Isothiazolinone Disrupts Reproductive Endocrinology by Targeting the G-Protein-Coupled Receptor Signaling.

The unintended exposure of humans and animals to isothiazolinones has led to an increasing concern regarding their health hazards. Isothiazolinones were previously found to disrupt reproductive endocrine homeostasis. However, the long-term reproductive toxicity and underlying mechanism remain unclear. In this study, life-cycle exposure of medaka to dichlorocthylisothiazolinone (DCOIT), a representative isothiazolinone, significantly stimulated the gonadotropin releasing hormone receptor (GnRHR)-mediated synthesis of follicle stimulating hormone and luteinizing hormone in the brain. Chem-Seq and proteome analyses revealed disturbances in the G-protein-coupled receptor, MAPK, and Ca2+ signaling cascades by DCOIT. The G protein αi subunit was identified as the binding target of DCOIT. Gαi bound by DCOIT had an enhanced affinity for the mitochondrial calcium uniporter, consequently changing Ca2+ subcellular compartmentalization. Stimulation of Ca2+ release from the endoplasmic reticulum and blockage of Ca2+ uptake into the mitochondria resulted in a considerably higher cytoplasmic Ca2+ concentration, which then activated the phosphorylation of MEK and ERK to dysregulate hormone synthesis. Overall, by comprehensively integrating in vivo, ex vivo, in silico, and in vitro evidence, this study proposes a new mode of endocrine disrupting toxicity based on isothiazolinones, which is expected to aid the risk assessment of the chemical library and favor the mechanism-driven design of safer alternatives.

A newly developed UPLC-MS/MS method for simultaneous quantitative analysis of ajuforrestin A, ajuforrestin B, ajugamacrin and 8-O-acetylharpagide derived from Ajuga plants in mice blood and the in vivo pharmacokinetics.

OBJECTIVE: To develop a sensitive and fast detection method via ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to assess the concentration of ajuforrestin A, ajuforrestin B, ajugamacrin and 8-O-Acetylharpagide primarily derived from Ajuga plants in mice blood and their pharmacokinetics. METHODS: Single protein precipitation with high-proportioned acetonitrile is chosen for sample clean-up. The UPLC HSS T3 (2.1 mm × 100 mm, 1.8 µm) column with a mobile phase in gradient elution mode at the flow rate of 0.4 mL/min was used for sample separation. Acetonitrile was selected as the organic phase solution and water containing 0.1% formic acid was chosen as the aqueous solution. A tandem mass spectrometer containing an electrospray ionization (ESI) source in the positive ionization mode was used to detect four compounds via multiple reaction monitoring (MRM). RESULTS: The calibration curves (5-1000 ng/mL) of four compounds were linear with correlation coefficients > 0.997. The matrix effects, accuracy, precision, and recovery were all within permissible scope. CONCLUSIONS: In this approach, the corresponding pharmacokinetic parameters were successfully clarified in mouse for the first time, which provided a theoretical basis for the improvement of the standard of Ajuga plants and the safety of clinical medication. Furthermore, this method may provide the UPLC-MS/MS evidence for the differentiation of the main close relative varieties of genus Ajuga according to these plants contain different mixtures of the four marker compounds.

Coupling harmful algae derived nitrogen and sulfur co-doped carbon nanosheets with CeO2 to enhance the photocatalytic degradation of isothiazolinone biocide.

