Soil microbial communities and degradation of pesticides in greenhouse effluent through a woodchip bioreactor.

Pesticides, including insecticides and fungicides, are major contaminants in the effluent from intensive agricultural systems, such as greenhouses. Because of their constant use and persistence, some pesticides can accumulate in soil and/or run off into adjacent waterways. Microbial communities in soil can degrade some pesticides, and bioreactors with enhanced microbial communities have the potential to facilitate decontamination before the effluent is released into the environment. In this study, we sampled the soil along a gradient from immediately below greenhouses, into, through and below a bioreactor. Multi-analyte pesticide screening was undertaken along with shotgun metagenomic sequencing, to assess microbial community taxonomic profiles and metabolic pathway responses for functional analysis. Two insecticides (imidacloprid and fipronil) and nine fungicides were identified in the soil samples, with a general decrease in most pesticides with increasing distance from the greenhouses. Diversity indexes of taxonomic profiles show changes in the microbial community along the gradient. In particular, microbial communities were significantly different in the bioreactor, with lower Shannon diversity compared to immediately below the greenhouses, in the channels leading into the bioreactor and further downstream. Metabolic pathway analysis revealed significant changes in a wide range of core housekeeping genes such as protein/amino acid synthesis and lipid/fatty acid biosynthesis among the sampling sites. The result demonstrates that the composition and potential functional pathways of the microbial community shifted towards an increased tendency for phytol and contaminant degradation in the bioreactor, facilitated by high organic matter content. This highlights the potential to use enhanced microbial communities within bioreactors to reduce contamination by some pesticides in sediment receiving run-off from greenhouses.

Semi-purified Antimicrobial Proteins from Oyster Hemolymph Inhibit Pneumococcal Infection.

Pneumococcal infections caused by Streptococcus pneumoniae are a leading cause of morbidity and mortality globally, particularly among children. The ability of S. pneumoniae to form enduring biofilms makes treatment inherently difficult, and options are further limited by emerging antibiotic resistance. The discovery of new antibiotics, particularly those with antibiofilm activity, is therefore increasingly important. Antimicrobial proteins and peptides (AMPs) from marine invertebrates are recognised as promising pharmacological leads. This study determined the in vitro antibacterial activity of hemolymph and unique protein fractions from an Australian oyster (Saccostrea glomerata) against multi-drug-resistant S. pneumoniae. We developed a successful method for hemolymph extraction and separation into 16 fractions by preparative HPLC. The strongest activity was observed in fraction 7: at 42 µg/mL protein, this fraction was bactericidal to S. pneumoniae and inhibited biofilm formation. Proteomic analysis showed that fraction 7 contained relatively high abundance of carbonic anhydrase, cofilin, cystatin B-like, and gelsolin-like proteins, while surrounding fractions, which showed lower or no antibacterial activity, contained these proteins in lower abundance or not at all. This work supports traditional medicinal uses of oysters and contributes to further research and development of novel hemolymph/AMP-based treatments for pneumococcal infections.

Pesticide occurrence in a subtropical estuary, Australia: Complementary sampling methods.

Monitoring pesticide run-off in the aquatic environment is ecologically important. Effective methods are required to detect the wide range of possible pesticides that enter estuaries from the surrounding catchment. Here, we investigate the occurrence of pesticides in the Richmond River estuary, Australia, and compare the effectiveness of using oysters and Chemcatcher® passive sampling devices against composite water samples. Samples were collected from six sites during two sampling periods: from January to March 2020 (4 weekly composite water samples and oyster collections) and from February to March 2021 (8 twice weekly composite water samples and Chemcatcher® deployment). Samples were analysed for up to 174 pesticides. A total of 21 pesticides were detected across all sites using all methods. The number of pesticides and mixture of pesticides detected in the 2020 sampling was higher in oyster samples than in water samples. In 2021, Chemcatcher® samplers detected more pesticides than in water samples. Herbicides were the most common in all samples. Insecticides and most fungicides were detected only in oysters and Chemcatcher®. Overall, the use of three complementary sampling approaches demonstrated a high level of pesticide input into the Richmond River estuary, highlighting the usefulness of oysters as biomonitors for some pesticides.

