Fine particulate matter (PM₂.₅) has been associated with risk of oral and respiratory diseases. However, the biological mechanisms of adverse oral and respiratory health response to PM₂.₅ fluctuation have not been well characterized. This study aims to explore the relationships of PM₂.₅ with airway inflammation, salivary biomarkers and buccal mucosa microbiota. We performed a panel study among 40 college students involving 4 follow-ups from August to October 2021 in Hefei, Anhui Province, China. Health outcomes included fractional exhaled nitric oxide (FeNO), salivary biomarkers [C-reactive protein (CRP), cortisol, lysozyme and alpha-amylase] and buccal mucosa microbial diversity. Linear mixed-effect models were applied to explore the cumulative impacts of PM₂.₅ on health indicators. PM₂.₅ was positively correlated with FeNO, CRP, cortisol and alpha-amylase, while negatively with lysozyme. Per 10-μg/m³ increase in PM₂.₅ was linked to maximum increments in FeNO of 10.71% (95%CI: 2.01%, 19.41%) at lag 0–24 h, in CRP of 7.10% (95%CI: 5.39%, 8.81%) at lag 0–24 h, in cortisol of 1.25% (95%CI: 0.44%, 2.07%) at lag 0–48 h, and in alpha-amylase of 2.12% (95%CI: 0.53%, 3.71%) at lag 0–24 h, while associated with maximum decrement in lysozyme of 0.53% (95%CI: 0.12%, 0.95%) at lag 0–72 h. Increased PM₂.₅ was linked to reduction in the richness and evenness of buccal microbe and o_Bacillales and o_Bacteroidales were identified as differential microbes after PM₂.₅ inhalation. Bio-information analysis indicated that immunity system pathway was the most important enriched abundant process altered by PM₂.₅ exposure. In summary, short-term PM₂.₅ exposure may impair oral and respiratory health by inducing inflammatory and stress responses, weakening immune function and altering buccal mucosa microbial diversity.

In the marine environment, microplastic contamination and acidification may occur simultaneously, this study evaluated the effects of ocean acidification and microplastics on oxidative stress responses and digestive enzymes in mussels. The thick shell mussels Mytilus coruscus were exposed to four concentrations of polystyrene microspheres (diameter 2 μm, 0, 10, 10⁴ and 10⁶ particles/L) under two pH levels (7.7 and 8.1) for 14 days followed by a 7-day recovery acclimation. Throughout the experiment, we found that microplastics and ocean acidification exerted little oxidative stress to the digestive gland. Only catalase (CAT) and glutathione (GSH) showed a significant increase along with increased microplastics during the experiment, but recovered to the control levels once these stressors were removed. No significant effects of pH and microplastics on glutathione peroxidase (GPx) and superoxide dismutase (SOD) were observed. The responses of digestive enzymes to both stressors were more pronounced than antioxidant enzymes. During the experiment, pepsin (PES), trypsin (TRS), alpha-amylase (AMS) and lipase (LPS) were significantly inhibited under microplastics exposure and this inhibition was aggravated by acidification conditions. Only PES and AMS tended to recover during the recovery period. Lysozyme (LZM) increased significantly under microplastic exposure conditions, but acidification did not exacerbate this effect. Therefore, combined stress of microplastics and ocean acidification slightly impacts oxidative responses but significantly inhibits digestive enzymes in mussels.

Microplastics (MPs) are small plastic particles less than 5 mm in diameter. MPs in the form of microfibers (MFs) are widely detected in aquatic habitats and are of high environmental concern. Despite many reports on the effects of MFs on marine animals, their effect on sea cucumbers is still unclear. In addition, our previous filed study has shown that MFs may transfer to the coelomic fluid of the sea cucumber Apostichopus japonicus (A. japonicus). Here, we show how MFs transfer to the coelomic fluid of the sea cucumber. We captured the MFs during their transfer from the water to the coelomic fluid through the respiratory tree. A. japonicus ingested in the MFs along with the water during respiration; the MFs got stuck in the respiratory tree or transferred to the coelomic fluid. The transferred MFs increased during 72 h of exposure and persisted for 72 h after the transfer to clean water. Among the immunity indices, lysozyme (LZM) levels increased in response to the transferred MFs, which confirms the defensive role of LZMs against strange substances. Additionally, non-significantly decreased levels of total antioxidant capacity (T-AOC), malondialdehyde (MDA), peroxidase (POD) and phenol oxidase (PPO) were observed at 24 h and 48 h post-exposure, suggesting minimal oxidative imbalance. Furthermore, there were no significant changes in the speed and the total distance moved by A. japonicus post MFs transfer. This study revealed that MFs transfer and accumulate in the coelomic fluid of A. japonicus.

