Unveiling the inhibition of chlortetracycline photodegradation and the increase of toxicity when coexisting with silver nanoparticles.

Silver nanoparticles (AgNPs) and antibiotics inevitably co-exist in water environment. Nonetheless, little is known regarding the interactions between AgNPs and antibiotics or the effects of AgNPs on environmental behavior of antibiotics, particularly on sunlight-driven transformation. In the present work, we found that AgNPs obviously inhibit the photochemical decay of chlortetracycline (CTC), and CTC boosts the dissolution of AgNPs. With the help of electron paramagnetic resonance (EPR) and quenching experiment, we ascertained that these results originated from the competition between AgNPs against CTC for capturing 1O2 generated from CTC photosensitization. 1O2 reacting with CTC contributed mostly to CTC photodegradation, while 1O2 as well reacting with AgNPs leads to release of Ag+. When compared to reaction of 1O2 with CTC, 1O2 is prone to react with AgNPs, based on lower Gibbs free energy of AgNPs reacting with 1O2. Therefore, upon CTC co-existing with AgNPs, the release of Ag+ was accelerated and the photodegradation of CTC was inhibited obviously. Furthermore, the accelerated release of Ag+ significantly increased their toxicity toward E. coli cells under simulate sunlight irradiation. Overall, the findings demonstrate how AgNPs interact with CTC and how these interactions affect the environmental behaviors of CTC or AgNPs, allowing more accurate assessments of the risk to ecosystems posed by AgNPs coexisting with antibiotics.

Application of the Aliivibrio fischeri bacterium bioassay for assessing single and mixture effects of antibiotics and copper.

The Aliivibrio fischeri bioassay was successfully applied in order to evaluate the acute effect of sulfamethoxazole (SMX), ciprofloxacin (CIP), chlortetracycline (CTC) and copper (Cu), alone or in binary, ternary, and overall mixture. The toxicity results are reported in terms of both effective concentrations, which inhibited 50% of the bacterium bioluminescence (EC50%), and in Toxic Units (TUs). The TUs were compared with predicted values obtained using the Concentration Addition model (CA). Finally, the toxicity of water extracts from a soil contaminated by the three antibiotics (7 mg Kg-1 each) in the presence/absence of copper (30 mg Kg-1) was also evaluated. Copper was the most toxic chemical (EC50: 0.78 mg L-1), followed by CTC (EC50: 3.64 mg L-1), CIP (96 mg L-1) and SMX (196 mg L-1). Comparing the TU and CA values of the mixtures, additive effects were generally found. However, a synergic action was recorded in the case of the CIP+Cu co-presence and antagonistic effects in the case of CTC+Cu and the ternary mixture (containing each antibiotic at 0.7 mg L-1), were identified. Soil water extracts did not show any toxicity, demonstrating the buffering ability of the soil to immobilize these chemicals.

Response of microbial interactions in activated sludge to chlortetracycline.

Chlortetracycline (CTC) has attracted increasing attention due to its potential environmental risks. However, its effects on bacterial communities and microbial interactions in activated sludge systems remain unclear. To verify these issues, a lab-scale sequencing batch reactor (SBR) exposed to different concentrations of CTC (0, 0.05, 0.5, 1 mg/L) was carried out for 106 days. The results showed that the removal efficiencies of COD, TN, and TP were negatively affected, and the system functions could gradually recover at low CTC concentrations (≤0.05 mg/L), but high CTC concentrations (≥0.5 mg/L) caused irreversible damage. CTC significantly altered bacterial diversity and the overall bacterial community structure, and stimulated the emergence of many taxa with antibiotic resistance. Molecular ecological network analysis showed that low concentrations of CTC increased network complexity and enhanced microbial interactions, while high concentrations of CTC had the opposite effect. Sub-networks analysis of dominant phyla (Bacteriodota, Proteobacteria, and Actionobacteriota) and dominant genera (Propioniciclava, a genus from the family Pleomorphomonadaceae and WCHB1-32) also showed the same pattern. In addition, keystone species identified by Z-P analysis had low relative abundance, but they were important in maintaining the stable performance of the system. In summary, low concentrations of CTC enhanced the complexity and stability of the activated sludge system. While high CTC concentrations destabilized the stability of the overall network and then caused effluent water quality deterioration. This study provides insights into our understanding of response in the bacteria community and their network interactions under tetracycline antibiotics in activated sludge system.

