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 278.91 mg/L at an exposure time of 12 hr. The long-term effects of CTC on the process were examined in a continuous-flow nitritation-anammox reactor. Within 14 days, the nitrogen removal rate significantly decreased from 0.61 to 0.25 kg N/m^3/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（An AOB）in the reactor, resulting in both a decreased amount of and an imbalance between AOB and An AOB. 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.

Photolysis of chlortetracycline in aqueous solution:Kinetics,toxicity and products

The aqueous photodegradation of the widely used antibiotic chlortetracycline（CTC） was investigated under simulated sunlight.The quantum yield of photodegradation increased from 3.3 × 10-4 to 8.5 × 10-3 within the pH range of 6.0 to 9.0.The presence of Ca2＋,Fe3＋,and NO3-enhanced the photodegradation of CTC,whereas Mg2＋,Mn2＋,and Zn2＋ inhibited the degradation with the order Mn2＋ Zn2＋ Mg2＋ at pH 7.3.The monovalent cations（Na＋ and K＋） had negligible effect on the photolysis of CTC.Fulvic acid（FA） decreased the photodegradation of CTC due to light screening effect.Hydrogen peroxide（H2O2） was formed concurrently with direct photodegradation of CTC.The generation rate of H2O2 increased from 0.027 to 0.086 μmol/（L.min） when the pH ranged from 6.0 to 9.0.The CTC solution was about three-fold more toxic to the Photobacterium phosphoreum bacteria after irradiation,suggesting that the photoproducts and H2O2 formed in the CTC solution exhibited high risk on the bacteria.By LC-ESI（＋）-MS,the photoproducts of CTC were identified.The direct photodegradation of CTC was involved in hydroxylation and N-demethyl/dedismethyl processes.The main photoproducts included the iso-CTC analog containing hydroxyl groups（m/z 511.4 and 495.4）,and the N-demethyl/dedismethyl products of the photoproduct m/z 495.4（m/z 481.3 and 467.4）.In addition,the photochemical dechlorination of CTC led to tetracycline（m/z 445.5）.

Natural capsicum extract replacing chlortetracycline enhances performance via improving digestive enzyme activities,antioxidant capacity,anti-inflammatory function,and gut health in weaned pigs

The objective of this study was to investigate the effects of natural capsicum extract(NCE,containing 2%natural capsaicin,the rest is carrier)replacing chlortetracycline(CTC)on performance,digestive enzyme activities,antioxidant capacity,inflammatory cytokines,and gut health in weaned pigs.A total of 108 weaned pigs(Duroc×[Landrace×Yorkshire],initial body weight-8.68±1.34 kg;weaned on d 28)were randomly allotted into 3 treatments with 6 replicate pens per treatment(3 barrows and 3 gilts per pen).The treatments include a corn-soybean meal basal diet as a control group(CON),a CTC group(basal diet+CTC at 75 mg/kg),and a NCE group(basal diet--NEC at 80 mg/kg).Compared with CON and CTC,NCE had increased(P<0.05)average daily gain in phase 2(d 15 to 28)and overall(d 1 to 28),and higher(P<0.05)apparent total tract digestibility of gross energy,dry matter,crude protein,and organic matter in phase 1(d 1 to 14).These pigs also had increased(P<0.05)pancrelipase activity in pancreas,α-amylase,lipase and protease activities in the jejunal mucosa,and lipase activity in the ileal mucosa on d 28.Moreover,NCE had increased(P<0.05)the contents of growth hormone,β-endorphin,5-hydroxytryptamine,total antioxidant capacity,total superoxide dismutase,catalase,and IL-10,as well as decreased(P<0.05)contents of malondialdehyde,tumor nuclear factor-a,interferon-y,and interleukin-6 in serum on d 28 compared with CON and CTC.NCE showed higher(P<0.05)propionic acid,butyric acid and total volatile fatty acids(VFA)contents,and increased(P<0.05)relative abundance of Faecalibacterium in colon,as well as higher(P<0.05)propionic acid and total volatile fatty acids in cecum on d 28 compared with CON.In conclusion,NCE replacing CTC could enhance performance via improving digestive enzyme activities,antioxidant capacity,anti-inflammatory function,gut VFA composition and microbiota community in weaned pigs,and it could be used as a potential target for the development of feed additives.

