Linking the removal of enrofloxacin to the extracellular polymers of microalgae in water bodies: A case study focusing on the shifts in microbial communities.

Microalgae can promote antibiotic removal, which has attracted growing attention. However, its synergistic removal performance with bacteria in antibiotic pollutants is still poorly understood. In this study, firstly, we selected two green algae (Dictyosphaerium sp. and Chlorella sp.) and exposed them to Enrofloxacin (ENR) to observe their extracellular polysaccharides (EPS) concentration dynamic and the removal of antibiotics. Secondly, EPS was extracted and added to in situ lake water (no algae) to investigate its combined effect with bacteria. The results indicate that both Dictyosphaerium sp. and Chlorella sp. exhibited high tolerance to ENR stress. When the biomass of microalgae was low, ENR could significantly stimulate algae to produce EPS. The removal rates of Dictyosphaerium sp. and Chlorella sp. were 15.8% and 10.5%, respectively. The addition of EPS can both alter the microbial community structure in the lake water and promote the removal of ENR. The LEfSe analysis showed that there were significant differences in the microbial marker taxa, which promoted the increase of special functional bacteria for decomposing ENR, between the EPS-added group and the control group. The EPS of Dictyosphaerium sp. increased the abundance of Moraxellaceae and Spirosomaceae, while the EPS of Chlorella sp. increased the abundance of Sphingomonadaceae and Microbacteriaceae. Under the synergistic effect, Chlorella sp. achieved a maximum removal rate of 24.2%, while Dictyosphaerium sp. achieved a maximum removal rate of 28.9%. Our study provides new insights into the removal performance and mechanism of antibiotics by freshwater microalgae in water bodies and contribute to the development of more effective water treatment strategies.

Effect of four fluoroquinolones on the viability of bladder cancer cells in 2D and 3D cultures.

Introduction: The anticancer properties of fluoroquinolones and the high concentrations they achieve in urine may help in bladder cancer therapy. This study aimed to analyze the properties of 4 fluoroquinolones as potential candidates for supportive treatment of bladder cancer. Methods: Comparative analyses were performed on the cytotoxic effects of norfloxacin, enrofloxacin, moxifloxacin, and ofloxacin on normal and cancer urothelial cell lines. In 2D culture, the cytotoxic properties of fluoroquinolones were evaluated using MTT assay, real-time cell growth analysis, fluorescence and light microscopy, flow cytometry, and molecular analysis. In 3D culture, the properties of fluoroquinolones were tested using luminescence assays and confocal microscopy. Results and Discussion: All tested fluoroquinolones in 2D culture decreased the viability of both tested cell lines in a dose- and timedependent manner. Lower concentrations did not influence cell morphology and cytoskeletal organization. In higher concentrations, destruction of the actin cytoskeleton and shrinkage of the nucleus was visible. Flow cytometry analysis showed cell cycle inhibition of bladder cancer cell lines in the G2/M phase. This influence was minimal in the case of normal urothelium cells. In both tested cell lines, increases in the number of late apoptotic cells were observed. Molecular analysis showed variable expression of studied genes depending on the drug and concentration. In 3D culture, tested drugs were effective only in the highest tested concentrations which was accompanied by caspase 3/7 activation and cytoskeleton degradation. This effect was hardly visible in non-cancer cell lines. According to the data, norfloxacin and enrofloxacin had the most promising properties. These two fluoroquinolones exhibited the highest cytotoxic properties against both tested cell lines. In the case of norfloxacin, almost all calculated LC values for bladder cancer cell lines were achievable in the urine. Enrofloxacin and norfloxacin can be used to support chemotherapy in bladder cancer patients.

Determination of residual enrofloxacin in eggs due to long term administration to laying hens. Analysis of the consumer exposure assessment to egg derivatives.

The use of the antibiotic enrofloxacin (ENR) in poultry is controversial. A high-performance liquid chromatography coupled to fast-scanning fluorescence detection (HPLC-FSFD) method for the determination of ENR in egg white, egg yolk, and lyophilized samples was developed. In a first analysis, the long-term administration of ENR (100 days) to laying hens was carried out to determine its presence in egg white, yolk, or both. The predominance of ENR was observed in egg white and variations in the weight of egg white and eggshell was evidenced, showing a potential problem in the industry. Eventually, the presence of ENR was confirmed in commercial lyophilized egg white samples in concentration values around 350 µg kg-1. The consumer exposure assessment was estimated for children, adolescents, and adults. The result displayed that, in an intake of lyophilized egg white with food-producing animals, the %ADI exceeds 100%, showing toxicological levels.

[Toxicological Effects of Enrofloxacin and Its Removal by Freshwater Micro-Green Algae Dictyosphaerium sp.]