Toward non-invasive collection methods for sampling the microbiome of diabetic foot ulcers.

Identifying the microbiome within chronic diabetic foot ulcers is essential if effective antimicrobial therapies are to be administered. Using culture and 16S rRNA gene sequencing, the aim of this study was to compare the microbiome of paired tissue scraping samples with swab samples, collected from participants during attendance at a high-risk foot clinic. The mean richness of cultured swab and tissue scraping samples was consistent, with anaerobes infrequently isolated from both sample types. Comparing percentage frequencies of detection of selected genera of known and potential pathogens namely Staphylococcus, Streptococcus, Enterococcus, Corynebacterium, Enterobacteriaceae and Pseudomonas from cultured and sequenced swab and tissue scrapings indicated that both collection methods captured varying percentages of all the selected genera. The mean abundance of sequenced samples was not significantly different between swabs and tissue scrapings. The mean richness or number of distinct operational taxonomic units (OTUs) and Shannon's H diversity index were not significantly different between the two collection methods. The mean relative abundance of the selected genera of known and potential pathogens, including anaerobes Anaerococcus and Finegoldia, was higher in swabs compared with tissue scrapings and significantly so in Staphylococcus and Pseudomonas genera. Multivariate analyses confirmed no significant differences between the bacterial community compositions of the paired samples. These results suggest that tissue scrapings and swabs can effectively capture the microbiome of chronic DFUs using culture and 16S rRNA gene sequencing.

Bioaccumulation of estuarine pollutants in leaf oysters (Isognomon ephippium) on the mid-north coast, New South Wales, Australia.

Filter feeding bivalves are useful bioindicators for the detection of biologically available pollutants. We investigated trace metals, metalloids, and pesticides in leaf oyster (Isognomon ephippium) soft tissue and shells and compared them to sediment in five estuaries in northern New South Wales, Australia. Concentrations of Pb, Cr, Mn, Ni, Fe and Al were higher in sediments, whereas Zn, Cd, Ag, Hg, Se and As bioaccumulated in the soft tissue. The amount of Cu, Hg and Ni in the sediment from Tweed and Richmond River estuaries exceeded the Australian national sediment quality guideline values. Only one pesticide, atrazine, was detected in leaf oyster soft tissue. Combinations of six elements in the soft tissue were the best predictors of oyster condition index and shell size, whereas sediment contaminants showed weak relationships. Overall, the bioaccumulation of metals and metalloids increases with leaf oyster size and reduces leaf oyster condition, suggesting these large bivalves are useful bioindicators for pollution in estuarine environments.

Impacts of seasonal temperatures, ocean warming and marine heatwaves on the nutritional quality of eastern school prawns (Metapenaeus macleayi).

Ocean warming and marine heatwaves significantly alter environmental conditions in marine and estuarine environments. Despite their potential global importance for nutrient security and human health, it is not well understood how thermal impacts could alter the nutritional quality of harvested marine resources. We tested whether short-term experimental exposure to seasonal temperatures, projected ocean-warming temperatures, and marine heatwaves affected the nutritional quality of the eastern school prawn (Metapenaeus macleayi). In addition, we tested whether nutritional quality was affected by the duration of exposure to warm temperatures. We show the nutritional quality of M. macleayi is likely to be resilient to short- (28 d), but not longer-term (56 d) exposure to warming temperatures. The proximate, fatty acid and metabolite compositions of M. macleayi were unchanged after 28 d exposure to simulated ocean warming and marine heatwaves. The ocean-warming scenario did, however, show potential for elevated sulphur, iron and silver levels after 28 d. Decreasing saturation of fatty acids in M. macleayi after 28 d exposure to cooler temperatures indicates homeoviscous adaptation to seasonal changes. We found that 11 % of measured response variables were significantly different between 28 and 56 d when exposed to the same treatment, indicating the duration of exposure time and time of sampling are critical when measuring this species' nutritional response. Further, we found that future acute warming events could reduce harvestable biomass, despite survivors retaining their nutritional quality. Developing a combined knowledge of the variability in seafood nutrient content with shifts in the availability of harvested seafood is crucial for understanding seafood-derived nutrient security in a changing climate.

