Characterization of the transcriptional effects of the plastic additive dibutyl phthalate alone and in combination with microplastic on the green-lipped mussel Perna canaliculus.

The presence and persistence of microplastics (MPs) in diverse aquatic environments are of global concern. Microplastics can impact marine organisms via direct physical interaction and the release of potentially harmful chemical additives incorporated into the plastic. These chemicals are physically bound to the plastic matrix and can leach out. The hazards associated with chemical additives to exposed organisms is not well characterized. We investigated the hazards of plastic additives leaching from plastic. We used the common plasticizer dibutyl phthalate (DBP) as a chemical additive proxy and the New Zealand green-lipped mussel (Perna canaliculus) as a model. We used early-adult P. canaliculus exposed to combinations of virgin and DBP-spiked polyvinyl chloride (PVC), MPs, and DBP alone for 7 days. Whole transcriptome sequencing (RNA-seq) was conducted to assess whether leaching of DBP from MPs poses a hazard. The differences between groups were evaluated using pairwise permutational multivariate analysis of variance (PERMANOVA), and all treatments were significantly different from controls. In addition, a significant difference was seen between DBP and PVC MP treatment. Transcriptome analysis revealed that mussels exposed to DBP alone had the most differentially expressed genes (914), followed by PVC MP + DBP (448), and PVC MP (250). Gene ontology functional analysis revealed that the most enriched pathway types were in cellular metabolism, immune response, and endocrine disruption. Microplastic treatments enriched numerous pathways related to cellular metabolism and immune response. The combined exposure of PVC MP + DBP appears to cause combined effects, suggesting that DBP is bioavailable to the exposed mussels in the PVC MP + DBP treatment. Our results support the hypothesis that chemical additives are potentially an important driver of MP toxicity. Environ Toxicol Chem 2024;43:1604-1614. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Biochar derivation at low temperature: A novel strategy for harmful resource usage of antibiotic mycelial dreg.

Antibiotic mycelial dreg (AMD) has been categorized as hazardous waste due to the high residual hazardous contaminants. Inappropriate management and disposal of AMD can cause potential environmental and ecological risks. In this study, the potential of pleuromutilin mycelial dreg (PMD) as a novel feedstock for preparing tetracycline hydrochloride (TC) adsorbent was explored to achieve safe management of PMD. The results suggested that residual hazardous contaminants were completely eliminated after pyrolysis. With the increase of pyrolysis temperature, the yields, H/C, O/C, (O + N)/C, and pore size in PMD-derived biochars (PMD-BCs) decreased, while BET surface area and pore volume increased, resulting in the higher stability of the PMD-BCs prepared from higher temperatures. The TC adsorption of the PMD-BCs increased from 27.3 to 46.9 mg/g with the increase of the pyrolysis temperature. Surprisingly, pH value had a strong impact on the TC adsorption, the adsorption capacity of BC-450 increased from 6.5 to 71.1 mg/g when the solution pH value increased from 2 to 10. Lewis acid-base interaction, pore filling, π-π interaction, hydrophobic interaction, and charge-assisted hydrogen bond (CAHB) are considered to drive the adsorption. This work provides a novel pathway for the concurrent detoxification and reutilization of AMD.

Effect-based monitoring of two rivers under urban and agricultural influence reveals a range of biological activities in sediment and water extracts.

