Border Biosecurity Research Articles

Efficient border biosecurity inspection leverages superspreading to reduce biological invasion risk.

Biological invasions are a growing threat to biodiversity, food security, and economies. Rising pressure from increased global trade requires improving border inspection efficiency. Here, we depart from the conventional consignment-by-consignment approach advocated in current inspection standards. Instead, we suggest a broader perspective: evaluating border inspection regimes based on their ability to reduce propagule pressure across entire pathways. Additionally, we demonstrate that most biosecurity pathways exhibit superspreading behavior, that is, consignments from the same pathway have varying infestation rates and contain rare right-tail events (also called overdispersion). We show that greater overdispersion leads to more pronounced diminishing returns, with consequences on the optimal allocation of sampling effort. We leverage these two insights to develop a simple and efficient border inspection regime that can significantly reduce propagule pressure compared to current standards. Our analysis revealed that consignment size is a key driver of biosecurity risk and that sampling proportional to the square root of consignment size is near optimal. In testing, our framework reduced propagule pressure by 31 to 38% compared to current standards. We also identified opportunities to further improve inspection efficiency by considering additional pathway characteristics (i.e., overdispersion parameters, zero inflation, relative risk, sampling cost, detectability) and developed solutions for these more complex scenarios. We anticipate our result will mitigate biological invasion risk with significant implications for biodiversity conservation, food security, and economiesworldwide.

Now you see me, now you don't: verifying the absence of alien invasive yellow crazy ant Anoplolepis gracilipes in South Africa.

Anoplolepis gracilipes is an invasive species that is a major threat to native ecosystems worldwide. It has been listed as one of the top 100 worst invasive species in the world and is well known for its negative impact on native arthropods and some vertebrates. This study aimed to confirm the presence or absence of A. gracilipes in some major South African harbours. We did so by surveying four harbours in the Western Cape and KwaZulu-Natal provinces, using pitfall trapping, yellow pan traps, and baiting. In addition, ant collections from Iziko Museums of South Africa (Cape Town, South Africa), University of KwaZulu-Natal (Pietermaritzburg campus, South Africa), Iimbovane Outreach Project (Stellenbosch University, South Africa), and AfriBugs CC (Pretoria, South Africa) were examined for specimens of A. gracilipes. The invasive species A. gracilipes was not detected from any of the sampled harbours during this study, nor in the main ant collections in South Africa. The only, and potentially erroneous published record of A. gracilipes in South Africa, is from Durban harbour and subsequent possibly erroneous citizen science observations are from other coastal sites such as Gansbaai, Knysna, Table Bay, and Kalk Bay. This is a positive outcome for conservation authorities as this species is highly invasive and, if introduced, will likely outcompete native fauna and result in ecosystem collapse. Although A. gracilipes was not detected in the samples from this study, early detection and eradication of this species should be prioritised. This can be achieved through existing pest monitoring programs at harbours, and continued border biosecurity measures.

A snapshot of online wildlife trade: Australian e-commerce trade of native and non-native pets

The international trade of non-domesticated pets impacts both conservation and biosecurity via the harvest and release of live animals beyond their native distributions. The extent to which individual countries mitigate these impacts via regulation of trade is inconsistent, as is their capacity to monitor internet facilitated trade. We investigated the online trade of vertebrate pets within Australia, a country with a reputation for relatively stringent pet-importation regulations and world-class border biosecurity. Using semi-automated data mining (i.e., webscraping) techniques, we collected online pet trade data over the course of 14 weeks from 12 Australian e-commerce platforms selected using an a priori set of search terms. We analysed spatial, temporal and taxonomic biases in trade and identified instances of high rates of trade in: (i) threatened species, (ii) non-native species, (iii) and species not permissible for live import. We identified over 100,000 individual live animals across 1192 species, including: 667 non-native species for sale within Australia from 03/12/2019 to 20/03/2020 (mammals were excluded from our analysis). Our findings constitute a much greater scale (in terms of abundance and richness) of non-native species trade than previously recorded in Australia. Substantial changes to legislative control of domestically traded pets are needed at the national level to reduce the volume of non-native pets that may contribute to the establishment of invasive species in Australia. We suggest that contemporary examples of permit systems applied to native taxa may provide a valuable template for the implementation of such changes.