Influence on denitrifying community performance by the long-term exposure to sulfamethoxazole and chlortetracycline in the continuous-flow EGSB reactors.

The response of the denitrification community to long-term antibiotic exposure requires further investigation. Here, the significantly altered denitrifying community structure and function were observed by continuous exposure to 1 mg/L sulfamethoxazole (SMZ) or chlortetracycline (CTC) for 180 d in the expanded granular sludge bed reactors. Thaurea, positively correlated with SMZ and NO3- removal efficiency (NrE), was highly enriched in the SMZ-added reactor, while, Comamons and Acinetobacter were largely inhibited. The acute inhibited and then gradual-recovered NrE (87.17-90.38 %) was observed with highly expressed narG, indicating the adaptability of Thaurea to SMZ. However, the abundance of Thaurea and Comamonas greatly decreased, while Melioribacter and Acinetobacter were largely enriched in the CTC-added reactor. CTC created more serious and continuous inhibition of NO3- reduction (NrE of 64.53-66.95 %), with lowly expressed narG. Improved NO2- reduction capacity was observed in both reactors (70.16-95.42 %) with highly expressed nirS and nosZ, revealing the adaptability of NO2- reduction populations to antibiotics.

Efficient biodegradation of chlortetracycline in high concentration from strong-acidity pharmaceutical residue with degrading fungi.

Chlortetracycline (CTC) pharmaceutical residue with strong acidity and in high CTC concentration is a hazardous solid waste. There is a huge attention but few studies on whether and how the CTC raw residue (CRR) can be degraded in microbiological way. In this study, three self-screened fungi, LJ245, LJ302 and LJ318, were used and thoroughly investigated to remove CTC, strong acidity and biotoxicity in CRR. The result disclosed that the concentration of CTC decreased rapidly in the first seven days and declined slowly subsequently, and the decreasing curve was similar to "L" shape. the corresponding degradation ratios of three strains were 95.73%, 98.53% and 98.07%, respectively. Meanwhile, numerous intermediates in degradation appeared in early days and gradually reduced, and eventually disappeared once the degradation time was long enough, among which eleven intermediates from CTC were identified. Moreover, the strong acidity of CRR declined dramatically using this biological method along with the CTC being metabolized, the pH value increased from 2.30 to 8.32 in the first 7 days. The toxicity of CRR was significantly reduced by LJ302 with inhibition rate from 96.02% to no inhibition effect to Micrococcus luteus. Therefore, CTC, strong acidity and biotoxicity of CRR could be effectively removed simultaneously through a biodegradation process driven with proposed strains.

Toxicity of chlortetracycline and oxytetracycline on Vallisneria natans (Lour.) Hare.

