Mesocosm study of PAC-modified clay effects on Karenia brevis cells and toxins, chemical dynamics, and benthic invertebrate physiology.

Modified clay compounds are used globally as a method of controlling harmful algal blooms, and their use is currently under consideration to control Karenia brevis blooms in Florida, USA. In 1400 L mesocosm tanks, chemical dynamics and lethal and sublethal impacts of MC II, a polyaluminum chloride (PAC)-modified kaolinite clay, were evaluated over 72 h on a benthic community representative of Sarasota Bay, which included blue crab (Callinectes sapidus), sea urchin (Lytechinus variegatus), and hard clam (Mercenaria campechiensis). In this experiment, MC II was dosed at 0.2 g L-1 to treat bloom-level densities of K. brevis at 1 × 106 cells L-1. Cell removal in MC II-treated tanks was 57% after 8 h and 95% after 48 h. In the water column, brevetoxin analogs BTx-1 and BTx-2 were found to be significantly higher in untreated tanks at 24 and 48 h, while in MC II-treated tanks, BTx-3 was found to be higher at 48 h and BTx-B5 was found to be higher at 24 and 48 h. In MC II floc, we found no significant differences in BTx-1 or BTx-2 between treatments for any time point, while BTx-3 was found to be significantly higher in the MC II-treated tanks at 48 and 72 h, and BTx-B5 was higher in MC II-treated tanks at 24 and 72 h. Among various chemical dynamics observed, it was notable that dissolved phosphorus was consistently significantly lower in MC II tanks after 2 h, and that turbidity in MC II tanks returned to control levels 48 h after treatment. Dissolved inorganic carbon and total seawater alkalinity were significantly reduced in MC II tanks, and partial pressure of CO2 (pCO2) was significantly higher in the MC II-only treatment after 2 h. In MC II floc, particulate phosphorus was found to be significantly higher in MC II tanks after 24 h. In animals, lethal and sublethal responses to MC II-treated K. brevis did not differ from untreated K. brevis for either of our three species at any time point, suggesting MC II treatment at this dosage has negligible impacts to these species within 72 h of exposure. These results appear promising in terms of the environmental safety of MC II as a potential bloom control option, and we recommend scaling up MC II experiments to field trials in order to gain deeper understanding of MC II performance and dynamics in natural waters.

Genome-wide sequencing reveals geographical variations in the thermal adaptation of an aquaculture species with frequent seedling introductions.

Climate change and human activity are essential factors affecting marine biodiversity and aquaculture, and understanding the impacts of human activities on the genetic structure to increasing high temperatures is crucial for sustainable aquaculture and marine biodiversity conservation. As a commercially important bivalve, the Manila clam Ruditapes philippinarum is widely distributed along the coast of China, and it has been frequently introduced from Fujian Province, China, to other regions for aquaculture. In this study, we collected four populations of Manila clams from different areas to evaluate their thermal tolerance by measuring cardiac performance and genetic variations using whole-genome resequencing. The upper thermal limits of the clams showed high variations within and among populations. Different populations displayed divergent genetic compositions, and the admixed population was partly derived from the Zhangzhou population in Fujian Province, implying a complex genomic landscape under the influence of local genetic sources and human introductions. Multiple single nucleotide polymorphisms (SNPs) were associated with the cardiac functional traits, and some of these SNPs can affect the codon usage and the structural stability of the resulting protein. This study shed light on the importance of establishing long-term ecological and genetic monitoring programs at the local level to enhance resilience to future climate change.

The long-lived deep-sea bivalve Acesta excavata is sensitive to the dual stressors of sediment and warming.

Human influence in the deep-sea is increasing as mining and drilling operations expand, and waters warm because of climate change. Here, we investigate how the long-lived deep-sea bivalve, Acesta excavata responds to sediment pollution and/or acute elevated temperatures. A. excavata were exposed to suspended sediment, acute warming, and a combination of the two treatments for 40 days. We measured O2 consumption, NH4+ release, Total Organic Carbon (TOC), and lysosomal membrane stability (LMS). We found suspended sediment and warming interacted to decrease O:N ratios, while sediment as a single stressor increased the release of TOC and warming increased NH4+ release in A. excavata. Warming also increased levels of LMS. We found A. excavata used protein catabolism to meet elevated energetic demands indicating a low tolerance to stress. A. excavata has limited capacity for physiological responses to the stressors of warming and sediment which may lead to decreased fitness of A. excavata.

