Differentiation of cellular responses to particulate and soluble constituents in sunscreen products.

Despite the growing awareness of potential human and environmental risks associated with sunscreens, identifying the specific constituents responsible for their potential toxicity is challenging. In this study, we applied three different types of sunscreens with contrasting compositions and compared the effects of their particulate and soluble fractions based on 15 cellular biomarkers of HaCaT cells. Multilinear regression analysis revealed that the internalized soluble fractions played a primary role in the overall cytotoxicity of sunscreen mixtures, which was primarily attributed to their biotransformation, generating metabolites with higher toxicity. The presence of plastic microspheres in sunscreens either inhibited the internalization of soluble fractions or led to their redistribution toward lysosomes. Conversely, subcellular toxicity resulting from the sunscreen mixture was predominantly influenced by particulates. Bio-transformable particulates such as ZnO dissolved in the organelles and induced higher subcellular toxicity compared to bioinert particulates such as microplastics. Subcellular biomarkers including lysosomal count, lysosomal size, mitochondrial count and mitochondrial shape emerged as the potential predictors of sunscreen presence. Our study provides important understanding of sunscreen toxicity by elucidating the differential impacts of particulate and soluble fractions in mixture contaminants.

Tracking the Cellular Degradation of Silver Nanoparticles: Development of a Generic Kinetic Model.

Understanding the degradation of nanoparticles (NPs) after crossing the cell plasma membrane is crucial in drug delivery designs and cytotoxicity assessment. However, the key factors controlling the degradable kinetics remain unclear due to the absence of a quantification model. In this study, subcellular imaging of silver nanoparticles (AgNPs) was used to determine the intracellular transfer of AgNPs, and single particle ICP-MS was utilized to track the degradation process. A cellular kinetic model was subsequently developed to describe the uptake, transfer, and degradation behaviors of AgNPs. Our model demonstrated that the intracellular degradation efficiency of AgNPs was much higher than that determined by mimicking testing, and the degradation of NPs was highly influenced by cellular factors. Specifically, deficiencies in Ca or Zn primarily decreased the kinetic dissolution of NPs, while a Ca deficiency also resulted in the retardation of NP transfer. The biological significance of these kinetic parameters was strongly revealed. Our model indicated that the majority of internalized AgNPs dissolved, with the resulting ions being rapidly depurated. The release of Ag ions was largely dependent on the microvesicle-mediated route. By changing the coating and size of AgNPs, the model results suggested that size influenced the transfer of NPs into the degradation process, whereas coating affected the degradation kinetics. Overall, our developed model provides a valuable tool for understanding and predicting the impacts of the physicochemical properties of NPs and the ambient environment on nanotoxicity and therapeutic efficacy.

Aging effects of titanium dioxide on Cu toxicity to Daphnia magna: Exploring molecular docking and significance of surface properties.

Cosmetics and personal care products containing titanium dioxide nanoparticles (TiO2 NPs) may enter aquatic environments, where the surface coatings of TiO2 NPs may change with aging due to environmental factors such as light, and potentially affect their bioaccumulation and toxicity. This study examined how aging impacted the physicochemical properties of three commercially available TiO2 NPs and subsequent influence on the bioaccumulation and toxicity of copper (Cu) in Daphnia magna (D. magna). We demonstrated that aging significantly affected the hydrophobicity of TiO2 NPs, which affected their binding to water molecules and adsorption of Cu. Changes of bioaccumulation of TiO2 NPs and Cu in D. magna ultimately affected the activities of intracellular antioxidant enzymes such as SOD, CAT, GSH-Px, and the transmembrane protein Na+/K+-ATPase. Molecular docking calculations demonstrated that changes of activities of these biological enzymes were due to the interaction between TiO2 NPs, Cu, and amino acid residues near the sites with the lowest binding energy and active center of the enzyme. Such effect was closely related to the hydrophobicity of TiO2 NPs. Our study demonstrated the close relationship between surface properties of TiO2 NPs and their biological effects, providing important evidence for understanding the behavior of nanomaterials in aquatic environments.

