Glutamicibacter nicotianae AT6: A new strain for the efficient biodegradation of tilmicosin.

The degradation of tilmicosin (TLM), a semi-synthetic 16-membered macrolide antibiotic, has been receiving increasing attention. Conventionally, there are three tilmicosin degradation methods, and among them microbial degradation is considered the best due to its high efficiency, eco-friendliness, and low cost. Coincidently, we found a new strain, Glutamicibacter nicotianae sp. AT6, capable of degrading high-concentration TLM at 100 mg/L with a 97% removal efficiency. The role of tryptone was as well investigated, and the results revealed that the loading of tryptone had a significant influence on TLM removals. The toxicity assessment indicated that strain AT6 could efficiently convert TLM into less-toxic substances. Based on the identified intermediates, the degradation of TLM by AT6 processing through two distinct pathways was then proposed.

Enhanced hydrolytic removal of tylosin in wastewater using polymer-based solid acid catalysts converted from polystyrene.

Antibiotic production wastewater usually contains high concentrations of antibiotic residues, which can cause instability and deterioration of biological wastewater treatment units and also domestication and proliferation of antibiotic-resistance bacteria. An effective pretreatment on antibiotics production wastewater is expected to selectively reduce the concentration of antibiotics and decrease the toxicity, rather than mitigate organic and other contaminants before further treatments. In this work, two polymer-based solid acids, PS-S and CPS-S bearing high concentrations of -SOH3 groups (up to 4.57 mmol/g), were prepared and successfully used for hydrolytic mitigation of 100 mg/L tylosin within 20 min. The co-existence of high concentrations of COD and humic substances did not affect the mitigation of tylosin obviously, while more than 500 mg/L of nitrogenous compounds suppressed the hydrolytic efficiency. Recycle and reuse experiments showed that the solid acids performed well in five cycles after regeneration. Three transformation products (P1, P2 and P3) were identified using UPLC-QTOF-MS/MS. Sugar moieties including mycarse, mycaminose, and mycinose detached and released simultaneously or in order from the 16-member lactone ring through desugarization, which led to a dramatic decrease in antibacterial activity as revealed by cytotoxicity evaluations using S. aureus. Ecotoxicity estimation indicated the acute toxicities of the hydrolyzed products to model species (e.g., fish, daphnid and green algae) were classified as "not harmful". This work suggested an effective and selective method to pretreat tylosin-contained production wastewater by using polymer-based solid acids. These results will shed light on effective elimination of antibiotics pollution from pharmaceutical industries through strengthening the pretreatments.

Integrated comparison of growth and oxidative stress induced by tylosin in two freshwater algae Chlorella vulgaris and Raphidocelis subcapitata.

Two model algae, Chlorella vulgaris (C. vulgaris) and Raphidocelis subcapitata (R. subcapitata), are commonly used in registration procedures to evaluate compounds with antimicrobial capacity. However, it has been found that these two algae show considerable differences in sensitivity when exposed to antibiotics. The selection of a suitable test species plays a crucial role in assessing the environmental hazards and risks of a compound, as the balance between oxidative stress and antioxidants is a key factor for alga growth. This study was conducted to investigate the status of oxidative stress and mechanism of antioxidant defense system of algae under antibiotic stress. Different tylosin (TYN) exposure-concentrations were used for the tests in this study. Oxidative stress biomarkers (malondialdehyde (MDA)), non-enzymatic antioxidants (reduced glutathione (GSH)), antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GP), glutathione S-transferase (GST)) and photosynthetic pigments were measured to determine the status of the antioxidant defense system. With increasing TYN concentration, the growth of R. subcapitata was significantly inhibited, while there was no effect on C. vulgaris. When the growth of R. subcapitata was inhibited, the content of MDA was significantly increased and the antioxidant system was activated, which indicated a significant increase in the activity of SOD and CAT.

Transcriptomic investigation reveals toxic damage due to tilmicosin and potential resistance against tilmicosin in primary chicken myocardial cells.