Chemical contaminants, such as pesticides, pharmaceuticals and industrial compounds are ubiquitous in surface water and sediment in areas subject to human activity. While targeted chemical analysis is typically used for water and sediment quality monitoring, there is growing interest in applying effect-based methods with in vitro bioassays to capture the effects of all active contaminants in a sample. The current study evaluated the biological effects in surface water and sediment from two contrasting catchments in Aotearoa New Zealand, the highly urbanised Whau River catchment in Tamaki Makaurau (Auckland) and the urban and mixed agricultural Koreti (New River) Estuary catchment. Two complementary passive sampling devices, Chemcatcher for polar chemicals and polyethylene (PED) for non-polar chemicals, were applied to capture a wide range of contaminants in water, while composite sediment samples were collected at each sampling site. Bioassays indicative of induction of xenobiotic metabolism, receptor-mediated effects, genotoxicity, cytotoxicity and apical effects were applied to the water and sediment extracts. Most sediment extracts induced moderate to strong estrogenic and aryl hydrocarbon (AhR) activity, along with moderate toxicity to bacteria. The water extracts showed similar patterns to the sediment extracts, but with lower activity. Generally, the polar Chemcatcher extracts showed greater estrogenic activity, photosynthesis inhibition and algal growth inhibition than the non-polar PED extracts, though the PED extracts showed greater AhR activity. The observed effects in the water extracts were compared to available ecological effect-based trigger values (EBT) to evaluate the potential risk. For the polar extracts, most sites in both catchments exceeded the EBT for estrogenicity, with many sites exceeding the EBTs for AhR activity and photosynthesis inhibition. Of the wide range of endpoints considered, estrogenic activity, AhR activity and herbicidal activity appear to be the primary risk drivers in both the Whau and Koreti Estuary catchments.

Monitoring the Health of Coastal Environments in the Pacific Region-A Review.

Coastal areas provide important ecological services to populations accessing, for example, tourism services, fisheries, minerals and petroleum. Coastal zones worldwide are exposed to multiple stressors that threaten the sustainability of receiving environments. Assessing the health of these valuable ecosystems remains a top priority for environmental managers to ensure the key stressor sources are identified and their impacts minimized. The objective of this review was to provide an overview of current coastal environmental monitoring frameworks in the Asia-Pacific region. This large geographical area includes many countries with a range of climate types, population densities and land uses. Traditionally, environmental monitoring frameworks have been based on chemical criteria set against guideline threshold levels. However, regulatory organizations are increasingly promoting the incorporation of biological effects-based data in their decision-making processes. Using a range of examples drawn from across the region, we provide a synthesis of the major approaches currently being applied to examine coastal health in China, Japan, Australia and New Zealand. In addition, we discuss some of the challenges and investigate potential solutions for improving traditional lines of evidence, including the coordination of regional monitoring programs, the implementation of ecosystem-based management and the inclusion of indigenous knowledge and participatory processes in decision-making.

Comparison between droplet digital PCR and reverse transcription-quantitative PCR methods to measure ecotoxicology biomarkers.

Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is currently the gold-standard technique for detecting and quantifying messenger RNA. However, without proper validation, the method may produce artefactual and non-reproducible cycle threshold values generating poor-quality data. The newer droplet digital PCR (ddPCR) method allows for the absolute quantification of targeted nucleic acids providing more sensitive and accurate measurements without requiring external standards. This study compared these two PCR-based methods to measure the expression of well-documented genes used in ecotoxicology studies. We exposed Mediterranean mussels (Mytilus galloprovincialis) to copper and analyzed gene expression in gills and digestive glands using RT-qPCR and ddPCR assays. A step-by-step methodology to optimize and compare the two technologies is described. After ten-fold serial complementary DNA dilution, both RT-qPCR and ddPCR exhibited comparable linearity and efficiency and produced statistically similar results. We conclude that ddPCR is a suitable method to assess gene expression in an ecotoxicological context. However, RT-qPCR has a shorter processing time and remains more cost-effective.

The relationships between specific dynamic action, nutrient retention and feed conversion ratio in farmed freshwater Chinook salmon (Oncorhynchus tshawytscha).