Modernization of National Border Biosecurity Defense System

Modernization of National Border Biosecurity Defense System

Prioritization of invasive alien species with the potential to threaten agriculture and biodiversity in Kenya through horizon scanning

Invasive alien species (IAS) rank among the most significant drivers of species extinction and ecosystem degradation resulting in significant impacts on socio-economic development. The recent exponential spread of IAS in most of Africa is attributed to poor border biosecurity due to porous borders that have failed to prevent initial introductions. In addition, countries lack adequate information about potential invasions and have limited capacity to reduce the risk of invasions. Horizon scanning is an approach that prioritises the risks of potential IAS through rapid assessments. A group of 28 subject matter experts used an adapted methodology to assess 1700 potential IAS on a 5-point scale for the likelihood of entry and establishment, potential socio-economic impact, and impact on biodiversity. The individual scores were combined to rank the species according to their overall potential risk for the country. Confidence in individual and overall scores was recorded on a 3-point scale. This resulted in a priority list of 120 potential IAS (70 arthropods, 9 nematodes, 15 bacteria, 19 fungi/chromist, 1 viroid, and 6 viruses). Options for risk mitigation such as full pest risk analysis and detection surveys were suggested for prioritised species while species for which no immediate action was suggested, were added to the plant health risk register and a recommendation was made to regularly monitor the change in risk. By prioritising risks, horizon scanning guides resource allocation to interventions that are most likely to reduce risk and is very useful to National Plant Protection Organisations and other relevant stakeholders.

Horizon scanning for prioritising invasive alien species with potential to threaten agriculture and biodiversity in Ghana

Invasive alien species (IAS) continue to shape the global landscape through their effects on biological diversity and agricultural productivity. The effects are particularly pronounced in Sub-Saharan Africa, which has seen the arrival of many IAS in recent years. This has been attributed to porous borders, weak cross border biosecurity, and inadequate capacity to limit or stop invasions. Prediction and early detection of IAS, as well as mechanisms of containment and eradication, are needed in the fight against this global threat. Horizon scanning is an approach that enables gathering of information on risk and impact that can support IAS management. A study was conducted in Ghana to establish two ranked lists of potential invasive alien plant pest species that could be harmful to agriculture, forestry, and the environment, and to rank them according to their potential threat. The ultimate objective was to enable prioritization of actions including pest risk analysis, prevention, surveillance and contingency plans. Prioritisation was carried out using an adapted version of horizon scanning and consensus methods developed for ranking IAS worldwide. Following a horizon scan of invasive alien species not yet officially present in Ghana, a total of 110 arthropod and 64 pathogenic species were assessed through a simplified pest risk assessment. Sixteen species, of which 14 were arthropods and two pathogens, had not been recorded on the African continent at the time of assessment. The species recorded in Africa included 19 arthropod and 46 pathogenic species which were already recorded in the neighbouring countries of Burkina Faso, Côte d’Ivoire, and Togo. The majority of arthropod species were likely to arrive as contaminants on commodities, followed by a sizable number which were likely to arrive as stowaways, while some species were capable of long distance dispersal unaided. The main actions suggested for species that scored highly included full pest risk analyses and, for species recorded in neighbouring countries, surveys to determine their presence in Ghana were recommended.