Exposure to multiple elements reduces the health of Saccostrea glomerata: An assessment of the Richmond River estuary, NSW, Australia.

This study investigated relationships between Sydney Rock Oyster (SRO) health and element concentrations in sediments and oysters from the Richmond River estuary. Six sites were sampled between November 2019 and May 2020. Multivariate permutational analysis of variance was used to compare oyster health parameters and element concentrations between sites, wet and dry conditions, and in oyster and sediment samples. Statistical analysis revealed significant spatial differences in oyster mortality, condition index, and size. Metal concentrations in oyster flesh significantly differed from metals in sediments. Most metals in sediments were below guideline values, except for Ni at some sites. Mortality, condition index, and weight correlated negatively with individual elements in oyster flesh (P, Zn, Mg, Al, Ni). BEST statistical models included various combinations of metals in sediment and flesh. This study highlights that spatial differences in SRO health tend to be related to site-specific metal compositions in sediment and oysters.

Ni accumulation and effects on a representative Cnidaria - Exaiptasia pallida during single element exposure and in combination with Mn.

Nickel (Ni) and manganese (Mn) are well known for the production of steel and alloys and are commonly found co-occurring in Ni ores. They are metals of environmental concern and contamination in the marine environment is problematic single exposures and in combination. Several studies have documented the effects of single metal exposure on the model anemone E. pallida, but research on the effects of metal mixtures is far less common. This novel study assesses the accumulation and stress effects of Ni and Mn over a 12-d exposure period. E. pallida were exposed in two separate experiments; Ni alone and Ni in combination with Mn, to assess accumulation, along with any effect on the density of symbionts and anemone tentacle length. Anemones were transferred to ambient seawater to assess depuration and recovery over 6 d. Anemone tissue accumulated Ni at a magnitude of five times higher in a mixture of 0.5 mg Ni/L with 2.5 mg Mn/L compared to the same concentration in a single Ni exposure experiment. In both experiments, Ni and Mn preferentially accumulated in the Symbiodinium spp. compared to the anemone tissue, but Ni depuration was more rapid in the mixture than Ni alone exposure. This study reveals a significant reduction in anemone Symbiodinium spp. density after exposure to Ni and Mn mixtures, but not with Ni exposure alone. A significant dose-dependent reduction in tentacle length was observed in anemones after 12 d of the Ni exposure both with and without Mn. The estimated sublethal concentration that causes tentacle retraction in 50% of test anemones (EC50) by Ni was 0.51 (0.25-0.73) mg/L, while in combination with Mn the EC50 was 0.30 mg Ni/L (confidence limits not calculatable). The present data reveals the importance of testing metal effects in combination before establishing safe limits for marine invertebrates.

Out of control: The need for standardised solvent approaches and data reporting in antibiofilm assays incorporating dimethyl-sulfoxide (DMSO).

Bacteria in biofilm formations are up to 1000 times less susceptible to antibiotics than their planktonic counterparts. Recognition of the role of biofilms in ∼80% of chronic infections, their contribution to bacterial tolerance and development of antimicrobial resistance, and thus the search for compounds with antibiofilm properties, has increased greatly in recent years. The need for robust experimental methods is therefore critical but currently undermined by inappropriate controls when dimethyl-sulfoxide (DMSO) is used to enhance test compound solubility. DMSO is known to have a limited effect on planktonic growth, but emerging data indicates that the solvent can affect biofilm formation even at low concentrations. Here, we present both a literature review on the application of DMSO in in vitro antibiofilm studies, as well as a series of experiments and Bayesian hormetic dose-response modelling to define the effects of DMSO alone and in combination with standard antibiotics using two clinically important biofilm-forming bacteria. DMSO has been used in 76 published studies to solubilise a wide variety of synthesised and natural products, including plant extracts, isolated secondary metabolites, modified lead molecules and proteins, in in vitro antibiofilm assays. DMSO solvent concentrations to which biofilms were exposed ranged between <1 and 100% but unfortunately, 35% of articles did not specify the DMSO concentrations used, 50% of articles did not include solvent controls and, of those that did, 26% did not specify control concentrations, 47% did not report or discuss control data, and 53% omitted media controls. In a further 12 studies, DMSO is used as a biofilm treatment, demonstrating the antibiofilm properties of this solvent at higher concentrations. We provide evidence that DMSO (between 0.03 and 25%) significantly inhibits biofilm formation in Pseudomonas aeruginosa, but not Streptococcus pneumoniae, and acts synergistically with standard antibiotics at very low concentrations (<1%). Interestingly, intermediate concentrations of DMSO (∼6%) strongly promote the growth of P. aeruginosa biofilms. As the research community strives to identify bioactive antimicrobial compounds, there is a need for increased scientific rigour when using DMSO as a solvent in antibiofilm assays.