Combined simultaneous transsphenoidal and transcranial regimen improves surgical outcomes in complex giant pituitary adenomas: A longitudinal retrospective cohort study.

BACKGROUND: Surgical treatment of complex giant pituitary adenomas (GPAs) presents significant challenges. The efficacy and safety of combining transsphenoidal and transcranial approaches for these tumors remain controversial. In this largest cohort of patients with complex GPAs, we compared the surgical outcomes between those undergoing a combined regimen and a non-combined regimen. We also examined the differences in risks of complications, costs, and logistics between the two groups, which might offer valuable information for the appropriate management of these patients. MATERIALS AND METHODS: This was a multicenter retrospective cohort study conducted at 13 neurosurgical centers. Consecutive patients who received a combined or non-combined regimen for complex GPAs were enrolled. The primary outcome was gross total resection, while secondary outcomes included complications, surgical duration, and relapse. A propensity score-based weighting method was used to account for differences between the groups. RESULTS: Out of 647 patients (298 [46.1%] women, mean age: 48.5 ± 14.0 years) with complex GPAs, 91 were in the combined group and 556 were in the non-combined group. Compared with the non-combined regimen, the combined regimen was associated with a higher probability of gross total resection (50.5% vs. 40.6%, odds ratio [OR]: 2.18, 95% confidence interval [CI]: 1.30-3.63, P = 0.003). The proportion of patients with life-threatening complications was lower in the combined group than in the non-combined group (4.4% vs. 11.2%, OR: 0.25, 95% CI: 0.08-0.78, P = 0.017). No marked differences were found between the groups in terms of other surgical or endocrine-related complications. However, the combined regimen exhibited a longer average surgery duration of 1.3 h (P < 0.001) and higher surgical costs of 22,000 CNY (approximate 3,000 USD, P = 0.022) compared with the non-combined approach. CONCLUSIONS: The combined regimen offered increased rates of total resection and decreased incidence of life-threatening complications, which might be recommended as the first-line choice for these patients.

Bioprospecting of Chlamydomonas reinhardtii for boosting biofuel-related products production based on novel aggregation-induced emission active extracellular polymeric substances nanoprobes.

Biofuel production from microalgae has been greatly restricted by low biomass productivity and long-term photosynthetic efficacy. Here, a novel strategy for selecting high-growing, stress-resistant algal strains with high photosynthetic capacity was proposed based on biocompatible extracellular polymeric substances (EPS) probes with aggregation-induced emission (AIE) properties. Specifically, AIE active EPS probes were synthesized for in-situ long-term monitoring of the EPS productivity at different algal growth stages. By coupling the AIE-based fluorescent techniques, algal cells were classified into four diverse populations based on their chlorophyll and EPS signals. Mechanistic studies on the sorted algal cells revealed their remarkable stress resistance and high expression of cell division, biopolymer production and photosynthesis-related genes. The sorted and subcultured algal cells consistently exhibited relatively higher growth rates and photosynthetic capacities, resulting in an increased (1.2 to 1.8-fold) algal biomass production, chlorophyll, and lipids. This study can potentially open new strategies to boost microalgal-based biofuel production.

Single-Cell RNA Sequencing Profiling Cellular Heterogeneity and Specific Responses of Fish Gills to Microplastics and Nanoplastics.