Tilmicosin is widely used to treat respiratory infections in animals and has been reported to induce cardiac damage and even sudden death. However, its exact mechanisms, especially in chickens, remain unclear. This study confirmed the dose-dependent damaging effect of tilmicosin on primary chicken myocardial cells. Primary chicken myocardial cells treated with tilmicosin (0.5 µg/mL) for 0 h, 12 h, and 48 h were subjected to RNA sequencing and bioinformatics analysis. Transcriptomic analysis revealed that cytokine-cytokine receptor interactions, calcium signaling pathway, peroxisomes, phagosomes, mitogen-activated protein kinase (MAPK) signaling pathway, and oxidative phosphorylation were significantly and differentially affected after 12 h or 48 h of tilmicosin treatment. Further evidence demonstrated consistently increased proinflammatory factors, peroxidation, and ferroptosis, and intracellular ion imbalance was caused by tilmicosin for 12 h, but this imbalance had recovered at 48 h. Meanwhile, intracellular resistance to tilmicosin-induced toxicity involved the active regulation of cyclooxygenase-1 and ATPase H+/K+-transporting beta subunit at 48 h, sustained activation of MAPK12, and downregulation of dual specificity phosphatase 10 at 12 h. In summary, this study suggests that tilmicosin exerts its cardiotoxicity in primary chicken myocardial cells through multiple mechanisms and finds several intracellular molecular targets to resist the toxicity.

Astragalus polysaccharides alleviate tilmicosin-induced toxicity in rats by inhibiting oxidative damage and modulating the expressions of HSP70, NF-kB and Nrf2/HO-1 pathway.

The present study evaluated the toxic effects of Tilmicosin (TIL) on adult rats. The rats received a single subcutaneous injection of TIL at different doses (10, 25, 50, 75 and 100 mg/kg bw). TIL altered the biochemical parameters including liver and kidney function markers, glucose level and lipid profile as well as resulted in histopathological lesions in liver and adrenal glands mostly in rats exposed to 75 and 100 mg/kg bw. Then the role of Astragalus polysaccharide (APS) at 100 and 200 mg/kg bw, in modulating the toxic effects induced by high dose of TIL was evaluated. Single injection of TIL at a dose of 75 mg/kg bw was found to increase the activity of ALT, AST and ALP enzymes, induce the generation of reactive oxygen species (ROS) and decrease the total antioxidant capacity (TAC). TIL upregulated the hepatic mRNA expression of heat shock protein 70 (HSP70) and nuclear factor kappa B (NF-kB) while blocked the Nrf2/HO-1 mediated response. These changes were also associated with increasing tumer necrosis factor-alpha (TNF-α), interlukin1-beta (IL-1ß) and nitric oxide levels. On the other hand, the results indicate that APS has a beneficial role particularly at high level in alleviating the stress and the hepatotoxic effects elicited by TIL injection in rats.

Contributions of the microbial community and environmental variables to antibiotic resistance genes during co-composting with swine manure and cotton stalks.

Understanding the main drivers that affect the spread of antibiotic resistance genes (ARGs) during the composting process is important for the removal of ARGs. In this study, three levels of tylosin (25, 50, and 75 mg kg-1 on a dry weight basis) were added to swine manure plus a control, which was composted with cotton stalks. Each treatment was repeated in triplicate and the ARG profiles were determined with different levels of tylosin. The top 35 genera and ARGs profiles were clustered together based on the composting time. Combined composting parameters (temperature, pH, NH4+-N, NO3-N, and moisture content) accounted for 78.4% of the total variation in the changes in the potential host bacteria. In addition, the selected five composting parameters and six phyla (including 25 potential host bacterial genera) explained 46.9% and 30.7% of the variation in the ARG profiles according to redundancy analysis, respectively. The variations in ARGs during the composting process were mainly affected by the dynamics of potential host bacteria rather than integrons and the selective pressure due to bio-Cu and bio-Zn.

Temporal succession of soil antibiotic resistance genes following application of swine, cattle and poultry manures spiked with or without antibiotics.

