Acute and chronic toxicity of veterinary antibiotics to Daphnia magna

The acute and chronic toxicity of nine antibiotics used both therapeutically and as growth promoters in intensive farming was investigated on the freshwater crustacean Daphnia magna. The effect of the antibiotics metronidazole (M), olaquindox (OL), oxolinic acid (OA), oxytetracycline (OTC), streptomycin (ST), sulfadiazine (SU), tetracycline (TC), tiamulin (TI) and tylosin (TY) was tested in accordance to the ISO (1989) and OECD (1996) standard procedures. The acute toxicities (48-h EC50 value, mg/l) in decreasing order were OA (4.6), TI (40), SU (221), ST (487), TY (680) and OTC (approximately 1000). NOECs were 340 mg/l for TC and 1000 mg/l for M and OL. Toxic effect on reproduction occurred generally at concentrations, which were one order of magnitude below the acute toxic levels. The chronic toxicity (EC50 values, mg/l) in the D. magna reproduction test in decreasing order were TI (5.4), SU (13.7), TC (44.8) and OTC (46.2). The NOECs (mg/l) obtained in the reproduction test with OA, ST, TY and M were 0.38 for OA, 32 for ST, 45 for TY and 250 for M. The observed toxicity of OA to D. magna indicates that this substance, which is a commonly used feed additive in fish farms, has a potential to cause adverse effects on the aquatic environment.     

Biological variation in sensitivity to N-heterocyclic PAHs; effects of acridine on seven species of micro-algae

The toxicity of the nitrogen (N) heterocyclic polyaromatic hydrocarbon (PAH) acridine was tested for seven species of microalgae: Scenedesmus acuminatus, Selenastrum capricornutum, Chlamydomonas eugametos, Staurastrum chaetoceras, Staurastrum manfeldtii, Navicula salinarum and Nitzschia sigma. The effect of acridine on the algae was studied in a 96-h growth test, in which growth rates were determined using cell numbers and biovolume. The obtained EC50 values (for growth rates based on cell numbers) ranged from 0.08 mg litre(-1) for N. sigma to 0.78 mg litre(-1) for C. eugametos and N. salinarum. Effect concentrations based on biovolume were slightly higher for most species. Metabolism of acridine was observed for one species (S. capricornutum), but this capacity did not result in a very different tolerance. Acridine toxicity was neither related to taxonomical background (green algae versus diatoms) nor to original habitat of the species (planktonic or benthic, eutrophic or oligo-mesotrophic). The presence of near-UV radiation during the incubation might explain the higher toxicity of acridine than is expected on basis of QSAR derived narcotic toxicity.     

Anaerobic digestion of livestock and poultry manures spiked with tetracycline antibiotics

Abstract

We investigated the anaerobic degradation of tetracycline antibiotics (tetracycline [TC], oxytetracycline [OTC] and chlortetracycline [CTC]) in swine, cattle, and poultry manures. The manures were anaerobically digested inside polyvinyl chloride batch reactors for 64?days at room temperature. The degradation rate constants and half-lives of the parent tetracyclines were determined following first-order kinetics. For CTC the fastest degradation rate was observed in swine manure (k?=?0.016?±?0.001 d^?1; half-life = 42.8?days), while the slowest degradation rate was observed in poultry litter (k?=?0.0043?±?0.001 d^?1; half-life = 161?days). The half-lives of OTC ranged between 88.9 (cattle manure) and 99.0?days (poultry litter), while TC persisted the longest of the tetracycline antibiotics studied with half-lives ranging from 92.4?days (cattle manure) to 330?days (swine manure). In general, the tetracyclines were found to degrade faster in cattle manure, which had the lowest concentrations of organic matter and metals as compared to swine and poultry manures. Our results demonstrate that tetracycline antibiotics persist in the animal manure after anaerobic digestion, which can potentially lead to emergence and persistence of antibiotic resistant bacteria in the environment when anaerobic digestion byproducts are land applied for crop production.     