Fish gills are highly sensitive organs for microplastic (MP) and nanoplastic (NP) invasions, but the cellular heterogeneity of fish gills to MPs and NPs remains largely unknown. We employed single-cell RNA sequencing to investigate the responses of individual cell populations in tilapia Oreochromis niloticus gills to MP and NP exposure at an environmentally relevant concentration. Based on the detected differentially expressed gene (DEG) numbers, the most affected immune cells by MP exposure were macrophages, while the stimulus of NPs primarily targeted T cells. In response to MPs and NPs, H+-ATPase-rich cells exhibited distinct changes as compared with Na+/K+-ATPase-rich cells and pavement cells. Fibroblasts were identified as a potential sensitive cell-type biomarker for MP interaction with O. niloticus gills, as evidenced by the largely reduced cell counts and the mostly detected DEGs among the 12 identified cell populations. The most MP-sensitive fibroblast subpopulation in O. niloticus gills was lipofibroblasts. Cell-cell communications between fibroblasts and H+-ATPase-rich cells, neurons, macrophages, neuroepithelial cells, and Na+/K+-ATPase-rich cells in O. niloticus gills were significantly inhibited by MP exposure. Collectively, our study demonstrated the cellular heterogeneity of O. niloticus gills to MPs and NPs and provided sensitive markers for their toxicological mechanisms at single-cell resolution.

Dynamic Regulation of Intracellular Labile Cu(I)/Cu(II) Cycle in Microalgae Chlamydomonas reinhardtii: Disrupting the Balance by Cu Stress.

The labile metal pool involved in intracellular trafficking and homeostasis is the portion susceptible to environmental stress. Herein, we visualized the different intracellular distributions of labile Cu(I) and Cu(II) pools in the alga Chlamydomonas reinhardtii. We first demonstrated that labile Cu(I) predominantly accumulated in the granules within the cytoplasmic matrix, whereas the labile Cu(II) pool primarily localized in the pyrenoid and chloroplast. The cell cycle played an integral role in balancing the labile Cu(I)/Cu(II) pools. Specifically, the labile Cu(II) pool primarily accumulated during the SM phase following cell division, while the labile Cu(I) pool dynamically changed during the G phase as cell size increased. Notably, the labile Cu(II) pool in algae at the SM stage exhibited heightened sensitivity to environmental Cu stress. Exogenous Cu stress disrupted the intracellular labile Cu(I)/Cu(II) cycle and balance, causing a shift toward the labile Cu(II) pool. Our proteomic analysis further identified a putative cupric reductase, potentially capable of reducing Cu(II) to Cu(I), and four putative multicopper oxidases, potentially capable of oxidizing Cu(I) to Cu(II), which may be involved in the conversion between the labile Cu(I) pool and labile Cu(II) pool. Our study elucidated a dynamic cycle of the intracellular labile Cu(I)/Cu(II) pools, which were accessible and responsive to environmental changes.

Toxic effects and action mechanism of metal-organic framework UiO-66-NH2 in Microcystisaeruginosa.

The zirconium metal-organic framework UiO-66-NH2 has garnered considerable attention for their potentials of removing environmental contaminants from water. The production and application of UiO-66-NH2 make their releases into the aquatic environment inevitable. Nevertheless, little information is available regarding its potential risk to the environment and aquatic organisms, thus limiting the evaluation of its safe and sustainable use. In this study, the ecotoxicity of UiO-66-NH2 was evaluated, specifically its impacts on growth, extracellular organic matter release, and metabolomic changes of the model phytoplankton Microcystis aeruginosa (M. aeruginosa). UiO-66-NH2 exhibited moderate effects on algal physiology including growth, viability, and photosynthetic system. At concentrations below 20 mg/L, UiO-66-NH2 induced negligible inhibition of algal growth, algal viability, and photosynthesis. In contrast, UiO-66-NH2 boosted the release of extracellular organic matter even at concentration as low as 0.02 mg/L. These findings indicated that, while no evident damage to algal cells was observed, UiO-66-NH2 was hazardous to the aquatic environment as it stimulated the release of algal toxins. Moreover, UiO-66-NH2 entered algal cells rather than adhering to the surface of M. aeruginosa as observed by the fluorescence imaging. Based on metabolic analysis, UiO-66-NH2 influenced the cyanobacteria mainly through interference with purine metabolism and ABC transporter. This study sheds light on the potential threat UiO-66-NH2 posing to microalgae, and has potential implications for its safe utilization in the environmental field.