Influencing intertidal food web: Implications of ocean acidification on the physiological energetics of key species the 'wedge' clam Donax faba.

Ocean acidification has become increasingly severe in coastal areas. It poses emerging threats to coastal organisms and influences ecological functioning. Donax faba, a dominant clam in the intertidal zone of the Bay of Bengal, plays an important role in the coastal food web. This clam has been widely consumed by the local communities and also acts as a staple diet for shorebirds and crustaceans. In this paper, we investigated how acidified conditions will influence the physiology, biochemical constituents, and energetics of Donax faba. Upon incubation for 2 months in lowered pH 7.7 ± 0.05 and control 8.1 ± 0.05 conditions, we found a delayed growth in the acidified conditions followed by decrease in calcium ions in the clam shell. Although not significant, we found the digestive enzymes showed a downward trend. Total antioxidant was significantly increased in the acidified condition compared to the control. Though not significant, the expression level of MDA and antioxidant enzymes (SOD, CAT, GST, GPX, and APX) showed increasing trend in acidified samples. Among nutrients such as amino acids and fatty acids, there was no significant difference between treatments, however, showed a downward trend in the acidified conditions compared to control. Among the minerals, iron and zinc showed significant increase in the acidified conditions. The above results suggest that the clam growth, and physiological energetics may have deleterious effects if exposed for longer durations at lowered pH condition thereby affecting the organisms involved in the coastal food web.

Clearance rates of sand-burrowed and laterally pressed unburrowed Pismo clam Tivela stultorum (Mawe 1823) in a laboratory open-flow system.

Pismo clam extraction is currently banned in Mexico to help the recovery of natural populations. Thus, the primary objective of this study was to gain insight on its basic biology and husbandry protocols. Growth and clearance rate (CR) of sand-burrowed and sediment-free, laterally pressed adult Pismo clams were quantified in the laboratory as a function of burrowing condition, flow, temperature, and microalgal concentration using open-flow chambers. After 40 days, clams remained healthy regardless of burrowing condition and showed a hyperbolic CR response pattern to increased flow, with CR directly proportional to flows lower than 1000 ml min-1. Maximal asymptotic CR values (300 to 400 ml min-1 org-1) were observed from 1000 to 2000 ml min-1. No significant CR differences were observed between burrowed and laterally pressed clams, yet microalgal concentration effects were detected, with constant maximal CRs of ∼250 ml min-1 in the range of 50 to 200 cells µl-1 and decline at higher concentrations. Maintenance protocols of laterally pressed organisms were validated in the laboratory with both weight and CR data. To our knowledge, this is the first study providing whole-body physiological data translated into effective husbandry protocols for Pismo clams. This approach represents a fresh perspective to traditional research areas, opening the possibility for continued experimentation under controlled conditions.

Stoichiometry of growth under variable scenarios of nutrient limitation: Differential homeostasis of body composition among growth phenotypes of the Manila clam.

Fast- and slow-growing phenotypes from two separate breeding families of the Manila clam (Ruditapes philippinarum) were alternatively fed two monoalgal diets with high and low N content (C:N ratios of 4.9 and 13.5, respectively). After 35 days of food conditioning, clams were sacrificed, and the soft body was dissected out into five different tissue fractions to determine the corresponding ponderal ratios (tissue wt./body wt.) and a separate analysis of the elemental composition of these tissues. Previously reported C and N balances performed with the same conditioning diets were integrated and compared with tissue composition of the same phenotypes in order to assess the efficacy of mechanisms elicited to compensate for N deficit. Broad differences in dietary N content resulted in only minor changes in whole-body C:N composition which suggests a noticeable degree of homeostatic regulation of nutrient balances. This regulation was found to be stricter in fast-compared to slow-growing phenotypes and differed among the various body tissues. Using the threshold element ratio approach, physiological mechanisms were identified that partly compensate for large stoichiometric mismatches between low-N food and body tissues.

Behavioral responses of intertidal clams to compound extreme weather and climate events.

Rapidly increasing concentration of carbon dioxide (CO2) in the atmosphere not only results in global warming, but also drives increasing seawater acidification. Infaunal bivalves play critical roles in benthic-pelagic coupling, but little is known about their behavioral responses to compound climate events. Here, we tested how heatwaves and acidification affected the behavior of Manila clams (Ruditapes philippinarum). Under acidified conditions, the clams remained capable of burrowing into sediments. Yet, when heatwaves attacked, significant decreases in burrowing ability occurred. Following two consecutive events of heatwaves, the clams exhibited rapid behavioral acclimation. The present study showed that the behavior of R. philippinarum is more sensitive to heatwaves than acidification. Given that the behavior can act as an early and sensitive indicator of the fitness of intertidal bivalves, whether, and to what extent, behavioral acclimation can persist under scenarios of intensifying heatwaves in the context of ocean acidification deserve further investigations.

