Tissue-Specific Uptake, Depuration Kinetics, and Suspected Metabolites of Three Emerging Per- and Polyfluoroalkyl Substances (PFASs) in Marine Medaka.

Restrictions on legacy per- and polyfluoroalkyl substances (PFASs) have led to the widespread use of emerging PFASs. However, their toxicokinetics have rarely been reported. Here, tissue-specific uptake and depuration kinetics of perfluoroethylcyclohexanesulfonate (PFECHS) and 6:2 and 8:2 chlorinated polyfluoroalkyl ether sulfonates (Cl-PFESAs) were studied in marine medaka (Oryzias melastigma). The fish were exposed to these substances for 28 days (0.2 µg/L), followed by a clearance period of 14 days. The depuration constant (kd) of PFECHS [0.103 ± 0.009 day-1 (mean ± standard deviation)] was reported for the first time. Among the six studied tissues, the highest concentrations of 6:2 Cl-PFESA, 8:2 Cl-PFESA, and PFECHS were found in the liver [1540, 1230, and 188 ng (g of wet weight)-1, respectively] on day 28 while the longest residence times were found in the eyes (t1/2 values of 21.7 ± 4.3, 23.9 ± 1.5, and 17.3 ± 0.8 days, respectively). No significant positive correlation was found between the bioconcentration factors of the studied PFASs and the phospholipid or protein contents in different tissues of the studied fish. Potential metabolites of Cl-PFESAs, i.e., their hydrogen-substituted analogues (H-PFESAs), were identified by time-of-flight mass spectrometry. However, the biotransformation rates were low (<0.19%), indicating the poor capacity of marine medaka to metabolize Cl-PFESAs to H-PFESAs.

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.

Microcystin-leucine arginine causes brain injury and functional disorder in Lithobates catesbeianus tadpoles by oxidative stress and inflammation.

Microcystin-leucine arginine (MC-LR) is a toxin commonly found in eutrophic waters worldwide, but its potential effects on amphibian brain toxicity and exposure mechanisms are unclear. In this study, Lithobates catesbeianus tadpoles were exposed to MC-LR for 30 days at realistic ambient concentrations (0, 0.5, and 2 µg/L) to reveal its effects on brain health. The MC-LR bioaccumulation in the brain increased in dependence on the concentration of MC-LR exposure. Exposure to 0.5 and 2 µg/L MC-LR resulted in a significant down-regulation of the expression of structural components of the blood-brain barrier (CLDN1), while the expression of genes associated with inflammation (NLRP3, TNF, IL-1ß, and CXCL12) was significantly up-regulated with increased number of eosinophils. In the hippocampal and hypothalamic regions, the number of vacuolated neuropils increased with increasing MC-LR exposure concentration, while the expression of genes associated with neuronal development (LGALS1, CACNA2D2, and NLGN4X) and neurotransmitter transmission (SLC6A13 and AChE) was significantly down-regulated. Moreover, the levels of neurotransmitters (5-HT, glutamate, GABA, and ACh) were significantly reduced. These results provide strong evidence that MC-LR exposure at realistic ambient concentrations of 0.5 and 2 µg/L can break the blood-brain barrier and raise the accumulation of MC-LR in the brain tissue, causing structural damage and functional disorder to brain neurons. Further, based on transcriptomic and biochemical analysis, it was revealed that MC-LR exposure induces DNA damage through oxidative stress and may be an important pathway causing brain structural damage and functional disorder. Overall, this study demonstrates the significant effects of MC-LR on the brain tissue of amphibians, highlighting the sensitivity of amphibians to MC-LR.

Application of aerobic denitrifier for simultaneous removal of nitrogen, zinc, and bisphenol A from wastewater.

High concentrations of heavy metals and other pollutants affect microbial activity in the wastewater treatment system and impede biological denitrification process. In this study, a novel Zn(II)-resistant aerobic denitrifier (Pseudomonas stutzeri KY-37) was isolated with potential in Bisphenol A (BPA) biodegradation and removal. The capability of this denitrifier in removing nitrogen, zinc, and BPA was tested. Using 56 mg/L nitrate as the sole nitrogen source, its removal efficiency achieved 98.5% in 12 h. This novel denitrifier had a strong auto-aggregation (maximum 65.8%), a high hydrophobicity rate (maximum 88.2%), and a massive amount (maximum 41.1 mg/g cell dry weight) of extracellular polymeric substances (EPS) production. Moreover, Zn(II) removal efficiency reached more than 95% with the initial high concentrations of 200 mg/L. The maximum BPA removal efficiency reached 88.8% with initial 10 mg/L. The removal mechanism of BPA was further explored in terms of microbial degradation, EPS adsorption, and intermediate degradation products.