Tetracyclines are frequently detected in water bodies due to their widespread use in aquaculture and animal husbandry. A hydroponic experiment was conducted to explore the phytotoxic effects of Vallisneria natans (Lour.) Hare exposed to various concentrations of chlortetracycline (CTC) and oxytetracycline (OTC) (0, 0.1, 1, 10, 30, 50, and 100 mg/L) for 7 days (7 D) and 14 days (14 D), respectively. The results showed that similar to OTC treatment for 7 D, the relative growth rates (RGR) and catalase (CAT) activity of V. natans, after 7 D of CTC exposure, decreased significantly at 10 mg/L and 30 mg/L, respectively. The content of soluble protein notably decreased when CTC ≥ 10 mg/L and OTC ≥ 30 mg/L. The hydrogen peroxide (H2O2) content was significantly stimulated when OTC ≥ 10 mg/L, while it hardly changed when exposed to CTC. After 14 D, the malondialdehyde (MDA) and H2O2 contents of V. natans were significantly higher than those of the control group under a high concentration of OTC (≥ 30 mg/L), but they did not change significantly under a high concentration of CTC. The activity of polyphenol oxidase (PPO), under CTC treatment after 14 D, showed first a significant increase then decreases; the maximum value (125% of the control) was noticed at 10 mg/L CTC, while it remained unchanged when exposed to OTC. The soluble protein content significantly decreased at 10 mg/L CTC and 0.1 mg/L OTC, respectively. The RGR, CAT, and peroxidase (POD) activities, similar to OTC treatment after 14 D, decreased evidently when CTC was 10 mg/L, 30 mg/L, and 0.1 mg/L, respectively. CTC and OTC harm the chlorophyll content of V. natans after 14 D, and the reductions of chlorophyll a and carotenoid were more pronounced than chlorophyll b. The results suggest that CTC and OTC both have a negative effect on the growth of V. natans, and OTC can cause oxidative damage in V. natans but CTC harms the metabolism process without inducing oxidative damage. Overall, the toxicity of OTC to V. natans is stronger than that of CTC.

Use of waste materials to prevent tetracycline antibiotics toxicity on the growth of soil bacterial communities.

The increase of concentrations of tetracycline antibiotics in agricultural soils worldwide is of special concern, due to its potential toxic effects on soil bacterial communities. In the present work, the reuse of two waste/by-product materials as soil amendments was tested as a preventive practice for reducing tetracycline antibiotics toxicity in soils. Pine bark (PB), with high percentage of organic carbon, and crushed mussel shell (CMS), a frequent natural liming material, were added to 4 soils in doses 0, 6, 12 and 48 g of by-product per kg-1 of soil (dry weight) of each one (separately). The soils and soil-waste mixtures were then spiked with tetracycline (TC), oxytetracycline (OTC) and chlortetracycline (CTC). After one day of incubation, the bacterial growth was estimated in soils and soil-mixtures using the leucine incorporation technique. The addition of PB to the soils showed two different behaviors, depending on the antibiotics. The toxicity of TC and OTC decreased with the addition of PB (toxicities going from 6 to 25% and from 5 to 36%, respectively). However, CTC toxicity did not change, or even increased in response to the PB amendment. Regarding soil amendment with CMS, it was not effective to prevent the toxicity of any of the three antibiotics studied.

The enhanced degradation and detoxification of chlortetracycline by Chlamydomonas reinhardtii.

Nowadays, numerous studies have focused on the newly developed technologies for the thorough removal of tetracyclines (TCs). However, it is often ignored that the parent TCs have limited stability in aquatic environments. Thus, this study selected green alga Chlamydomonas reinhardtii with high chlorophyll content to rapidly degrade chlortetracycline (CTC) into products with low toxicity. As the results shown, the half-life times of CTC (1 × 10-6 mol/L) decreased from 10.35 h to 2.55 h by the presence of C. reinhardtii at 24±1 °C with 12/12 h dark/light cycle. The main transformation products were iso-chlortetracycline (ICTC), 4-epi-iso-chlortetracycline (EICTC), and other degradation products with lower molecular weight. The toxicity evaluation shows that the negative effects of CTC on growth rate and soluble protein content of green algae were significantly alleviated after the enhanced degradation treatment, while the generation of reactive oxygen species (ROS) and antioxidant response in algal cells returned to normal levels. The chlorophyll of algae played an important role of photosensitizer, which catalyzed the photo-induced electron/energy transfer of CTC degradation. The ROS generation of algae also was also inseparable from the enhanced degradation of CTC, especially when the chlorophyll was damaged at the high CTC concentration. Based on these results, we can better select suitable algal species to further strengthen the degradation of antibiotics and effectively reduce the environmental risk of CTC in aqueous system.