Pesticide occurrence in an agriculturally intensive and ecologically important coastal aquatic system in Australia.

Coastal agricultural practices are often located in catchments upstream of ecologically important aquatic systems. Here, we investigate the occurrence of pesticides in a coastal creek flowing into a habitat-protected area within the Solitary Islands Marine Park, Australia. Water samples were collected from six sites along a creek transect during three sampling periods. Samples were analysed for 171 pesticide analytes, including organochlorines, organophosphates, herbicides, and fungicides. Five insecticides, two herbicides, and two fungicides were detected. The neonicotinoid imidacloprid was detected at 5 out of 6 sites, with concentrations reaching 294 µg L-1, the highest yet detected in Australian waterways. The organophosphate insecticide dimethoate was detected at 4 sites, which occurred at the 2nd highest detected concentration in the study (12.8 µg L-1). The presence of these pesticides in the aquatic environment downstream of horticulture in this and other regions may have serious implications for stream biota and ecologically important marine ecosystems.

Water quality and the health of remnant leaf oyster (Isognomon ephippium) populations in four Australian estuaries.

Leaf oysters (Isognomon ephippium) are large intertidal bivalves that form shellfish reefs. They have a patchy and restricted distribution in estuaries in northern New South Wales, Australia, where the water quality is impacted by a range of anthropogenic stressors from coastal agriculture, urbanisation, industry and recreational activities, along with natural stochastic events such as flooding. Little, however, is currently known about the tolerance of leaf oysters to poor water quality. This study investigated the condition of leaf oyster populations in four estuaries, by assessing the density, size-frequency and condition index. These biological parameters were modelled against habitat water quality parameters including nutrient loads, organic and inorganic carbon, algal biomass, sedimentation and total alkalinity. The water quality assessments were replicated during dry conditions and after rain in two estuaries. Leaf oyster density, size, and body condition significantly differed among the four estuaries. Density ranged from as few as 0.04 oysters per m2 in the heavily impacted Richmond River, up to 267 oysters per m2 in the Tweed River. Overall water quality was also significantly different among the four estuaries, and significant declines in water quality were observed after rain. Distance based linear models revealed significant correlations between biological indicators of leaf oyster reef health and a range of water quality parameters. Leaf oysters with higher condition occur in locations with high dissolved oxygen and salinity levels, whereas low pH and high nutrient run-off were associated with low density and poorer oyster condition. Nevertheless, dense populations were found to persist in areas with pH below 7.9, in high turbidity (over 27 NTU) and areas dominated by fine silt. Their ability to form dense reefs on estuarine soft muddy bottom habitats with low water flow, suggests that leaf oysters could be used to complement rock oysters in future intertidal shellfish reef restoration programs.

The nutritional and sensory quality of seafood in a changing climate.

Climate change is impacting living marine resources, whilst concomitantly, global reliance on seafood as a source of nutrition is increasing. Here we review an emerging research frontier, identifying significant impacts of climate-driven environmental change on the nutritional and sensory quality of seafood, and implications for human health. We highlight that changing ocean temperature, pH and salinity can lead to reductions in seafood macro and micronutrients, including essential nutrients such as protein and lipids. However, the nutritional quality of seafood appears to be more resilient in taxa that inhabit naturally variable environments such as estuaries and shallow near-coastal habitats. We develop criteria for assessing confidence in categorising the nutritional quality of seafood as vulnerable or resilient to climate change. The application of this criteria to a subset of seafood nutritional studies demonstrates confidence levels are generally low and could be improved by more realistic experimental designs and research collaboration. We highlight knowledge gaps to guide future research in this emerging field.

