Distribution characteristics of antibiotic resistance genes and microbial diversity in the inshore aquaculture area of Wenchang, Hainan, China.

The rapid development of marine aquaculture has led to the increased use and release of antibiotics into the marine environment, consequently contributing to the emergence of antibiotic resistance. Information on antibiotic resistance in nearshore marine aquaculture areas remains limited, and research on the microbial composition and potential hosts of antibiotic resistance genes (ARGs) in marine aquaculture areas is scarce. This study used SmartChip real-time fluorescent quantitative PCR and qPCR to quantitatively analyze 44 ARGs and 10 mobile genetic elements (MGEs) genes in 12 sampling points in the nearshore aquaculture area of Wenchang. High-throughput sequencing of 16S rRNA was used to study microbial diversity in the study area, to clarify the correlation between ARGs, MGEs, and microbial diversity, and to determine the possible sources and potential hosts of ARGs. The results showed that a total of 37 ARGs and 8 MGEs were detected in the study area. The detection rate of 9 ARGs (aac(6')-Ib(aka aacA4)-02, catA1, cmlA, cfr, sul1, sul2, sulA/folP-01, tetC, tetX) was 100 %. The absolute abundance of ARGs in the 12 sampling points ranged from 2.75 × 107 to 3.79 × 1010 copies·L-1, and the absolute abundance of MGEs was 1.30 × 105 to 2.54 × 107 copies·L-1, which was relatively high compared to other research areas. ARGs and MGEs were significantly correlated, indicating that MGEs play an important role as a mediator in the spread of ARGs. At the phylum level, Proteobacteria and Cyanobacteria were the dominant bacteria in the study area, with HIMB11 and unidentifiedChloroplast being the dominant levels, respectively. Network analysis of ARGs and microorganisms (genus level) revealed that Cognatishimia, Thalassobius, Aestuariicoccus, Thalassotalea, and Vibrio were significantly correlated with multiple ARGs and were the main potential hosts of ARGs in the nearshore waters of Wenchang.

Mechanistic toxicity and growth abnormalities mediated by subacute exposure to environmentally relevant levels of benzophenone-3 in clown anemonefish (Amphiprion ocellaris).

Benzophenone-3 (BP-3) is a UV filter that is ubiquitously present in the environment due to its photostability and degradation resistance and has wide applications in personal care products. BP-3 will eventually be discharged into the ocean. Studies shows BP-3 interferes with endocrine system of aquatic organisms, especially fish. However, the toxicity and mechanisms of subacute exposure of the coral reef fish to BP-3 remain elusive. Here, we exposed the one-month-old clown anemonefish to BP-3 at 1 and 10 µg/L for 14 and 28 days, respectively. After chronic exposure, the effects of BP-3 on the growth of clown anemonefish were investigated in terms of growth-related hormones, immune enzyme activity, digestive enzyme activity, transcriptional profiling of feeding- and obesity-related genes and digital RNA sequencing. The body weight in the BP-3 groups were abnormally increased (1 µg/L group in 14 days treatment and all groups in 28 days treatment), altered insulin content (28 days exposure), immune-related and digestive-related enzymatic activities. At the molecular level, BP-3 interferes with the expression of feeding- and obesity-related genes. Digital RNA sequencing analysis showed that BP-3 interferes with Kyoto encyclopedia of genes and genomes (KEGG) pathways related to growth, social behavior (learning behavior), Mitogen-Activated Protein Kinase (MAPK) signaling pathway, PI3K-Akt signaling pathway, and insulin secretion. Notably, in the insulin secretion, BP-3 induced Ca2+ up-regulation that may damage ß cells. Growth abnormalities and social behavior (learning behavior) KEGG pathway disturbances may have potential impacts on populations of clown anemonefish. Our results reveal the toxicological effects of subacute exposure to BP-3, and provides insight into the effects and mechanisms of BP-3 on clown anemonefish growth.

Occurrence and Distribution of Antibiotics in a Tropical Mariculture Area of Hainan, China: Implications for Risk Assessment and Management.