Idea paper: Monitoring and databasing nonnative species to manage establishment debt in aquatic ecosystems

AbstractIn the era of globalization, biological invasions are one of the most serious social issues. Thus, managing its impact is an urgent task. It is essential to control nonnative species before they become established. However, it is insufficient to address establishment debt, which occurs when a nonnative species has been introduced into an area but has not yet been established in the wild. In particular, unintentionally introduced or contaminated organisms of the aquatic ornamental pet trade are referred to as “hitchhikers” and have not received much attention in the context of establishment debt. To understand the nature of establishment debt, including that of aquatic hitchhikers, I propose the monitoring of nonnative species inhabiting artificial isolated waters, such as indoor aquariums, and the construction of a database using environmental DNA metabarcoding. This idea would be an effective nonregulatory management approach when implemented broadly, at the country level. Furthermore, implementation of this strategy in combination with border biosecurity and field monitoring may promote accurate prioritization, rapid species identification, and effective invasion pathway assessment.

Considering unseen arrivals in predictions of establishment risk based on border biosecurity interceptions.

Assessing species establishment risk is an important task used for informing biosecurity activities aimed at preventing biological invasions. Propagule pressure is a major contributor to the probability of invading species establishment; however, direct assessment of numbers of individuals arriving is virtually never possible. Inspections conducted at borders by biosecurity officials record counts of species (or higher-level taxa) intercepted during inspections, which can be used as proxies for arrival rates. Such data may therefore be useful for predicting species establishments, though some species may establish despite never being intercepted. We present a stochastic process-based model of the arrival-interception-establishment process to predict species establishment risk from interception count data. The model can be used to estimate the probability of establishment, both for species that were intercepted and species that had no interceptions during a given observation period. We fit the stochastic model to data on two insect families, Cerambycidae and Aphididae, that were intercepted and/or established in the United States or New Zealand. We also explore the effects of variation in model parameters and the inclusion of an Allee effect in the establishment probability. Although interception data sets contain much noise due to variation in inspection policy, interception effort and among-species differences in detectability, our study shows that it is possible to use such data for predicting establishments and distinguishing differences in establishment risk profile between taxonomic groups. Our model provides a method for predicting the number of species that have breached border biosecurity, including both species detected during inspections but also "unseen arrivals" that have never been intercepted, but have not yet established a viable population. These estimates could inform prioritization of different taxonomic groups, pathways or identification effort in biosecurity programs.

Biosecurity risk mapping and gap analysis in South East Asia

IntroductionIn today’s globalized world where travel is commonplace, a threat in one region can easily spread throughout the whole world. It is, therefore, essential for multi-disciplinary risk assessment, risk mitigation, and collaborative strategies to take place among various stakeholders to mitigate this. Any strategic plan to deal with biosecurity, therefore, needs to be a complete loop, top to bottom and bottom to top. This paper describes the results of Project 62, which involved mapping and biosecurity risk assessment in South East Asia. Materials and methodsThe mapping and biosecurity risk assessment activities for the participating partner countries was carried out in two phases. The first phase involved risk assessment by six partner-country bio-experts for their own countries and the second phase involved conducting a joint Strengths, Weaknesses, Opportunities, and Threats (SWOT) analysis and risk assessment for the South East Asian region as whole, together with international bio-experts at a workshop. Results and discussionThe following key recommendations of the project show that biosecurity needs to be addressed through stakeholder engagement at multiple levels starting from the top echelons of the government to the worker who needs to recognize and understand the threats they might face: 1.Systematic analysis of existing information from BWC ISU Article 10, IHR capacities, JEE, UN Security Council Resolution 1540 Action Plan, EU CBRN NAP, and other relevant sources to develop a common understanding of the definition of biosecurity for all stakeholders in the region.2.Creation of programs for awareness building in biosecurity, not limiting them to laboratory biosecurity but using an all-inclusive approach to include border biosecurity, pandemic response, etc.3.Creation of a country-specific list of high-risk biological materials.4.Compulsory standardized biosecurity training, including responsible conduct in biosecurity research.5.Framework for one-health and security interface addressing biosecurity threats.6.Framework for ensuring information security in relation to biosecurity.7.Involvement of local/regional experts in the implementation of EU-CBRN CoE projects.

First report of luminous stimuli eliciting sound production in weevils.