Manganese uptake and partitioning between the tissue of the anemone host Exaiptasia pallida and Symbiodinium spp., including assessment of stress and recovery.

Manganese (Mn) is essential for global steel and Mn-iron (Fe) alloy production. The human health effects of elevated Mn concentrations have been well established, but studies on its impact on marine invertebrates are limited. This study is the first to investigate Mn uptake in the sea anemone Exaiptasia pallida after chronic exposure (0.5, 1, 10, and 100 mg/L) for 24-d. Following exposure, E. pallida were transferred to ambient seawater for 6-d to assess Mn depuration. Mn accumulation and partitioning in host tissue and symbionts (Symbiodinium spp.), tentacle retraction, and symbiont cell density were measured during exposure and depuration. Mn concentrations were substantially higher in symbionts than tissue in all treatments after 24-d. No significant difference was observed for symbiont cell density after Mn exposure. Tentacle retractions were significantly higher in all Mn exposed treatments than controls at all time points. Mn depuration was observed for both tissue and symbionts but was more rapid in symbionts. This study reveals that Symbiodinium spp. can play a role in Mn uptake and depuration in anemones, but Mn loading does not affect cell density. These results help understand metal uptake and depuration in complex relationships between Symbiodinium spp. and other host taxa like corals.

Assessment of acetylcholinesterase, catalase, and glutathione S-transferase as biomarkers for imidacloprid exposure in penaeid shrimp.

Shrimp aquaculture is a valuable source of quality seafood that can be impacted by exposure to insecticides, such as imidacloprid. Here, adult black tiger shrimp (Penaeus monodon) were used to evaluate the activity of acetylcholinesterase (AChE), catalase (CAT), and glutathione S-transferase (GST) in abdominal, head, gill, and hepatopancreas tissue as biomarkers for imidacloprid exposure. Adult P. monodon were continuously exposed to imidacloprid in water (5 µgL-1 and 30 µgL-1) or feed (12.5 µg g-1 and 75 µg g-1) for either 4 or 21 days. The imidacloprid concentration in shrimp tissues was determined using liquid chromatography-mass spectrometry after QuEChER extraction, and AChE, CAT, and GST activities were estimated by spectrophotometric assay. Imidacloprid exposure in shrimp elevated the activity of biomarkers, and the enzymatic activity was positively correlated to tissue imidacloprid accumulation, although the effects varied in a tissue-, dose- and time-dependent manner. AChE activity was correlated to imidacloprid concentration in the abdominal tissue of shrimp and was likely related to neural tissue distribution, while the activity of CAT and GST confirmed a generalised anti-oxidant stress response. AChE, CAT, and GST were valuable biomarkers for assessing shrimp response to imidacloprid exposure from dietary or water sources, and the abdominal tissue was the most reliable for exposure assessment. An elevated response in each of these biomarkers during routine monitoring could provide an early warning of shrimp stress, suggesting that investigating potential contamination by neonicotinoid pesticides would be worthwhile.

Mollusc-Derived Brominated Indoles for the Selective Inhibition of Cyclooxygenase: A Computational Expedition.