Land application of animal manure is a common agricultural practice potentially leading to dispersal and propagation of antibiotic resistance genes (ARGs) in environmental settings. However, the fate of resistome in agro-ecosystems over time following application of different manure sources has never been compared systematically. Here, soil microcosm incubation was conducted to compare effects of poultry, cattle and swine manures spiked with or without the antibiotic tylosin on the temporal changes of soil ARGs. The high-throughput quantitative PCR detected a total of 185 unique ARGs, with Macrolide-Lincosamide-Streptogramin B resistance as the most frequently encountered ARG type. The diversity and abundance of ARGs significantly increased following application of manure and manure spiked with tylosin, with more pronounced effects observed in the swine and poultry manure treatments than in the cattle manure treatment. The level of antibiotic resistance gradually decreased over time in all manured soils but was still significantly higher in the soils treated with swine and poultry manures than in the untreated soils after 130 days' incubation. Tylosin-amended soils consistently showed higher abundances of ARGs than soils treated with manure only, suggesting a strong selection pressure of antibiotic-spiked manure on soil ARGs. The relative abundance of ARGs had significantly positive correlations with integrase and transposase genes, indicative of horizontal transfer potential of ARGs in manure and tylosin treated soils. Our findings provide evidence that application of swine and poultry manures might enrich more soil ARGs than cattle manure, which necessitates the appropriate treatment of raw animal manures prior to land application to minimise the spread of environmental ARGs.

Size-selective toxicity effects of the antimicrobial tylosin on estuarine phytoplankton communities.

The purpose of this study was to determine the lethal and sublethal effects of the antimicrobial tylosin on natural estuarine phytoplankton communities. Bioassays were used in experimental treatments with final concentrations of 5 to 1000 µg tylosin l(-1). Maximum percent inhibition ranged from 57 to 85% at concentrations of 200-400 µg tylosin l(-1). Half maximum inhibition concentrations of tylosin were ca. 5x lower for small phytoplankton (<20 µm) relative to larger phytoplankton (>20 µm) and suggests that small phytoplankton are more sensitive to tylosin exposure. Sublethal effects occurred at concentrations as low as 5 µg tylosin l(-1). Environmental concentrations of tylosin (e.g., 0.2-3 µg l(-1)) may have a significant sublethal effect that alters the size structure and composition of phytoplankton communities. The results of this study highlight the potential importance of cell size on toxicity responses of estuarine phytoplankton.

Tylosin effect on methanogenesis in an anaerobic biomass from swine wastewater treatment.

The effect of different concentrations of tylosin on methane production was investigated: first methanogenesis in a biomass without contact with the antibiotic, and later the ability of the sludge to adapt to increasing concentrations of tylosin. Results showed that, for biomass that had no contact with the antibiotic, the presence of tylosin inhibits the generation of methane even at concentrations as small as 0.01 mg L(-1), and samples at concentrations above 0.5 mg L(-1) produced practically no methane, whereas, in the digesters acclimated in the presence of tylosin at a concentration of 0.01 to 0.065 mg L(-1), methanogenesis is not inhibited in the presence of antibiotic and the generation of methane is improved. This behaviour suggests the microorganisms have developed not only resistance to the antibiotic but also an ability to metabolize it.

Synergistic protective role of mirazid (Commiphora molmol) and ascorbic acid against tilmicosin-induced cardiotoxicity in mice.

Tilmicosin (TIL) is a long-acting macrolide antibiotic approved for the treatment of cattle with Bovine Respiratory Disease. However, overdose of TIL has been reported to induce cardiotoxicity. The purpose of our experiment was to evaluate the protective effects of Commiphora molmol (mirazid (MRZ); myrrh) and (or) ascorbic acid (AA) against TIL-induced cardiotoxicity in mice. MRZ and AA were orally administered using stomach gavage, either alone or in combination for 5 consecutive days, followed with a single TIL overdose. TIL overdose induced a significant increase in serum levels of cardiac damage biomarkers (AST, LDH, CK, CK-MB, and cTnT), as well as cardiac lipid peroxidation, but cardiac levels of antioxidant biomarkers (GSH, SOD, CAT, and TAC) were decreased. Both MRZ and AA tended to normalize the elevated serum levels of cardiac injury biomarkers. Furthermore, MRZ and AA reduced TIL-induced lipid peroxidation and oxidative stress parameters. MRZ and AA combined produced a synergistic cardioprotective effect. We conclude that myrrh and (or) vitamin C administration minimizes the toxic effects of TIL through their free-radical-scavenging and potent antioxidant activities.

Preparation and evaluation of tilmicosin-loaded hydrogenated castor oil nanoparticle suspensions of different particle sizes.