Development of antibiotic resistance genes in soils with ten successive treatments of chlortetracycline and ciprofloxacin.

Antibiotic contamination caused by the long-term use of organic manure (OM) in greenhouse agricultural soils poses potential detrimental effects to the soil environment. By applying OM containing chlortetracycline (CTC) and/or ciprofloxacin (CIP) ten times in soil under laboratory conditions, we investigated the dissipation and accumulation characteristics of CTC and CIP in the soil, the changes in the microbial pollution-induced community tolerance (PICT), and the diversity and abundance of antibiotic resistance genes (ARGs) in the soil microbiome. The dissipation of CTC was rapid while CIP was accumulated in repeatedly treated soils; further, CIP could inhibit the dissipation of CTC. Meanwhile, the PICT to CTC and/or CIP significantly increased up to 15.0-fold after ten successive treatments compared to that in the first treatment. As the treatment frequency increased, significant upward trends in the abundances of tetracycline resistance genes tetA(G), tetX2, tetX, tetG, tetA(33), tetA, tetW, and tetA(P), fluoroquinolone resistance gene qnrA6, and multiple resistance gene mexF were revealed by both metagenomic and qPCR analyses. The findings demonstrated that repeated treatments with CTC and/or CIP can alter the dissipation rate, promote an increase in PICT to CTC and/or CIP, and increase the ARGs abundance in steps.

Environmental Concentrations of Antibiotics May Diminish Wolbachia infections in Aedes aegypti (Diptera: Culicidae).

Wolbachia-infected Aedes aegypti (L.) mosquitoes for control of dengue transmission are being released experimentally in tropical regions of Australia, south-east Asia, and South America. To become established, the Wolbachia Hertig (Rickettsiales: Rickettsiaceae) strains used must induce expression of cytoplasmic incompatibility (CI) in matings between infected males and uninfected females so that infected females have a reproductive advantage, which will drive the infection through field populations. Wolbachia is a Rickettsia-like alphaproteobacterium which can be affected by tetracycline antibiotics. We investigated whether exposure of Wolbachia-infected mosquitoes to chlortetracycline at environmentally relevant levels during their aquatic development resulted in loss or reduction of infection in three strains, wAlbB, wMel, and wMelPop. Wolbachia density was reduced for all three strains at the tested chlortetracycline concentrations of 5 and 50 µg/liter. Two of the strains, wMel and wMelPop, showed a breakdown in CI. The wAlbB strain maintained CI and may be useful at breeding sites where tetracycline contamination has occurred. This may include drier regions where Ae. aegypti can utilize subterranean water sources and septic tanks as breeding sites.

Lipid metabolism disorder induced by up-regulation of miR-125b and miR-144 following ß-diketone antibiotic exposure to F0-zebrafish (Danio rerio).

ß-Diketone antibiotics (DKAs) are widely used in human and veterinary medicine to prevent and treat a large variety of infectious diseases. Long-term DKA exposure to zebrafish can result in lipid metabolism disorders and liver function abnormalities. Based on our previous miRNA-seq analyses, miR-144 and miR-125b were identified as target genes regulating lipid metabolism. DKA-exposure at 12.5 and 25 mg/L significantly increased the expressions of miR-144 and miR-125b. The expression levels for the two miRNAs exhibited an inverse relationship with their lipid-metabolism-related target genes (ppardb, bcl2a, pparaa and pparda). Over-expression and inhibition of miR-144 and miR-125b were observed by micro-injection of agomir-144, agomir-125b, antagomir-144 and antagomir-125b. The over-expression of miR-144 and miR-125b enhanced lipid accumulation and further induced lipid-metabolism-disorder syndrome in F1-zebrafish. The expression of ppardb and bcl2a in whole-mount in situ hybridization was in general agreement with results from qRT-PCR and was concentration-dependent. Oil red O and H&E staining, as well as related physiological and biochemical indexes, showed that chronic DKA exposure resulted in lipid-metabolism-disorder in F0-adults, and in F1-larvae fat accumulation, increased lipid content, abnormal liver function and obesity. The abnormal levels of triglyceride (TG) and total cholesterol (TCH) in DKA-exposed zebrafish increased the risk of hyperlipidemia, atherosclerosis and coronary heart disease. These observations improve our understanding of mechanisms leading to liver disease from exposure to environmental pollution, thereby having relevant practical significance in health prevention, early intervention, and gene therapy for drug-induced diseases.