Inflammation plays an important role in different chronic diseases. Brominated indoles derived from the Australian marine mollusk Dicathais orbita (D. orbita) are of interest for their anti-inflammatory properties. This study evaluates the binding mechanism and potentiality of several brominated indoles (tyrindoxyl sulfate, tyrindoleninone, 6-bromoisatin, and 6,6'-dibromoindirubin) against inflammatory mediators cyclooxygenases-1/2 (COX-1/2) using molecular docking, followed by molecular dynamics simulation, along with physicochemical, drug-likeness, pharmacokinetic (pk), and toxicokinetic (tk) properties. Molecular docking identified that these indole compounds are anchored, with the main amino acid residues, positioned in the binding pocket of the COX-1/2, required for selective inhibition. Moreover, the molecular dynamics simulation based on root mean square deviation (RMSD), radius of gyration (Rg), solvent accessible surface area (SASA), and root mean square fluctuation (RMSF) analyses showed that these natural brominated molecules transit rapidly to a progressive constant configuration during binding with COX-1/2 and seem to accomplish a consistent dynamic behavior by maintaining conformational stability and compactness. The results were comparable to the Food and Drug Administration (FDA)-approved selective COX inhibitor, aspirin. Furthermore, the free energy of binding for the compounds assessed by molecular mechanics-Poisson-Boltzmann surface area (MM-PBSA) confirmed the binding capacity of indoles towards COX-1/2, with suitable binding energy values except for the polar precursor tyrindoxyl sulfate (with COX-1). The physicochemical and drug-likeness analysis showed zero violations of Lipinski's rule, and the compounds are predicted to have excellent pharmacokinetic profiles. These indoles are projected to be non-mutagenic and free from hepatotoxicity, with no inhibition of human ether-a-go-go gene (hERG) I inhibitors, and the oral acute toxicity LD50 in rats is predicted to be similar or lower than aspirin. Overall, this work has identified a plausible mechanism for selective COX inhibition by natural marine indoles as potential therapeutic candidates for the mitigation of inflammation.

Marine heatwaves have minimal influence on the quality of adult Sydney rock oyster flesh.

Marine heatwaves (MHWs) are impacting marine biodiversity, including fisheries and aquaculture. However, it is largely unknown which species will be able to endure MHWs and at what price. Here, we applied elevated temperature (2 °C above ambient) and two different heatwave scenarios to adults of the economically important Sydney rock oyster (SRO, Saccostrea glomerata), and evaluated the impact on nutritional properties, gene expression profiles and immune health indicators. We found that elevated temperature (23 °C) and a variable heatwave (VHW) during winter caused some significant differences in the micronutrient and trace elements levels in oyster flesh. There was an increase of lead under VHW and a decrease in chromium, barium and aluminium under elevated temperature. Conversely, gene expression profiles and other physiological parameters, including flesh protein, fatty acid profiles and hemocyte numbers, were not affected by MHWs. These results indicate that adult SRO are reasonably resilient, and should continue to provide high-quality seafood, under near-future ocean warming and moderate heatwave scenarios.

Where three snail species attach while emersed in relation to heterogenous substrate temperatures underneath intertidal boulders.

Mobile intertidal gastropods can employ behavioural thermoregulation to mitigate thermal stress, which may include retreating under boulders when emersed. However, little is known about how gastropod occupancy of under-boulder habitats is associated with any variations in substrate temperature that exist under boulders. Thermal imagery was used to measure the temperature of boulder lower surfaces and investigate how three snail species were associated at low tide with the maximum and average temperatures underneath grey siltstone and quartzite. Lower boulder surfaces had heterogeneous temperatures, with grey siltstone having temperature gradients and quartzite temperature showing mosaics. Temperature differences between the hottest and coolest gradient or mosaic locations were >5 °C; thus there was a range of temperatures that snails could interact with. All three snail species occupied cooler parts of temperature mosaics or gradients, avoiding the hottest areas. Stronger associations were detected on the hotter grey siltstone and for the more-thermally sensitive Nerita atramentosa and Diloma concameratum. Even though snails were associated with cooler areas, some individuals were still exposed to extreme substratum heat (>50 °C). These results suggest that gastropod thermoregulatory behaviour is far more complex than simply retreating underneath boulders at low tide, as there is also a range of under-boulder temperatures that they interact with. Untangling interactions between intertidal gastropods and heterogenous substrate temperatures is important given rocky seashores already represent a thermally-variable and potentially-stressful habitat, which may be exacerbated further given predictions of warming temperatures associated with climate change.

Rocks of different mineralogy show different temperature characteristics: implications for biodiversity on rocky seashores.