Three tilmicosin-loaded hydrogenated castor oil nanoparticle (TMS-HCO-NP) suspensions of different particle sizes were prepared with different polyvinyl alcohol surfactant concentrations using a hot homogenization and ultrasonic technique. The in vitro release, in vitro antibacterial activity, mammalian cytotoxicity, acute toxicity in mice, and stability study were conducted to evaluate the characteristics of the suspensions. The in vitro tilmicosin release rate, antibacterial activity, mammalian cytotoxicity, acute toxicity in mice, and stability of the suspensions were evaluated. When prepared with polyvinyl alcohol concentrations of 0.2%, 1%, and 5%, the mean diameters of the nanoparticles in the three suspensions were 920±35 nm, 452±10 nm, and 151±4 nm, respectively. The three suspensions displayed biphasic release profiles similar to that of freeze-dried TMS-HCO-NP powders, with the exception of having a faster initial release. Moreover, suspensions of smaller-sized particles showed faster initial release, and lower minimum inhibitory concentrations and minimum bactericidal concentrations. Time-kill curves showed that within 12 hours, the suspension with the 151 nm particles had the most potent bactericidal activity, but later, the suspensions with larger-sized particles showed increased antibacterial activity. None of the three suspensions were cytotoxic at clinical dosage levels. At higher drug concentrations, all three suspensions showed similar concentration-dependent cytotoxicity. The suspension with the smallest-sized particle showed significantly more acute toxicity in mice, perhaps due to faster drug release. All three suspensions exhibited good stability at 4°C and at room temperature for at least 6 months. These results demonstrate that TMS-HCO-NP suspensions can be a promising formulation for tilmicosin, and that nanoparticle size can be an important consideration for formulation development.

Sublethal effects of the antibiotic tylosin on estuarine benthic microalgal communities.

Pharmaceuticals are common chemical contaminants in estuaries receiving effluent from wastewater and sewage treatment facilities. The purpose of this research was to examine benthic microalgal (BMA) community responses to sublethal exposures to tylosin, a common and environmentally persistent antibiotic. Bioassays, using concentrations of 0.011-218 µmol tylosin l(-1), were performed on intertidal muddy sediments from North Inlet Estuary, SC. Exposure to tylosin resulted in a reduction in total BMA biomass and primary productivity. Furthermore, exposure seemed to retard diatom growth while having a minimal effect on cyanobacteria biomass. Estuarine systems receiving chronic inputs of trace concentrations of tylosin, as well as other antibiotics, may experience significant reductions in BMA biomass and primary productivity. Given the well-documented role of BMA in the trophodynamics of estuaries, these impacts will likely be manifested in higher trophic levels with possible impairments of the structure and function of these sensitive systems.

Toxic effect of the combined antibiotics ciprofloxacin, lincomycin, and tylosin on two species of marine diatoms.

The role that antibiotics and other "emerging contaminants" play in shaping environmental microbial communities is of growing interest. The use of the prokaryotic metabolic inhibitors tylosin (T), lincomycin (L), and ciprofloxacin (C) in livestock and humans is both global and extensive. Each of these antibiotic compounds exhibits an affinity for sediment particles, increasing the likelihood of their deposition in the benthos of aquatic systems and each are often present in environmental samples. The purpose of this study was to determine if T, L, and C and their mixtures exhibit significant toxicity to two species of marine diatoms, an algal class comprised of ubiquitous eukaryotic primary producers. Subpopulations from laboratory cultures of Cylindrotheca closterium and Navicula ramosissima were reared in 24-well microtiter plates in the presence of single or combined antibiotics in dilution series. Population growth rates were assessed via epifluorescent microscopic cell counts, from which the half-max inhibitory concentrations (IC(50)) were calculated and used as part of a toxic unit (TU) method for assessing mixture interactions. The single-compound IC(50)'s were, for C. closterium: T = 0.27 mg L(-1), L = 14.16 mg L(-1), C = 55.43 mg L(-1), and for N. ramosissima: T = 0.99 mg L(-1), L = 11.08 mg L(-1), C = 72.12 mg L(-1). These values were generally higher than similar metrics for freshwater species. Mixture IC(50)'s were generally synergistic against C. closterium and additive for N. ramosissima. Both single and combined treatments reduced or eliminated diatom motility. Monochemical responses were similar between species and were not useful for predicting mixture interactions. Mixtures had compound-specific and species-specific effects, favoring N. ramosissima. These results suggest that anthropogenic antibiotics may play a significant role in the ecology of environmental benthic microbial communities. They also suggest single-compound/species studies do not yield useful predictions of the ecological impact of anthropogenic pharmaceuticals.

