Isolation and characterization of 8 microsatellite loci for the ‘‘killer shrimp’’, an invasive Ponto-Caspian amphipod Dikerogammarus villosus (Crustacea: Amphipoda)

Dikerogammarus villosus is a freshwater amphipod of the Ponto-Caspian origin recognized as one of the 100 worst alien species in Europe, having negative impact on biodiversity and functioning of the invaded aquatic ecosystems. The species has a wide ecophysiological tolerance and during the last 20 years it has rapidly spread throughout European inland waters. In consequence, it presents a major conservation management problem. We describe eight polymorphic microsatellite loci developed for D. villosus by combining a biotin-enrichment protocol and new generation 454GS-FLX Titanium pyrosequencing technology. When genotyped in 64 individuals from two locations, the loci exhibited a mean diversity of 4.87 alleles per locus (2–13). The mean observed and expected heterozygosities were, respectively, 0.439 (0.091–0.844) and 0.468 (0.089–0.843). Gametic disequilibrium was not detected for any pair of loci. The microsatellite markers will be a valuable tool in assessing the demographic processes associated with invasion of the killer shrimp from a genetic point of view

‘Candidatus Aquirickettsiella gammari’ (Gammaproteobacteria: Legionellales: Coxiellaceae): A bacterial pathogen of the freshwater crustacean Gammarus fossarum (Malacostraca: Amphipoda)

Invasive and non-native species can pose risks to vulnerable ecosystems by co-introducing bacterial pathogens. Alternatively, co-introduced bacterial pathogens may regulate invasive population size and invasive traits. We describe a novel candidate genus and species of bacteria (‘Candidatus Aquirickettsiella gammari’) found to infect Gammarus fossarum, from its native range in Poland. The bacterium develops intracellularly within the haemocytes and cells of the musculature, hepatopancreas, connective tissues, nervous system and gonad of the host. The developmental cycle of ‘Candidatus Aquirickettsiella gammari’ includes an elementary body (496.73 nm ± 37.56 nm in length, and 176.89 nm ± 36.29 nm in width), an elliptical, condensed spherical stage (737.61 nm ± 44.51 nm in length and 300.07 nm ± 44.02 nm in width), a divisional stage, and a spherical initial body (1397.59 nm ± 21.26 nm in diameter). We provide a partial genome for ‘Candidatus Aquirickettsiella gammari’, which clades phylogenetically alongside environmental 16S rRNA sequences from aquatic habitats, and bacterial symbionts from aquatic isopods (Asellus aquaticus), grouping separately from the Rickettsiella, a genus that includes bacterial pathogens of terrestrial insects and isopods. Increased understanding of the diversity of symbionts carried by G. fossarum identifies those that might regulate host population size, or those that could pose a risk to native species in the invasive range. Identification of ‘Candidatus Aquirickettsiella gammari’ and its potential for adaptation as a biological control agent is explored

Histopathological screening of Pontogammarus robustoides (Amphipoda), an invader on route to the United Kingdom

Biological invasions may act as conduits for pathogen introduction. To determine which invasive non-native species pose the biggest threat, we must first determine the symbionts (pathogens, parasites, commensals, mutualists) they carry, via pathological surveys that can be conducted in multiple ways (i.e., molecular, pathological, and histological). Whole animal histopathology allows for the observation of pathogenic agents (virus to Metazoa), based on their pathological effect upon host tissue. Where the technique cannot accurately predict pathogen taxonomy, it does highlight pathogen groups of importance. This study provides a histopathological survey of Pontogammarus robustoides (invasive amphipod in Europe) as a baseline for symbiont groups that may translocate to other areas/hosts in future invasions

Establishment of a taxonomic and molecular reference collection to support the identification of species regulated by the Western Australian Prevention List for Introduced Marine Pests

Introduced Marine Pests (IMP, = non-indigenous marine species) prevention, early detection and risk-based management strategies have become the priority for biosecurity operations worldwide, in recognition of the fact that, once established, the effective management of marine pests can rapidly become cost prohibitive or impractical. In Western Australia (WA), biosecurity management is guided by the “Western Australian Prevention List for Introduced Marine Pests” which is a policy tool that details species or genera as being of high risk to the region. This list forms the basis of management efforts to prevent introduction of these species, monitoring efforts to detect them at an early stage, and rapid response should they be detected. It is therefore essential that the species listed can be rapid and confidently identified and discriminated from native species by a range of government and industry stakeholders. Recognising that identification of these species requires very specialist expertise which may be in short supply and not readily accessible in a regulatory environment, and the fact that much publicly available data is not verifiable or suitable for regulatory enforcement, the WA government commissioned the current project to collate a reference collection of these marine pest specimens. In this work, we thus established collaboration with researchers worldwide in order to source representative specimens of the species listed. Our main objective was to build a reference collection of taxonomically vouchered specimens and subsequently to generate species-specific DNA barcodes suited to supporting their future identification. To date, we were able to obtain specimens of 75 species (representative of all but four of the pests listed) which have been identified by experts and placed with the WA Government Department of Fisheries and, where possible, in accessible museums and institutions in Australasia. The reference collection supports the fast and reliable taxonomic and molecular identification of marine pests in WA and constitutes a valuable resource for training of stakeholders with interest in IMP recognition in Australia. The reference collection is also useful in supporting the development of a variety of DNA-based detection strategies such as real-time PCR and metabarcoding of complex environmental samples (e.g. biofouling communities). ThePrevention List is under regular review to ensure its continued relevance and that it remains evidence and risk-based. Similarly, its associated reference collection also remains to some extent a work in progress. In recognition of this fact, this report seeks to provide details of this continually evolving information repository publicly available to the biosecurity management community worldwid