Both microplastics and persistent organic pollutants (POPs) are ubiquitously present in natural water environment, posing a potential threat to aquatic organisms. While it has been suggested that the immune responses of aquatic organisms could be hampered by exposure to microplastics and POPs, the synergistic immunotoxic impact of these two types of pollutants remain poorly understood. In addition, little is known about the mechanism behind the immunotoxic effect of microplastics. Therefore, in the present study, the immunotoxicity of microplastics and two POPs, benzo[a]pyrene (B[a]P) and 17β-estradiol (E2), were investigated alone or in combination in a bivalve species, Tegillarca granosa. Evident immunotoxicity, as indicated by alterations of haemocyte count, blood cell composition, phagocytic activity, intracellular content of ROS, concentration of Ca²⁺ and lysozyme, and lysozyme activity, was revealed for both microplastics and the two POPs examined. In addition, the expression of six immune-, Ca²⁺ signalling-, and apoptosis-related genes was significantly altered by exposure of clams to the contaminants studied. Furthermore, the toxicity of POPs was generally aggravated by smaller microplastics (500 nm) and mitigated by larger ones (30 μm). This size dependent effect on POP toxicity may result from size dependent interactions between microplastics and POPs. Data obtained in this study also indicate that similar to exposure to B[a]P and E2, exposure to microplastics may hamper the immune responses of clams through a series of interdependent physiological and molecular processes.

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) has been detected in various environmental media and has been implicated as a weak mutagen or carcinogen, but whether TDCIPP can promote the progression of liver tumor remains unclear. In this study, krasⱽ¹² genetically modified zebrafish, Tg(fabp10:rtTA2s-M2; TRE2:EGFP-krasᴳ¹²ⱽ), a model system in which liver tumors can be induced by doxycycline (DOX), was used to evaluate the liver tumor promotion potential of TDCIPP. Briefly, krasⱽ¹² transgenic females were exposed to 0.3 mg/L TDCIPP, 20 mg/L DOX or a binary mixture of 0.3 mg/L TDCIPP with 20 mg/L DOX, and liver size, histopathology, and transcriptional profiles of liver were determined. Treatment with TDCIPP resulted in increased liver size and caused more aggressive hepatocellular carcinoma (HCC). Compared with the exposure to DOX, TDCIPP in the presence of DOX up-regulated the expression of genes relevant with salmonella infection and the toll-like receptor signaling pathway. These results implied an occurrence of inflammatory reaction, which was sustained by the increase in the amount of infiltrated neutrophils in the liver of Tg(lyz:DsRed2) transgenic zebrafish larvae whose neutrophils were labelled by red fluorescent protein under the lysozyme C promoter. Furthermore, compared with the binary exposure of DOX and TDCIPP, treatment with a ternary mixture of TDCIPP, DOX and inflammatory response inhibitor (ketoprofen) significantly decrease the liver size and the amounts of neutrophils in the livers of kras and lyz double transgenic zebrafish larvae. Collectively, our results suggested that TDCIPP could promote the liver tumor progression by induction of hepatic inflammatory responses.

Gabapentin (GPT) has become an emerging contaminant in aquatic environments due to its wide application in medical treatment all over the world. In this study, embryos of zebrafish were exposed to gabapentin at realistically environmental concentrations, 0.1 μg/L and 10 μg/L, so as to evaluate the ecotoxicity of this emergent contaminant. The transcriptomics profiling of deep sequencing was employed to illustrate the mechanisms. The zebrafish (Danio rerio) embryo were exposed to GPT from 12 hpf to 96 hpf resulting in 136 and 750 genes differentially expressed, respectively. The results of gene ontology (GO) analysis and the Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis illustrated that a large amount of differentially expressed genes (DEGs) were involved in the antioxidant system, the immune system and the nervous system. RT-qPCR was applied to validate the results of RNA-seq, which provided direct evidence that the selected genes involved in those systems mentioned above were all down-regulated. Acetylcholinesterase (AChE), lysozyme (LZM) and the content of C-reactive protein (CRP) were decreased at the end of exposure, which is consistent with the transcriptomics results. The overall results of this study demonstrate that GPT simultaneously affects various vital functionalities of zebrafish at early developmental stage, even at environmentally relevant concentrations.