Acute toxicity study of tilmicosin-loaded hydrogenated castor oil-solid lipid nanoparticles.

BACKGROUND: Our previous studies demonstrated that tilmicosin-loaded hydrogenated castor oil solid lipid nanoparticles (Til-HCO-SLN) are a promising formulation for enhanced pharmacological activity and therapeutic efficacy in veterinary use. The purpose of this work was to evaluate the acute toxicity of Til-HCO-SLN. METHODS: Two nanoparticle doses were used for the study in ICR mice. The low dose (766 mg/kg.bw) with tilmicosin 7.5 times of the clinic dosage and below the median lethal dose (LD(50)) was subcutaneously administered twice on the first and 7th day. The single high dose (5 g/kg.bw) was the practical upper limit in an acute toxicity study and was administered subcutaneously on the first day. Blank HCO-SLN, native tilmicosin, and saline solution were included as controls. After medication, animals were monitored over 14 days, and then necropsied. Signs of toxicity were evaluated via mortality, symptoms of treatment effect, gross and microscopic pathology, and hematologic and biochemical parameters. RESULTS: After administration of native tilmicosin, all mice died within 2 h in the high dose group, in the low dose group 3 died after the first and 2 died after the second injections. The surviving mice in the tilmicosin low dose group showed hypoactivity, accelerated breath, gloomy spirit and lethargy. In contrast, all mice in Til-HCO-SLN and blank HCO-SLN groups survived at both low and high doses. The high nanoparticle dose induced transient clinical symptoms of treatment effect such as transient reversible action retardation, anorexy and gloomy spirit, increased spleen and liver coefficients and decreased heart coefficients, microscopic pathological changes of liver, spleen and heart, and minor changes in hematologic and biochemical parameters, but no adverse effects were observed in the nanoparticle low dose group. CONCLUSIONS: The results revealed that the LD50 of Til-HCO-SLN and blank HCO-SLN exceeded 5 g/kg.bw and thus the nanoparticles are considered low toxic according to the toxicity categories of chemicals. Moreover, HCO-SLN significantly decreased the toxicity of tilmicosin. Normal clinic dosage of Til-HCO-SLN is safe as evaluated by acute toxicity.

Development and validation of a potentiometric biosensor assay for tylosin with demonstrated applicability for the detection of two other antimicrobial growth-promoter compounds in feedstuffs.

A potentiometric biosensor assay based on a commercially available polyclonal antibody was developed to detect tylosin residues in animal feed. The method can be used as a rapid (less than 45 min) laboratory-based procedure or as a portable field-test for the simultaneous measurement of up to 12 different samples. For both procedures the qualitative detection capability (CCß) for tylosin was determined as 0.2 mg kg(-1) in a range of animal feeds with a measurement repeatability at concentrations between 0.2 and 4 mg kg(-1) of ≤13% coefficient of variation (%CV). The field-test format was capable of detecting tylosin residues at operating (external air) temperatures ranging between +4 and 37°C, although some reduction in signal was observed at the lower temperatures. The laboratory-based tylosin assay was evaluated using 16 medicated and 22 non-medicated feeds and was found to give comparable data with a confirmatory method based upon liquid chromatography-tandem mass spectrometry (LC-MS/MS). The potential to develop a multi-probe format assay for the simultaneous detection of tylosin, spiramycin and virginiamycin was also demonstrated. Cross-validation in a second laboratory showed the assay to be transferable, reliable and robust.

Comparing the sensitivity of algal, cyanobacterial and bacterial bioassays to different groups of antibiotics.

Antibiotics may affect both primary producers and decomposers, potentially disrupting ecosystem processes. Hence, it is essential to assess the impact of antibiotics on aquatic ecosystems. The aim of the present study was therefore to evaluate the potential of a recently developed test for detecting antibiotics in animal tissue, the Nouws Antibiotic Test (NAT), as a sensitive bioassay to assess the effects of antibiotics in water. To this purpose, we determined the toxicity of sulphamethoxazole, trimethoprim, flumequine, tylosin, streptomycin, and oxytetracycline, using the NAT adapted for water exposure. The sensitivity of the NAT was compared to that of bioassays with bacteria (Microtox), cyanobacteria and green algae. In the Microtox test with Vibrio fischeri as test organism, no effects were observed for any of the test compounds. For three of the six antibiotics tested, the cyanobacteria were more vulnerable than the green algae when using photosynthetic efficiency as an endpoint. The lowest EC50 values for four out of six tested antibiotics were obtained using the NAT bacterial bioassay. The bacterial plate system responded to antibiotics at concentrations in the microgL(-1) and lower mgL(-1) range and, moreover, each plate proved to be specifically sensitive to the antibiotics group it was designed for. It is concluded that the NAT bioassay adapted for water exposure is a sensitive test to determine the presence of antibiotics in water. The ability of this test to distinguish five major antibiotic groups is a very strong additional value.