Removing the algae from water bodies is an effective treatment toward the worldwide frequently occurred harmful algae blooms (HAB), but processing the salvaged algae waste without secondary pollution places another burden on the economy and environment. Herein, a green hydrothermal process without any chemical addition was developed to resource the HAB algae (Microcystis sp.) into autogenous nitrogen and sulfur co-doped carbon nanosheet materials C-CNS and W-CNS, whose alga precursors were collected from pure culture and a wild bloom pond, respectively. After coupling with CeO2, the obtained optimal C-CNS/CeO2 and W-CNS/CeO2 composites photocatalytically degraded 95.4% and 88.2% of the marine pollutant 4,5-Dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) in 90 min, significantly higher than that of pure CeO2 (63.15%). DCOIT degradation on CNS/CeO2 was further conducted under different conditions, including pH value, coexisting cations and anions, and artificial seawater. Although different influences were observed, the removal efficiencies were all above 76%. Along with the ascertained good stability and reusability in five consecutive runs, the great potential of CNS/CeO2 for practical application was validated. UV-vis DRS showed the increased light absorption of CNS/CeO2 in comparison to pure CeO2. PL spectra and photoelectrochemical measurements suggested the lowered charge transfer resistance and thereby inhibited charge recombination of CNS/CeO2. Meanwhile, trapping experiments and electron paramagnetic resonance (EPR) detection verified the primary roles of hydroxyl radical (OH) and superoxide radical (O2-) in DCOIT degradation, as well as their notably augmented generation by CNS. Consequently, a mechanism of CNS enhanced photocatalytic degradation of DCOIT was proposed. The intermediates involved in the reaction were identified by LC-QTOF-MS, giving rise to a deduced degradation pathway for DCOIT. This study offers a new approach for resourceful utilization of the notorious HAB algae waste. Besides that, photocatalytic degradation has been explored as an effective measure to remove DCOIT from the ocean.

Significant Variability in the Developmental Toxicity of Representative Perfluoroalkyl Acids as a Function of Chemical Speciation.

Current toxicological data of perfluoroalkyl acids (PFAAs) are disparate under similar exposure scenarios. To find the cause of the conflicting data, this study examined the influence of chemical speciation on the toxicity of representative PFAAs, including perfluorooctanoic acid (PFOA), perfluorobutane carboxylic acid (PFBA), and perfluorobutanesulfonic acid (PFBS). Zebrafish embryos were acutely exposed to PFAA, PFAA salt, and a pH-negative control, after which the developmental impairment and mechanisms were explored. The results showed that PFAAs were generally more toxic than the corresponding pH control, indicating that the embryonic toxicity of PFAAs was mainly caused by the pollutants themselves. In contrast to the high toxicity of PFAAs, PFAA salts only exhibited mild hazards to zebrafish embryos. Fingerprinting the changes along the thyroidal axis demonstrated distinct modes of endocrine disruption for PFAAs and PFAA salts. Furthermore, biolayer interferometry monitoring found that PFOA and PFBS acids bound more strongly with albumin proteins than did their salts. Accordingly, the acid of PFAAs accumulated significantly higher concentrations than their salt counterparts. The present findings highlight the importance of chemical forms to the outcome of developmental toxicity, calling for the discriminative risk assessment and management of PFAAs and salts.

Legacy and Emerging Per- and Polyfluoroalkyl Substances Surveillance in Bufo gargarizans from Inlet Watersheds of Chaohu Lake, China: Tissue Distribution and Bioaccumulation Potential.

Amphibians are sensitive biomonitors of environmental pollutants but reports regarding per- and polyfluoroalkyl substances (PFAS), a class of synthetic organofluorine substances, are limited. In this study, samples of water and Chinese toads (Bufo gargarizans) were collected in Chaohu Lake, China. Tissue-specific bioaccumulation characteristics of 39 PFAS, including 19 perfluoroalkyl acids (PFAAs), 8 emerging PFAS, and 12 PFAA precursors, were investigated, and the levels of some biochemical indicators were determined. The highest PFAS concentrations were found in the liver [215.97 ng/g dry weight (dw)] of Chinese toads, followed by gonads (135.42 ng/g dw) and intestine (114.08 ng/g dw). A similar tissue distribution profile was found between legacy and emerging PFAS in the toads, and the occurrence of two emerging PFAS, 2,3,3,3-tetrafluoro-2-propanoate (HFPO-DA) and 6:2 hydrogen-substituted polyfluorooctane ether sulfonate (6:2 H-PFESA) in the amphibians were for the first time reported. Field-based bioaccumulation factors of HFPO-DA were higher than perfluorooctanoic acid, indicating the higher bioaccumulation potential of this emerging PFAS than the legacy C8 compound. Males had significantly higher gonad PFAS levels than females while estradiol levels in gonads increased with increasing concentrations of certain PFAS (e.g., 6:2 H-PFESA), implying that PFAS may trigger estrogenic effects in the toads, especially for male toads.