Gabapentin-lactam (GBP-L) is a transformation product (TP) of gabapentin (GBP), a widely used anti-epileptic pharmaceutical. Due to its high persistence, GBP-L has been frequently detected in the surface water. However, the effects of GBP-L on aquatic organisms have not been thoroughly investigated. In the present study, zebrafish (Danio rerio) embryos as a model organism were used to study the impacts of GBP-L in terms of embryos LC₅₀, spontaneous movement at 24 hpf (hours post fertilization), heartbeat rates at 48 hpf, and body length at 72 hpf, with the concentrations of GBP-L down to 0.01 μg/L, covering its environmental concentrations. Various biomarkers from nervous, antioxidant and immune systems of zebrafish larvae were analyzed, including acetylcholinesterase, acetylcholine, dopamine, gamma-aminobutyric acid, superoxide dismutase, catalase, glutathione S-transferase, C reactive protein, and lysozyme, to assess its toxicity on these systems. RT-qPCR was then used to further verify the results and explain the toxicological mechanism at the gene level. The results demonstrated that GBP-L is much more toxic than its parent compound, and could lead to adverse impacts on the aquatic organisms even at every low concentrations.

Glyphosate (GLY) is one of the most used herbicide worldwide. Considering that information concerning the impact of GLY on bivalves is scarce, in this study we evaluated for the first time the effects of environmentally realistic concentrations of GLY (10, 100 and 1000 μg/L) to the mussel Mytilus galloprovincialis. Mussels were exposed for 7, 14 and 21 days and several biomarkers were measured in haemocytes/haemolymph (total haemocyte counts, haemocyte diameter and volume, haemolymph pH, haemolymph lactate dehydrogenase activity, haemocyte lysate lysozyme and acid phosphatase activities), as well as in gills and digestive gland (antioxidant enzyme and acetylcholinesterase activities). The concentrations of GLY and its main metabolite aminomethylphosphonic acid in the experimental tanks were also measured. The MANOVA analysis demonstrated that the experimental variables considered (exposure concentration, exposure duration, and their interaction) affected significantly biomarker responses. In addition, the two-way ANOVA analysis indicated that GLY was able to affect most of the cellular parameters measured, whereas antioxidant enzyme activities resulted to be influenced moderately. Interestingly, exposure to GLY reduced significantly acetylcholinesterase activity in gills. Although preliminary, the results of this study demonstrated that GLY can affect both cellular and biochemical parameters in mussels, highlighting a potential risk for aquatic invertebrates.

Several biomarkers were measured to evaluate the effects of Trimethoprim (TMP; 300, 600 and 900 ng/L) in the clam Ruditapes philippinarum after exposure for 1, 3 and 7 days. The actual TMP concentrations were also measured in the experimental tanks. The total haemocyte count significantly increased in 7 day-exposed clams, whereas alterations in haemocyte volume were observed after 1 and 3 days of exposure. Haemocyte proliferation was increased significantly in animals exposed for 1 and 7 days, whereas haemocyte lysate lysozyme activity decreased significantly after 1 and 3 days. In addition, TMP significantly increased haemolymph lactate dehydrogenase activity after 3 and 7 days. Regarding antioxidant enzymes, only a significant time-dependent effect on CAT activity was recorded. This study demonstrated that environmentally realistic concentrations of TMP affect haemocyte parameters in clams, suggesting that haemocytes are a useful cellular model for the assessment of the impact of TMP on bivalves.

Robust antimicrobial capability is crucial for marine organisms survival in complex ocean environments. Although the detrimental impacts of emergent pollutants on cellular immune response of marine bivalve mollusks were increasingly documented, the effects of bisphenol A (BPA) and microplastics (MPs) on humoral immune response and hemocyte chemotactic activity remain unclear. Therefore, in this study, the toxicities of BPA and MPs, alone or in combination, to the antimicrobial ability, humoral immune response, and hemocyte chemotactic activity were investigated in the blood clam Tegillarca granosa. Our data demonstrated that exposure of blood clams to BPA, MPs, and BPA-MPs for 2 weeks lead to significant reductions in their survival rates upon pathogenic bacterial challenge, indicating evident impairment of antimicrobial ability. Compared to control, the plasma of pollutant-incubated blood clams exhibited significantly less antimicrobial activity against the growth of V. harveyi, suggesting significant reduction in humoral immune effectors including defensin, lysozyme (LZM), and lectin. Moreover, hemocytes migration across the polycarbonate membrane to the serum containing chamber was markedly arrested by 2-week exposure to BPA, MPs, and BPA-MPs, suggesting a hampered chemotactic activity. In addition, the intracellular contents of ROS and protein carbonyl in hemocytes were markedly induced whereas the expression levels of key genes from the MAPK and actin cytoskeleton regulation pathways were significantly suppressed upon exposure. In this study, it was also found that BPA-MP coexposure was significantly more toxic than single exposures. In summary, our findings revealed that exposure to the pollutants tested possibly impair the antimicrobial ability of blood clam through (1) reducing the inhibitory effect of plasma on bacterial growth, the contents of humoral immune effectors, and the chemotactic activity of hemocytes, (2) interrupting IL-17 activation of MAPK signal pathway, (3) inducing intracellular ROS, elevating protein carbonylation levels, and disrupting actin cytoskeleton regulation in hemocytes.