Effect of a high dose of three antibiotics on the reproduction of a parthenogenetic strain of Folsomia candida (Isotomidae: Collembola).

Folsomia candida Willem (Isotomidae: Collembola) is an edaphic parthenogenetic species commonly used in ecotoxicity studies. We exposed F. candida to a high dose of three antibiotics, tylosin, ampicillin, and oxytetracycline, that target different bacterial groups. Possible toxic effects were assessed through egg production, hatching, and body size. All three antibiotics caused toxic effects. Treatment with oxytetracycline proved the most toxic. This group showed the smallest body size and lowest number of eggs laid, likely the result of a combination of antibiotic toxicity and avoidance of the antibiotic spiked food. Active toxin avoidance by F. candida in toxicological assays may play a role in minimizing their exposure to toxic compounds. Despite the administration of high doses of oxytetracycline, F. candida individuals remained infected with the intracellular bacteria Wolbachia indicating that this strain is resistant to this antibiotic or that the host or its gut flora had detoxified the compound. An increase in percent egg hatch with time was seen in the ampicillin and oxytetracycline treatments, indicating a possible accommodation of the host and/or gut-flora to these antibiotics.

Hydrogenated castor oil nanoparticles as carriers for the subcutaneous administration of tilmicosin: in vitro and in vivo studies.

Tilmicosin-loaded solid lipid nanoparticles (SLN) were prepared with hydrogenated castor oil (HCO) by o/w emulsion-solvent evaporation technique. The nanoparticle diameters, surface charges, drug loadings and encapsulation efficiencies of different formulations were 90 approximately 230 nm, -6.5 approximately -12.5 mV, 40.3 approximately 59.2% and 5.7 approximately 11.7% (w/w), respectively. In vitro release studies of the tilmicosin-loaded nanoparticles showed a sustained release and the released tilmicosin had the same antibacterial activity as that of the free drug. Pharmacokinetics study after subcutaneous administration to Balb/c mice demonstrated that a single dose of tilmicosin-loaded nanoparticles resulted in sustained serum drug levels (>0.1 microg/mL) for 8 days, as compared with only 5 h for the same amount of tilmicosin phosphate solution. The time to maximum concentration (Tmax), half-life of absorption (T(1/2) ab) and half-life of elimination (T(1/2) el) of tilmicosin-loaded nanoparticles were much longer than those of tilmicosin phosphate solution. Tissue section showed that drug-loaded nanoparticles caused no inflammation at the injection site. Cytotoxicity study in cell culture and acute toxicity test in mice demonstrated that the nanoparticles had little or no toxicity. The results of this exploratory study suggest that the HCO-SLN could be a useful system for the delivery of tilmicosin by subcutaneous administration.

Effects of six selected antibiotics on plant growth and soil microbial and enzymatic activities.

The potential impact of six antibiotics (chlortetracycline, tetracycline and tylosin; sulfamethoxazole, sulfamethazine and trimethoprim) on plant growth and soil quality was studied by using seed germination test on filter paper and plant growth test in soil, soil respiration and phosphatase activity tests. The phytotoxic effects varied between the antibiotics and between plant species (sweet oat, rice and cucumber). Rice was most sensitive to sulfamethoxazole with the EC10 value of 0.1 mg/L. The antibiotics tested inhibited soil phosphatase activity during the 22 days' incubation. Significant effects on soil respiration were found for the two sulfonamides (sulfamethoxazole and sulfamethazine) and trimethoprim, whereas little effects were observed for the two tetracyclines and tylosin. The effective concentrations (EC10 values) for soil respiration in the first 2 days were 7 mg/kg for sulfamethoxazole, 13 mg/kg for sulfamethazine and 20 mg/kg for trimethoprim. Antibiotic residues in manure and soils may affect soil microbial and enzyme activities.