Can denticle morphology help identify southeastern Australian elasmobranchs?

Elasmobranchs are covered in scale-like structures called dermal denticles, comprising dentine and enameloid. These structures vary across the body of an individual and between species, and are frequently shed and preserved in marine sediments. With a good understanding of denticle morphology, current and historical elasmobranch diversity and abundance might be assessed from sediment samples. Here, replicate samples of denticles from the bodies of several known (deceased) shark species were collected and characterized for morphology before being assigned morphotypes. These data were used to expand the established literature describing denticles and to investigate intra- and interspecific variability, with the aim of increasing the viability of using sediment samples to assess elasmobranch diversity and abundance. Denticle morphology was influenced more by life-history traits than by species, where demersal species were largely characterized by generalized function and defense denticles, whereas pelagic and benthopelagic species were characterized by drag-reduction denticles. Almost all species possessed abrasion strength or defense denticles on the snout, precluding their utility for separating species. In a separate manipulative experiment, samples of denticles were collected from sediments in two aquaria with known elasmobranchs to determine their utility for reliably separating species. Visual examination of denticles, morphometric measurements, scaled photographs, and reference collections allowed for some precise identification, but not always to the species level. Ongoing work to develop denticle reference collections could help to identify past and present families and, in some cases, species.

Integrating isotopic and nutritional niches reveals multiple dimensions of individual diet specialisation in a marine apex predator.

Dietary specialisations are important determinants of ecological structure, particularly in species with high per-capita trophic influence like marine apex predators. These species are, however, among the most challenging in which to establish spatiotemporally integrated diets. We introduce a novel integration of stable isotopes with a multidimensional nutritional niche framework that addresses the challenges of establishing spatiotemporally integrated nutritional niches in wild populations, and apply the framework to explore individual diet specialisation in a marine apex predator, the white shark Carcharodon carcharias. Sequential tooth files were sampled from juvenile white sharks to establish individual isotopic (δ-space; δ13 C, δ15 N, δ34 S) niche specialisation. Bayesian mixing models were then used to reveal individual-level prey (p-space) specialisation, and further combined with nutritional geometry models to quantify the nutritional (N-space) dimensions of individual specialisation, and their relationships to prey use. Isotopic and mixing model analyses indicated juvenile white sharks as individual specialists within a broader, generalist, population niche. Individual sharks differed in their consumption of several important mesopredator species, which suggested among-individual variance in trophic roles in either pelagic or benthic food webs. However, variation in nutrient intakes was small and not consistently correlated with differences in prey use, suggesting white sharks as nutritional specialists and that individuals could use functionally and nutritionally different prey as complementary means to achieve a common nutritional goal. We identify how degrees of individual specialisation can differ between niche spaces (δ-, p- or N-space), the physiological and ecological implications of this, and argue that integrating nutrition can provide stronger, mechanistic links between diet specialisation and its intrinsic (fitness/performance) and extrinsic (ecological) outcomes. Our time-integrated framework is adaptable for examining the nutritional consequences and drivers of food use variation at the individual, population or species level.

Non-lethal sampling does not misrepresent trophic level or dietary sources for Sagmariasus verreauxi (eastern rock lobster).

