A protocol for screening potentially invasive non-native species using Weed Risk Assessment-type decision-support tools.

There is increasing use worldwide of electronic decision-support tools to identify potentially invasive non-native species so as to inform policy and management decisions aimed at preventing or mitigating the environmental and socio-economic impacts of biological invasions. This study reviews the analytical approaches used to calibrate scores generated by the Weed Risk Assessment and subsequent adaptations thereof and provides a protocol for: (i) the identification of the assessor(s) who will carry out the screenings; (ii) the definition of the risk assessment area; (iii) the criteria for selection of the species for screening; and (iv) the a priori categorisation of the species into invasive or non-invasive necessary to compute the thresholds by which to distinguish between high-risk and medium-risk non-native species. This analytical approach represents an evidence-based and statistically robust means with which to inform decision-makers and stakeholders about policy and management of potentially invasive species and is expected to serve as a general reference of forthcoming screening applications of Weed Risk Assessment-type toolkits.

A global-scale screening of non-native aquatic organisms to identify potentially invasive species under current and future climate conditions.

The threat posed by invasive non-native species worldwide requires a global approach to identify which introduced species are likely to pose an elevated risk of impact to native species and ecosystems. To inform policy, stakeholders and management decisions on global threats to aquatic ecosystems, 195 assessors representing 120 risk assessment areas across all six inhabited continents screened 819 non-native species from 15 groups of aquatic organisms (freshwater, brackish, marine plants and animals) using the Aquatic Species Invasiveness Screening Kit. This multi-lingual decision-support tool for the risk screening of aquatic organisms provides assessors with risk scores for a species under current and future climate change conditions that, following a statistically based calibration, permits the accurate classification of species into high-, medium- and low-risk categories under current and predicted climate conditions. The 1730 screenings undertaken encompassed wide geographical areas (regions, political entities, parts thereof, water bodies, river basins, lake drainage basins, and marine regions), which permitted thresholds to be identified for almost all aquatic organismal groups screened as well as for tropical, temperate and continental climate classes, and for tropical and temperate marine ecoregions. In total, 33 species were identified as posing a 'very high risk' of being or becoming invasive, and the scores of several of these species under current climate increased under future climate conditions, primarily due to their wide thermal tolerances. The risk thresholds determined for taxonomic groups and climate zones provide a basis against which area-specific or climate-based calibrated thresholds may be interpreted. In turn, the risk rankings help decision-makers identify which species require an immediate 'rapid' management action (e.g. eradication, control) to avoid or mitigate adverse impacts, which require a full risk assessment, and which are to be restricted or banned with regard to importation and/or sale as ornamental or aquarium/fishery enhancement.

Unstable intraguild predation causes establishment failure of a globally invasive species.

Biotic resistance is often posited, but rarely known, to be the cause of invasion failure. Competition and predation are the most frequently identified processes that may prevent or limit the establishment of nonnative species. Interactions between nonnative and native species that involve intraguild predation (IGP) are very common in nature, although theory predicts most IGP systems should be unstable and lead to extinction. If this prediction is true, the frequency of invasion failures due to IGP may be underappreciated because of their fleeting nature and, thus, studies of unstable IGP systems are lacking, despite the opportunities they offer for understanding the factors affecting their unstable dynamics. We investigated a failed invasion involving an IGP relationship. In Florida, the guppy (Poecilia reticulata), a worldwide invader, fails to establish in the presence of eastern mosquitofish (Gambusia holbrooki). We tested whether and how resident mosquitofish cause guppy invasion failure using replicated mesocosm and aquarium trials. Both the predator and competitor components of the IGP relationship were strongly asymmetrical, with large impacts on guppies. We identified two effects, direct consumption of neonates and aggressive interference competition, that limited survival and recruitment. The highly unstable nature of this IGP relationship is the primary cause of the failure of the guppy to establish in Florida. Our study shows that the transient nature of an ephemeral IGP relationship can yield important insights into the underlying causes of invasion failure, including the role of strong biotic resistance.

Coastal restoration evaluated using dominant habitat characteristics and associated fish communities.

