic New Zealand weta is an enigmatic insect. Although the insect is well known by its distinctive name, considerable size, and morphology, many basic aspects of weta biology remain unknown. Here, we employed cultivation-independent enumeration techniques and rRNA gene sequencing to investigate the gut microbiota of the Auckland tree weta (Hemideina thoracica). Fluorescence in situ hybridisation performed on different sections of the gut revealed a bacterial community of fluctuating density, while rRNA gene-targeted amplicon pyrosequencing revealed the presence of a microbial community containing high bacterial diversity, but an apparent absence of archaea. Bacteria were further studied using full-length 16S rRNA gene sequences, with statistical testing of bacterial community membership against publicly available termite- and cockroach-derived sequences, revealing that the weta gut microbiota is similar to that of cockroaches. These data represent the first analysis of the weta microbiota and provide initial insights into the potential function of these microorganisms.

Modelling the impacts of an invasive species across landscapes: a step-wise approach.

We estimate the extent of ecological impacts of the invasive Asian paper wasp across different landscapes in New Zealand. We used: (i) a baseline distribution layer (modelled via MaxEnt); (ii) Asian paper wasp nest density (from >460 field plots, related to their preferences for specific land cover categories); and (iii) and their foraging intensity (rates of foraging success, and the time available to forage on a seasonal basis). Using geographic information systems this information is combined and modelled across different landscapes in New Zealand in a step-wise selection process. The highest densities of Asian paper wasps were in herbaceous saline vegetation, followed closely by built-up areas, and then scrub and shrubland. Nest densities of 34 per ha, and occupancy rates of 0.27 were recorded for herbaceous saline vegetation habitats. However, the extent of impacts of the Asian paper wasp remains relatively restricted because of narrow climate tolerances and spatial restriction of preferred habitats. A step-wise process based on geographic information systems and species distribution models, in combination with factors such as distribution, density, and predation, create a useful tool that allows the extent of impacts of invasive species to be assessed across large spatial scales. These models will be useful for conservation managers as they provide easy visual interpretation of results, and can help prioritise where direct conservation action or control of the invader are required.

Benthic and pelagic pathways of methylmercury bioaccumulation in estuarine food webs of the northeast United States.

Methylmercury (MeHg) is a contaminant of global concern that bioaccumulates and bioamagnifies in marine food webs. Lower trophic level fauna are important conduits of MeHg from sediment and water to estuarine and coastal fish harvested for human consumption. However, the sources and pathways of MeHg to these coastal fisheries are poorly known particularly the potential for transfer of MeHg from the sediment to biotic compartments. Across a broad gradient of human land impacts, we analyzed MeHg concentrations in food webs at ten estuarine sites in the Northeast US (from the Hackensack Meadowlands, NJ to the Gulf of Maine). MeHg concentrations in water column particulate material, but not in sediments, were predictive of MeHg concentrations in fish (killifish and Atlantic silversides). Moreover, MeHg concentrations were higher in pelagic fauna than in benthic-feeding fauna suggesting that MeHg delivery to the water column from methylation sites from within or outside of the estuary may be an important driver of MeHg bioaccumulation in estuarine pelagic food webs. In contrast, bulk sediment MeHg concentrations were only predictive of concentrations of MeHg in the infaunal worms. Our results across a broad gradient of sites demonstrate that the pathways of MeHg to lower trophic level estuarine organisms are distinctly different between benthic deposit feeders and forage fish. Thus, even in systems with contaminated sediments, transfer of MeHg into estuarine food webs maybe driven more by the efficiency of processes that determine MeHg input and bioavailability in the water column.

Molecular identification of the prey range of the invasive Asian paper wasp.