Novel Near-Infrared-II In Vivo Visualization Revealed Rapid Calcium Intestine Turnover in Daphnia magna with Delayed Impact by Cadmium and Acidification.

Calcium is a highly demanded metal, and its transport across the intestine of Daphnia magna remains a significant unresolved question. Due to technical constraints, the visualization of the kinetic process of Ca passage through D. magna has been challenging. Here, we developed the second near-infrared Ca sensor (NIR-II Ca) and conducted real-time in vivo imaging of Ca in daphnids with a high signal-to-noise ratio, deep tissue penetration, and minimal damage. Through the utilization of the NIR-II Ca sensor, we for the first time visualized and quantified the kinetic process of Ca passage in the intestine in real time. The results revealed that trophically available Ca passed through the intestines in 24 h, whereas waterborne Ca required only 35 min. This rapid "flushing through" mechanism established waterborne Ca as the primary source of Ca absorption. However, environmental stressors such as water acidification and cadmium significantly delayed the Ca passage and absorption. The development of NIR imaging and sensors allows for real-time dynamic visualization of contaminants/nutrients in organisms and holds great potential as a powerful tool for future studies into material kinetic processes in living animals.

Reducing Gut Dissolution of Zinc Oxide Nanoparticles by Secondary Microplastics with Consequent Impacts on Barnacle Larvae.

The environmental impact of sunscreen is a growing concern, yet the combined effects of its components on marine animals are poorly understood. In this study, we investigated the combined effects of sunscreen-extracted zinc oxide nanoparticles (nZnO) and microplastics (MPs) on the development of barnacle larvae, focusing on the different roles played by primary microplastics (PMPs) and secondary microplastics (SMPs) generated through the phototransformation of PMPs. Our findings revealed that a lower concentration of nZnO (50 µg/L) enhanced molting and eye development in barnacle larvae, while a higher concentration (500 µg/L) inhibited larval growth. Co-exposure to PMPs had no significant effect on larval development, whereas SMPs mitigated the impact of nZnO by restricting the in vivo transformation to ionic Zn. Accumulated SMPs reduced gut dissolution of nZnO by up to 40%, lowering gut acidity by 85% and buffering the in vivo dissolution of nZnO. We further identified a rough-surfaced Si-5 fragment in SMPs that damaged larval guts, resulting in decreased acidity. Another Si-32 resisted phototransformation and had no discernible effects. Our study presented compelling evidence of the impacts of SMPs on the bioeffect of nZnO, highlighting the complex interactions between sunscreen components and their combined effects on marine organisms.

Microplastics and nanoplastics induced differential respiratory damages in tilapia fish Oreochromis niloticus.

With the increasing micro(nano)plastics (MNPs) pollution in aquatic environments, fish respiration is encountering a huge threat. Herein, polystyrene (PS) MNPs with three sizes (80 nm, 2 µm, and 20 µm) were exposed to tilapia Oreochromis niloticus at an environmentally relevant concentration of 100 µg/L for 28 days and their impacts on respiratory function were investigated. Based on the results of oxygen consumption and histological analysis, all the three treatments could induce respiratory damages and such impacts were more severe for the 2 µm and 20 µm treatments than for the 80 nm treatment. These results were explained by the more significant upregulation of egln3 and nadk, and the downregulation of isocitrate. Transcriptomics and metabolomics further revealed that TCA cycle played a key role in respiratory dysfunction induced by micro-sized PS particles, and cytokine and chemokine related functions were simultaneously enriched. Although nano-sized PS particles had the potential to penetrate the respiratory epithelium and reached the internal structure of the O. niloticus gills, they were easily expelled through the blood circulation. Our results highlighted the serious threat of MNPs to fish respiration and provided insights into the differential toxicological mechanisms between micro-sized and nano-sized particles, thus assisting in ecological risk assessments.

Metagenomics-Based Microbial Ecological Community Threshold and Indicators of Anthropogenic Disturbances in Estuarine Sediments.