Variability in fecal metabolome depending on age, PFBS pollutant, and fecal transplantation in zebrafish: A non-invasive diagnosis of health.

To protect the wellbeing of research animals, certain non-invasive measures are in increasing need to facilitate an early diagnosis of health and toxicity. In this study, feces specimen was collected from adult zebrafish to profile the metabolome fingerprint. Variability in fecal metabolite composition was also distinguished as a result of aging, perfluorobutanesulfonate (PFBS) toxicant, and fecal transplantation. The results showed that zebrafish feces was very rich in a diversity of metabolites that belonged to several major classes, including lipid, amino acid, carbohydrate, vitamin, steroid hormone, and neurotransmitter. Fecal metabolites had functional implications to multiple physiological activities, which were characterized by the enrichment of digestion, absorption, endocrine, and neurotransmission processes. The high richness and functional involvement of fecal metabolites pinpointed feces as an abundant source of diagnostic markers. By comparison between young and aged zebrafish, fundamental modifications of fecal metabolomes were caused by aging progression, centering on the neuroactive ligand-receptor interaction pathway. Exposure of aged zebrafish to PFBS pollutant also significantly disrupted the metabolomic structure in feces. Of special concern were the changes in fecal hormone intermediates after PFBS exposure, which was concordant with the in vivo endocrine disrupting effects of PFBS. Furthermore, it was intriguing that transplantation of young zebrafish feces efficiently mitigated the metabolic perturbation of PFBS in aged recipients, highlighting the health benefits of therapeutic strategies based on gut microbiota manipulation. In summary, the present study provides preliminary clues to evidence the non-invasive advantage of fecal metabolomics in the early diagnosis and prediction of physiology and toxicology.

Exposure to methylparaben at environmentally realistic concentrations significantly impairs neuronal health in adult zebrafish.

Methylparaben (MeP) is an emerging aquatic pollutant that is found to impact neural functions. However, it still lacks a comprehensive understanding about its neurotoxicology. The present study exposed adult zebrafish to environmentally realistic concentrations (0, 1, 3, and 10 µg/L) of MeP for 28 days, with objectives to elucidate the neurotoxic effects and mechanisms. Proteomic profiling found that MeP pollutant induced distinct mechanism of neurotoxicity as a function of sex. MeP pollutant appeared to preferentially target the neurotransmission cascade via synapse junctions. In male brain, glutamatergic neural signaling was enhanced by 10 µg/L of MeP in characteristics of higher glutamate neurotransmitter content (by 61.9%) and up-regulated glutamate receptor expression by 2.6-fold relative to the control. In MeP-exposed female brain, biomarker proteins of synapse formation and regeneration had significantly lower abundance, accounting for the blockage of synaptic neurotransmission. Furthermore, under the stress of MeP pollutant, both male and female zebrafish initiated a negative feedback mechanism along stress neuroendocrine axis by down-regulating the transcriptions of corticotropin-releasing hormone and its binding protein, which subsequently decreased blood cortisol concentrations. MeP subchronic exposure also disturbed innate immune function. In particular, significant increases in lipopolysaccharide (LPS) content by 15.6% were caused by MeP exposure in male brain, thereby inducing the synthesis of pro-inflammatory cytokines. In contrast, female brain was able to adaptively up-regulate the protein expression of blood brain barrier to inhibit the infiltration of LPS endotoxin into brain. Overall, the present findings pinpoint the potent neurotoxicity of MeP pollutant even at environmentally realistic concentrations.

Metabolomic profiles in zebrafish larvae following probiotic and perfluorobutanesulfonate coexposure.