RATIONALE: Isotope analysis can be used to investigate the diets of predators based on assimilation of nitrogen and carbon isotopes from prey. Recent work has shown that tissues taken from legs, antennae or abdomen of lobsters can give different indications of diet, but this has never been evaluated for Sagmariasus verreauxi (eastern rock lobster). Work is now needed to prevent erroneous conclusions being drawn about lobster food webs, and undertaking this work could lead to developing non-lethal sampling methodologies. Non-lethal sampling for lobsters is valuable both ethically and for areas of conservation significance such as marine reserves. METHOD: We evaluated this by dissecting 76 lobsters and comparing δ13 C and δ15 N isotope values in antennae, leg and abdomen tissue from the same individuals ranging from 104 to 137 mm carapace length. Stable isotope values were determined using a Europa EA GSL elemental analyser coupled with Hydra 20-20 Isoprime IRMS. RESULTS: We found the abdomen δ13 C values to be lower than other tissues by 0.3 ± 0.2‰ for antennae tissue and 0.1 ± 0.2‰ δ13 C for leg tissues, whereas for δ15 N, no significant difference between tissues was observed. There was no significant effect of lobster size or sex, though we did observe interactions between month and tissue type, indicating that differences may be seasonal. Importantly, the detected range of isotopic variability between tissues is within the range of uncertainty used for discrimination factors in isotopic Bayesian modelling of 0‰-1.0‰ for δ13 C and 3.0‰-4.0‰ for δ15 N. CONCLUSIONS: We show that S. verreauxi can be sampled non-lethally with mathematical corrections applied for δ13 C, whereas any tissue is suitable for δ15 N. Our results indicate that a walking leg is most favourable and would also be the least intrusive for the lobster. The application of non-lethal sampling provides avenues for the contribution of citizen science to understanding lobster food webs and to undertake fieldwork in ecologically sensitive areas such as marine reserves.

Elevated estuary water temperature drives fish gut dysbiosis and increased loads of pathogenic vibrionaceae.

Marine water temperatures are increasing globally, with eastern Australian estuaries warming faster than predicted. There is growing evidence that this rapid warming of coastal waters is increasing the abundance and virulence of pathogenic members of the Vibrionaceae, posing a significant health risk to both humans and aquatic organisms. Fish disease, notably outbreaks of emerging pathogens in response to environmental perturbations such as heatwaves, have been recognised in aquaculture settings. Considerably less is known about how rising sea surface temperatures will impact the microbiology of wild fish populations, particularly those within estuarine systems that are more vulnerable to warming. We used a combination of Vibrio-specific quantitative PCR and amplicon sequencing of the 16S rRNA and hsp60 genes to examine seawater and fish (Pelates sexlineatus) gut microbial communities across a quasi-natural experimental system, where thermal pollution from coal-fired power stations creates a temperature gradient of up to 6 °C, compatible with future predicted temperature increases. At the warmest site, fish hindgut microbial communities were in a state of dysbiosis characterised by shifts in beta diversity and a proliferation (71.5% relative abundance) of the potential fish pathogen Photobacterium damselae subsp. damselae. Comparable patterns were not identified in the surrounding seawater, indicating opportunistic proliferation within estuarine fish guts under thermal stress. A subsequent evaluation of predicted future warming-related risk due to pathogenic Vibrionaceae in temperate estuarine fish demonstrated that warming is likely to drive opportunistic pathogen increases in the upper latitudinal range of this estuarine fish, potentially impacting adaptations to future warming. These findings represent a breakthrough in our understanding of the dynamics of emerging pathogens in populations of wild aquatic organisms within environments likely to experience rapid warming under future climate change.

Diving into the diet of provisioned smooth stingrays using stable isotope analysis.

Recreational fishing waste, produced from processing catches at shore-based fish cleaning facilities and discarded into adjacent waters, is foraged by various aquatic species. However, the potential alterations to the diet of consumers of these resources are poorly studied. Smooth stingrays (Bathytoshia brevicaudata) are a large demersal mesopredatory ray species and common scavenger of recreational fishing discards around southern Australia. Due to their attraction to fish cleaning sites, they are also common targets of unregulated 'stingray feeding' tourism where they are fed commercially produced baits (e.g., pilchards). This study provides a preliminary assessment of the diet of smooth stingrays provisioned recreational fishing discards and baits at two sites in southern New South Wales, Australia (Discard Site: recreational fishing discards only; Provisioning Site: recreational fishing discards and commercial baits) using stable isotope analysis of carbon (δ13 C) and nitrogen (δ15 N), and Bayesian stable isotope mixing models. Our results indicate that at both sites invertebrates, considered a main part of the natural diet of smooth stingrays, made a limited contribution to the diets of provisioned stingrays, while a benthic teleost fish that is a common recreational catch was the dominant contributor. As the assessed teleost is potentially a natural prey item for smooth stingrays, it remains unclear whether the contribution came from recreational fishing discards or natural foraging. However, due to smooth stingrays' typically opportunistic foraging strategy, we expected a greater mixture of resources from low to high trophic level prey than was observed. These results suggest that smooth stingrays have either lower reliance on invertebrates as a result of utilizing provisioned resources or higher reliance on teleost fishes than previously thought. Commercial bait products fed to stingrays at the Provisioning Site were not a major contributor to the diets of smooth stingrays, suggesting this activity has a low impact on their nutrition.