Toxic effects of chlortetracycline on maize growth,reactive oxygen species generation and the antioxidant response

The toxicity of chlortetracycline （CTC） on maize （Zea mays L.） growth and reactive oxygen species （ROS） generation was studied.The root and shoot lengths and fresh weights of maize seedlings were inhibited by CTC treatment （p 〈 0.05）.Root length was more sensitive than other parameters with the EC 10 value of 0.064 mg/L.The spin trapping technique followed by electron paramagnetic resonance （EPR） analysis was used to quantify the ROS production.The ROS generated in maize roots after exposure to CTC was identified as hydroxyl radical （·OH）.The EPR signal intensity correlated positively with the logarithm of CTC concentrations exposed （p 〈 0.05）.The dynamic changes of malondialdehyde （MDA） contents and the antioxidative enzyme activities in maize roots were also determined.As compared to the control group,CTC was found to significantly increase MDA content.Treatment of maize roots with the ·OH scavenger sodium benzoate （SB） reduced the MDA content and enhanced the antioxidative enzyme activities.The results demonstrated the harmfulness of CTC at high dose to maize in the early developmental stage,and clarified that the inducement of ·OH is one of the mechanisms of CTC toxicity.

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 2 L anaerobic digestion reactors were established including reactor A（control）, B（CTC spiked by 0.5 g/kg dry weight, dw）, C（Cu spiked by 5 g/kg dw） and D（combination of CTC, 0.5 g/kg dw, and Cu, 5 g/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 by21.6% 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.

Effect of the chlortetracycline addition method on methane production from the anaerobic digestion of swine wastewater

Effects of antibiotic residues on methane production in anaerobic digestion are commonly studied using the following two antibiotic addition methods：（1） adding manure from animals that consume a diet containing antibiotics, and（2） adding antibiotic-free animal manure spiked with antibiotics. This study used chlortetracycline（CTC） as a model antibiotic to examine the effects of the antibiotic addition method on methane production in anaerobic digestion under two different swine wastewater concentrations（0.55 and 0.22 mg CTC/g dry manure）. The results showed that CTC degradation rate in which manure was directly added at 0.55 mg CTC/g（HSPIKE treatment） was lower than the control values and the rest of the treatment groups. Methane production from the HSPIKEtreatment was reduced（p 〈 0.05） by 12% during the whole experimental period and 15% during the first 7 days. The treatments had no significant effect on the pH and chemical oxygen demand value of the digesters, and the total nitrogen of the0.55 mg CTC/kg manure collected from mediated swine was significantly higher than the other values. Therefore, different methane production under different antibiotic addition methods might be explained by the microbial activity and the concentrations of antibiotic intermediate products and metabolites. Because the primary entry route of veterinary antibiotics into an anaerobic digester is by contaminated animal manure, the most appropriate method for studying antibiotic residue effects on methane production may be using manure from animals that are given a particular antibiotic, rather than adding the antibiotic directly to the anaerobic digester.

From illite/smectite clay to mesoporous silicate adsorbent for efficient removal of chlortetracycline from water

A series of mesoporous silicate adsorbents with superior adsorption performance for hazardous chlortetracycline（CTC） were sucessfully prepared via a facile one-pot hydrothermal reaction using low-cost illite/smectite（IS） clay,sodium silicate and magnesium sulfate as the starting materials.In this process,IS clay was ＂teared up＂ and then ＂rebuilt＂ as new porous silicate adsorbent with high specific surface area of 363.52 m^2/g（about 8.7 folds higher than that of IS clay） and very negative Zeta potential（- 34.5 mV）.The inert Si- O- Si（Mg,Al） bonds in crystal framework of IS were broken to form Si（Al）- O^- groups with good adsorption activity,which greatly increased the adsorption sites served for holding much CTC molecules.Systematic evaluation on adsorption properties reveals the optimal silicate adsorbent can adsorb 408.81 mg/g of CTC（only 159.7 mg/g for raw IS clay） and remove 99.3%（only 46.5%for raw IS clay） of CTC from 100 mg/L initial solution（pH 3.51;adsorption temperature 30℃;adsorbent dosage,3 g/L）.The adsorption behaviors of CTC onto the adsorbent follows the Langmuir isotherm model,Temkin equation and pseudo second-order kinetic model.The mesopore adsorption,electrostatic attraction and chemical association mainly contribute to the enhanced adsorption properties.As a whole,the high-efficient silicate adsorbent could be candidates to remove CTC from the wastewater with high amounts of CTC.