Assessing the impacts of cumulative anthropogenic disturbances on estuarine ecosystem health is challenging. Using spatially distributed sediments from the Pearl River Estuary (PRE) in southern China, which are significantly influenced by anthropogenic activities, we demonstrated that metagenomics-based surveillance of benthic microbial communities is a robust approach to assess anthropogenic impacts on estuarine benthic ecosystems. Correlational and threshold analyses between microbial compositions and environmental conditions indicated that anthropogenic disturbances in the PRE sediments drove the taxonomic and functional variations in the benthic microbial communities. An ecological community threshold of anthropogenic disturbances was identified, which delineated the PRE sediments into two groups (H and L) with distinct taxa and functional traits. Group H, located nearshore and subjected to a higher level of anthropogenic disturbances, was enriched with pollutant degraders, putative human pathogens, fecal pollution indicators, and functional traits related to stress tolerance. In contrast, Group L, located offshore and subjected to a lower level of anthropogenic disturbances, was enriched with halotolerant and oligotrophic taxa and functional traits related to growth and resource acquisition. The machine learning random forest model identified a number of taxonomic and functional indicators that could differentiate PRE sediments between Groups H and L. The identified ecological community threshold and microbial indicators highlight the utility of metagenomics-based microbial surveillance in assessing the adverse impacts of anthropogenic disturbances in estuarine sediments, which can assist environmental management to better protect ecosystem health.

Lysosomal Cu(I)/Cu(II) Dependence of Antimicrobial Ability of Oyster Hemocytes and Regulation of Phagolysosomal System.

Copper (Cu) is hyperaccumulated in oyster hemocytes and is an essential trace metal indispensable for diverse innate immune functions. However, the roles of Cu in oyster immune defense are still unclear. In this study, Cu exposure enhanced the phagocytosis of zymosan by increasing the number and length of filopodia, as well as mitochondrial ROS (mitoROS) production mainly in granulocytes, followed by semigranulocytes and agranulocytes. The intracellular calcium level increased to promote the phagosome-lysosome fusion after Cu exposure. The enhancement of phagosomal acidification and mitochondrion-phagosome juxtaposition were also found in granulocytes after Cu exposure. These results indicated that Cu could regulate the phagolysosomal system to enhance the antimicrobial ability of oyster hemocytes with the assistance of mitoROS. Furthermore, Cu(I) and Cu(II) were predominately located in lysosomes, and degranulation may provide a mechanism for exposing Cu to bacteria to prevent their survival and proliferation. Specifically, we showed that the newly formed Cu(I) arising from lysosomal Cu(II) moved to lysosomes and mitochondria in activated hemocytes to induce strong immune responses. The ability of the transformation of Cu(I) from Cu(II) followed granulocytes > semigranlocytes > agranulocytes, indicating that granulocytes played important roles in immune functions of oysters. Our results provided new insights into the understanding of antimicrobial effects of Cu in oyster hemocytes.

Tracking Nano- and Microplastics Accumulation and Egestion in a Marine Copepod by Novel Fluorescent AIEgens: Kinetic Modeling of the Rhythm Behavior.

Nano- and microplastics (NMPs) are now prevalent in the marine environment. This study quantified the uptake and depuration kinetics of spherical polystyrene NMPs of different particle sizes (200 nm/30 µm) and functional groups (-NH2/-COOH) in a temperate calanoid copepod Calanus sinicus (C. sinicus), which exhibited rhythmic feeding patterns in natural environments. Aggregated-induced emission (AIE) fluorescent probes were employed to track and quantify the kinetics of NMPs with excellent photostability and biocompatibility. The results showed that C. sinicus consumed all NMPs types, with preference of NMPs to small size and amino group. Increased diatom concentrations also inhibited the bioaccumulation of NMPs. Influenced by rhythmic behavior, the bioaccumulation of NMPs by C. sinicus was nonstationary during the 6 h uptake phase. After 1-3 h of rapid uptake, the body burden peaked and then slowly declined. During the 3 h depuration phase, C. sinicus rapidly and efficiently removed NMPs with a mean half-life of only 0.23 h. To further quantify the body burden of C. sinicus under the influence of rhythmic feeding behavior, a biokinetic model was established, and the Markov chain Monte Carlo method was used to estimate the parameter distribution. Our results highlighted that copepods exhibited unique rhythmic feeding behavior under environmentally relevant concentrations of NMPs exposure, which may influence the bioaccumulation, trophic transfer, and environmental fate of NMPs.