不同质量浓度黄莒蒲和狭叶香蒲对铜绿微囊藻的化感作用

分别采用黄菖蒲（IrispseudacorusL．）、狭叶香蒲（TyphaangustifoliaL．）和铜绿微囊藻（Microcystisaeruginosa）共生培养的实验方法研究了不同质量浓度黄菖蒲、狭叶香蒲对铜绿微囊藻的化感作用。结果表明,黄菖蒲在质量浓度大于10g／L时对初始密度为1．0×10^7ind／mL的铜绿微囊藻具有较好的抑制作用,表现为黄菖蒲质量浓度为10、20和40g／L时,第15天对铜绿微囊藻的抑制率分别为30．1％、51．8％和84．0％;狭叶香蒲在质量浓度大于20g／L时对铜绿微囊藻有明显的抑制作用,表现为狭叶香蒲质量浓度为20g／L和40g／L时,第15天对铜绿微囊藻的抑制率分别为34．2％和77．7％,实验过程中,铜绿微囊藻叶绿素a含量逐渐减少,而藻密度、SOD活性及MDA含量先增加后逐渐降低,表明经过一段时间持续地化感胁迫,黄菖蒲和狭叶香蒲可以诱导铜绿微囊藻产生氧化胁迫,导致细胞结构严重损伤和叶绿素大量分解,从而强烈抑制铜绿微囊藻的生长。     

Comparative toxicity of effluents processed by different treatments in V79 fibroblasts and the algae Selenastrum capricornutum

The efficacy of ozonation and of photocatalysis processing in the treatment of pulp mill ECF (elementary chlorine free) bleaching and textile effluents was evaluated by determining total organic carbon reduction (TOC) and the toxicity. The chronic toxicity of the effluents was evaluated by the ability to inhibit the growth of algae Selenastrum capricornutum. Cultured hamster V79 fibroblasts were used to assess the cytotoxicity of effluents submitted to different detoxification processes. Two endpoints were measured in V79 cells: 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide (MTT) reduction and neutral red uptake (NRU). Both treatment processes were able to reduce the TOC, although ozonization was less effective for pulp mill ECF bleaching. The pulp mill ECF bleaching and textile effluents reduced the growth of S. capricornutum by 39% and 27%, respectively. However, at the highest concentration tested, the textile effluents treated by photochemical process for 60 min showed increased cytotoxicity in V79 cells compared to the untreated effluent when assessed by the NRU and MTT reduction assays (increases of 30% and 40%, respectively). Pulp mill ECF bleaching effluent treated by ozonization had a similar cytotoxicity to that of untreated effluent in the NRU assay. In contrast, the MTT reduction assay indicated that effluents treated with ozone were around 20% more cytotoxic than untreated effluents. These results show that cultured fibroblasts may be useful for studying cellular responses to pollutants and may be included in tests to monitor the efficiency of effluent detoxification processes.     

超声波降解四环素类抗生素废水

采用超声波降解模拟废水中四环素类抗生素(TCs):土霉素(OTC)、四环素(TC)和金霉素(CTC),考查废水初始浓度、初始pH、超声波输入功率密度、曝气量和自由基清除剂(正丁醇)对降解效率的影响。结果表明,超声波可以快速高效地降解废水中的TCs,CTC最易被降解,TC和OTC次之;在OTC、TC和CTC初始浓度0.25 mg/L,初始pH=8.2,声功率密度1.2 W/mL,气水体积比30∶1的条件下,超声波辐照20 min后去除率分别达到76.8%、84.0%和94.4%。在实验研究条件下,TCs去除率均随初始浓度(0.25~2.00 mg/L)的增大而降低,但总去除浓度升高;碱性条件有利于TCs降解,去除率随初始pH(6~11)升高而升高;功率密度(0.24~1.96 W/mL)越高,TCs去除率也越高;曝气后TCs的去除率明显升高,随着气水比(10∶1~60∶1)的增大,去除率逐渐上升;正丁醇有效抑制TCs超声波降解,可推断TCs主要降解途径为自由基氧化。TCs超声波降解过程符合伪一级动力学反应特征。     

Evaluation of different algal species sensitivity to mercury and metolachlor by PAM-fluorometry.