Probiotic supplements are able to attenuate the developmental toxicity of perfluorobutanesulfonate (PFBS) pollutant. However, the underlying mechanisms remain unexplored. To this end, the present study acutely exposed zebrafish larvae for 4 days to 0 and 10 mg/L of PFBS, with or without the addition of probiotic Lactobacillus rhamnosus in the rearing water. The metabolomics approach was used to reveal the combined effects of PFBS and probiotics on metabolic dynamics, based on which gene transcriptions, enzymatic activities, and behavioral endpoints were further examined. The results showed that probiotic supplements were the major driver of the metabolomic fingerprints in coexposed zebrafish larvae. The addition of probiotic bacteria significantly decreased the methylation potential whilst up-regulating the demethylation process of genomic DNA, which may globally stimulate the gene expression to improve somatic growth. Acute exposure to PFBS significantly increased the cortisol concentration in zebrafish larvae, subsequently inducing stress response and hyperactive behavior. In contrast, probiotic supplementation promoted the degradation of cortisol, thus alleviating the stressful state. Antagonistic action of probiotics against PFBS developmental toxicity was also noted regarding the locomotor behavior. In addition, gut microbiota-mediated production of secondary bile acids was remarkably enhanced by probiotic supplements regardless of PFBS exposure. Overall, the present study underlines the efficacy of probiotic bacteria to protect zebrafish larvae from the metabolic disturbances of PFBS, thereby providing more evidence to support the application of probiotics in aquaculture and fishery as an environmentally-friendly choice.

Disturbed glucose metabolism by perfluorobutanesulfonate pollutant and benefit of young fecal transplantation in aged zebrafish.

Perfluorobutanesulfonate (PFBS) is an environmental pollutant of emerging concern, which significantly impacts the metabolism and health of animals. Because of the loss of functional capacity, the aged animals are regarded more susceptible to the toxicity of environmental pollutants. In the present study, aged zebrafish were employed as the toxicological animal and transplanted with the feces collected from young donors for 14 days, after which the acclimated elderly were exposed to PFBS at environmentally relevant concentrations (0 and 100 µg/L) for another 14 days. When the exposure was concluded, glucose metabolic disturbances of PFBS in the aged and efficacy of young fecal transplant to mitigate the toxicity of PFBS were explored along the gut-liver axis. The results showed that PFBS exposure significantly inhibited the enzymatic activity of α-amylase in the gut, but increased the alanine aminotransferase (ALT) activity in the blood of the aged zebrafish, suggesting the impairment of intestinal digestive functions of carbohydrates and the induction of liver damage by PFBS. However, young fecal transplantation successfully ameliorated the toxicity of PFBS on α-amylase and ALT, underlining the benefits conveyed to the health of the elderly. In addition, transplantation of young feces was efficient to alleviate the hyperglycemia symptom in the elderly via stimulating the secretion of insulin. PFBS exposure increased blood glucagon level, disrupted insulin receptor transcription, and depleted hepatic glycogen store, which were all mitigated by young fecal transplant. Hepatic proteomic analysis also found dynamic interactions between young fecal transplantation and PFBS pollutant on the metabolic pathways of glucose and glycogen, involving glycolysis, gluconeogenesis, glycogenesis, and glycogenolysis. Overall, the present findings highlighted the beneficial effects of young fecal transplantation to protect the aged from the glucose metabolism toxicity of PFBS, thus providing a plausible measure to improve the health aging status.

Enhanced photocatalytic degradation of paraben preservative over designed g-C3N4/BiVO4 S-scheme system and toxicity assessment.