Light-based stimuli elicited acoustic responses in male Hylesinus aculeatus Say (Curculionidae: Scolytinae: Hylesinina) instantaneously, with 100% reliability. Stridulations were elicited with a white light beam in a dark environment and recorded with an ultrasonic microphone. Acoustic responses were consistent, and, when compared with sounds produced under stressful conditions (i.e. physical stimulation), no significant differences were found. Hylesinus aculeatus possess an elytro-tergal stridulatory organ and acoustic communication is only present in males. This is also the first report of acoustic communication for this species. Instantaneous light-elicited acoustic communication has potential applications in the development of electronic traps and real-time acoustic detection and identification of beetles, border biosecurity, and noise-reduction in acoustic data collection.

Volatile release, mobility, and mortality of diapausing Halyomorpha halys during simulated shipping movements and temperature changes

The brown marmorated stink bug, Halyomorpha halys, is an agricultural and urban pest that has become widely established as an invasive species of major concern in the USA and throughout Europe. The insect forms large aggregations before entering diapause, and it is often these aggregations that are found by biosecurity officials conducting inspections of internationally shipped freight. Here, we aimed to gain insights into the potential volatile emissions, mobility, and mortality rates of the insects after undergoing a ship voyage across the equator (i.e. from northern hemisphere winter to southern hemisphere summer). Experiments were performed to simulate the effects on aggregations of diapausing H. halys of two variables associated with trans-Pacific voyages, i.e. ship and container movement and temperature changes within containers. Thus, H. halys aggregations were exposed to simulated shipping movement, using a 6-axis VS-6577G-B Denso robot arm, and emission of defence volatiles was measured repeatedly during the “voyage.” We also simulated temperature changes as they occur during a 26-day voyage from Baltimore, northeast USA, to Auckland, New Zealand, and assessed the effects on the mortality and mobility of the bugs. Aggregations that experienced movement were not found to be any more likely to release volatiles than stationary aggregations and neither did the movement affect bug mobility. Temperature changes had a significant effect on mobility. However, towards the end of the temperature simulation, most H. halys had died, probably from a lack of food and moisture. These findings are highly relevant for border biosecurity risk assessment and pathway risk management.

An Australian dog diagnosed with an exotic tick-borne infection: should Australia still be considered free from Hepatozoon canis?

Recent molecular and sero-surveillance studies of the tick-borne pathogen Hepatozoon canis have identified new hosts, potential vector species, and have revealed that H. canis is more widespread than previously thought. We report the first diagnosed case of canine hepatozoonosis in Australia from a Maremma Sheepdog in Sarina, Queensland. Hepatozoon canis was detected with blood smear examination and 18S rRNA sequencing. It is unknown when or how the organism was introduced into Australia, which raises questions about border biosecurity policies and the H. canis infection status of its potential vectors and hosts in Australia. Surveillance for this pathogen is required to determine whether H. canis has established in Australia.