Metal(loid) accumulation in the leaves of the grey mangrove (Avicennia marina): Assessment of robust sampling requirements and potential use as a bioindicator.

This study assessed the appropriate sampling design required for quantifying variability in metal accumulation in the leaf tissues of A. marina, a dominant mangrove inhabiting Australian estuaries, by applying a hierarchical nested sampling design to sample mangroves at various levels of biological and spatial hierarchies (leaf, branch, tree, site). It was revealed that most variation in metal accumulation occurred among trees and branches, with insignificant variation between sites and among leaves. We also examined the accumulation of metal (loid)s in the leaf tissues collected from six locations across the Georges River estuary in southern Sydney, which differ in metal contamination history. Prospect Creek and Salt Pan Creek were the most contaminated locations, which exceeded sediment quality guideline values for Cu (66.71 ± 2.18 µg/g), Zn (317.14 ± 46.14 µg/g) and Pb (81.02 ± 2.79 µg/g). All metal(loid) concentrations in leaf tissues were much lower than their concentrations in sediment, but essential metals exhibited greater mobility. Out of 10 metal(loid)s, Mn, Co and Pb in leaves showed linear relationships (R2 = 0.28-0.47) with sediment, indicating that mangrove leaves may be used as a bioindicator of environmental loads for these metals.

Patterns of mother-embryo isotope fractionation in batoids vary within and between species.

Patterns of mother-embryo fractionation of 13 C and 15 N were assessed for their predictability across three species of batoids caught as by-catch in south-eastern Australia. Stable isotope analysis of 24 mothers and their litters revealed that isotope ratios of embryos were significantly different from their corresponding mothers and that the scale and direction of the difference varied within and across species. The range of variation across species was 3.5‰ for δ13 C and 4‰ for δ15 N, equivalent to a difference in trophic level. In one species (Urolophus paucimaculatus) litters could be significantly enriched or depleted in 13 C and 15 N relative to their mothers' isotope signatures. These results suggest that patterns of mother-embryo isotope fractionation vary within and between species and that these patterns may not be explained only by developmental mode. Contrasting patterns of fractionation between and within species make it difficult to adjust mother-embryo fractionation with broad-scale correction factors.

Diet-tissue discrimination and turnover of δ13 C and δ15 N in muscle tissue of a penaeid prawn.

RATIONALE: Stable isotopes are used to study trophic and movement ecology in aquatic systems, as they provide spatially distinct, time-integrated signatures of diet. Stable isotope ecology has been used to quantify species-habitat relationships in many important fisheries species (e.g., penaeid prawns), with approaches that typically assume constant values for diet-tissue discrimination and diet-tissue steady state, but these can be highly variable. Here we provide the first report of these processes in Metapenaeus macleayi (eastern school prawn). METHODS: Here we explicitly measure and model carbon (δ13 C) and nitrogen (δ15 N) diet-tissue discrimination and turnover in eastern school prawn muscle tissue as a function of experimental time following a change in diet to an isotopically distinct food source. RESULTS: Diet-tissue discrimination factors were 5 and 0.6‰ for δ13 C and δ15 N, respectively. Prawn muscle tissue reached an approximate steady state after approximately 50 and 30 days for δ13 C and δ15 N. Half-lives indicated faster turnover of δ15 N (~8 days) than δ13 C (~14 days). CONCLUSIONS: Our diet-tissue discrimination factors deviate from 'typical' values with larger values for carbon than nitrogen isotopes, but are generally similar to those measured in other crustaceans. Similarly, our estimates of isotopic turnover align with those in other penaeid species. These findings confirm muscle tissue as a reliable indicator of long-term diet and movement patterns in eastern school prawn.