Increased seawater temperature triggers thermal, oxidative and metabolic response of Ostrea edulis, leading to anaerobiosis.

Bivalves are among the marine organisms most influenced by climate change. Despite the flat oyster's Ostrea edulis high economic value, its culture is developed on a very small scale, since this species possesses a strong susceptibility to abiotic stressors. Due to climate change, temperature is one of the most critical environmental parameters for the welfare of the Mediterranean basin's marine inhabitants. The present study's purpose was to investigate the physiological performance of the Mediterranean's native O. edulis as it faces exposure to different temperatures. Since juveniles are more susceptible to abiotic stressors, this experimental procedure was focused on young individuals. The seawater temperatures studied included a standard control temperature of 21 °C (often observed in several marine areas throughout the Mediterranean), as well as increased seawater temperatures of 25 °C and 28 °C, occasionally occurring in shallow Mediterranean waters inhabited by bivalve spat. These were selected since the tissues of O. edulis becomes partly anaerobic in temperatures exceeding 26 °C, while cardiac dysfunction (arrhythmia) emerges at 28 °C. The results demonstrate that temperatures above 25 °C trigger both the transcriptional upregulation of hsp70 and hsp90, and the antioxidant genes Cu/Zn sod and catalase. Enhancement of thermal tolerance and increased defense against increased ROS production during thermal stress, were observed. As the intensity and duration of thermal stress increases, apoptotic damage may also occur. The increased oxidative and thermal stress incurred at the highest temperature of 28 °C, seemed to trigger the switch from aerobic to anaerobic metabolism, reflected by higher pepck mRNA expressions and lower ETS activity.

Effects of cadmium exposure on haemocyte immune function of clam Ruditapes philippinarum at different temperatures.

Haemocytes are crucial for the immune defence of mollusks. It is important to explore the immune performance of haemocytes of mollusks under the stress of heavy metals with global warming. In order to study the effects of cadmium (Cd) exposure and temperature stress on the haemocyte immune function of clam Ruditapes philippinarum, clams were exposed to different Cd concentrations (0.05, 0.10 and 0.25 mg/L) at 20 °C, 25 °C and 30 °C respectively. Haemocyte mortality, reactive oxygen species (ROS) and superoxide dismutase (SOD) activity were measured at day 1, day 3, day 5 and day 7. The results showed that the changes of the three indexes were not obvious when exposed to 0.05 mg/L of Cd at 20 °C, while significant differences were observed with the increase of temperature, Cd concentration and exposure time. Under a condition of relative high temperature coupling with high concentration of Cd, the clams were significantly influenced, showing an obvious synergistic effect. Selected indexes reflect the clam's response to the combined stress of temperature and Cd. Moreover, R. philippinarum might be an ideal biological index species to the Cd pollution.

A global synthesis of predation on bivalves.

Predation is a dominant structuring force in ecological communities. In aquatic environments, predation on bivalves has long been an important focal interaction for ecological study because bivalves have central roles as ecosystem engineers, basal components of food webs, and commercial commodities. Studies of bivalves are common, not only because of bivalves' central roles, but also due to the relative ease of studying predatory effects on this taxonomic group. To understand patterns in the interactions of bivalves and their predators we synthesised data from 52 years of peer-reviewed studies on bivalve predation. Using a systematic search, we compiled 1334 studies from 75 countries, comprising 61 bivalve families (N = 2259), dominated by Mytilidae (29% of bivalves), Veneridae (14%), Ostreidae (8%), Unionidae (7%), and Dreissenidae and Tellinidae (6% each). A total of 2036 predators were studied, with crustaceans the most studied predator group (34% of predators), followed by fishes (24%), molluscs (17%), echinoderms (10%) and birds (6%). The majority of studies (86%) were conducted in marine systems, in part driven by the high commercial value of marine bivalves. Studies in freshwater ecosystems were dominated by non-native bivalves and non-native predator species, which probably reflects the important role of biological invasions affecting freshwater biodiversity. In fact, while 81% of the studied marine bivalve species were native, only 50% of the freshwater species were native to the system. In terms of approach, most studies used predation trials, visual analysis of digested contents and exclusion experiments to assess the effects of predation. These studies reflect that many factors influence bivalve predation depending on the species studied, including (i) species traits (e.g. behaviour, morphology, defence mechanisms), (ii) other biotic interactions (e.g. presence of competitors, parasites or diseases), and (iii) environmental context (e.g. temperature, current velocity, beach exposure, habitat complexity). There is a lack of research on the effects of bivalve predation at the population and community and ecosystem levels (only 7% and 0.5% of studies respectively examined impacts at these levels). At the population level, the available studies demonstrate that predation can decrease bivalve density through consumption or the reduction of recruitment. At the community and ecosystem level, predation can trigger effects that cascade through trophic levels or effects that alter the ecological functions bivalves perform. Given the conservation and commercial importance of many bivalve species, studies of predation should be pursued in the context of global change, particularly climate change, acidification and biological invasions.