Paraben preservatives have been listed as emerging pollutants due to their ubiquity in various environmental matrices, especially the water bodies. How to efficiently and practically eliminate these paraben pollutants is therefore of great importance. Herein, a designed S-scheme heterojunction photocatalyst, consisting of graphitic carbon nitride (g-C3N4) and monoclinic bismuth vanadate (BiVO4), was fabricated by a facile hydrothermal synthesis and employed to treat benzyl-paraben (BzP). TEM and XPS analysis testified the intimate interaction between g-C3N4 and BiVO4, and the consequently smoothed interfacial charge transfer rendered the feasible recombination of the photoexcited electrons (from BiVO4) and holes (from g-C3N4). The as-established S-scheme system enabled the left g-C3N4 electrons and BiVO4 holes to maintain the high redox abilities and accelerated the charge separation concurrently. In particular, the g-C3N4/BiVO4 composite generated much higher photocurrent response as compared with pure g-C3N4 and BiVO4, highlighting the improved separation of photoinduced charges. Therefore, under visible light and natural solar light irradiation, the g-C3N4/BiVO4 composite showed the significantly enhanced photocatalytic degradation of BzP, which was further optimized with 5 wt% g-C3N4 in the composite. According to the Mott-Schottky plots and identified active species, the mechanism of the g-C3N4/BiVO4 S-scheme heterojunction system was illustrated. In addition, during the photocatalytic degradation process, the acute toxicity of the reaction solutions on zebrafish embryos was notably reduced. In conclusion, the demonstrated strategy to enhance the photocatalytic performance by designing S-scheme heterostructure may provide more insights into the development of high-efficiency photocatalyst towards the solar energy utilization and environmental treatment. Furthermore, photocatalytic degradation had been proved to be an efficient method for eliminating the ecological risk of paraben pollutants, warranting more attention in future work.

Microcystin-leucine arginine induces skin barrier damage and reduces resistance to pathogenic bacteria in Lithobates catesbeianus tadpoles.

Despite the importance of the skin mucosal barrier and commensal microbiota for the health of amphibians, the potential of environmental contaminants to disrupt the skin mucosal barrier and microbiota have rarely been studied in toxicology. In this study, tadpoles (Lithobates catesbeianus) were exposed to 0, 0.5, and 2 µg/L of microcystin-leucine arginine (MC-LR) for 30 days to explore the impacts of environmentally realistic MC-LR concentrations on the physical skin barrier, immune barrier, commensal microbiota, and skin resistance to pathogenic bacterial invasion. MC-LR exposure significantly reduced the collagen fibrils in the dermis of skin tissues and down-regulated tight junction and stratum corneum-related gene transcriptions, suggesting the damage caused by MC-LR to the physical barrier of the skin. Increased skin eosinophils and upregulated transcriptions of inflammation-related genes in the exposed tadpoles underline the development of skin inflammation resulting from MC-LR exposure even at environmentally realistic concentrations. Comparative transcriptome and immunobiochemical analyses found that antimicrobial peptides (Brevinin-1PLc, Brevinin-2GHc, and Ranatuerin-2PLa) and lysozyme were down-regulated in the exposed groups, while complement, pattern recognition receptor, and specific immune processes were up-regulated. However, the content of endotoxin lipopolysaccharide produced by bacteria increased in a dose-dependent pattern. The disc diffusion test showed a reduced ability of skin supernatant to inhibit pathogenic bacteria in the exposed groups. Analysis of microbial 16 S rRNA gene by high-throughput sequencing revealed that MC-LR interfered with the abundance, composition, and diversity of the skin commensal microbiota, which favored the growth of pathogen-containing genera Rhodococcus, Acinetobacter, and Gordonibacter. In summary, the current study provides the first clues about the impact of MC-LR on the integrity and function of skin barrier of amphibians. These new toxicological evidences can facilitate a more comprehensive evaluation of the ecological risk of MC-LR to amphibians.

Disturbances in Microbial and Metabolic Communication across the Gut-Liver Axis Induced by a Dioxin-like Pollutant: An Integrated Metagenomics and Metabolomics Analysis.