As some intertidal biota presently live near their upper tolerable thermal limits when emersed, predicted hotter temperatures and an increased frequency of extreme-heat events associated with global climate change may challenge the survival and persistence of such species. To predict the biological ramifications of climate change on rocky seashores, ecologists have collected baseline rock temperature data, which has shown substrate temperature is heterogenous in the rocky intertidal zone. A multitude of factors may affect rock temperature, although the potential roles of boulder surface (upper versus lower), lithology (rock type) and minerology have been largely neglected to date. Consequently, a common-garden experiment using intertidal boulders of six rock types tested whether temperature characteristics differed among rock types, boulder surfaces, and whether temperature characteristics were associated with rock mineralogy. The temperature of the upper and lower surfaces of all six rock types was heterogeneous at the millimetre to centimetre scale. Three qualitative patterns of temperature difference were identified on boulder surfaces: gradients; mosaics; and limited heterogeneity. The frequency of occurrence of these temperature patterns was heavily influenced by cloud cover. Upper surfaces were generally hotter than lower surfaces, plus purple siltstone and grey siltstone consistently had the hottest temperatures and white limestone and quartzite the coolest. Each rock type had unique mineralogy, with maximum temperatures correlated with the highest metallic oxide and trace metal content of rocks. These baseline data show that rock type, boulder surface and mineralogy all contribute to patterns of heterogenous substrate temperature, with the geological history of rocky seashores potentially influencing the future fate of species and populations under various climate change scenarios.

Acute toxicity, accumulation and sublethal effects of four neonicotinoids on juvenile Black Tiger Shrimp (Penaeus monodon).

Neonicotinoid pesticides have been detected in aquatic habitats, and exposure may impact the health of aquatic organisms such as commercially-important crustaceans. Black Tiger Shrimp (Penaeus monodon) is a broadly distributed and high-value shrimp species that rely on estuaries for early life stages. Differences in the acute toxicity and accumulation of different neonicotinoids in tissues of commercial crustaceans have not been widely investigated. This study compared acute toxicity, uptake, and depuration of four neonicotinoids; thiamethoxam, clothianidin, acetamiprid, and imidacloprid, on juvenile P. monodon and their effects on enzyme biomarkers. Acute toxicity (48-h LC50) was determined as 190 µg L-1 (clothianidin), 390 µg L-1 (thiamethoxam), 408 µg L-1 (imidacloprid), and >500 µg L-1(acetamiprid). To assess uptake and elimination, shrimp were exposed to a fixed 5 µg L-1 water concentration for eight days (uptake) or four days of exposure followed by four days of depuration (elimination). Neonicotinoid water and tissue concentrations were measured by liquid chromatography-mass spectrometry following solid-phase extraction and QuEChER extraction respectively. The lower toxicity associated with acetamiprid could be associated with lower accumulation in the tissue, with concentrations remaining below 0.01 µg g-1. The activity of acetylcholinesterase, catalase and glutathione S-transferase in abdominal tissues was determined by spectrophotometric assay, with significant sublethal effects detected for all four neonicotinoids. Depuration reduced the tissue concentration of the active ingredient and reduced the activity of oxidative stress enzymes. Given acetamiprid showed no acute toxicity and reduced impact on the enzymatic activity of P. monodon, it may be an appropriate alternative to other neonicotinoids in shrimp producing areas.

Impacts of Neonicotinoids on Molluscs: What We Know and What We Need to Know.

The broad utilisation of neonicotinoids in agriculture has led to the unplanned contamination of adjacent terrestrial and aquatic systems around the world. Environmental monitoring regularly detects neonicotinoids at concentrations that may cause negative impacts on molluscs. The toxicity of neonicotinoids to some non-target invertebrates has been established; however, information on mollusc species is limited. Molluscs are likely to be exposed to various concentrations of neonicotinoids in the soil, food and water, which could increase their vulnerability to other sources of mortality and cause accidental exposure of other organisms higher in the food chain. This review examines the impacts of various concentrations of neonicotinoids on molluscs, including behavioural, physiological and biochemical responses. The review also identifies knowledge gaps and provides recommendations for future studies, to ensure a more comprehensive understanding of impacts from neonicotinoid exposure to molluscs.