In this study, the pulse-amplitude-modulation (PAM)-fluorometric method was used to evaluate the difference in the sensitivity to mercury (Hg) and metolachlor of six algal species: Ankistrodesmus falcatus, Selenastrum capricornutum, Chlorella vulgaris, Nannoplankton (PLS), Microcystis aeruginosa and Pediastrum biwae. We found that the fluorescence parameters (phiM, the maximal photosystem II (PSII) quantum yield, phi'M, the operational PSII quantum yield at steady state of electron transport, Q(P), the photochemical quenching value, and Q(N), the non-photochemical quenching value) were appropriate indicators for inhibitory effects of mercury but only phi'M and Q(N) were useful for metolachlor. The examined algal species showed very different levels of sensitivity to the effect of Hg and of metolachlor. The most sensitive species to Hg and metolachlor were respectively M. aeruginosa and A. falcatus, while the least sensitive were C. vulgaris and P. biwae. We interpreted these differences by the action mode of pollutants and by the different metabolism properties and morphological characteristics between algal species. These results related to fluorescence parameters may offer useful tool to be used in bioassay for different pollutants. Heterogeneous algal sensitivity to the same pollutant suggests the need to use a battery of species to evaluate the effects of mixtures of pollutants in aquatic systems.     

Preliminary ecotoxicological characterization of a new energetic substance, CL-20

A new energetic substance hexanitrohexaazaisowurtzitane (or CL-20) was tested for its toxicities to various ecological receptors. CL-20 (epsilon-polymorph) was amended to soil or deionized water to construct concentration gradients. Results of Microtox (15-min contact) and 96-h algae growth inhibition tests indicate that CL-20 showed no adverse effects on the bioluminescence of marine bacteria Vibrio fischeri and the cell density of freshwater green algae Selenastrum capricornutum respectively, up to its water solubility (ca. 3.6 mg l(-1)). CL-20 and its possible biotransformation products did not inhibit seed germination and early seedling (16-19 d) growth of alfalfa (Medicago sativa) and perennial ryegrass (Lolium perenne) up to 10,000 mg kg(-1) in a Sassafras sandy loam soil (SSL). Indigenous soil microorganisms in SSL and a garden soil were exposed to CL-20 for one or two weeks before dehydrogenase activity (DHA) or potential nitrification activity (PNA) were assayed. Results indicate that up to 10,000 mg kg(-1) soil of CL-20 had no statistically significant effects on microbial communities measured as DHA or on the ammonium oxidizing bacteria determined as PNA in both soils. Data indicates that CL-20 was not acutely toxic to the species or microbial communities tested and that further studies are required to address the potential long-term environmental impact of CL-20 and its possible degradation products.     

Interactions between algae (Selenastrum capricornutum) and pesticides: implications for managing constructed wetlands for pesticide removal

This laboratory study examined the interactions between an algal species found in wetlands (Selenastrum capricornutum) and two agricultural pesticides (atrazine and lindane). Pesticide additions had a positive effect on the chlorophyll a concentrations of the treatments. The presence of algae decreased the aqueous persistence of both pesticides. It is speculated that the algae either provided sites for pesticide sorption or facilitated pesticide degradation.     

Surfactant (Genapol OX-80) toxicity to Selenastrum capricornutum

A non-ionic surfactant (Genapol OX-80) was proposed as a means of reducing destructive crayfish activity in rice. In order to study the toxicity of this product on algae, a test in accordance to the ISO 8692 protocol (1989) was performed. The growth inhibiting effect of the pure formulation of Genapol OX-80 was studied on Selenastrum capricornutum in the concentration range of 0.01-1.0 mg/l. The result indicated a 72 h EC50-value of 0.5 mg/l. The suggested field concentration of approximately 50 mg/l is therefore two orders of magnitude above the algal EC50-value. From our findings it is expected that the impact on the rice field algae will not be negligible.     