Inhibitive effects of chlortetracycline on performance of the nitritation-anaerobic ammonium oxidation (anammox) process and strategies for recovery.

The short- and long-term effects of chlortetracycline (CTC) on the nitritation- anaerobic ammonium oxidation (anammox) process were evaluated. The half maximal inhibitory concentration of CTC in the batch tests of the nitritation-anammox process was 278.91mg/L at an exposure time of 12hr. The long-term effects of CTC on the process were examined in a continuous-flow nitritation-anammox reactor. Within 14days, the nitrogen removal rate significantly decreased from 0.61 to 0.25kgN/m3/day with 60 mg/L CTC in the influent. The performance suppressed by CTC barely recovered, even after CTC was removed from the influent. Furthermore, the inhibition of CTC also reduced the relative abundance of ammonium oxidizing bacteria (AOB) and anaerobic ammonium oxidizing bacteria (AnAOB) in the reactor, resulting in both a decreased amount of and an imbalance between AOB and AnAOB. When fresh anammox sludge was reseeded into the nitritation-anammox reactor, the nitrogen removal rate recovered to 0.09 ± 0.03 kg N/m3/day.

Effects of chlortetracycline, Cu and their combination on the performance and microbial community dynamics in swine manure anaerobic digestion.

Swine manure was typical for the combined pollution of heavy metals and antibiotics. The effects of widely used veterinary antibiotic chlortetracycline (CTC), Cu and their combination on swine manure anaerobic digestion performance and microbial community have never been investigated. Thus, four 2L anaerobic digestion reactors were established including reactor A (control), B (CTC spiked by 0.5g/kg dry weight, dw), C (Cu spiked by 5g/kg dw) and D (combination of CTC, 0.5g/kg dw, and Cu, 5g/kg dw), and dynamics of bacterial and archaeal community structure was investigated using high throughput sequencing method. Results showed that addition of CTC and Cu separately could increase the total biogas production by 21.6% and 15.8%, respectively, while combination of CTC and Cu severely inhibited anaerobic digestion (by 30.3%). Furthermore, corresponding to different stages and reactors, four kinds of microbes including bacteria and archaea were described in detail, and the effects of CTC, Cu and their combination mainly occurred at hydrolysis and acidification phases. The addition of Cu alone changed the dynamics of archaeal community significantly. It was genus Methanomassiliicoccus that dominated at the active methane production for A, B and D, while it was genus Methanobrevibacter and Methanoculleus for C.

Magnetic chitosan-based adsorbent prepared via Pickering high internal phase emulsion for high-efficient removal of antibiotics.

A novel magnetic chitosan-g-poly(2-acrylamide-2-methylpropanesulfonic acid) (CTS-g-AMPS) porous adsorbent was prepared by grafting the AMPS onto the CTS in the Fe3O4 stabilized Pickering high internal phase emulsions (Pickering-HIPEs) and used for the adsorptive removal of the antibiotics tetracycline (TC) and chlorotetracycline (CTC). The results of the structure characterization showed that porous structure of the adsorbent can be tuned easily by altering amount of Fe3O4-MNPs-M and the electrostatic attraction of between SO3- and CTC, TC was the main adsorption driving force. The adsorption capacities of the adsorbent for TC and CTC can be reached to 806.60 and 876.60mg/g in a wide pH ranged from 3.0 to 11.0, respectively. And the adsorption equilibrium can be reached within 90min for TC and 50min for CTC. The magnetic porous adsorbent had good reusability, which can still attain a high adsorption capacity of 759.82 and 842.99mg/g for TC and CTC after five consecutive adsorption cycles, respectively. Therefore, the as-prepared CTS-g-AMPS magnetic adsorbent is potential to be used for adsorption removal of antibiotics from water.