Eyeless razor clam Sinonovacula constricta discriminates light spectra through opsins to guide Ca2+ and cAMP signaling pathways.

Phototransduction is based on opsins that drive distinct types of Gα cascades. Although nonvisual photosensitivity has long been known in marine bivalves, the underlying molecular basis and phototransduction mechanism are poorly understood. Here, we introduced the eyeless razor clam Sinonovacula constricta as a model to clarify this issue. First, we showed that S. constricta was highly diverse in opsin family members, with a significant expansion in xenopsins. Second, the expression of putative S. constricta opsins was highly temporal-spatio specific, indicating their potential roles in S. constricta development and its peripheral photosensitivity. Third, by cloning four S. constricta opsins with relatively higher expression (Sc_opsin1, 5, 7, and 12), we found that they exhibited different expression levels in response to different light environments. Moreover, we demonstrated that these opsins (excluding Sc_opsin7) couple with Gαq and Gαi cascades to mediate the light-dependent Ca2+ (Sc_opsin1 and 5) and cAMP (Sc_opsin12) signaling pathways. The results indicated that Sc_opsin1 and 5 belonged to Gq-opsins, Sc_opsin12 belonged to Gi-opsins, while Sc_opsin7 might act as a photo-isomerase. Furthermore, we found that the phototransduction function of S. constricta Gq-opsins was dependent on the lysine at the seventh transmembrane domain, and greatly influenced by the external light spectra in a complementary way. Thus, a synergistic photosensitive system mediated by opsins might exist in S. constricta to rapidly respond to the transient or subtle changes of the external light environment. Collectively, our findings provide valuable insights into the evolution of opsins in marine bivalves and their potential functions in nonvisual photosensitivity.

Morphological description of the glochidia of Buldowskia suifunica (Bivalvia: Unionidae).

Freshwater mussels of the genus Buldowskia (Bivalvia, Unionidae) are distributed from the Amur River basin in Russia and China southward to the Korean Peninsula and some Japanese islands. This work is an integrative morphological study of Buldowskia suifunica glochidia from locations in the Primorsky Territory, the Russian Far East. Glochidia of B. suifunica, taken from the same gill have asynchronous development. The external and internal morphology of its shell has been characterized. The morphology of its sensory system, within three stages of larval development (immature, intermediate and mature glochidia), consists of hair cells as well as nonhair cells. Their muscle system is composed of massive adductor and minor muscle bundles. The FMRFamid-ergic nervous system turned out to be a complex system includes basal cells (neurons), their neurites and anterior neurons. FMRFamide and tubulin was found in all neurons. Glochidia of B. suifunica have only four 5-HT-lir neurons. We concluded that B. suifunica glochidial nervous system differs from those of the larval systems of planktotrophic marine mollusks.

A dynamic biointerface controls mussel adhesion.

The mussel-adherent secreta interface reveals how nonliving material can be compatible with tissue.

Environmental influence on summer survival of Manila Clam Ruditapes philippinarum: A case study in an aquaculture bay.