Comparative use of bacterial, algal and protozoan tests to study toxicity of azo- and anthraquinone dyes

Toxicity of two azo dyes (Reactive Orange 16 (RO16); Congo Red (CR)) and two anthraquinone dyes (Remazol Brilliant Blue R (RBBR); Disperse Blue 3 (DB3)) were compared using bacterium Vibrio fischeri, microalga Selenastrum capricornutum and ciliate Tetrahymena pyriformis. The following respective endpoints were involved: acute toxicity measured as bacterial luminescence inhibition, algal growth inhibition, and the effects on the protozoa including viability, growth inhibition, grazing effect and morphometric effects. In addition, mutagenicity of the dyes was determined using Ames test with bacterium Salmonella typhimurium His(-). DB3 dye was the most toxic of all dyes in the bacterial, algal and protozoan tests. In contrast to other dyes, DB3 exhibited mutagenic effects after metabolic activation in vitro in all S. typhimurium strains used. Of the methods applied, the algal test was the most sensitive to evaluate toxicity of the dyes tested.     

Zinc acclimation and its effect on the zinc tolerance of Raphidocelis subcapitata and Chlorella vulgaris in laboratory experiments.

The effect of zinc acclimation of Raphidocelis subcapitata (syn. Selenastrum capricornutum) and Chlorella vulgaris on their sensitivity towards this metal was examined in a series of laboratory experiments. These two commonly used algal species were acclimated to 65 microg Zn/l and changes in zinc tolerance were monitored using standard growth inhibition tests. The chemically defined ISO medium was used as a control culture medium. Both species demonstrated a maximum increase in zinc tolerance of a factor of 3 after 100 days of acclimation. Shifts in the shape of the concentration-response curve due to acclimation were observed for R. subcapitata. Compared to non-acclimated algae, acclimated R. subcapitata exhibited higher growth rates in all zinc treatments as well as in the controls. This suggests that the use of ISO-medium results in sub-optimal growth due to zinc deficiency. These effects could not be demonstrated for C. vulgaris. The zinc tolerance of both species decreased significantly one week after returning the acclimated algae to control (ISO) medium. 72hEC50 values based on growth rate were two to four times higher than those calculated using biomass measurements. Algal toxicity test results, particularly if used for metal risk assessments, must not be conducted using nutrient deficient media.     

Enhanced removal of tetracycline via advanced oxidation of sodium persulfate and biochar adsorption

Advanced oxidation of antibiotic tetracycline (TC) is becoming an accessible and efficient technology. The removal of TC from the complex wastewater needs to be lucubrated. In this study, a TC removal system involving degradation and adsorption was established. TC degradation was accomplished by enhanced advanced oxidation via the addition of sodium persulfate (SP) and biochar into simulated wastewater containing Mn²⁺ and TC wastewater. The adsorption of TC and its derivatives was removed by biochar. The results indicate that the optimized reaction parameters were 3.0 g/L of biochar prepared at 600 °C (B600) and 400 mg/L of SP under acidic condition, and the removal percentage of TC was 87.48%, including 74.23% of degradation and 13.28% of adsorption; the anions Cl^?, NO₃^?, and H₂PO₄^? had negligible effects on the removal of TC in this Mn²⁺/B600/SP system. The system also functioned well with an aqueous solution with a high chemical oxygen demand (COD) concentration. Electron paramagnetic resonance (EPR) analysis indicated that ·OH and SO₄^? free radicals were present in the Mn²⁺/B600/SP system. Based on the testing and analysis results, a removal mechanism and potential TC degradation pathway for this system were proposed. TC can be degraded by ·OH and SO₄^? via three degradation pathways. Mn²⁺ can be precipitated as MnO₂, and a part of the TC and its derivatives can be adsorbed on the biochar surface. The Mn²⁺/B600/SP system also performed satisfactorily for a complex aqueous solution with various cations and antibiotics.

     

On the uptake and release of zinc (65Zn) in the growing alga Selenastrum capricornutum Printz