Improving the feed conversion ratio (FCR; the amount of feed consumed relative to the amount of weight gain) can reduce both production costs and environmental impacts of farmed fish. The aim of this study was to investigate what drives FCR to understand how nutrients are retained, as well as the amount of oxygen consumed for digestion, absorption and assimilation (a metabolic process known as specific dynamic action, SDA). Feed-efficient and inefficient Chinook salmon (Oncorhynchus tshawytscha) in fresh water were identified using ballotini beads and X-radiography that tracked individual feed intake across three assessment periods under satiated feeding. This allowed a comparison of physiological traits and body composition between the two FCR phenotypes over two time points as Chinook salmon grew from 305 to 620 g. Fish with higher daily feed intake (DFI) had higher daily weight gain (DWG) as expected. Nonetheless, the relationship between FCR and DFI as well as FCR and DWG was variable between time points. FCR and DWG were not correlated at the first time point and were negatively correlated at the second time point. In contrast, FCR and DFI were positively correlated at the first time point but not the second. Despite this, efficient fish ate smaller meals and retained more protein, lipid and energy in their body tissues. There was no detectable difference in metabolism between the two FCR phenotypes with respect to minimal resting metabolic rate, maximum metabolic rate, aerobic scope, or SDA parameters. In conclusion, FCR is not consistently associated with growth and metabolic differences in freshwater Chinook salmon, but FCR-efficient fish retain more nutrients and consume smaller meals.

Does plastic type matter? Insights into non-indigenous marine larvae recruitment under controlled conditions.

Marine plastic debris (MPD) are a global threat to marine ecosystems. Among countless ecosystem impacts, MPD can serve as a vector for marine 'hitchhikers' by facilitating transport and subsequent spread of unwanted pests and pathogens. The transport and spread of these non-indigenous species (NIS) can have substantial impacts on native biodiversity, ecosystem services/functions and hence, important economic consequences. Over the past decade, increasing research interest has been directed towards the characterization of biological communities colonizing plastic debris, the so called Plastisphere. Despite remarkable advances in this field, little is known regarding the recruitment patterns of NIS larvae and propagules on MPD, and the factors influencing these patterns. To address this knowledge gap, we used custom-made bioassay chambers and ran four consecutive bioassays to compare the settlement patterns of four distinct model biofouling organisms' larvae, including the three notorious invaders Crassostrea gigas, Ciona savignyi and Mytilus galloprovincialis, along with one sessile macro-invertebrate Spirobranchus cariniferus, on three different types of polymers, namely Low-Linear Density Polyethylene (LLDPE), Polylactic Acid (PLA), Nylon-6, and a glass control. Control bioassay chambers were included to investigate the microbial community composition colonizing the different substrates using 16S rRNA metabarcoding. We observed species-specific settlement patterns, with larvae aggregating on different locations on the substrates. Furthermore, our results revealed that C. savignyi and S. cariniferus generally favoured Nylon and PLA, whereas no specific preferences were observed for C. gigas and M. galloprovincialis. We did not detect significant differences in bacterial community composition between the tested substrates. Taken together, our results highlight the complexity of interactions between NIS larvae and plastic polymers. We conclude that several factors and their potential interactions influenced the results of this investigation, including: (i) species-specific larval biological traits and ecology; (ii) physical and chemical composition of the substrates; and (iii) biological cues emitted by bacterial biofilm and the level of chemosensitivity of the different NIS larvae. To mitigate the biosecurity risks associated with drifting plastic debris, additional research effort is critical to effectively decipher the mechanisms involved in the recruitment of NIS on MPD.

Enhancing consumption responsibility to address global plastic pollution.

Plastic pollution is a global crisis, especially in the marine environment. Excessive consumption and unsound disposal are responsible for the constant accumulation of plastic waste, resulting in plastic litter and microplastic contamination on a global scale. Establishing a new global framework is regarded as a promising tool to address plastic pollution, including marine plastic litter. However, there is a need to raise awareness of the role of consumers at individual and national levels in reducing the use of unnecessary plastics and increasing the recycling of plastic waste. The global framework should incorporate aspects of the importance of consumption responsibility in solutions addressing the issue of plastic pollution.

Disentangling the influence of microplastics and their chemical additives on a model detritivore system.