Effects of three veterinary antibiotics and their binary mixtures on two green alga species.

The individual and combined toxicities of chlortetracycline (CTC), oxytetracycline (OTC) and enrofloxacin (ENF) have been examined in two green algae representative of the freshwater environment, the international standard strain Pseudokichneriella subcapitata and the native strain Ankistrodesmus fusiformis. The toxicities of the three antibiotics and their mixtures were similar in both strains, although low concentrations of ENF and CTC + ENF were more toxic in A. fusiformis than in the standard strain. The toxicological interactions of binary mixtures were predicted using the two classical models of additivity: Concentration Addition (CA) and Independent Action (IA), and compared to the experimentally determined toxicities over a range of concentrations between 0.1 and 10 mg L-1. The CA model predicted the inhibition of algal growth in the three mixtures in P. subcapitata, and in the CTC + OTC and CTC + ENF mixtures in A. fusiformis. However, this model underestimated the experimental results obtained in the OTC + ENF mixture in A. fusiformis. The IA model did not predict the experimental toxicological effects of the three mixtures in either strain. The sum of the toxic units (TU) for the mixtures was calculated. According to these values, the binary mixtures CTC + ENF and OTC + ENF showed an additive effect, and the CTC + OTC mixture showed antagonism in P. subcapitata, whereas the three mixtures showed synergistic effects in A. fusiformis. Although A. fusiformis was isolated from a polluted river, it showed a similar sensitivity with respect to P. subcapitata when it was exposed to binary mixtures of antibiotics.

Effects of chlortetracycline on the fate of multi-antibiotic resistance genes and the microbial community during swine manure composting.

Excessive use of antibiotics in breeding industry leads to accumulation of antibiotic residuals and antibiotic resistance genes (ARGs) in environment from improperly treated livestock excrements. Four commonly used veterinary antibiotics including chlortetracycline (CTC), sulfamerazine (SMZ), enrofloxacin (ENR) and erythromycin (ERY) were monitored in the swine manure composting. Co-resistance and cross-resistance effects among relative ARGs, correlations between ARGs and bacterial community under multiple antibiotics residual during the composting were investigated in this research. With CTC addition up to 20 mg/kg, more than 99% of CTC removal was achieved after composting, and most of the other antibiotics can be thoroughly removed as well. The variations in ARGs during the composting were strongly correlated to the compositions of the microbial community, Bacteroides and Sporosarcina were main ARGs carriers in the thermophlic phase. Clostridium Ⅺ, Clostridium sensu stricto, and Pseudoxanthomonas, might spread ARGs in cooling and maturing stage. Most of the tested ARGs in swine manure can be effectively reduced through composting, thus makes the compost products safe for soil fertilization.

Toxicity and Genotoxicity of Three Antimicrobials Commonly Used in Veterinary Medicine.

The toxicity of chlortetracycline (CTC), oxytetracycline (OTC) and enrofloxacin (ENF) was tested on two green algal species: the international standard Pseudokirchneriella subcapitata and the native Argentine species Ankistrodesmus fusiformis. All three antibiotics inhibited the algal growth. The most sensitive species was P. subcapitata, for which the EC50 for CTC, OTC and ENF were 1.19 ± 0.53, 0.92 ± 0.30 and 5.18 ± 3.80 mg L-1, respectively. The EC50 for A. fusiformis, were 3.23 ± 0.53, 7.15 ± 2.69 and 10.6 ± 1.28 mg L-1, respectively. The genotoxicity of these veterinary antibiotics was also assessed using chromosome aberration (CA) and micronuclei (MN) induction in Allium cepa roots. Three concentrations were tested (0.1, 1 and 10 mg L-1). Only ENF at 1 and 10 mg L-1 showed any significant MN induction. These data revealed that CTC, OTC and ENF could cause toxicity on green algae, whereas ENF could cause genotoxicity on A. cepa plants.