New Zealand Plant Protection Medal 2017

This medal is awarded by the New Zealand Plant Protection Society to honour those who have made exceptional contributions to plant protection in New Zealand in the widest sense. The medal is awarded for outstanding services to plant protection, whether through research, education, implementation or leadership.
 In 2017, the New Zealand Plant Protection Medal was awarded Prof David Maxwell (Max) Suckling. In his 35+ years of research, Max has been pivotal in bringing odour-based technologies to New Zealand. In particular, his research on insect pheromones has enabled integrated pest management to be realised in this country. As a result, New Zealand plant-based industries can access premium overseas markets due to the low pest prevalence and low pesticide residue on primary produce. It has also reduced grower exposure to pesticides.
 The acknowledgement of his broad knowledge of risks and benefits that new organisms and substances can pose to New Zealand has been exemplified by his position of Chair of the Environmental Risk Management Authority Hazardous Substances and New Organisms Committee where he presided over decisions from determining which organisms are new to New Zealand through to whether the benefits of the release of new organisms outweighed the risks. His unique knowledge and ability to make sound judgements based on the evidence presented also led to two invitations back to the Environmental Protection Authority after he had finished as a special member on the Committee so that New Zealand could safely continue to assess the use of biological control agents.
 Since 2004, in his role as Science Group Leader of the Biosecurity group at The New Zealand Institute for Plant and Food Research Ltd (PFR), Max has been instrumental in developing tools to improve detection sensitivity and socially acceptable eradication options for new pests that threaten New Zealand, such as the Queensland fruit fly and the painted apple moth.
 Max has been a member of the New Zealand Plant Protection Society for many years and served as President from 1999 to 2001. He was nominated for the Medal because of the passion he has displayed towards developing and making available socially acceptable pest eradication and management tools in New Zealand. The work that he and the chemical ecology team he has built and led, has had a large impact in many sectors from horticulture to biosecurity.
 This use of socially acceptable tools for the productive sectors naturally led to Max’s involvement in the pest surveillance and eradication space. He led the Eradication and Response Theme in the Better Border Biosecurity collaboration for over ten years, co-ordinating research among Crown Research Institutes to achieve their goals and the goals of New Zealand’s biosecurity practitioners. He has gone beyond odour-based technologies and branched into sound, vision and sterile-insect technologies for managing pests, sticking with the social acceptance theme.
 Max is an innovative thinker, testing novel approaches for pest management, and can bring quite separate groups together to achieve a goal. For example, he combined an irradiator used to sterilise medical equipment and insect rearing to achieve a boutique insect-sterilisation programme against the painted apple moth. By pushing the envelope, he is seen as a world leader in his field of using socially acceptable tools, with numerous invitations as a keynote speaker at international meetings, which has allowed him to return to New Zealand with some of the latest scientific ideas.
 He has served on working groups of the sterile-insect technique for the joint division of the Food and Agriculture Organization/International Atomic Energy Agency. He was recently made a professor when he was made a joint appointment at PFR and the University of Auckland, and has supervised and co-supervised a number of PhD and MSc students.
 His outstanding collaboration and mentoring skills enable him to work across different fields, secure new knowledge and tools for novel pest-management approaches, bring together people from different organisations, and mentor ‘thinking-out-of-the-square’ scientists for the future.
 His desire to protect New Zealand’s flora, fauna and people, make him a worthy recipient of the New Zealand Plant Protection Medal.
 
 NZPP Medal recipients for the previous five years:
 2016: Rob Beresford
 2015: Gary Barker
 2014: -
 2013: Andrew Hodson
 2012: Margaret Dick

Establishment patterns of non-native insects in New Zealand

Insects comprise the majority of non-native animal species established around the world. However, geographic biases in knowledge hamper an overall understanding of biological invasions globally. A dataset of accidentally introduced non-native insect species established in New Zealand was compiled from databases, entomological literature, and examination of specimens in the New Zealand Arthropod Collection. For each non-native species, the first recorded location and first recorded date of detection was obtained. Excluding intentionally introduced species, there are 1477 non-native insect species successfully established in New Zealand across 16 orders, 234 families and 1017 genera. Four orders (Coleoptera, Hemiptera, Hymenoptera and Diptera) contributed 77.5% of all established insect species. Herbivores represented the largest feeding guild (47.7%), comprised of polyphagous (48.3%) or oligophagous (39.7%) species. The majority of these species originated in the Australasian (36.7%) and Palearctic regions (24.8%). Regression trees, using a binary recursive partitioning approach, found the number of international tourist arrivals, exotic vegetation cover, and regional gross domestic product were the main factors explaining spatial patterns of recently established species. Gross domestic product best explained temporal patterns of establishment over the last century. Our findings demonstrate that broad-scale analyses of non-native species have important applications for border biosecurity by providing insight into the extent of invasions. In New Zealand, the current trajectory indicates fewer non-native species are establishing annually, suggesting biosecurity efforts are being effective at reducing rates of establishment.