Near-Infrared-II In Vivo Visualization and Quantitative Tracking of Micro/Nanoplastics in Fish.

As emerging contaminants, micro/nanoplastics (MNPs) are widely present in aquatic environments and are often ingested by aquatic organisms. However, the in vivo trafficking and fate of MNPs remain largely unknown. Here, we developed near-infrared (NIR) aggregated-induced emission (AIE) fluorophore-labeled microplastics (2 µm) and nanoplastics (100 nm) as models of MNPs. This model was based on the NIR-AIE technique with strong emission at the second near-infrared (NIR-IIII) window, which overcomes the interference of autofluorescence and observation artifacts in the detection of commercial fluorescent-labeled particles. Due to its deep tissue penetration and noninvasiveness, the dynamic process of accumulation and transport of MNPs in individuals can be tracked with NIR imaging. We then directly visualized and quantified the uptake and depuration processes of MPs and NPs in zebrafish. The results showed that the MPs and NPs were mainly accumulated in the fish gut, and the distribution was heterogeneous. MPs tended to accumulate more in the fore and mid areas of the gut compared with NPs. Besides, both MPs and NPs could accumulate in large quantities locally in the gut and might cause intestinal obstruction. MNPs accumulated slowly during the initial exposure followed by rapid and sustained accumulation in gut. Based on these kinetic accumulation and depuration, we developed a refined toxicokinetic (TK) model to describe the dynamic changes in the uptake and depuration of MNPs. Overall, this study proposed a MNP model based on the NIR-AIE technique, which provided a reliable tracer technology for the visualization, tracking and quantification of MNPs in vivo.

Infiltration of freshwater food chain by nanoplastics: An examination of trophic transfer and biological impact.

The contamination of freshwater by nanoplastics (NPs) poses a significant threat to various organisms. However, the influence of food factors on the NPs toxicity and the associated risks to water safety remains poorly understood. In this study, we employed highly fluorescent NPs incorporating aggregation-induced emission fluorogens to quantify and visualize the interactions of NPs with freshwater phytoplanktonic alga Chlamydomonas reinhardtii and zooplankton Daphnia magna, including the uptake of NPs by algae and daphnids, as well as gut environment responses through both waterborne and foodborne routes. NPs at environmentally relevant concentrations were capable of aggregating and internalizing in planktonic algae, thus permeating the food chain and potentially causing ecological risks. Notably, positively charged NPs exhibited greater infiltration than the negatively charged NPs. Our findings indicated that NPs had a greater tendency to accumulate in Daphnia via the food chain rather than through direct exposure to water. This was attributed to the loss of selectivity, suggesting a potential mechanism for NP accumulation in aquatic food chains. However, daphnids exposed to NPs through waterborne route exhibited more gut pH acidification, esterase activity, and gut inflammation compared to those exposed via the foodborne route. Moreover, chronic exposure to NPs through waterborne routes significantly disrupted the growth, molting, and reproduction of Daphnia compared to the foodborne group. These findings provided valuable insights into the complexities of plastic pollution on primary trophic levels in aquatic environments.

Identification of Transformation Products of Organic UV Filters by Photooxidation and Their Differential Estrogenicity Assessment.