To determine how the aryl hydrocarbon receptor (AhR) signaling acts along the gut-liver axis, we employed an integrated metagenomic and metabolomic approach to comprehensively profile the microbial and metabolic networks. Adult zebrafish were exposed to a model agonist of the AhR: polychlorinated biphenyl (PCB) 126. The metagenomic analysis showed that PCB126 suppressed microbial activities related to primary bile acid metabolism in male intestines. Accordingly, a suite of primary bile acids consistently showed higher concentrations, suggesting that bacterial conversion of primary bile acids was blocked. PCB126 also disturbed bacterial metabolism of bile acids in female intestines, as revealed by higher concentrations of primary bile acids (e.g., chenodeoxycholic acid) and activation of the nuclear farnesoid X receptor signaling. In addition, PCB126 exposure impaired the metabolism of various essential vitamins (e.g., retinol, vitamin B6, and folate). Degradation of vitamin B6 by bacterial enzymes was inhibited in male intestines, resulting in its intestinal accumulation. However, PCB126 suppressed the bacterial metabolism of vitamins in female intestines, causing systematic deficiency of essential vitamins. Overall, we found that PCB126 exposure dysregulated gut microbial activities, consequently interrupting bile acid and vitamin metabolism along the gut-liver axis. The findings provided an insight of the AhR action in microbe-host metabolic communication related to PCBs.

Nonalcoholic Fatty Liver Disease Development in Zebrafish upon Exposure to Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate, a Novel Brominated Flame Retardant.

Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH), a novel brominated flame retardant, can potentially cause lipid metabolism disorder; however, its biological effects on lipid homeostasis remain unknown. We investigated its ability to cause nonalcoholic fatty liver disease (NAFLD) in zebrafish. Female zebrafish were fed a high-fat diet (HFD, 24% crude fat) or normal diet (ND, 6% crude fat), and exposed to TBPH (0.02, 2.0 µM) for 2 weeks. Consequently, HFD-fed fish showed a higher measured concentration of TBPH than ND-fed fish. Further, TBPH-treated fish in the HFD group showed higher hepatic triglyceride levels and steatosis. In comparison to ND-fed fish, treating HFD-fed fish with TBPH led to an increase in the concentration of several proinflammatory markers (e.g., TNF-α, IL-6); TBPH exposure also caused oxidative stress. In addition, the mRNA levels of genes encoding peroxisome proliferator-activated receptors were increased, and the transcription of genes involved in lipid synthesis, transport, and oxidation was upregulated in both ND- and HFD-fed fish. Both the ND and HFD groups also showed demethylation of the peroxisome proliferator-activated receptor-γ coactivator 1-α gene promoter, accompanied by the upregulation of tet1 and tet2 transcription. To summarize, we found that TBPH amplified the disruption of lipid homeostasis in zebrafish, leading to the enhancement of diet-induced NAFLD progression.

Shift in skin microbiota and immune functions of zebrafish after combined exposure to perfluorobutanesulfonate and probiotic Lactobacillus rhamnosus.

Dysbiosis of fish skin microbiome and immunity by environmental pollutants are rarely studied in toxicological research in spite of their importance for fish health. In the present study, adult zebrafish were exposed to 0 and 10 µg/L of perfluorobutanesulfonate (PFBS) for 40 days, with or without the supplementation of probiotic Lactobacillus rhamnosus, with objectives to explore the interaction between PFBS pollutant and probiotic bacteria on skin mucosal microbiota and immune response. Amplicon sequencing analysis found that PFBS alone significantly disturbed the microbial community composition and abundance on the skin, favoring the growth of stress-tolerant bacteria (e.g., Deinococcus and Enhydrobacter genera). However, the administration of probiotic inhibited the dysbiosis of PFBS and shaped the skin microbiome in the combined exposure group. PFBS single exposure also promoted the production of mucus on the skin of male zebrafish, which may be related to the growth of Limnobacter bacteria. In contrast, probiotic supplements remarkably improved the immune functions in male skin mucus from the combined group, as evidenced by the consistent increases in lysozyme activity, immunoglobulin concentrations and peroxidase activity. Overall, the present study provides the first clue about the singular and combined effects of PFBS and probiotic on skin microbiota and immunity, highlighting the beneficial action of probiotic L. rhamnosus against PFBS stress.

Interaction between probiotic additive and perfluorobutanesulfonate pollutant on offspring growth and health after parental exposure using zebrafish.