Increasing coastal populations and urban development have led to the loss of estuarine habitats for fish and wildlife. Specifically, a decline in complexity and heterogeneity of tidal marshes and creeks is thought to negatively impact fish communities by altering the function of nursery grounds, including predator refuge and prey resources. To offset these impacts, numerous agencies are restoring degraded habitats while also creating new ones where habitat has been lost. To improve understanding of what contributes to a successful restoration, six quarterly sampling events using two gear types to collect small- and large-bodied fishes were conducted to compare the fish community structure and habitat characteristics at three natural, three restored, and three impacted (i.e. ditched) areas along the coast of Tampa Bay, Florida. Overall, impacted sites had significantly lower small-bodied and juvenile fish diversity than natural and restored areas, while restored sites harbored a greater number of fish species than impacted sites for both large- and small-bodied fish. Habitat features such as shoreline slope differentiated impacted and restored from natural areas. Although we did not find a direct correlation, habitat heterogeneity likely played a role in structuring fish communities. These findings provide guidance for future coastal restoration or modification of existing projects. Specifically, the habitat mosaic approach of creating a geographically compact network of heterogenous habitat characteristics is likely to support fish diversity, while decreasing shoreline slope in a greater amount of area within coastal wetland restorations would more closely mimic natural areas.

Why did the invasive walking catfish cross the road? Terrestrial chemoreception described for the first time in a fish.

Clarias batrachus (walking catfish) is an invasive species in Florida, renowned for its air-breathing and terrestrial locomotor capabilities. However, it is unknown how this species orients in terrestrial environments. Furthermore, while anecdotal life history information is widespread for this species in its nonnative range, little of this information exists in the literature. The goals of this study were to identify sensory modalities that C. batrachus use to orient on land, and to describe the natural history of this species in its nonnative range. Fish (n = 150) were collected from around Ruskin, FL, and housed in a greenhouse, where experiments took place. Individual catfish were placed in the center of a terrestrial arena and were exposed to nine treatments: two controls, L-alanine, quinine, allyl isothiocynate, sucrose, volatile hydrogen sulphide, pond water and aluminium foil. These fish exhibited significantly positive chemotaxis toward alanine and pond water, and negative chemotaxis away from volatile hydrogen sulphide, suggesting chemoreception - both through direct contact and through the air - is important to their terrestrial orientation. Additionally, 88 people from Florida wildlife-related Facebook groups who have personal observations of C. batrachus on land were interviewed for information regarding their terrestrial natural history. These data were combined with observations from 38 YouTube videos. C. batrachus appear to emerge most frequently during or just after heavy summer rains, particularly from stormwater drains in urban areas, where they may feed on terrestrial invertebrates. By better understanding the full life history of C. batrachus, we can improve management of this species.

Characterizing the US trade in lionfishes.

Invasive lionfishes Pterois volitans and Pterois miles have spread throughout the tropical western Atlantic Ocean, Gulf of Mexico, and Greater Caribbean. Beyond these two invaders, additional species within the subfamily Pteroinae are regularly imported into the United States. We evaluated the trade of lionfishes as a surrogate measure for propagule pressure, an important component of invasion success. Proactive evaluation of marine ornamental fishes in trade is vital, particularly for those sharing characteristics with known invaders. We utilized one year of import records from the U.S. Fish and Wildlife Service's Law Enforcement Management Information System database and two domestic databases to capture the trade of all lionfishes in the US, the invasive complex in its invaded range in Florida, and two Hawaiian endemic lionfishes. Retail surveys were completed to assess lionfish availability across 10 coastal states. Compared to species diversity within the subfamily, the number of traded species was low and just two species were traded at moderate to high volume, including P. volitans and Dendrochirus zebra. At the retail level, fewer species are available to consumers. The trade in lionfishes is consolidated because most lionfishes originate from two Indo-Pacific countries and arrive through the port of Los Angeles. The volume and diversity of traded lionfishes presents some risk of introduction for lionfishes which are not established, and secondary introductions of the invasive P. volitans. In combination with rapid risk screening, this research can be applied to a proactive risk management framework to identify risky species prior to introduction and establishment.

Assessing environmental DNA detection in controlled lentic systems.