Seasonal and developmental diet shifts in sympatric and allopatric intertidal gobies determined by stomach content and stable isotope analysis.

Resource partitioning facilitates the coexistence of sympatric species through spatial, temporal and/or trophic strategies. Fishes living in the intertidal zone demonstrate highly adaptive plastic behaviour, including resource partitioning, through spatial and temporal shifts in diet and microhabitat. Although intertidal fish assemblages are influenced by inter- and intraspecific competition, few studies have compared the extent of resource partitioning between sympatric species in the context of trophic niche plasticity. Here we used complementary approaches, stomach content and stable isotope (δ13 C and δ15 N) analyses, to evaluate seasonal and developmental shifts in trophic niche position in two sympatric (Favonigobius lentiginosus and Bathygobius krefftii) and one allopatric (Bathygobius cocosensis) species of intertidal goby. The results indicate that resource partitioning in the two sympatric species varied with season, with almost no trophic niche overlap in summer to about ~30% overlap in winter. Also, evidence of dietary changes was found in B. cocosensis, which is likely associated with a shift in microhabitat and intraspecific competition. The findings highlight the temporal range of behavioural plasticity in trophic niche position of intertidal gobies, which likely has high adaptive value in the dynamic intertidal zone.

Resource use of great hammerhead sharks (Sphyrna mokarran) off eastern Australia.

Great hammerhead sharks Sphyrna mokarran are the largest member of Sphyrnidae, yet the roles of these large sharks in the food webs of coastal ecosystems are still poorly understood. Here we obtained samples of muscle, liver and vertebrae from large S. mokarran (234-383 cm total length; LT ) caught as by-catch off eastern Australia and used stable-isotope analyses of δ15 N, δ13 C and δ34 S to infer their resource use and any associated ontogenetic patterns. The results indicated large S. mokarran are apex predators primarily relying on other sharks and rays for their diet, with a preference for benthic resources such as Australian cownose rays Rhinoperon neglecta during the austral summer. Teleosts, cephalopods and crustaceans were not significant components of S. mokarran diets, though some conspecifics appeared to rely on more diverse resources over the austral summer. Ontogenetic shifts in resource use were detected but trajectories of the increases in trophic level varied among individuals. Most S. mokarran had non-linear trajectories in ontogenetic resource-use shifts implying size was not the main explanatory factor. Stable isotope values of δ13 C and δ34 S in muscle suggest S. mokarran span coastal, pelagic and benthic food webs in eastern Australia.

Spatial patterns in host-associated and free-living bacterial communities across six temperate estuaries.

A major goal of microbial ecology is to establish the importance of spatial and environmental factors in driving community variation. Their relative importance likely varies across spatial scales, but focus has primarily been on free-living communities within well-connected aquatic environments rather than less connected island-like habitats such as estuaries, and key host-associated communities within these systems. Here we sampled both free-living (seawater and sediment) and host-associated (estuarine fish hindgut microbiome, Pelates sexlineatus) communities across six temperate Australian estuaries spanning ∼500 km. We find that spatial and environmental factors have different influences on these communities, with seawater demonstrating strong distance-decay relationships (R = -0.69) and significant associations with a range of environmental variables. Distance-decay relationships were weak for sediment communities but became stronger over smaller spatial scales (within estuaries, R = -0.5), potentially reflecting environmental filtering across biogeochemical gradients or stochastic processes within estuary sediments. Finally, P. sexlineatus hindgut microbiome communities displayed weak distance-decay relationships (R = -0.36), and limited variation explained by environmental variables, indicating the significance of host-related factors in driving community variation. Our findings provide important ecological insights into the spatial distributions and driving forces of both free-living and host-associated bacterial patterns across temperate estuarine systems.

eDNA metabarcoding reveals shifts in sediment eukaryote communities in a metal contaminated estuary.