A new and highly effective male lure for the guava fruit fly Bactrocera correcta

Bactrocera correcta is an economically important and invasive fruit fly pest. As part of the quest to develop new chemical attractants for potential control of this species, we had previously detected a large amount of β-caryophyllene (CAR) in the pheromone glands of wild B. correcta males. This compound was found in addition to the metabolites from consumed methyl eugenol (ME), a common male lure used for many Bactrocera species. We conducted bioassays to compare the attractiveness of CAR and ME to male B. correcta, determine the behavioural and chemo-ecological basis of the attraction and to monitor their trapping efficacy. Males of B. correcta responded strongly to CAR at a much younger age before sexual maturity compared to ME. CAR feeding increased the male mating success in comparison with lure-deprived males, and CAR-fed males were equally competitive as ME-fed males when competing for conspecific females in semi-field trials. Field trapping data also revealed that ME-baited traps captured both B. dorsalis and B. correcta, while CAR-baited traps brought in only B. correcta males. Numbers of total flies trapped per trap per day (FTD) were also significantly higher in traps baited with CAR compared to ME-baited traps, capturing up to seven times more B. correcta males. This study demonstrated that CAR is a more specific and potent male lure than ME for B. correcta. This discovery may contribute to improve the tools currently available for pest management and border biosecurity against B. correcta.

Biocontrol agent risk assessment: A surprise find ...


 
 
 A Better Border Biosecurity (B3)-funded project investigating the interactions between the self-introduced generalist hymenopteran parasitoid Meteorus pulchricornis and native parasitoids of lepidopteran larvae throughout Auckland involved three years of eld work. The last eld trip was to Te Hauturu-o-Toi (Little Barrier Island) in December 2016 to survey some of the least-modi ed native forest in the North Island. Forty Cleora scriptaria moth larvae were collected by hand from Piper excelsum (kawakawa) host plants into ziplock bags, with a small amount of leaf material to support feeding. They were transported to the laboratory and reared to fate in a temperature-controlled room. Of the 40 larvae collected, twelve were parasitised by M. pulchricornis. This is the rst record of M. pulchricornis from Te Hauturu-o-Toi. In addition, nine larvae were parasitised by an undescribed parasitoid (Casinaria sp. 3). This new species may be endemic only to Te Hauturu-o-Toi, but it may have been out-competed on the more modi ed habitat of mainland Auckland, and found refuge in a more pristine environment. Further surveys of island and mainland parasitoid complexes would provide better baseline risk-assessment data prior to border invasions or importing biocontrol agents.
 
 

Methods and approaches for the management of arthropod border incursions

Biological invasions are increasing and are strongly associated with negative agricultural, economic and ecological impacts. It is increasingly recognized that the primary focus in minimizing biological invasions should be to prevent initial entry of alien species. However, exclusion of terrestrial arthropods such as insects and mites is difficult, in part because of their relatively small size, cryptic habits, broad physiological tolerances and close association with various internationally traded goods. Here we discuss methods, approaches, management and intervention systems used by border biosecurity agencies to prevent entry of inadvertently transported arthropods. We examine the at-border systems that exist for the detection and identification of and response to alien arthropods, and discuss the constraints and challenges present in these systems. We critically review current border biosecurity systems and discuss their relative efficacy. We then discuss additional measures and key areas that could be addressed that may further improve these systems. These include: (1) the application of appropriate sampling strategies; (2) employment of suitable inspection methods adequate to detect small and hidden arthropods; and (3) thorough recording of methods, organisms detected and both negative and positive results of inspections. We emphasize that more research is needed on taxonomy, biology, genetics, distribution, host and disease associations, impacts and pathways of introductions for invasive arthropods. Of critical importance is the compilation of complete and accurate invasive species lists and high-risk species watch-lists. The adoption of these recommendations will contribute to improved biosecurity systems for the exclusion of alien, invasive and pest arthropods.