Perfluorobutanesulfonate (PFBS) pollutant and probiotic bacteria can interact to affect the reproductive outcomes of zebrafish. However, it is still unexplored how the growth and health of offspring are modulated by the combination of PFBS and probiotic. In the present study, adult zebrafish were exposed to 0 and 10 µg/L PFBS for 40 days, with or without dietary supplementation of probiotic Lactobacillus rhamnosus. After parental exposure, the development, growth and viability of offspring larvae were examined, with the integration of molecular clues across proteome fingerprint, growth hormone/insulin-like growth factor (GH/IGF) axis, calcium homeostasis, hypothalamic-pituitary-adrenal (HPA) axis and nutrient metabolism. Parental probiotic supplementation significantly increased the body weight and body length of offspring larvae. Despite the spiking of PFBS, larvae from the combined exposure group still had longer body length. RNA processing and ribosomal assembly pathways may underlie the enhancement of offspring growth by probiotic bacteria. However, the presence of PFBS remarkably increased the concentrations of cortisol hormone in offspring larvae as means to cope with the xenobiotic stress, which required more energy production. As evidenced by the proteomic analysis, the addition of probiotic bacteria likely alleviated the energy metabolism disorders of PFBS, thus allocating more energy for the larval offspring growth from the combined group. It was noteworthy that multiple molecular disturbances caused by PFBS were antagonized by probiotic additive. Overall, the present study elucidated the intergenerational interaction between PFBS and probiotic on offspring growth and health after parental exposure.

Fingerprinting fecal DNA and mRNA as a non-invasive strategy to assess the impact of polychlorinated biphenyl 126 exposure on zebrafish.

In toxicological studies, experimental animals are generally subjected to dissection to obtain the tissues of concern, which causes great harm to the animals. In this regard, it is necessary to test and develop a non-invasive strategy to prevent the animals from anthropic injury when achieving scientific objectives. Therefore, zebrafish fecal DNA and mRNA pools were assessed by using metagenomic and transcriptomic analyses based on their potential to diagnose toxicological impairment of polychlorinated biphenyl (PCB) 126, a model persistent organic pollutant. The results showed that there was abundant zebrafish DNA and mRNA in the feces, which were, however, associated with contrasting profiles of physiological activities. As compared to DNA fragments, fecal mRNA provided a better representation of zebrafish physiological status. PCB126 exposure dramatically shifted the composition of fecal zebrafish DNA and mRNA as a function of sex. The differential mRNA caused by PCB126 clearly identified the toxicological fingerprint of PCB126. In summary, this study provides preliminary clues about the potential of fecal genes (mRNA in particular) in the development of non-invasive toxicological approaches. In the future, it is expected that more works will be conducted to screen sensitive diagnostic biomarkers from feces to increase the rate and reduce the cost of ecological risk assessment.

Unexpected Observations: Probiotic Administration Greatly Aggravates the Reproductive Toxicity of Perfluorobutanesulfonate in Zebrafish.

The present study exposed adult zebrafish to 0, 10, and 100 µg/L perfluorobutanesulfonate (PFBS) with or without dietary supplement of probiotic Lactobacillus rhamnosus. Interaction between probiotic and PFBS on sex endocrine and reproduction was investigated. It was striking to find that PFBS and probiotic coexposures almost ceased the fecundity, which was accompanied by disturbances in sex hormones and oocyte maturation in females. In contrast, probiotic additive efficiently antagonized the estrogenic activity of PFBS in males. For the first time, this study reported that probiotic heavily depended on sex to modulate the endocrine disruption and reproductive toxicity of aquatic pollutants.

Probiotic Modulation of Lipid Metabolism Disorders Caused by Perfluorobutanesulfonate Pollution in Zebrafish.

