Zhenyang Yu

Occurrence, distribution and seasonal variation of antibiotics in the Huangpu River, Shanghai, China

Water samples were collected from 19 sampling sites along the Huangpu River in June and December 2009. The occurrence, distribution and seasonal variation of 22 antibiotics, including four tetracyclines, three chloramphenicols, two macrolides, six fluoroquinolones, six sulfonamides and trimethoprim were investigated. It was found that all 19 sampling sites were contaminated by antibiotics. Four antibiotics (sulfamerazine, norfloxacin, fleroxacin and sarafloxacin) were not detected. The detection frequencies of the other 18 antibiotics were in the range of 5.3-100%. The median concentrations of the detected antibiotics ranged from quantification limits to 36.71 ng L(-1) (tetracycline) in June and to 313.44 ng L(-1) (sulfamethazine) in December. The number of detected antibiotics and the overall antibiotic concentrations were higher in December than in June due to the different river flow conditions. Different dominant antibiotics were observed for each group of antibiotics between June and December. Higher total concentrations of veterinary antibiotics such as tetracyclines were observed in suburban sampling sites than in urban sites, indicating the role of livestock and agricultural activities as an important source of antibiotic contamination.

Combined effects of titanium dioxide and humic acid on the bioaccumulation of cadmium in Zebrafish.

The combined effects of titanium dioxide (TiO2) nanoparticles and humic acid (HA) on the bioaccumulation of cadmium (Cd) in Zebrafish were investigated. Experimental data on the equilibrium Cd bioaccumulation suggest that only the dissolved Cd effectively contributed to Cd bioaccumulation in HA solutions whereas both the dissolved and TiO2 associated Cd were accumulated in TiO2 or the mixture of HA and TiO2 solutions, due likely to the additional intestine uptake of the TiO2-bound Cd. The equilibrium Cd bioaccumulation in the mixed system was comparable to that in the corresponding HA solutions, and significantly lower than that in the corresponding TiO2 solutions (n=3, p<0.05). The presence of either HA or TiO2 (5-20 mg L(-1)) in water slightly increased the uptake rate constants of Cd bioaccumulation whereas combining HA and TiO2 reduced the uptake rate constants.

Time-dependent hormetic effects of 1-alkyl-3-methylimidazolium bromide on Vibrio qinghaiensis sp.-Q67: luminescence, redox reactants and antioxidases.

The green credentials of ionic liquids (ILs) are being challenged due to the increasing evidence of their toxicity. The hormetic effects further raised their ecological concern. However, it remained poorly studied on the time-dependent changes of the hormetic effects and the mechanisms. In this study, we investigated the time-dependent hormetic effects of four 1-alkyl-3-methylimidazolium bromide ([amim]Br), including 1-ethyl ([emim]Br), -butyl ([bmim]Br), -hexyl ([hmim]Br) and -octyl ([omim]Br), on the luminescence of Vibrio qinghaiensis sp.-Q67. The results showed that [amim]Br with shorter side chains, [emim]Br and [bmim]Br, caused obvious hormetic time-dependent toxicities. The effective concentration (EC) values for the hormetic effects of [emim]Br and [bmim]Br increased with time. [amim]Br with longer side chains, [hmim]Br and [omim]Br, produced sigmoid concentration-dependent inhibitions on the luminescence, and the EC50 values almost unchanged. To illustrate the mechanism, we subsequently examined the responses of redox reactants and antioxidases. [emim]Br and [bmim]Br significantly induced FMN (flavin mononucleotide), NADH (reduced nicotinamide adenine dinucleotide), SOD (superoxide dismutase) and CAT (catalase), and the inductions increased with time, which is similar to the time-dependent changes of their hormetic effects on Q67. Meanwhile, [hmim]Br and [omim]Br did not cause significant effects on the redox reactants and antioxidases. In conclusion, the hormetic effects of [amim]Br on the luminescence, redox reactants and antioxidases showed the dependence on both exposure time and side chains. Our findings provided insights into the time-dependent biological process of the hormetic effects of [emim]Br and [bmim]Br on the photobacterium and its biochemical indicators.

Transgenerational effects of heavy metals on L3 larva of Caenorhabditis elegans with greater behavior and growth inhibitions in the progeny.