Inadvertent biological control: an Australian thrips killing an invasive New Zealand tree in California

Transport hubs of international trade and tourism are sites of unprecedented long-distance dispersal of species and novel ecological interactions. In cases of invasive plants released from their specialist natural enemies, novel interactions with both resident enemies and new arrivals can accumulate and potentially reduce weed competitiveness. I present here one dramatic example of this, where an invasive woody weed in southern California is being rapidly controlled by an accidentally introduced genus-specialist herbivorous insect. The New Zealand native shrub/small tree, Myoporum laetum, is a long-time popular ornamental plant in California and has become an invasive woody weed. In 2005, a Myoporum-specific thrips, Klambothrips myopori, was discovered (and described) in California feeding on M. laetum leaves. Several searches have failed to find K. myopori in New Zealand and a population has recently been discovered in Tasmania, Australia, feeding on Myoporum insulare. In 5 years, K. myopori has killed off about half of southern Californian M. laetum with almost all surviving individuals being gradually defoliated. Inadequate border biosecurity has resulted in inadvertent biological control, in a rapid timeframe, caused by a novel enemy. Unfortunately, K. myopori has subsequently been accidentally transported from California to Hawaii where it is now killing off Hawaiian native Myoporum sandwicense. Transport hubs can both connect weeds with natural enemies and disperse those enemies more widely.

Border biosecurity and the use of species identification techniques in the ornamental fish trade: The case of the South American freshwater pufferfishes

The international aquarium trade reached an annual trade volume estimated at U$15–25 billion, being by far the largest exchange route for several exotic plant, invertebrate, and vertebrate species. In this sense, the poorly regulated international trade, mainly related with ornamental fishes, is a source of major risks to both biodiversity and local economy. In such scenario, the species identification still is one of the major obstacles for efficient law enforcement by the border security officials and reliable tools, protocols, and databases for species identification are still needed. In the present contribution we aimed to compare the academic approach of species identification, using the DNA barcoding methods, with the forensic approach, and the use of Insertion-Deletion polymorphisms for species determination, in a study case including the identification of a new species of Amazonian freshwater pufferfish.

Barcoding and border biosecurity: identifying cyprinid fishes in the aquarium trade.

BackgroundPoorly regulated international trade in ornamental fishes poses risks to both biodiversity and economic activity via invasive alien species and exotic pathogens. Border security officials need robust tools to confirm identifications, often requiring hard-to-obtain taxonomic literature and expertise. DNA barcoding offers a potentially attractive tool for quarantine inspection, but has yet to be scrutinised for aquarium fishes. Here, we present a barcoding approach for ornamental cyprinid fishes by: (1) expanding current barcode reference libraries; (2) assessing barcode congruence with morphological identifications under numerous scenarios (e.g. inclusion of GenBank data, presence of singleton species, choice of analytical method); and (3) providing supplementary information to identify difficult species.Methodology/Principal FindingsWe sampled 172 ornamental cyprinid fish species from the international trade, and provide data for 91 species currently unrepresented in reference libraries (GenBank/Bold). DNA barcodes were found to be highly congruent with our morphological assignments, achieving success rates of 90–99%, depending on the method used (neighbour-joining monophyly, bootstrap, nearest neighbour, GMYC, percent threshold). Inclusion of data from GenBank (additional 157 spp.) resulted in a more comprehensive library, but at a cost to success rate due to the increased number of singleton species. In addition to DNA barcodes, our study also provides supporting data in the form of specimen images, morphological characters, taxonomic bibliography, preserved vouchers, and nuclear rhodopsin sequences. Using this nuclear rhodopsin data we also uncovered evidence of interspecific hybridisation, and highlighted unrecognised diversity within popular aquarium species, including the endangered Indian barb Puntius denisonii.Conclusions/SignificanceWe demonstrate that DNA barcoding provides a highly effective biosecurity tool for rapidly identifying ornamental fishes. In cases where DNA barcodes are unable to offer an identification, we improve on previous studies by consolidating supplementary information from multiple data sources, and empower biosecurity agencies to confidently identify high-risk fishes in the aquarium trade.