To determine whether and how probiotic supplement can alter gut microbiota dysbiosis and lipid metabolism disorders caused by perfluorobutanesulfonate (PFBS), the present study exposed adult zebrafish to 0, 10, and 100 µg/L PFBS for 28 days, with or without dietary administration of probiotic Lactobacillus rhamnosus. Regarding intestinal health and gut microbiota, probiotic supplement altered the innate toxicities of PFBS, depending on exposure concentration and the sex of the fish. Lactobacillus genus correlated positively (P < 0.001; r > 0.5) with other beneficial bacteria in the gut microbiota, thereby indirectly regulating host metabolic activities. In female fish, the PFBS and probiotic combination enhanced fatty acid synthesis and ß-oxidation, but mitigated the accumulation of cholesterol in the blood compared with PFBS single exposure, highlighting the benefits of the probiotic to host health. In male zebrafish, probiotic administration antagonized the PFBS-induced disturbances of bile acid metabolism, presumably via farnesoid X receptor signaling. However, coexposure to PFBS and probiotic caused significant accumulation of triglyceride in male livers (2.6-fold relative to the control), implying the induction of hepatic steatosis. Overall, the present study underlined the potential of probiotics to modulate gut microbial dysbiosis and lipid metabolism disorders caused by PFBS exposure, which could provide implications to the application of probiotics in aquaculture.

Metabolic and transcriptional changes in seminal plasma of asthenozoospermia patients.

This study was designed to investigate the metabolic and transcriptional alterations in seminal fluid caused by asthenozoospermia (AS). To address these issues, a method of metabonomics based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and real-time quantitative PCR (RT-qPCR) was performed to identify some crucial biomarkers and transcription levels of the enzymes in seminal fluid. Seminal fluid samples were collected from 87 AS patients and 73 healthy males with normozoospermia. The quantitative analysis by UPLC-MS/MS showed that 19 metabolites in seminal plasma were associated with AS, and they were involved in several metabolic pathways, such as energy metabolism, purine metabolism, methionine cycle, and branched chain amino acid metabolism. Among these metabolites, the levels of citric acid, malic acid, succinic acid, and pyruvic acid, which are related to energy metabolism, were collectively reduced in the AS group, whereas the lactic acid level was enhanced. These results indicated that lesser energy source (adenosine triphosphate) was produced through the anaerobic glycolysis pathway rather than via aerobic catabolism of suger and tricarboxylic acid cycle, resulting in reduced power of sperms. Meanwhile, partial least squares discriminant analysis showed significant differences in metabolic profiles between the AS and control groups. In addition, RT-qPCR results revealed that the expression levels of four genes encoding fructokinase citrate synthase, succinate dehydrogenase, and spermine synthase, which were related to energy metabolism, were decreased in the AS group. The 23 descriptors with differential expression in AS may be valuable for the diagnosis and sequential study on AS. These results will help highlight the role of sperm inactivity in AS pathogenesis.

Metagenomic Analysis of Zinc Surface-Associated Marine Biofilms.

Biofilms are a significant source of marine biofouling. Marine biofilm communities are established when microorganisms adhere to immersed surfaces. Despite the microbe-inhibiting effect of zinc surfaces, microbes can still attach to the surface and form biofilms. However, the diversity of biofilm-forming microbes that can attach to zinc surfaces and their common functional features remain elusive. Here, by analyzing 9,000,000 16S rRNA gene amplicon sequences and 270 Gb of metagenomic data, we comprehensively explored the taxa and functions related to biofilm formation in subtidal zones of the Red Sea. A clear difference was observed between the biofilm and adjacent seawater microbial communities in terms of the taxonomic structure at phylum and genus levels, and a huge number of genera were only present in the biofilms. Saturated alpha-diversity curves suggested the existence of more than 14,000 operational taxonomic units in one biofilm sample, which is much higher than previous estimates. Remarkably, the biofilms contained abundant and diverse transposase genes, which were localized along microbial chromosomal segments and co-existed with genes related to metal ion transport and resistance. Genomic analyses of two cyanobacterial strains that were abundant in the biofilms revealed a variety of metal ion transporters and transposases. Our analyses revealed the high diversity of biofilm-forming microbes that can attach to zinc surfaces and the ubiquitous role of transposase genes in microbial adaptation to toxic metal surfaces.