Metal contamination is a global issue impacting biodiversity in urbanised estuaries. Traditional methods to assess biodiversity are time consuming, costly and often exclude small or cryptic organisms due to difficulties with morphological identification. Metabarcoding approaches have been increasingly recognised for their utility in monitoring, however studies have focused on freshwater and marine systems despite the ecological significance of estuaries. We targeted estuarine eukaryote communities within the sediments of Australia's largest urbanised estuary, where a history of industrial activity has resulted in a metal contamination gradient. We identified specific eukaryote families with significant correlations with bioavailable metal concentrations, indicating sensitivity or tolerance to specific metals. While polychaete families Terebellidae and Syllidae demonstrated tolerance to the contamination gradient, members of the meio- and microfaunal communities including diatoms, dinoflagellates and nematodes displayed sensitivities. These may have high value as indicators but are frequently missed in traditional surveys due to sampling limitations.

Tracking the impacts of nutrient inputs on estuary ecosystem function.

Estuaries are one of the most impacted coastal environments globally, subjected to multiple stressors from urban, industry and coastal development. With increasing anthropogenic activity surrounding estuarine systems, sewage inputs have become a common concern. Stable isotope analysis provides a well-established tool to investigate the incorporation of nitrogen into marine organisms and identify major nutrient sources. Benthic macroinvertebrate communities are often used as bioindicators in ecological studies as they typically display predictable responses to anthropogenic pressures, however have a suite of limitations and costs associated with their use. 16S rDNA amplicon sequencing techniques allow for investigation of the microbial communities inhabiting complex environmental samples, with potential as a tool in the ecological assessment of pollution. These communities have not yet been adequately considered for ecological studies and biomonitoring, with a need to better understand interactions with environmental stressors and implications for ecosystem function. This study used a combination of stable isotope analysis to trace the uptake of anthropogenic nitrogen in biota, traditional assessment of benthic macroinvertebrate communities, and 16S rDNA genotyping of benthic microbial communities. Stable isotope analysis of seagrass and epiphytes identified multiple treated and untreated sewage inputs, ranges of 5.2-7.2‰ and 1.9-4.0‰ for δ15N respectively, as the dominant nitrogen source at specific locations. The benthic macroinvertebrate community reflected these inputs with shifts in dominant taxa and high abundances of polychaetes at some sites. Microbial communities provided a sensitive indication of impact with a breadth of information not available using traditional techniques. Composition and predicted function reflected sewage inputs, particularly within sediments, with the relative abundance of specific taxa and putative pathogens linked to these inputs. This research supports the growing body of evidence that benthic microbial communities respond rapidly to anthropogenic stressors and have potential as a monitoring tool in urban estuarine systems.

Legacy metal contamination is reflected in the fish gut microbiome in an urbanised estuary.

Estuaries are critical habitats subject to a range of stressors requiring effective management. Microbes are gaining recognition as effective environmental indicators, however, the response of host associated communities to stressors remains poorly understood. We examined microbial communities from seawater, sediments and the estuarine fish Pelates sexlineatus, in Australia's largest urbanised estuary, and hypothesised that anthropogenic contamination would be reflected in the microbiology of these sample types. The human faecal markers Lachno3 and HF183 were not detected, indicating negligible influence of sewage, but a gradient in copy numbers of the class 1 integron (intI-1), which is often used as a marker for anthropogenic contamination, was observed in sediments and positively correlated with metal concentrations. While seawater communities were not strongly driven by metal contamination, shifts in the diversity and composition of the fish gut microbiome were observed, with statistical links to levels of metal contamination (F2, 21 = 1.536, p < 0.01). Within the fish gut microbiome, we further report increased relative abundance of amplicon sequence variants (ASVs; single inferred DNA sequences obtained in sequencing) identified as metal resistant and potentially pathogenic genera, as well as those that may have roles in inflammation. These results demonstrate that microbial communities from distinct habitats within estuarine systems have unique response to stressors, and alterations of the fish gut microbiome may have implications for the adaptation of estuarine fish to legacy metal contamination.