A Zn- and pH-buffered medium was used to study the accumulation of Zn in batch-cultured algae (Selenastrum capricornutum Printz.). All experiments were carried out using (65)Zn radiotracers, which were measured by gamma-ray spectrometry. Zinc was applied in the presence of nitrilotriacetic acid (NTA), leading to free Zn(2+) concentrations ranging from 4.2 x 10(-11) to 1.6 x 10(-7) mol litre(-1). During the 75-h experiments, the rates of Zn influx and efflux were determined, the latter with full consideration for algal growth rates. Algal growth stopped completely at a free Zn(2+) concentration of 1.6 x 10(-8) mol litre(-1). Initial Zn binding was determined and, based on free Zn(2+) concentrations, presented by the apparent dissociation constant K(diss) and capacity C as 4.6 x 10(-9) mol litre(-1) and 19.8 x 10(6) mol gDW(-1), respectively. The rate constant of Zn efflux could be calculated as a constant 0.021 +/- 0.003 h(-1), irrespective of the Zn concentrations applied. The rates of Zn influx were expressed by the V(max) and K(M) values as 3.3 x 10(-10) mol gDW(-1) s(-1) and 6.3 x 10(-9) mol litre(-1), respectively. These values, which are based on applied free Zn(2+) concentrations, are compatible with the operation of a high-affinity low-concentration carrier mechanism. The results suggest that, under the conditions applied, free Zn(2+) may be the most relevant Zn species for Zn uptake; furthermore, relative growth rate may be regarded as a relatively sensitive signal for Zn stress circumstances.     

Polystyrene nanoplastics affect growth and microcystin production of Microcystis aeruginosa

Nanoplastics are widely distributed in freshwater environments, but few studies have addressed their effects on freshwater algae, especially on harmful algae. In this study, the effects of polystyrene (PS) nanoplastics on Microcystis aeruginosa (M. aeruginosa) growth, as well as microcystin (MC) production and release, were investigated over the whole growth period. The results show that PS nanoplastics caused a dose-dependent inhibitory effect on M. aeruginosa growth and a dose-dependent increase in the aggregation rate peaking at 60.16% and 46.34%, respectively, when the PS nanoplastic concentration was 100 mg/L. This caused significant growth of M. aeruginosa with a specific growth rate up to 0.41 d^?1 (50 mg/L PS nanoplastics). After a brief period of rapid growth, the tested algal cells steadily grew. In addition, the increase in PS nanoplastics concentration promoted the production and release of MC. When the PS nanoplastic concentration was 100 mg/L, the content of the intracellular (intra-) and extracellular (extra-) MC increased to 199.1 and 166.5 μg/L, respectively, on day 26, which was 31.4% and 31.1% higher, respectively, than the control. Our results provide insights into the action mechanism of nanoplastics on harmful algae and the potential risks to freshwater environments.

     

溶藻放线菌改性制剂对铜绿微囊藻的控藻能力研究

采用喷雾干燥法制备了一种溶藻放线菌粉剂,此粉剂按1∶10 000的质量比投加4 d后对铜绿微囊藻的溶藻效率可达90%,该粉剂经过壳聚糖改性处理后,在1∶20 000的质量投加比下,0.5 h即可絮凝去除90%的藻细胞,4 d后的溶藻效率仍可达80%。由于既能快速絮凝除藻,又能长效溶藻,使得改性溶藻放线菌粉剂具有更为广阔的应用前景。     

The effects of residual tetracycline on soil enzymatic activities and plant growth

A pot trial was carried out to investigate the adverse effects of tetracycline (TC) on soil microbial communities, microbial activities, and the growth of ryegrass (Lolium perenne L). The results showed that the presence of TC significantly disturbed the structure of microbial communities and inhibited soil microbial activities in terms of urease, acid phosphatase and dehydrogenase (p < 0.05). Plant biomass was adversely influenced by TC, especially the roots with a reduction of 40% when compared with the control. Furthermore, TC decreased the assimilation of phosphorus by the plant although the concentration of phosphorus was increased by 20% due to decreased plant biomass. TC seemed to increase the concentration of dissolved organic carbon (by 20%) in soil. The findings implied that the agricultural use of animal manure or fishpond sediment containing considerable amounts of antibiotics may give rise to ecological risks.     