Methanogenic toxicity evaluation of chlortetracycline hydrochloride

Background Anaerobic digestion is a technology applied successfully to converting organic matter into biogas. However, the presence of inhibitory compounds such as antibiotics can adversely affect methane production. The aim of this study is to evaluate the toxic effect of chlortetracycline hydrochloride (CLOR) on the methanogenic bacteria. In order to study the methanogenic toxicity of CLOR, different concentrations of CLOR (10, 50, 100, 200 mg L- 1) were evaluated by methanogenic toxicity assays using three feedings. Results Maximum methane production was obtained for the assays with 10 mg CLOR L- 1, the values obtained were 277 ± 4.07; 193 ± 11.31 and 166 ± 7.07 mL for the first, second and third feedings, respectively. The average values for acetic, propionic and butyric acid at start of the experiments were 2104 ± 139; 632 ± 7.6; 544 ± 26 mg L- 1, respectively. The VFA values obtained finally of the experiment were dependent on the evaluated antibiotic concentrations, indicating that the efficiency of methanogenesis is directly affected by the CLOR concentration. Conclusions CLOR is an effective methanogenic bacteria inhibitor. Moreover, the results show that CLOR has a bactericidal effect on methanogenic activity given that methane production did not recover during the third feeding. This study shows that the 50% inhibitory concentration (IC50) for methanogenic bacteria in 10 mg L- 1.

Physiological effects of tetracycline antibiotic pollutants on non-target aquatic Microcystis aeruginosa.

This study aimed to evaluate the aquatic toxicity of three typical tetracycline antibiotics, including tetracycline, oxytetracycline, and chlortetracycline, on the cyanobacterium Microcystis aeruginosa. The cell density, chlorophyll a content, protein content, and enzymatic antioxidant activities were determined. The results showed that the cell growth was significantly inhibited by the three compounds at a low concentration. The chlorophyll a and protein content decreased significantly after exposure to 0.05 mg L(-1) of each compound for 9 d. When exposed to 0.2-1 mg L(-1) of tetracycline, the superoxide dismutase (SOD) activity increased, but peroxidase (POD) and catalase (CAT) activities decreased. In contrast, when exposed to oxytetracycline and chlortetracycline at different concentrations ranging from 0.2 to 1 mg L(-1) and from 0.01 to 0.05 mg L(-1), the SOD activity decreased, but the POD and CAT activities increased. These findings indicate that tetracycline antibiotics influence cell growth and protein synthesis, and they also induce oxidative stress in M. aeruginosa at environmentally similar concentrations. Thus, this study may provide further insights into the toxic effects of tetracycline antibiotics and the controlled use of antibiotics.

The combined and second exposure effect of copper (II) and chlortetracycline on fresh water algae, Chlorella pyrenoidosa and Microcystis aeruginosa.

In the experiment, Chlorella pyrenoidosa and Microcystis aeruginosa were chosen to test the individual, combined and second exposure effect of Cu(2+) and chlortetracycline (CTC). The 96 h EC50s of each test were calculated, with the ranges of 0.972-15.6 µmol/L (Cu(2+)), 29.5-102.5 µmol/L (CTC), 14.4-78.9 µmol/L (mixture). The combined toxicities were evaluated with toxicity units (TU) method. The toxicity of complex of Cu(2+) and chlortetracycline was analyzed using concentration addition (CA) model. In the initial test, the combined effect of the two substances was partly additive to C. pyrenoidosa and antagonistic to M. aeruginosa, while in the second exposure test, the combined effect was synergistic to both algae. The biochemical indicators measured in the experiment included chlorophyll fluorescence (Fv/Fm), MDA content, SOD activity and content of soluble proteins. When under combined stress, the biochemical features of both algae were significantly different between the initial test and the second exposure test.