Effects of reduced pH on an estuarine penaeid shrimp (Metapenaeus macleayi).

Acid sulfate soils are a major problem in modified coastal floodplains and are thought to have substantial impacts on estuarine species. In New South Wales, Australia, acid sulfate soils occur in every estuary and are thought to impact important fisheries species, such as Eastern School Prawn (Metapenaeus macleayi). These fisheries have experienced declining productivity over the last ten years and increasing occurrence of catchment-derived stressors in estuaries contribute to this problem. We evaluated the effect of pH 4-7.5 on School Prawn survival at two salinities (27 and 14.5), pH 5, 6 and 7.5 on the predation escape response (PER) speed at two salinities (27 and 14.5), and pH 4 and 7.5 on respiration rates. While mortality appeared to be greater in the high salinity treatment, there was no significant relationship between proportional survival and pH for either salinity treatment. Respiration was significantly slower under acidic conditions and the average PER was almost twice as fast at pH 7.5 compared to pH 5 (p < 0.05), indicating prawns may fall prey to predation more easily in acidic conditions. These findings confirm the hypothesised impacts of acidic water on penaeid prawns. Given that the conditions simulated in these experiments reflect those encountered in estuaries, acidic runoff may be contributing to bottlenecks for estuarine species and impacting fisheries productivity.

Detrimental effects of urbanization on the diet, health, and signal coloration of an ecologically successful alien bird.

Theory suggests that overcrowding and increased competition in urban environments might be detrimental to individual condition in avian populations. Unfavourable living conditions could be compounded by changes in dietary niche with additional consequences for individual quality of urban birds. We analysed the isotopic signatures, signal coloration, body condition, parasitic loads (feather mites and coccidia), and immune responsiveness of 191 adult common (Indian) mynas (Acridotheres tristis) captured in 19 localities with differing levels of urbanization. The isotopic signature of myna feathers differed across low and high urbanized habitats, with a reduced isotopic niche breadth found in highly urbanized birds. This suggests that birds in high urban environments may occupy a smaller foraging niche to the one of less urbanized birds. In addition, higher degrees of urbanization were associated with a decrease in carotenoid-based coloration, higher ectoparasite loads and higher immune responsiveness. This pattern of results suggests that the health status of mynas from more urbanized environments was poorer than mynas from less modified habitats. Our findings are consistent with the theory that large proportions of individual birds that would otherwise die under natural conditions survive due to prevailing top-down and bottom-up ecological processes in cities. Detrimental urban ecological conditions and search for more favourable, less crowded habitats offers the first reasonable explanation for why an ecological invader like the common myna continues to spread within its global invasive range.

Overcoming multi-year impacts of maternal isotope signatures using multi-tracers and fast turnover tissues in juvenile sharks.

Stable isotopes are often used to determine the ecological role of different age classes of animals, but particularly for young animals this approach may be compromised. During gestation and or incubation body tissues of the young are derived directly from the mother. In neonates or post hatching, there is a period of transformation as the young grow and forage independently, but during this period different organs will continue to reflect the maternal isotopic signature as a function of their turnover rate. How long this maternal hangover persists remains poorly understood. We applied a multi-tracer approach (δ15N, δ13C and δ34S) to stable isotope signatures in juvenile bull sharks (Carcharhinus leucas) up to 6.5 years post parturition. We found that maternal provisioning was detectable for up to 3.5 years after birth in muscle but only detectable in young-of-the-year for liver. Inclusion of sulphur revealed when maternal signatures disappeared from low-turnover tissue, while also identifying the spatial and trophic ecology patterns from fast-turnover tissue. These results reveal the importance of sampling fast turnover tissues to study the trophic ecology of juvenile elasmobranchs, and how the use of only δ15N and δ13C isotopes is likely to make maternal patterns more difficult to detect.