Comparison of spontaneous antibiotic resistance frequency of Salmonella Typhimurium growth in glucose amended continuous culture at slow and fast dilution rates

The objective of the study was to determine the frequency of spontaneous acquisition of resistance to select antibiotics by Salmonella Typhimurium (ST) when grown in glucose amended continuous flow culture at slow (D = 0.025 h(-1)) or fast (D = 0.27 h(-1)) dilution rates. The bacterium was grown in LB minimal medium (pH 6.25) containing no antibiotics. Upon achieving steady state, samples were plated to tryptic soy agar (TSA) alone or supplemented (per ml) with 2 and 16 microg oxytetracycline, 4 and 16 microg tetracycline, 2 and 64 microg kanamycin, and 0.25 and 2 microg enrofloxacin. Regardless of growth rate, CFU of resistant ST from the TSA containing antibiotics was less than 2 x 10(1) except for 2 microg kanamycin and 0.25 microg enrofloxacin treatments (higher than 1 x 10(9) and 4 x 10(7) CFU of resistant ST for trials 1 and 2, respectively). Frequency of recovering resistant ST from the TSA containing the higher antibiotic concentrations was less than 1 in 10(9) for all antibiotics, but was higher on the media containing 2 microg kanamycin and 0.25 microg enrofloxacin at both slow and fast growth rates. In general, minimal susceptibility differences were detected for isolates from slow and fast dilution rates.     

Oxidative transformations of environmental pharmaceuticals by Cl₂, ClO₂, O₃, and Fe(VI): kinetics assessment

Several pharmaceuticals have been detected globally in surface water and drinking water, which indicate their insufficient removal from water and wastewater using conventional treatment methods. This paper reviews the kinetics of oxidative transformations of pharmaceuticals (antibiotics, lipid regulators, antipyretics, anticonvulsants, and beta-blockers) by Cl(2), ClO(2), O(3), and ferrate(VI) (Fe(VI)O(4)(2-),Fe(VI)) under treatment conditions. In the chlorination of sulfonamide antibiotics, HOCl is the major reactive Cl(2) species whereas in the oxidation by Fe(VI), HFeO(4)(-) is the dominant reactive species. Both oxidation processes can oxidize sulfonamides in seconds at a neutral pH (t(1/2)≤ 220 s; 1 mg L(-1) HOCl or K(2)FeO(4)). The reactivity of O(3) with pharmaceuticals is generally higher than that of HOCl (k(app,pH 7) (O(3))=1-10(7)M(-1)s(-1); k(app,pH 7) (HOCl)=10(-2)-10(5)M(-1)s(-1)). Ozone selectively oxidizes pharmaceuticals and reacts mainly with activated aromatic systems and non-protonated amines. Oxidative transformation of most pharmaceuticals by O(3) occurs in seconds (t(1/2)≤ 100 s; 1 mg L(-1) O(3)) while half-lives for oxidations by HOCl differ by at least two orders of magnitude. Ozone appears to be efficient in oxidizing pharmaceuticals in aquatic environments. The limited work on Fe(VI) shows that it can also potentially transform pharmaceuticals in treatment processes.     

Comparative study of the effects of MCPA, butylate, atrazine, and cyanazine on Selenastrum capricornutum

The herbicides MCPA, butylate, atrazine and cyanazine are extensively used in Canadian agriculture and information regarding their effects on indigenous biota is scarce. Phytotoxicity assessments were conducted in the laboratory on the common green alga Selenastrum capricornutum using both the active ingredient of the herbicides and their formulated products (for MCPA and butylate). Endpoints determined after the 96 h exposure included algal population growth inhibition (IC50-cell counts), percent lethality (LC50-flow cytometry derived) and photosynthetic electron transport inhibition (EC50-fluorescence induction). Pesticide formulations had greater toxic effects than the active ingredient alone. The 96 h IC50 (50% Inhibition Concentration) and LOEC (Lowest Observable Effects Concentration) using cell counts of S. capricornutum were 18.4 and 8.9 mg l(-1) respectively for MCPA-active ingredient and for MCPA-formulated, these were 0.62 and 0.0062 mg l(-1) respectively. Those for butylateactive ingredient were 61.0 and 8.3 mg l(-1) and for butylate-formulated 1.46 and 0.17 mg l(-1), respectively. The triazines active ingredient, which are photosynthetic inhibitors, had greater effects than either the MCPA or butylate. The IC50 for cyanazine and atrazine were 0.059 and 0.026 mg l(-1), respectively. By comparing the IC50 and LC50 values for the tested active ingredients, it was found that the effects of atrazine were algicidal, whereas those of cyanazine, butylate and MCPA were algistatic.