With the rapid global demand for mariculture products in recent years, the use of antibiotics has increased intensively in the mariculture area. Current research on antibiotic residues in mariculture environments is limited, and less information is available on the presence of antibiotics in tropical waters, limiting a comprehensive understanding of their environmental presence and risk. Therefore, this study investigated the environmental occurrence and distribution of 50 antibiotics in the near-shore aquaculture waters of Fengjia Bay. A total of 21 antibiotics were detected in 12 sampling sites, including 11 quinolones, 5 sulfonamides, 4 tetracyclines, and 1 chloramphenicol; the quinolones pyrimethamine (PIP), delafloxacin (DAN), flurofloxacin (FLE), ciprofloxacin (CIP), norfloxacin (NOR), pefloxacin (PEF), enrofloxacin (ENO), and minocycline (MNO) of the tetracycline class were detected in all sampling points. The total antibiotic residue concentrations in the study area ranged from 153.6 to 1550.8 ng/L, the tetracycline antibiotics were detected in the range of 10 to 1344.7 ng/L, and the chloramphenicol antibiotics were detected in the range of 0 to 106.9 ng/L. The detected concentrations of quinolones ranged from 81.3 to 136.1 ng/L, and the residual concentrations of sulfonamide antibiotics ranged from 0 to 313.7 ng/L. The correlation analysis with environmental factors revealed that pH, temperature, conductivity, salinity, NH3--N, and total phosphorus had a strong correlation with antibiotics. Based on PCA analysis, the main sources of antibiotic pollution in the area were determined to be the discharge of farming wastewater and domestic sewage. The ecological risk assessment indicated that the residual antibiotics in the water environment of the near-shore waters of Fengjiawan had certain risks to the ecosystem. Among them, CIP, NOR, sulfamethoxazole (TMP), ofloxacin (OFL), enrofloxacin (ENO), sulfamethoxazole (SMX), and FLE showed medium to high risk. Therefore, it is recommended to regulate the use of these antibiotics and the discharge and treatment of culturing wastewater, and measures should be taken to reduce the environmental pollution caused by antibiotics and to monitor the long-term ecological risk of antibiotics in the region. Overall, our results provide an important reference for understanding the distribution and ecological risk of antibiotics in Fengjiawan.

Environmental concentrations of benzophenone-3 disturbed lipid metabolism in the liver of clown anemonefish (Amphiprion ocellaris).

Benzophenone-3 (BP-3) often used as a UV filter in various products and an endocrine disruptor. In this work, we exposed the clown anemonefish to 10 µg/L and 50 µg/L BP-3 for 7 and 14 days. Liver histological, biochemical analysis, and transcriptome sequencing were used to explore the mechanism of the lipid metabolism disorder in the liver of three-month-old clown anemonefish treated with BP-3. The histological and biochemical analysis showed that BP-3 induces morphological changes and lipid droplet accumulation, and the lipid content, lipase, and antioxidant enzyme activity were abnormal. After treatment with 10 µg/L and 50 µg/L BP-3 for 7 days, the transcriptome analysis further demonstrated that the KEGG analysis revealed that the differentially expressed genes (DEGs) were mainly associated with fat digestion and absorption, PPAR signaling pathway, circadian rhythm, and mineral absorption pathways; After 10 µg/L and 50 µg/L of BP-3 exposure for 14 days, the KEGG analysis were mainly associated with circadian rhythm, circadian rhythm-fly, protein processing in the endoplasmic reticulum, and beta-alanine metabolism pathways. Several key genes were involved in the process of liver lipid metabolism, including CD36, APoA-Ⅰ, FABP, LPL, ACS, and PEPCK. The qRT-PCR validation results showed that eight genes (CYP8B1, FABP1, LPL, MGAT, PEPCK, PER1, PSMB4, PSME2) were significantly down-regulated, and the other two genes (Fbxl3, RXR) were significantly up-regulated after 7 days of BP-3 exposure. Similarly, eleven genes (AMPK, ARNTL, Bmal1, CASP3, CYC, CYP2J, CYP2U1, GSK3A, PEPCK, RAC1, RORA) were significantly up-regulated, and the other four genes (NR1D1, PER1, PTGDS, HLF) were significantly down-regulated after 14 days of BP-3 exposure. In conclusion, our results elucidate the physiological and molecular responses to BP-3 exposure in the liver lipid metabolism of clown anemonefish, and these findings reveal that the regulation of lipid metabolism is disturbed when clown anemonefish is exposed to UV filters.