Microplastics (MPs) can negatively impact freshwater organisms via physical effects of the polymer itself and/or exposure to chemicals added to plastic during production to achieve desired characteristics. Effects on organisms may result from direct exposure to plastic particles and/or chemical additives or effects may manifest as indirect effects through ecological interactions between organisms (e.g., reduced food availability that impairs a consumer). To disentangle these issues, we used a simplified freshwater food web interaction comprising microbes and macroinvertebrate detritivores to evaluate the toxicity of 1) polyvinyl chloride (PVC) MPs without added chemicals (virgin), 2) the common chemical additive dibutyl phthalate (DBP), and 3) PVC MPs with incorporated DBP. Exposure to virgin PVC MPs (0.33 and 3.3 mg/L) caused negligible ecological effect with the exception of reduced macroinvertebrate feeding rates at 3.3 mg/L. Exposure to DBP (1 mg/L) both individually and when incorporated into the PVC MPs negatively impacted all tested endpoints, including microbial and macroinvertebrate respiration, feeding rate and assimilation efficiency. DBP leached rapidly from the MPs into the water, and also accumulated in macroinvertebrates and their food, providing multiple routes of exposure. Our findings suggest that additives which are intentionally incorporated into MPs could play a key role in MP toxicity and contribute to the disruption of key ecological interactions underpinning ecosystem processes, such as leaf litter decomposition.

Key Challenges to the Effective Management of Pollutants in Water and Sediment.

The intensification of human activities is placing increasing pressure on the ecosystems of riverine, estuarine, and coastal waters, as these compartments are sinks for many anthropogenic contaminants [...].

Characterization of the effects of triclosan on sperm and embryos of Mytilus and Perna mussel species.

The Greenshell™ mussel (GSM), Perna canaliculus, is a culturally and commercially important species in New Zealand. Declines in spat settlement of GSM have been observed in important growing areas and the cause(s) have not been identified. One hypothesis is that chemical contaminants could be a contributing factor. The aim to this study was to investigate the effects of acute exposure on early life stages using the anti-microbial triclosan (TCS) as a benchmark toxicant and the blue mussel (BM), Mytilus galloprovincialis, as a reference species. Sperm and embryos of BM and GSM were exposed to TCS for 1 h and 48 h, respectively. Following exposures, a range of parameters were investigated including spermatozoa cellular characteristics via flow cytometry, fertilization success, larval mortality and size. Exposure to TCS negatively impacted functional parameters of sperm, reduced the fertilization success and larval size, and increased larval mortality in both BM and GSM with LC5048h of 94.3 and 213 µg L-1, respectively. Triclosan increased sperm ROS production in both species, which could cause destabilisation of mitochondrial and other cellular membranes, resulting in reduced mitochondrial membrane potential (BM) and increased sperm size (GSM), leading to apoptosis in both species. Fertilization success of GSM was only affected at the highest TCS concentration tested (391 µg L-1), but development of larvae derived from exposed sperm was affected from the lowest concentrations tested (0.5 and 5.2 µg L-1) in both species. This highlights the importance of assessing the sensitivity of contaminants across developmental stages. Results of this study confirm that TCS causes oxidative stress and has membranotropic effects, and that early life stages of the endemic GSM are suitable to assess ecotoxicity of contaminants such as TCS.

First transcriptome of the copepod Gladioferens pectinatus subjected to chronic contaminant exposures.

Contaminants are often at low concentrations in ecosystems and their effects on exposed organisms can occur over long periods of time and across multiple generations. Alterations to subcellular mechanistic pathways in response to exposure to contaminants can provide insights into mechanisms of toxicity that methods measuring higher levels of biological may miss. Analysis of the whole transcriptome can identify novel mechanisms of action leading to impacts in exposed biota. The aim of this study was to characterise how exposures to copper, benzophenone and diclofenac across multiple generations altered molecular expression pathways in the marine copepod Gladioferens pectinatus. Results of the study demonstrated differential gene expression was observed in cultures exposure to diclofenac (569), copper (449) and benzophenone (59). Pathways linked to stress, growth, cellular and metabolic processes were altered by exposure to all three contaminants with genes associated with oxidative stress and xenobiotic regulation also impacted. Protein kinase functioning, cytochrome P450, transcription, skeletal muscle contraction/relaxation, mitochondrial phosphate translocator, protein synthesis and mitochondrial methylation were all differentially expressed with all three chemicals. The results of the study also suggested that using dimethyl sulfoxide as a dispersant influenced the transcriptome and future research may want to investigate it's use in molecular studies. Data generated in this study provides a first look at transcriptomic response of G. pectinatus exposed to contaminants across multiple generations, future research is needed to validate the identified biomarkers and link these results to apical responses such as population growth to demonstrate the predictive capacity of molecular tools.