The prey range of the invasive Asian paper wasp, Polistes chinensis antennalis, was studied using molecular diagnostics. Nests of paper wasps were collected from urban residential and salt marsh habitats, larvae were removed and dissected, and DNA in the gut of the paper wasp larvae was amplified and sequenced with cytochrome c oxidase subunit I (COI). Seventy percent of samples (211/299) yielded medium-to high-quality sequences, and prey identification was achieved using BLAST searches in BOLD. A total of 42 taxa were identified from 211 samples. Lepidoptera were the majority of prey, with 39 taxa from 91% of samples. Diptera was a relatively small component of prey (three taxa, 19 samples). Conclusive species-level identification of prey was possible for 67% of samples, and genus-level identification, for another 12% of samples. The composition of prey taken was different between the two habitats, with 2.5× more native prey species being taken in salt marsh compared with urban habitats. The results greatly extend the prey range of this invasive species. The technique is a more effective and efficient approach than relying on the collection of "prey balls", or morphological identification of prey, for the study of paper wasps.

Experimental and natural warming elevates mercury concentrations in estuarine fish.

Marine food webs are the most important link between the global contaminant, methylmercury (MeHg), and human exposure through consumption of seafood. Warming temperatures may increase human exposure to MeHg, a potent neurotoxin, by increasing MeHg production as well as bioaccumulation and trophic transfer through marine food webs. Studies of the effects of temperature on MeHg bioaccumulation are rare and no study has specifically related temperature to MeHg fate by linking laboratory experiments with natural field manipulations in coastal ecosystems. We performed laboratory and field experiments on MeHg accumulation under varying temperature regimes using the killifish, Fundulus heteroclitus. Temperature treatments were established in salt pools on a coastal salt marsh using a natural temperature gradient where killifish fed on natural food sources. Temperatures were manipulated across a wider range in laboratory experiments with killifish exposed to MeHg enriched food. In both laboratory microcosms and field mesocosms, MeHg concentrations in killifish significantly increased at elevated temperatures. Moreover, in field experiments, other ancillary variables (salinity, MeHg in sediment, etc.) did not relate to MeHg bioaccumulation. Modeling of laboratory experimental results suggested increases in metabolic rate as a driving factor. The elevated temperatures we tested are consistent with predicted trends in climate warming, and indicate that in the absence of confounding factors, warmer sea surface temperatures could result in greater in bioaccumulation of MeHg in fish, and consequently, increased human exposure.

Do low-mercury terrestrial resources subsidize low-mercury growth of stream fish? Differences between species along a productivity gradient.

Low productivity in aquatic ecosystems is associated with reduced individual growth of fish and increased concentrations of methylmercury (MeHg) in fish and their prey. However, many stream-dwelling fish species can use terrestrially-derived food resources, potentially subsidizing growth at low-productivity sites, and, because terrestrial resources have lower MeHg concentrations than aquatic resources, preventing an increase in diet-borne MeHg accumulation. We used a large-scale field study to evaluate relationships among terrestrial subsidy use, growth, and MeHg concentrations in two stream-dwelling fish species across an in-stream productivity gradient. We sampled young-of-the-year brook trout (Salvelinus fontinalis) and Atlantic salmon (Salmo salar), potential competitors with similar foraging habits, from 20 study sites in streams in New Hampshire and Massachusetts that encompassed a wide range of aquatic prey biomass. Stable isotope analysis showed that brook trout used more terrestrial resources than Atlantic salmon. Over their first growing season, Atlantic salmon tended to grow larger than brook trout at sites with high aquatic prey biomass, but brook grew two-fold larger than Atlantic salmon at sites with low aquatic prey biomass. The MeHg concentrations of brook trout and Atlantic salmon were similar at sites with high aquatic prey biomass and the MeHg concentrations of both species increased at sites with low prey biomass and high MeHg in aquatic prey. However, brook trout had three-fold lower MeHg concentrations than Atlantic salmon at low-productivity, high-MeHg sites. These results suggest that differential use of terrestrial resource subsidies reversed the growth asymmetry between potential competitors across a productivity gradient and, for one species, moderated the effect of low in-stream productivity on MeHg accumulation.