The aquaculture of Manila clam (Ruditapes philippinarum) is under threat due to its high mortality in summer. To investigate the environmental influence on mortality of the species, we have conducted field surveys in the aquaculture area in Laizhou Bay, during the summers of 2019 and 2020. Environmental and biological data were collected. The results of data analysis have shown that in 2020, large-scale mortality events were not recorded and spatial variations of environmental variables were no significant within the survey area. However, in August 2019, significant variations of environmental variables were detected in the middle tidal area. This area was charactered with relatively high-water temperature (with maximum temperature of 31.49 °C and mean value of 26.17 ± 1.62 °C), elevated hydrogen sulfide concentrations (with a maximum concentration of 24.72 µmol/L and a mean concentration of 7.89 ± 4.14 µmol/L), and low dissolved oxygen concentrations (with a minimum concentration of 3.35 mg/L and a mean concentration of 6.27 ± 0.42 mg/L). Correspondingly, high mortality of the clam was recorded in the area, the abundance of live Manila clams has decreased by 80%. No significant abnormalities of environmental variables and clam growth were observed in other regions in 2019. Glycogen content and condition index of the clam were analyzed to investigate the relationship between environmental stress and the health of the clam. Compared to the condition index, glycogen content is a more sensitive indicator of the health status of the clam because changes in glycogen content appeared earlier than changes in condition index and mortality. A principal component analysis further indicated that the high mortality of the clam corresponds with a few environmental variables, including elevated temperature, hypoxia and the concentration of hydrogen sulfide. The simultaneous presence of these multiple environmental stressors could have triggered alterations in the physiological responses of the clam.

Sex-specific responses of Ruditapes philippinarum to ocean acidification following gonadal maturation.

Ocean acidification (OA) can seriously affect marine bivalves at different levels of biological organization, generating widespread consequences on progeny recruitment and population maintenance. Yet, few effort has been devoted to elucidating whether female and male bivalves respond differentially to OA in their reproductive seasons. Here, we estimated differences in physiological responses of female and male Manila clams (Ruditapes philippinarum) to OA during gonadal maturation. In comparison to OA-stressed male clams, females significantly depressed activities in enzymes related to energy metabolism (NKA, T-ATP), antioxidant defence (SOD and MDA), and non-specific immune function (ACP), and downregulated expression of AMPK that plays a key role in cellular metabolism, indicating that sex did significantly affect responses of R. philippinarum to OA. Such sex-based differences can be likely couched in energetic terms, given the much more energetically expensive cost of egg production than that of sperms. These results indicate that sex-specific responses to OA during reproductive seasons do exist in marine bivalves, and therefore accounting for such sex specificity is of paramount importance when projecting population sustainability and formulating conservation strategies in an acidifying ocean.

Influence of sea-ice dynamics on coastal Antarctic benthos: A case study on lantern clams (Laternula elliptica) in Adélie Land.

Polar regions are warming faster than the world average and are profoundly affected by changes in the spatio-temporal dynamics of sea ice, with largely unknown repercussions on the functioning of marine ecosystems. Here, we investigated the impacts of interannual sea-ice variability on coastal benthic communities in Antarctica, focusing on a close-to-pristine area (Adélie Land). We investigated shell growth of the circum-Antarctic bivalve Laternula elliptica, considered a key species in these soft bottom benthic communities. Chondrophores of live-collected clams were prepared using standard sclerochronological methods to study the interannual variability of shell growth from 1996 to 2015. Our results show that the master chronology varied with sea-ice dynamics. When sea ice breaks up too early, sympagic algae do not have time to accumulate sufficiently high biomass, thus strongly limiting the energy input to the benthos. This negatively affects the physiological performance of L. elliptica, thereby altering their population dynamics and hence the functioning of these soft-bottom ecosystems.

Physiological and biochemical responses of clams to recurrent marine heatwaves.

In the past decade, the frequency, intensity and duration of marine heatwaves (MHWs) in the South China Sea have been increasing strikingly, resulting in serious impacts on intertidal bivalves and their ecosystems. The Manila clam, Ruditapes philippinarum, is one of the most ecologically and economically important bivalve species in the South China Sea, yet very little is known about its fate under intensifying MHWs events. Here, we examined how R. philippinarum responded to two consecutive scenarios of MHWs, with each composed of 4 °C and 8 °C rises of seawater temperatures, respectively. Up to 87% of Manila clams survived recurrent MHWs events, and significant increases in standard metabolic rate occurred predominantly under extreme conditions (+8 °C), indicating that the clams could trigger compensatory mechanisms to mitigate MHWs-induced thermal stress. Following acute and repeated exposures to MHWs, Manila clams showed similar responses in enzymes underpinning energy metabolism (NKA, CMA, and T-ATP), antioxidant defence (SOD, CAT, and MDA), and biomineralization (AKP and ACP), most of which exhibited significantly increasing and then decreasing trends with the intensification of MHWs. Of eight genes associated with physiological tolerance and fitness, ATAD3A, PFK, SOD, and C3 were significantly down-regulated in response to recurrent MHWs events, demonstrating the certain resistance to MHWs. These findings provide a better understanding that marine bivalves hold the potential to acclimate simulated MHWs events from the physiological and molecular processes.