Organic ultraviolet filters (OUVFs) are extensively released into aquatic environments, where they undergo complex phototransformation. However, there is little knowledge regarding their transformation products (TPs) and associated endocrine disruption potentials. In the present study, we characterized the chemical and toxicological profiles of TPs for two common OUVFs, oxybenzone (BP3) and ethylhexyl methoxycinnamate (EHMC), by photooxidation under environmentally relevant conditions. It is hypothesized that TPs of the tested OUVFs will show varied estrogenicity at different reaction times. High-resolution liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) identified 17 TPs of 7 m/z for BP-3 and 13 TPs of 8 m/z for EHMC at confidence levels ≤2. Five novel TPs of 2 m/z were reported for the first time with structure-diagnostic MS/MS spectra. Estrogenicity assessment using the MCF-7-luc cell line showed discrepant estrogenic activities exhibited by OUVF-TPs over time. Specifically, BP3-TPs exhibited significantly greater estrogenicity than the parent at several reaction times, whereas EHMC-TPs displayed fluctuating estrogenicity with a declining trend. Correlation analysis coupled with molecular docking simulations further suggested several TPs of BP3 as potential endocrine disruptive compounds. These findings underscore the necessity of considering mixtures during chemical testing and risk assessment and highlight the potentially greater risks associated with post-transformation cocktails.

Physiological and immune profiling of tilapia Oreochromis niloticus gills by high-throughput single-cell transcriptome sequencing.

The physiological and immune functions of fish gills are largely recognized, but their following functional heterogeneity at the single cell scale has been rarely reported. Here, we performed single cell RNA sequencing (scRNA-seq) on the gills of tilapia fish Oreochromis niloticus. We identified a total of 12 cell populations and analyzed their functional heterogeneity. To investigate the physiological function of O. niloticus gills, expression patterns of genes encoding ion transporters were selected from the identified H+-ATPase-rich cells (HR cells), Na+/K+-ATPase-rich cells (NaR cells), and pavement cells. Specific enrichment of ca4a, slc9a1a, and LOC100692482 in the HR cells of O. niloticus gills explained their functions in acid-base regulation. Genes encoding Ca2+ transporters, including atp2b1, LOC100696627, and LOC 100706765, were specifically expressed in the NaR cells. Pavement cells were presumably the main sites responsible for ammonia and urea transports in O. niloticus gills with specific enrichment of Rhbg and LOC100693008, respectively. The expression patterns of the four immune cell subtypes varied greatly, with B cells being enriched with the most immune-related GO terms. KEGG enrichment analysis showed that MAPK signaling pathway was the most enriched pathway among the four types of immune cells in O. niloticus gills. Our results are important in understanding the physiological and immune responses of fish gills at the cellular resolution.

Reproductive characteristics of the hermaphroditic four-finger threadfin, Eleutheronema tetradactylum (Shaw, 1804), in tropical coastal waters.

This study investigated the reproductive traits of the hermaphroditic four-finger threadfin, Eleutheronema tetradactylum, along the coasts of Thailand during January to December 2021. Fish samples were collected from Pattani Bay, Thailand to assess the sex ratio, gonadosomatic index (GSI), maturity stage and fecundity. Additional fish samples were also collected from other areas to evaluate the length and weight at first sex change (Ls50 and Ws50) and length at first maturity (Lm50). The overall sex ratio for male and female was 1:0.69 with male being predominant throughout the year. Threadfin fish spawn the whole year round with peaks during moderate rainy and heavy rainy seasons. Histological examination confirmed its protandrous hermaphrodite posing multiple spawning habits. The average fecundity was 1.85 × 105 ± 1.05 × 105 eggs and positively related with standard length, body weight, gonad weight, and egg diameter (p < 0.05). The Ls50 and Ws50 were 27.58 cm and 419.39 g, and 29.71 cm and 457.28 g, for fish from Pattani Bay and Samut Prakan province, respectively. The Lm50 of male from Pattani Bay and Samut Prakan province were 25.78 cm and 25.56 cm, respectively, which were larger than those from Satun and Nakhon Sri Thammarat provinces. The Lm50 of females from Pattani Bay was smaller than that from Samut Prakan province. This study provided fundamental information on the reproductive characteristics of E. tetradactylum, which can be implemented to support management of natural fish stock and aquaculture development.