Investigating the Ecotoxicity of Select Emerging Organic Contaminants Toward the Marine Copepod Gladioferens pectinatus.

Estuarine ecosystems are recipients of anthropogenic stressors released from land-based activities. The aim of the present study was to investigate the ecotoxicological hazards of organic contaminants toward the estuarine copepod Gladioferens pectinatus using acute and chronic testing. Most chemicals demonstrated acute toxicity and influenced development of the copepods. Further research should be conducted to investigate these chemicals and their mixtures using long-term, multigenerational testing to characterize mechanisms of toxicity. Environ Toxicol Chem 2022;41:792-799. © 2022 SETAC.

Trace Metal Residues in Marine Mussels: A Global Survey.

Pressures from anthropogenic activities are causing degradation of estuarine and coastal ecosystems around the world. Trace metals are key pollutants that are released and can partition in a range of environmental compartments, to be ultimately accumulated in exposed biota. The level of pressure varies with locations and the range and intensity of anthropogenic activities. The present study measured residues of trace metals in Mytilus mussel species collected from a range of locations around the world in areas experiencing a gradient of anthropogenic pressures that we classified as low, moderate, or high impact. The data showed no grouping/impact level when sampling sites in all countries were incorporated in the analysis, but there was significant clustering/impact level for most countries. Overall, high-impact areas were characterized by elevated concentrations of zinc, lead, nickel, and arsenic, whereas copper and silver were detected at higher concentrations in medium-impact areas. Finally, whereas most metals were found at lower concentrations in areas classified as low impact, cadmium was typically elevated in these areas. The present study provides a unique snapshot of worldwide levels of coastal metal contamination through the use of Mytilus species, a well-established marine biomonitoring tool. Environ Toxicol Chem 2021;40:3434-3440. © 2021 SETAC.

Towards more ecologically relevant investigations of the impacts of microplastic pollution in freshwater ecosystems.

Microplastic pollution is a major environmental concern and the subject of a rapidly growing body of research. Much of this research has focused on the direct effects of microplastics on single species and there is limited information on how microplastics affect different functional groups of organisms, multi-species interactions, and ecosystem processes. We focused on freshwater systems and reviewed 146 studies of microplastic effects on freshwater biota and recorded features including particle characteristics, study designs, functional types of species tested and ecotoxicological endpoints measured. Study species were categorized based on their ecosystem role/functional feeding group rather than taxonomy. We found that most studies were conducted on single species (95%) and focused on a narrow range of functional groups of organisms (mostly filter feeders, 37% of studies). Very few studies have investigated multi-species interactions and ecosystem processes. In many studies, certain characteristics of microplastics, such as polymer type were not well matched with the feeding and habitat ecology of test species, potentially reducing their ecological relevance. Median laboratory study test concentrations were 5-6 orders of magnitude higher than those reported in the field and few studies considered the effects of chemical additives in plastics (6%). We recommend that studies addressing the ecological effects of microplastics need to address neglected functional groups of organisms, design experiments to better match the ecology of test species, and increase in experimental scale and complexity to identify any indirect effects on species interactions and ecosystem processes. We suggest that examining microplastics through an ecological lens that better integrates the feeding and habitat ecology of test organisms will advance our understanding of the effects microplastics have in the environment.

Effects of pristine microplastics and nanoplastics on soil invertebrates: A systematic review and meta-analysis of available data.