Heavy metals are ubiquitous environmental pollutants, and their toxic effects have been widely studied. However, their transgenerational effects between parent and progeny at environmental relevant concentrations need further investigations. Currently, L3 stage of Caenorhabditis elegans was exposed to aqueous metals (Cd, Cu, Pb and Zn) at environmentally realistic concentrations for 96 h. The whole exposure time covered the formation of sperm, ovum and eggs. Subsequently the behavior and growth indicators were measured. The parent nematodes were then bleached to gain synchronized eggs, which were cultured under non-toxic conditions to L3 stage when the same indicators were measured in the progeny. The parent suffered concentration-dependent inhibitions on behavior and growth. Based on the median effective concentration (EC(50)) values, body bending frequency showed relatively higher sensitivity than other behavior indicators. The inhibitions on growth and behavior of progeny were more severe than those of the parent, based on their respective EC(50) values. Interestingly, Cd was not the most toxic metal in either parent or progeny according to EC(50) values, but its EC(50) ratios between parent and progeny (EC(50, parent)/EC(50, progeny)) were the most significant, indicating its greatest transgenerational effects. The results demonstrated the higher sensitivity of L3 larva stage of C. elegans in the transgenerational effect studies than other life stages used before. Our findings suggested that parental exposure to heavy metals can multiply their harmful effects in following generations.

Behavior toxicity to Caenorhabditis elegans transferred to the progeny after exposure to sulfamethoxazole at environmentally relevant concentrations

Sulfamethoxazole (SMX) is one of the most common detected antibiotics in the environment. In order to study whether SMX can affect behavior and growth and whether these effects could be transferred to the progeny, Caenorhabditis elegans was exposed at environmentally relevant concentrations for 24, 48, 72 and 96 hr, respectively. After exposure, the exposed parent generation (P0) was measured for behavior and growth indicators, which were presented as percentage of controls (POC). Then their corresponding unexposed progeny (F1) was separated and measured for the same indicators. The lowest POC for P0 after 96 hr-exposure at 100 mg/L were 37.8%, 12.7%, 45.8% and 70.1% for body bending frequency (BBF), reversal movement (RM), Omega turns (OT) and body length (BL), respectively. And F1 suffered defects with the lowest POC as 55.8%, 24.1%, 48.5% and 60.7% for BBF, RM, OT and BL, respectively. Defects in both P0 and F1 showed a time- and concentration-dependent fashion and behavior indicators showed better sensitivity than growth indicator. The observed effects on F1 demonstrated the transferable properties of SMX. Defects of SMX at environmental concentrations suggested that it is necessary to perform further systematical studies on its ecological risk in actual conditions.

Toxic and recovery effects of copper on Caenorhabditis elegans by various food-borne and water-borne pathways

Copper pollutions are typical heavy metal contaminations, and their ability to move up food chains urges comprehensive studies on their effects through various pathways. Currently, four exposure pathways were prescribed as food-borne (FB), water-borne plus clean food (WCB), water-food-borne (WFB) and water-borne (WB). Caenorhabditiselegans was chosen as the model organism, and growth statuses, feeding abilities, the amounts of four antioxidant enzymes, and corresponding recovery effects under non-toxic conditions with food and without food were investigated. Based on analysis results, copper concentrations in exposure were significantly influenced by the presence of food and its uptake by C.elegans. Both exposure and recovery effects depended on exposure concentrations and food conditions. For exposure pathways with food, feeding abilities and growth statuses were generally WFB<WCB≤FB (p<0.05). The antioxidant activities were up-regulated in the same order. Meanwhile, the exposure pathway without food (WB) caused non-up-regulated antioxidant activities, and had the best growth statuses. For recoveries with food, growth statuses, feeding abilities and the inductions of the antioxidant enzymes were all WB≈WFB<WCB<FB (p<0.05). For recoveries without food, the order of growth statuses remained WB>FB>WCB>WFB (p<0.05), while the antioxidant activities were all inhibited in a concentration-dependent fashion. In conclusion, contaminated food was the primary exposure pathway, and various pathways caused different responses of C.elegans.

Inhibitions on the behavior and growth of the nematode progeny after prenatal exposure to sulfonamides at micromolar concentrations.

Sulfonamides are one typical antibiotic which is an emerging hazardous material to the ecological stability due to their continuously application and biological effects to non-target organisms. The parent-progeny transgenerational effects need investigations to indicate their long-term consequences. Currently, we tested the transgenerational effects of sulfadiazine (SD), sulfapyridine (SP) and sulfamethazine (SMZ) on L3 larva of Caenorhabditis elegans. The nematodes were exposed to aqueous sulfonamides at micromolar concentrations for 96 h, and then the effects on the behavior and growth in the exposed parent and unexposed progeny were measured. Results showed that SD, SP and SMZ inhibited three behavior indicators including body bending frequency (BBF), reversal movement (RM) and Omega turn (OT), and the growth indicator (body length, BL). Behavior indicators showed higher sensitivities than the growth indicator, and BBF had the highest sensitivity among the behavior indicators. Moreover, the effects of sulfonamides were also observed in the unexposed progeny with partially rescued or more severe inhibitions on the indicators. The behavior also showed higher sensitivity than the growth in the progeny. The transgenerational effects of sulfonamides indicated that parental exposure can multiply the harmful effects of antibiotic pollution in following generations and their potential ecological risks at environmental concentrations were further raised.