Little consideration has been given to environmental DNA (eDNA) sampling strategies for rare species. The certainty of species detection relies on understanding false positive and false negative error rates. We used artificial ponds together with logistic regression models to assess the detection of African jewelfish eDNA at varying fish densities (0, 0.32, 1.75, and 5.25 fish/m3). Our objectives were to determine the most effective water stratum for eDNA detection, estimate true and false positive eDNA detection rates, and assess the number of water samples necessary to minimize the risk of false negatives. There were 28 eDNA detections in 324, 1-L, water samples collected from four experimental ponds. The best-approximating model indicated that the per-L-sample probability of eDNA detection was 4.86 times more likely for every 2.53 fish/m3 (1 SD) increase in fish density and 1.67 times less likely for every 1.02 C (1 SD) increase in water temperature. The best section of the water column to detect eDNA was the surface and to a lesser extent the bottom. Although no false positives were detected, the estimated likely number of false positives in samples from ponds that contained fish averaged 3.62. At high densities of African jewelfish, 3-5 L of water provided a >95% probability for the presence/absence of its eDNA. Conversely, at moderate and low densities, the number of water samples necessary to achieve a >95% probability of eDNA detection approximated 42-73 and >100 L, respectively. Potential biases associated with incomplete detection of eDNA could be alleviated via formal estimation of eDNA detection probabilities under an occupancy modeling framework; alternatively, the filtration of hundreds of liters of water may be required to achieve a high (e.g., 95%) level of certainty that African jewelfish eDNA will be detected at low densities (i.e., <0.32 fish/m3 or 1.75 g/m3).

Revisions of the Fish Invasiveness Screening Kit (FISK) for its application in warmer climatic zones, with particular reference to peninsular Florida.

The initial version (v1) of the Fish Invasiveness Scoring Kit (FISK) was adapted from the Weed Risk Assessment of Pheloung, Williams, and Halloy to assess the potential invasiveness of nonnative freshwater fishes in the United Kingdom. Published applications of FISK v1 have been primarily in temperate-zone countries (Belgium, Belarus, and Japan), so the specificity of this screening tool to that climatic zone was not noted until attempts were made to apply it in peninsular Florida. To remedy this shortcoming, the questions and guidance notes of FISK v1 were reviewed and revised to improve clarity and extend its applicability to broader climatic regions, resulting in changes to 36 of the 49 questions. In addition, upgrades were made to the software architecture of FISK to improve overall computational speed as well as graphical user interface flexibility and friendliness. We demonstrate the process of screening a fish species using FISK v2 in a realistic management scenario by assessing the Barcoo grunter Scortum barcoo (Terapontidae), a species whose management concerns are related to its potential use for aquaponics in Florida. The FISK v2 screening of Barcoo grunter placed the species into the lower range of medium risk (score = 5), suggesting it is a permissible species for use in Florida under current nonnative species regulations. Screening of the Barcoo grunter illustrates the usefulness of FISK v2 as a proactive tool serving to inform risk management decisions, but the low level of confidence associated with the assessment highlighted a dearth of critical information on this species.

Effects of metomidate hydrochloride sedation on blood glucose and marketability of transported threespot gourami Trichogaster trichopterus.

Our objectives were to determine whether sedation with metomidate hydrochloride (hereafter, "metomidate") during transportation of threespot gourami Trichogaster trichopterus would prevent an increase in blood glucose levels and improve fish marketability (i.e., based on appearance and behavior) in comparison with unsedated controls. Threespot gourami are obligate air-breathers that possess a labyrinth organ, enabling the fish to respire air above the water surface; these fish should be lightly sedated during transport. Fish were transported for approximately 24 h via truck and domestic airline. Blood was sampled at 0, 2, 6, and 12 h posttransport ation, and appearance and behavior were observed at 0, 2, 4, 6, and 12 h and 7 d posttransportation. Metomidate concentrations tested were 0.0 (control), 0.1, 0.2, 0.3, and 0.4 mg/L. At the concentrations tested, metomidate neither inhibited elevations in blood glucose nor improved marketability. Fish that were transported with 0.3-mg/L metomidate were less marketable based on behavioral indices, and fish that were transported with 0.4-mg/L metomidate had higher glucose levels than control fish. Use of metomidate as a transport sedative for threespot gourami should be considered with caution and may be problematic at the concentrations tested; however, further research examining additional indices of stress may clarify metomidate use for this species.