Effects of straw addition on increased greenhouse vegetable yield and reduced antibiotic residue in fluvo-aquic soil

Organic manure application is an important measure for high yield and good quality vegetable production, whereas organic manure is also a main source of residual antibiotic in soils. A 3-yr experiment was conducted on a fluvo-aguic soil in Tianjin of northern China. The objective of this study was to investigate the effects of different fertilization patterns on yield of six-sea- son vegetables with celery and tomato rotation, and dynamic change of tetracyclines residues in the soil during the sixth growing season （tomato season）. The field experiment comprised six treatments depending on the proportion of nitrogen of each type of fertilizer： 4/4 CN （CN, nitrogen in chemical fertilizer）, 3/4 CN＋1/4 MN （MN, nitrogen in pig manure）, 2/4 CN＋2/4 MN, 1/4 CN＋3/4 MN, 2/4 CN＋1/4 MN＋I/4 SN （SN, nitrogen in corn straw）, and CF （conventional fertilization, the amounts of nitrogen application were 943 and 912 kg N ha-1 for celery and tomato season, respectively）. In addition to CF treatment, the amount of nitrogen application in other treatments was greatly reduced and equal （450 and 450 kg N ha-1 for celery and tomato season, respectively）. Results showed that the combined application of 3/4 CN＋1/4 MN achieved the highest yield and economic benefit in the first four seasons, but addition of straw （2/4 CN＋1/4 MN＋I/4 SN treatment） performed better in the subsequent two seasons, and the average yields of 2/4 CN＋1/4 MN＋I/4 SN treatment were respectively higher by 9.9 and 12.8% than those of 4/4 CN treatment, and by 5.6 and 10.5% than those of CF treatment. The residual chlortet- racycline （CTC） in manure-amended soil for three consecutive years increased along with the increase of applied amount of pig manure. Under the same amount of pig manure application, content of CTC in straw-amended soil was obviously decreased compared with no straw-amended soil （3/4 CN＋1/4 MN treatment）, and averagely decreased by 41.9% for four sampling periods in the sixth season. Addition of crop straw facilitated the degradation of CTC in manure-amended soil. As a whole, the conventional fertilization was not the desirable pattern based on yield, economic benefit and environment, the optimal fertilization pattern with the highest yield and profit and the least soil chlortetracycline residue was the treatment of 2/4 CN＋1/4 MN＋I/4 SN under this experimental condition.

Green remediation of tetracyclines in soil-water systems

The presence of tetracyclines in soil and surface water is an emerging concern. The present study was undertaken to investigate remediation of tetracylines (tetracycline (TC), oxytetracycline (OTC) and chlortetracycline (CTC)) from aqueous solution using vetiver grass, water lettuce, and sunflower and root exudates of water lettuce, sunflower and from soil by tomato, Indian mustard and carrot plant. The data of this study denote that vetiver grass, water lettuce, sunflower remedy tetracyclines from water. The remediation % after 63 days of treatment was 87-61 for TC;88-68 for OTC and 87-68 for CTC. The remediation of tetracyclines at lower concentration of antibiotics in presence of root exudates of water lettuce and sunflower was more than 99% and remediation was faster than water lettuce or sunflower. The remediation of tetracyclines from aqueous solution may be due to oxidation of-OH group(s) of tetracycline through a process that is thought to involve reactive oxygen intermediates and/or role of peroxidase enzyme. The plant crops viz., tomato, Indian mustard and carrot can remedy 41%-72% of amended tetracyclines. The maximum bioaccumulation of TC and CTC was in Indian mustard and OTC was maximally bioaccumulated in tomato.

A Study of Fluorescence System of Eu-Chlorotetracycline-Benzoylacetone-CTMAB Multibasic Complex

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A New Evaluation Method for Antibiotic-Resistant Bacterial Groups in Environment

In the present manuscript it was presented whether spreading of antibiotic resistant bacterial groups in environment could be monitored by our newly developed method by enumerating antibiotic resistant bacterial groups in various biological wastes and composts. Although the numbers were not so high, diverse kinds of colistin resistant bacteria (25 mg·L-1) were included in row cattle feces (1.78 × 104 MPN g-1) and cattle feces manure (>3.84 × 104 MPN g-1). Compost originated from leftover food (>44.8 × 104 MPN g-1) and shochu lee (>320 × 104 MPN g-1) included higher numbers of chlortetracycline resistant Pseudomonas sp., (25 mg·L-1), and row cattle feces included higher numbers of chlortetracycline resistant Enterobacteriacea (15.7 × 104 MPN g-1), which mostly consisted from Pantoea sp. or Xenorhobdus doucetiae. Numbers of multi drug resistant bacteria, resistant to 25 mg·L-1 of ciprofloxacin, streptomycin, chloramphenicol, and ampicillin, were the highest in row cattle feces (>143.6 × 104 MPN g-1), followed by cattle feces manure (4.19 × 104 MPN g-1), and shochu lee (0.36 × 104 MPN g-1), which included diverse kinds of bacterial group. The present results indicated that higher numbers of multi drug resistant bacteria were typically found in row cattle feces, and the method was found suitable to enumerate and identify them. These results suggested that the method might become their environmental risk evaluation method.