Enrofloxacin (ENR), a fluoroquinolones antibiotic, is widely used in the medical and aquaculture fields. Its residues in surface waters in China are high. However, few studies have evaluated both its toxicity to phytoplankton and the degradation or removal by microalgae. In this study, the growth, photosynthetic activity, and exopolysaccharides (EPS) of freshwater micro-green algae Dictyosphaerium sp. and the dynamics of ENR concentrations (0, 5, 25, 50, and 100 mg·L-1) were studied through an exposure experiment for 12 days. Results showed that the biomass and photosynthetic pigment content of Dictyosphaerium sp. increased with increasing exposure time in each treatment; however, it showed a significant inhibitory effect on the growth and pigment accumulation of Dictyosphaerium sp. compared with the control group (P<0.01). The LC50 of ENR to Dictyosphaerium sp. was (241.29±7.33) mg·L-1 after 96-h exposure, indicating that Dictyosphaerium sp. could adapt to the stress conditions of high concentration ENR. Meanwhile, when the concentration of enrofloxacin was<5 mg·L-1, it was found to promote the maximum photosynthetic rate (Fv/Fm) of Dictyosphaerium sp. On the contrary, when the concentration of enrofloxacin was>5 mg·L-1, photosynthetic inhibition was observed (P<0.01). The actual photosynthetic rate (Yield) and the maximum electron transfer rate (ETRmax) showed a trend of initially decreasing and then increasing in 12 days. It can gradually adapt to the stress conditions and recover certain photosynthetic activity after 6 days' exposure. In addition, ENR can also stimulate the EPS (RPS and CPS) release. At the end of the experiment, the removal rates of ENR in the four control groups (no algae addition groups) (5, 25, 50, and 100 mg·L-1ENR) were 7.27%, 5.56%, 5.30%, and 4.88%, respectively, while the removal rates of the treatment groups were 3.21, 3.01, 2.69, and 2.83 times of the no algae groups, indicating that Dictyosphaerium sp. had a significant promoting effect on the removal of ENR (P<0.01). Overall, our results can provide new insights for the understanding of the ecological toxicity of fluoroquinolone antibiotics to primary producers in the aquatic system and also provide new ideas for the ecological removal of antibiotic residues in water bodies and the biological resource utilization of freshwater microalgae.

[Nanoscale Zero-valent Copper-Activated Molecular Oxygen for the Degradation of Enrofloxacin in Water].

Systematically studied the oxidation of enrofloxacin (ENR) in a nanoscale zero-valent copper (nZVC)-activated molecular oxygen system. The results show that nanoscale copper powder has a higher surface area than microscale copper powder, non-porous structure, and rough surface and exists in form of agglomerates. Nanoscale ZVC shows a superior activated performance toward molecular oxygen compared with microscale ZVC, which is due to its larger specific area and the fact that it corrodes easier. The H2O2 generated from the activation of molecular oxygen and the Cu+ released from surface corrosion form a novel Fenton-like system in which hydroxyl radicals are continuously produced, resulting in high-efficiency removal of ENR from water. The superoxide radicals produced during the reaction promote the reduction of Cu2+ to Cu+, thus speeding up the removal of ENR. The reaction conditions have a certain effect on the ENR degradation in nZVC-activated molecular oxygen systems. A higher nZVC dosage, lower ENR concentration, higher reaction temperature, and strong acidic conditions are favorable for the ENR removal.

Determination of residual enrofloxacin in food samples by a sensitive method of chemiluminescence enzyme immunoassay.

A chemiluminescence enzyme immunoassay (CLEIA) based on the HRP-luminol-H2O2 chemiluminescence system for highly sensitive detection of enrofloxacin (ENR) was proposed in this study. Key factors that affect the precision and accuracy for the determination of ENR residues were optimised. Under the optimal conditions, the proposed method showed an excellent performance. The linearity range for method developed for determination of ENR was 0.35-1.0 ng/mL with a correlation coefficient greater than 0.994. The limit of detection was 0.03 ng/mL and the relative standard deviations (RSDs) were less than 9.4% and 13.0% for intra-day and inter-day assays. The proposed method was satisfactorily applied to determine ENR in milk, eggs, and honey samples at three spiked levels (0.4, 0.7, and 1.0 ng/mL) and the recoveries ranged from 92.4% to 104.2% for milk, 93.8% to 103.2% for eggs and 94.1% to 105.0% for honey, respectively. Compared the results of CLEIA with those of ELISA and HPLC, the advantages of the CLEIA were further confirmed. Moreover, one 96-well microtiter plate coated with anti-ENR can be used to detect multiple samples at the same time, which indicated that the CLEIA using HRP-luminol-H2O2 system was a sensitive, high throughput and real-time method for ENR residues analysis.