Evaluation of emamectin benzoate for the control of experimentally induced infestations of Argulus sp. in goldfish and koi carp.

The effect of 0.2% emamectin benzoate (SLICE; Intervet/ Schering-Plough Animal Health, Roseland, New Jersey) administered in top-dressed, pelleted commercial fish feed was evaluated for control of freshwater Argulus sp. in goldfish Carassius auratus and koi carp, a variant of common carp Cyprinus carpio, in freshwater aquaria at 24-25 degrees C. Sixteen individually housed goldfish were each exposed to 37 Argulus. The number of fish lice attached to each fish at the start of the experiment was not determined; however, the total number of motile fish lice in each aquarium (on fish and in the water) was determined at the start and end of each experiment. Eight goldfish were fed the control diet (0 microg x kg fish biomass(-1) x d(-1)) and eight were fed the medicated diet (50 microg x kg fish biomass(-1) x d(-1)) for seven consecutive days. After treatment, fish louse infestation in controls was 20.5 +/- 1.5 (mean +/- SE) lice per fish. No Argulus were found on fish in the treated group. In a separate experiment, 10 individually housed koi were each exposed to 128 Argulus. Five koi were fed the control diet and five were fed a low-dose medicated diet (5 microg x kg fish biomass(-1) x d(-1)) for 7 d. After treatment, fish louse infestation among the controls was 14.6 +/- 3.8 lice per koi. No Argulus were found on koi in the treated group. Hence, a 7-d regimen of oral emamectin benzoate controlled experimental infestation of Argulus when administered to goldfish at 50 microg x kg fish biomass(-1) x d(-1) and to koi at 5 microg x kg fish biomass(-1) x d(-1).

Investigational use of metomidate hydrochloride as a shipping additive for two ornamental fishes.

During shipping, ornamental fish can be stressed due to handling, high stocking densities, and deteriorating water quality. Adding sedatives, such as metomidate hydrochloride, to shipping water may improve fish survival rates and the percentage of fish in saleable condition. Although the effects of metomidate hydrochloride on the stress response in fish have been studied, its application as a shipping additive has not been well investigated, particularly for tropical ornamental fishes shipped under industry conditions. Convict cichlids Cichlasoma nigrofasciatum and black mollies Poecilia sphenops were evaluated for 7 d after a 24-h period of exposure (including ground and air transport) to one of four metomidate hydrochloride concentrations: 0.0, 0.2, 0.5, and 1.0 mg/L. Immediate posttransport and cumulative mortality data, as well as 12-h and 7-d posttransport appearance and behavior scores, were generated. In convict cichlids, the highest dose of metomidate hydrochloride (1.0 mg/L) reduced mortality (0% compared with cumulative means of 5.5-9.2% in other groups) and increased the percentage of saleable fish (91.7% were immediately saleable compared with 12.5-50% in other groups). No effect was detected in black mollies at any concentration tested. Metomidate hydrochloride showed promise as a shipping additive for convict cichlids, but further studies are warranted to evaluate species-specific responses in other ornamental species.

Use of a novel induced spawning technique for the first reported captive spawning of Tetraodon nigroviridis.

The spotted green pufferfish (Tetraodon nigroviridis) is an important genetics model animal due to its small, well-mapped genome. However, only wild-caught juveniles and adults are available to researchers. A lack of gametes, fertilized eggs, developing embryos, and other early life stages hampers development of the full potential of T. nigroviridis as a model research species. We report on successful spawning trials using a novel induced spawning technique, ovarian lavage. Chorulon(®) (human chorionic gonadotropin, hCG) was injected into a catheter inserted into the oviduct at a rate of 3 µl/g body weight. In one trial, a female paired with a male spawned in an aquarium at about 72 h post-treatment. In other trials, females were hand-stripped of eggs at 36 h post-treatment. There were 3680 eggs/g of eggs and females produced up to 24% of their body weight in eggs. Hatch resulted from all trials on the 4th day post-fertilization. Ovarian lavage is a simple method for administering spawning hormones, uses a catheter technique similar to that frequently performed to determine egg maturity in broodstock, and eliminates the need for injection.