The combinational effects between sulfonamides and metals on nematode Caenorhabditis elegans.

As emerging pollutants, antibiotic sulfonamides are continuously emitted into the environment and encounter those already-existing contaminants, e.g., heavy metals, which may cause toxicity interactions in polluted habitats. So far, the sulfonamide mixture effects and the combinational effects between sulfonamides and metals have been seldom studied. In this study, lifespan, lethality (24 and 120 h), locomotion behavior and growth (96 h) of Caenorhabditis elegans were measured after exposure to mixtures containing sulfonamides (sulfadiazine, sulfapyridine, sulfamethoxazole and sulfamethazine as representatives) and/or metals (cadmium, copper, lead and zinc as representatives) at environmental concentrations. Results showed that sulfonamides did not cause acute (24 h) lethality at chosen concentrations, but they decreased the lifespan in a concentration dependent fashion. Moreover, sulfonamide mixtures caused synergisms at higher concentrations but antagonisms at lower concentrations on the subacute (120 h) lethal effects. The toxicity interactions of sulfonamide mixtures were addition action on body bending frequency, and antagonism on reversal movement and body length. In sulfonamide and metal mixtures, the toxicity interactions were different in acute and subacute lethal results, indicating the influence of the exposure time. According to the comparison among effects of mixtures containing sulfonamides and/or metals, subacute lethality of sulfonamides was enhanced by metals based on the synergistic mixture effects, while their inhibitions on the growth and behavior were weakened by metals based on the antagonistic mixture effects. Our findings highlighted studies on combinational effects between emerging and common contaminants for more accurate environmental risk evaluation, and also urged further mechanism studies.

Multigenerational Effects of Heavy Metals on Feeding, Growth, Initial Reproduction and Antioxidants in Caenorhabditis elegans

Earlier studies showed that toxicities of excessive metals lasted over generations. Yet, these studies mainly employed one-generation exposure, and the effects of multigenerational challenges need further studies. Presently, Caenorhabditis elegans were exposed to cadmium, copper, lead and zinc for four consecutive generations (G1 to G4) at environmental concentrations. The feeding, growth, initial reproduction, superoxide dismutase (SOD) and catalase (CAT) were determined. All data were represented in the percentage of that in control (POC), and POC in the control was normalized to 100%. In G1 and G2, the POC values in feeding, growth and initial reproduction were generally within 10% of the control (100%), indicating non-significant effects. The POC values in SOD and CAT were significantly higher than 100%, showing stimulatory effects. In G3 and G4, the POC values in feeding, growth and initial reproduction were significantly lower than 100%, showing inhibitory effects which were more severe in G4 than in G3. Meanwhile, SOD and CAT continuously showed stimulatory effects, and the stimulatory effects on SOD increased from G1 to G4. The effects with multigenerational challenges were different from those in one-generation exposure. The effects in later generations demonstrated the importance of multigenerational challenges in judging long-term influences of metals.

Biochemical and gene expression effects of 1-alkyl-3-methylimidazolium tetrafluoroborate on Vibrio qinghaiensis sp.-Q67

Applications of 1-alkyl-3-methylimidazolium salts ([amim]X) in pharmaceuticals call for studies on the biological effects. Previously, [amim]Cl and [amim]Br caused time-dependent stimulatory effects on Vibrio qinghaiensis sp.-Q67 (Q67). Further studies are still needed to answer the question whether [amim]X with other anions have similar effects, and to explain such stimulatory effects. Here, effects of four [amim]BF4 with different alkyl side chains on Q67 were investigated. At the apical level, they inhibited luminescence at high concentrations and the toxicities decreased with time increasing from 0.25 to 12h. Meanwhile, they stimulated luminescence at low concentrations, and the stimulations increased over time. The maximum stimulations followed an order of [emim]BF4>[bmim]BF4>[hmim]BF4>[omim]BF4. Compared with [emim]BF4, imidazole caused lower stimulations, while NaBF4 caused significantly higher ones. Overview on effects of [amim]BF4, imidazole and NaBF4 indicated significant anionic contributions. At biochemical and molecular levels, favin mono-nucleotide (FMN) and nicotinamide adenine dinucleotide (NADH) increased over time, and expressions of luxA and luxB (encoding luciferase that catalyzes FMN:NADH reaction) in Q67 showed up-regulations, suggesting an overall enhancement of the bioactivities. Moreover, superoxide dismutase and catalase showed inductions with up-regulations on their encoding genes, indicating improved antioxidant responses.