In laboratory studies, microplastics and/or nanoplastics (MPs/NPs) have been shown to cause a variety of ecotoxicological effects on soil invertebrates. Existing data on the effects of these plastic debris on biological functions and physiological systems, showed a great variability among studies. Thus, how soil invertebrates respond to different types, shapes, sizes and concentrations of pristine MPs/NPs remains to be further characterized. The present work is an up-to-date review on quantitative and qualitative data on the effects of pristine MPs/NPs on soil invertebrates in laboratory conditions. Research priorities are also discussed. Out of a total of 1061 biological endpoints investigated in 56 studies, 49% were significantly affected after exposed to pristine MPs/NPs. The polymers with chloro and phenyl groups had more negative impacts on soil invertebrates than other polymers. Most studies used earthworm and nematode species as model organisms. For nematodes, the impact of MPs/NPs seemed to be concentration-dependent and higher concentrations of pristine MPs/NPs appeared to have more adverse impacts on biological functions and physiological systems, but this trend was not confirmed in earthworms. Meta-analysis revealed that pristine MP/NP concentrations higher than 1 g kg-1 (in soil) may decrease growth and survival of earthworms, while a concentration higher than 1 µg L-1 (in water) may affect nematode reproductive fitness.

Plastic additives: challenges in ecotox hazard assessment.

The risk of plastic debris, and specifically micro(nano)plastic particles, to ecosystems remains to be fully characterized. One particular issue that warrants further characterization is the hazards associated with chemical additives within micro(nano)plastic as they are not chemically bound within the polymers and can be persistent and biologically active. Most plastics contain additives and are therefore potential vectors for the introduction of these chemicals into the environment as they leach from plastic, a process that can be accelerated through degradation and weathering processes. There are knowledge gaps on the ecotoxicological effects of plastic additives and how they are released from parent plastic materials as they progressively fragment from the meso to micro and nano scale. This review summarizes the current state of knowledge of the ecotoxicity of plastic additives and identifies research needs to characterize the hazard they present to exposed biota. The potential ecological risk of chemical additives is of international concern so key differences in governance between the European Union and New Zealand to appropriately characterize their risk are highlighted.

Biosecurity implications of drifting marine plastic debris: Current knowledge and future research.

The introduction and spread of marine non-indigenous species (NIS) and pathogens into new habitats are a major threat to biodiversity, ecosystem services, human health, and can have substantial economic consequences. Shipping is considered the main vector for marine biological invasions; less well understood is the increased spread of marine NIS and pathogens rafting on marine plastic debris (MPD). Despite an increasing research interest and recent progress in characterizing the plastisphere, this manuscript highlights critical knowledge gaps and research priorities towards a better understanding of the biosecurity implications of MPD. We advocate for future research to (i) investigate plastisphere community succession and the factors influencing NIS propagules and pathogens recruitment through robust experimental investigations; (ii) combine microscopy and molecular approaches to effectively assess the presence of specific taxa; (iii) include additional genetic markers to thoroughly characterize the biodiversity associated with MPD and explore the presence of specific marine pests.

Assessing the Efficacy of a Sediment Remediation Program Using Benthic and Pelagic Copepod Bioassays.

Tributyltin is an organotin chemical that has been commonly used in ship antifouling paints. Despite the global total prohibition of tributyltin-based paint in 2008, tributyltin continues to be found at toxic levels in areas of high maritime traffic such as ports and harbors. A remediation program was conducted at a New Zealand port to reduce tributyltin and copper concentrations to acceptable values. The present study assessed the efficacy of the program using a combination of chemical analyses and copepod bioassays. Sediment and water samples were collected at 3 locations along a spatial gradient within the port, and concentrations of various organotin compounds and trace metal levels were measured pre- and postremediation. The toxicity of sediment and elutriate samples was estimated by benthic and pelagic copepod bioassays. Although acute toxicity in sediment samples was reduced following remediation, reproductive success was still affected for the benthic copepod. This approach combining chemical analysis and bioassays is promising for assessing the efficacy of remediation processes at contaminated marine sites. Environ Toxicol Chem 2020;39:492-499. © 2019 SETAC.