Impact of Sea Warming and 17-α-Ethinylestradiol Exposure on the Lipid Metabolism of Ruditapes philippinarum Clams.

This paper reports on an NMR metabolomics study of lipophilic extracts of Ruditapes philippinarum clams exposed to the hormonal contaminant 17-α-ethinylestradiol (EE2), at 17 °C and 21 °C. The results reveal that exposure at 17 °C triggers a weak response at low EE2 concentrations, suggestive of a slight increase in membrane rigidity, followed by lipid metabolic stability at higher EE2 concentrations. On the other hand, at 21 °C, lipid metabolism begins to respond at 125 ng/L EE2, with antioxidant docosahexaenoic acid (DHA) helping to tackle high-oxidative-stress conditions, in tandem with enhanced storage of triglycerides. Exposure to 625 ng/L EE2 (highest concentration) enhances phosphatidylcholine (PtdCho) and polyunsaturated fatty acid (PUFA) levels, their direct intercorrelation suggesting PUFA incorporation in new membrane phospholipids. This should lead to increased membrane fluidity, probably aided by a decrease in cholesterol. PUFA levels, considered a measure of membrane fluidity, were strongly (and positively) correlated to intracellular glycine levels, thus identifying glycine as the main osmolyte entering the cells under high stress. Membrane fluidity also seems to elicit the loss of taurine. This work contributes to the understanding of the mechanisms of response of R. philippinarum clams to EE2 in tandem with warming while unveiling novel potential markers of stress mitigation, namely high levels of PtdCho, PUFAs (or PtdCho/glycerophosphocholine and PtdCho/acetylcholine ratios) and linoleic acid and low PUFA/glycine ratios.

CO2-Induced Ocean Acidification Alters the Burrowing Behavior of Manila Clam Ruditapes philippinarum by Reversing GABAA Receptor Function.

Biological burrowing behavior is an important driver shaping ecosystems that is being threatened by CO2-induced ocean acidification; however, the effects of ocean acidification on burrowing behavior and its neurological mechanism remain unclear. This study showed that elevated pCO2 significantly affected the burrowing behaviors of the Manila clam Ruditapes philippinarum, such as increased foot contraction, burrowing time, and intrabottom movement and decreased burrowing depth. Delving deeper into the mechanism, exposure to elevated pCO2 significantly decreased extracellular pH and increased [HCO3-]. Moreover, an indicator GABAA receptor, a neuroinhibitor for movement, was found to be closely associated with behavioral changes. In situ hybridization confirmed that the GABAA receptor was widely distributed in ganglia and foot muscles, and elevated pCO2 significantly increased the mRNA level and GABA concentration. However, the increase in GABAA receptor and its ligand did not suppress the foot movement, but rather sent "excitatory" signals for foot contraction. The destabilization of acid-base homeostasis was demonstrated to induce an increase in the reversal potential for GABAA receptor and an alteration in GABAA receptor function under elevated pCO2. This study revealed that elevated pCO2 affects the burrowing behavior of Manila clams by altering GABAA receptor function from inhibitory to excitatory.

Toxicity of microplastics and triclosan, alone and in combination, to the fertilisation success of a broadcast spawning bivalve Tegillarca granosa.

Since most marine invertebrates adopted external fertilisation, their fertilisation process is particularly vulnerable to aquatic pollutants. Both antimicrobial ingredients and microplastics (MPs) are ubiquitous in aquatic environments; however, their synergistic effects on the fertilisation of marine invertebrates remain unclear. Therefore, in this study, the fertilisation toxicity of MPs and triclosan (TCS), alone and in combination, was investigated in the broadcast spawning bivalve Tegillarca granosa. Results showed that MPs and TCS significantly suppressed the fertilisation success of T. granosa. As the fertilisation success of broadcast spawning invertebrates depends on successful gamete collisions, gamete fusion, and egg activation, sperm swimming velocity, viability, gamete collision probability, ATP status, and ion-transport enzyme activities were also analysed to further ascertain the underlying toxicity mechanisms. In summary, our findings indicate that the presence of MPs may enhance the fertilisation toxicity of TCS by hampering sperm-egg collision probability, reducing gamete fusion efficiency, and restricting Ca2+ oscillation formation.