Coastal wetlands can be saved from sea level rise by recreating past tidal regimes.

Climate change driven Sea Level Rise (SLR) is creating a major global environmental crisis in coastal ecosystems, however, limited practical solutions are provided to prevent or mitigate the impacts. Here, we propose a novel eco-engineering solution to protect highly valued vegetated intertidal ecosystems. The new 'Tidal Replicate Method' involves the creation of a synthetic tidal regime that mimics the desired hydroperiod for intertidal wetlands. This synthetic tidal regime can then be applied via automated tidal control systems, "SmartGates", at suitable locations. As a proof of concept study, this method was applied at an intertidal wetland with the aim of restabilising saltmarsh vegetation at a location representative of SLR. Results from aerial drone surveys and on-ground vegetation sampling indicated that the Tidal Replicate Method effectively established saltmarsh onsite over a 3-year period of post-restoration, showing the method is able to protect endangered intertidal ecosystems from submersion. If applied globally, this method can protect high value coastal wetlands with similar environmental settings, including over 1,184,000 ha of Ramsar coastal wetlands. This equates to a saving of US$230 billion in ecosystem services per year. This solution can play an important role in the global effort to conserve coastal wetlands under accelerating SLR.

Lethal and sub-lethal effects of environmentally relevant levels of imidacloprid pesticide to Eastern School Prawn, Metapenaeus macleayi.

Pesticides are frequently employed to enhance agricultural production. Neonicotinoid pesticides (including imidacloprid) are often used to control sucking insects but have been shown to impact aquatic crustaceans. Imidacloprid is highly water soluble and has been detected in estuaries where it has been applied in adjacent catchments. We examined the impact of environmentally relevant concentrations of imidacloprid on Eastern School Prawn (Metapenaeus macleayi), an important exploited crustacean in Australia. Prawns were held for 8 days in estuarine water containing 0-4 µg L-1 of imidacloprid to assess potential lethal and non-lethal impacts. There was a non-linear relationship between exposure concentration and tissue concentration, with tissue concentrations peaking at exposures of 1.4 µg L-1 (1.16 to 1.64 µg L-1, 90% C.I.). There was no evidence for direct mortality associated with imidacloprid exposure, but exposure did influence the organism metabolome which likely reflects alterations in metabolic homeostasis, such as changes in the fatty acid composition which indicate a shift in lipid homeostasis. There was a positive correlation between exposure concentration and moulting frequency. Shedding of the exoskeleton may represent a mechanism through which prawns can expel the contaminant from their bodies. These results indicate that prawns experience several different sub-lethal effects when exposed to these pesticides, which may have implications for the health of populations.

Meta-Transcriptomic Identification of Divergent Amnoonviridae in Fish.

Tilapia lake virus (TiLV) has caused mass mortalities in farmed and wild tilapia with serious economic and ecological consequences. Until recently, this virus was the sole member of the Amnoonviridae, a family within the order Articulavirales comprising segmented negative-sense RNA viruses. We sought to identify additional viruses within the Amnoonviridae through total RNA sequencing (meta-transcriptomics) and data mining of published transcriptomes. Accordingly, we sampled marine fish species from both Australia and China and discovered several segments of two new viruses within the Amnoonviridae, tentatively called Flavolineata virus and Piscibus virus, respectively. In addition, by mining vertebrate transcriptome data, we identified nine additional virus transcripts matching to multiple genomic segments of TiLV in both marine and freshwater fish. These new viruses retained sequence conservation with the distantly related Orthomyxoviridae in the RdRp subunit PB1, but formed a distinct and diverse phylogenetic group. These data suggest that the Amnoonviridae have a broad host range within fish and that greater animal sampling will identify additional divergent members of the Articulavirales.