Parasites, pathogens and commensals in the “low-impact” non-native amphipod host Gammarus roeselii

Background: Whilst vastly understudied, pathogens of non-native species (NNS) are increasingly recognised as important threats to native wildlife. This study builds upon recent recommendations for improved screening for pathogens in NNS by focusing on populations of Gammarus roeselii in Chojna, north-western Poland. At this location, and in other parts of continental Europe, G. roeselii is considered a well-established and relatively ‘low-impact’ invader, with little understanding about its underlying pathogen profile and even less on potential spill-over of these pathogens to native species. Results: Using a combination of histological, ultrastructural and phylogenetic approaches, we define a pathogen profile for non-native populations of G. roeselii in Poland. This profile comprised acanthocephalans (Polymorphus minutus Goese, 1782 and Pomphorhynchus sp.), digenean trematodes, commensal rotifers, commensal and parasitic ciliated protists, gregarines, microsporidia, a putative rickettsia-like organism, filamentous bacteria and two viral pathogens, the majority of which are previously unknown to science. To demonstrate potential for such pathogenic risks to be characterised from a taxonomic perspective, one of the pathogens, a novel microsporidian, is described based upon its pathology, developmental cycle and SSU rRNA gene phylogeny. The novel microsporidian Cucumispora roeselii n. sp. displayed closest morphological and phylogenetic similarity to two previously described taxa, Cucumispora dikerogammari Ovcharenko, 2010 and Cucumispora ornata Bojko, 2015. Conclusions: In addition to our discovery extending the host range for the genus Cucumispora Ovcharenko, 2010 outside of the amphipod host genus Dikerogammarus Stebbing, we reveal significant potential for the co-transfer of (previously unknown) pathogens alongside this host when invading novel locations. This study highlights the importance of pre-invasion screening of low-impact NNS and, provides a means to document and potentially mitigate the additional risks posed by previously unknown pathogens

Alien pathogens on the horizon: opportunities for predicting their threat to wildlife

According to the Convention on Biological Diversity, by 2020 invasive alien species (IAS) should be identified and their impacts assessed, so that species can be prioritized for implementation of appropriate control strategies and measures put in place to manage invasion pathways. For one quarter of the IAS listed as the “100 of the world's worst” environmental impacts are linked to diseases of wildlife (undomesticated plants and animals). Moreover, IAS are a significant source of “pathogen pollution” defined as the human-mediated introduction of a pathogen to a new host or region. Despite this, little is known about the biology of alien pathogens and their biodiversity impacts after introduction into new regions. We argue that the threats posed by alien pathogens to endangered species, ecosystems, and ecosystem services should receive greater attention through legislation, policy, and management. We identify 10 key areas for research and action, including those relevant to the processes of introduction and establishment of an alien pathogen and to prediction of the spread and associated impact of an alien pathogen on native biota and ecosystems. The development of interdisciplinary capacity, expertise, and coordination to identify and manage threats was seen as critical to address knowledge gaps

What doesn’t kill you doesn’t make you stronger: Parasites modify interference competition between two invasive amphipods

We used a freshwater amphipod-microsporidian model (Ponto-Caspian hosts: Dikerogammarus villosus and D. haemobaphes, parasite: Cucumispora dikerogammari) to check whether parasites affect biological invasions by modulating behaviour and intra- and interspecific interactions between the invaders. We tested competition for shelter in conspecific and heterospecific male pairs (one or both individuals infected or non-infected). In general, amphipods of both species increased their shelter occupancy time when accompanied by infected rather than non-infected conspecifics and heterospecifics. Infected amphipods faced lower aggression from non-infected conspecifics. Moreover, D. villosus was more aggressive than D. haemobaphes and more aggressive towards conspecifics vs. heterospecifics. In summary, infection reduced the intra- and interspecific competitivity of amphipods, which became less capable of defending their shelters, despite their unchanged need for shelter occupancy. Dikerogammarus haemobaphes, commonly considered as a weaker competitor, displaced by D. villosus from co-occupied locations, was able to compete efficiently for the shelter with D. villosus when microsporidian infections appeared on the scene. This suggests that parasites may be important mediators of biological invasions, facilitating the existence of large intra- and interspecific assemblages of invasive alien amphipods