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Sample records for chlortetracycline

  1. 6-Iso-chlortetracycline or keto form of chlortetracycline? Need for clarification for relevant monitoring of chlortetracycline residues in food.

    Science.gov (United States)

    Gaugain, Murielle; Gautier, Sophie; Bourcier, Sophie; Jacques, Anne-Marie; Laurentie, Michel; Abjean, Jean-Pierre; Hurtaud-Pessel, Dominique; Verdon, Eric

    2015-01-01

    Chlortetracycline (CTC) is a broad-spectrum antibiotic used in veterinary medicine for pulmonary or digestive infections and having a regulatory maximum residue limit (MRL) necessitating an official analytical control method. The purpose of this study was to clarify the identification of different forms of CTC observed in standard solution, in spiked muscle samples and in naturally incurred muscle samples of pigs analysed by LC-MS/MS and to demonstrate the in vivo formation of 6-iso-chlortetracycline and 4-epi-6-iso-CTC as a metabolite of CTC and 4-epi-CTC in muscle. The six following forms were identified, all being isobaric with a protonated molecule at m/z 479 (precursor ion): the keto-enol forms of CTC and the keto-enol forms of 4-epi-chlortetracycline (4-epi-CTC), 6-iso-chlortetracycline (6-iso-CTC) and 4-epi-6-iso-chlortetracycline (4-epi-6-iso-CTC). The 6-iso-CTC and 4-epi-6-iso-CTC were observed only in incurred pig samples so were identified for the first time as metabolites of CTC and 4-epi-CTC. Identification of the different forms was obtained by comparing incurred muscle samples with standard solutions and with spiked samples. Then the differences between the features of the chromatograms obtained by LC-TQ-MS and the fragmentation study of the different forms of CTC obtained by LC-Q-TOF-MS helped us to support this identification. The extraction steps and the LC-MS/MS conditions developed to analyse muscle tissue samples are described. This clarification concerning the rigorous identification of chromatographic peaks allowed us to evaluate the relevance of our monitoring method with regard to the regulations in place in the European Union and could be of help to laboratories involved in official control of antibiotic residues in food of animal origin. Additional results are also presented highlighting the transformation of the CTC when prepared in a mixture with other antibiotics.

  2. Adsorption of Chlortetracycline from Water by Rectories

    Institute of Scientific and Technical Information of China (English)

    吕国诚; 吴丽梅; 王晓龙; 廖立兵; 王小雨

    2012-01-01

    The removal of antibiotics from water by clay minerals has become the focus of research due to their strong adsorptive ability. In this study, adsorption of chlortetracycline (CTC) onto rectories was conducted and the effects of time, concentration, temperature and pH were investigated. Experimental results showed that adsorption equilibrium was reached in 8 h. Based on the Langmuir model, the maximum adsorption capacity of CTC on rectories was 177.7 mg·g 1 at room temperature. By the study on adsorption dynamics, it is found that the kinetic date fit the pseudo-second-order model well. The adsorption of CTC by rectories is endothermic and the free energy is in the range of 10 to 30 kJ·mol 1 . The pH value of solution has significant effects on adsorption and the optimal pH is at acidity (pH 2-6). At concentration of 2500 mg·L 1 , the intercalated CTC produces an interlayer space with a height of 1.38 nm, which is 1.12 nm in raw rectories, suggesting that the adsorption occurs between layers of rectories.

  3. Tetracycline resistance genes persist in soil amended with cattle feces independently from chlortetracycline selection pressure

    NARCIS (Netherlands)

    Kyselkova, Martina; Kotrbova, Lucie; Bhumibhamon, Gamonsiri; Chronakova, Alica; Jirout, Jiri; Vrchotova, Nadezda; Schmitt, Heike; Elhottova, Dana

    2015-01-01

    Antibiotic residues and antibiotic resistance genes originating from animal waste represent environmental pollutants with possible human health consequences. In this study, we addressed the question whether chlortetracycline (CTC) residues in soils can act as selective pressure enhancing the persist

  4. Tetracycline, oxytetracycline and chlortetracycline determination by flow injection potentiometry.

    Science.gov (United States)

    Couto, C M; Lima, J L; Conceição, M; Montenegro, B S; Reis, S

    1998-12-01

    This paper describes tetracycline (TCH), oxytetracycline (OTCH) and chlortetracycline (CTCH) determination by flow injection potentiometry. In the flow system proposed TC samples are inserted in a carrier solution and converged with a Cu(II) solution of known concentration; the Cu(II) decrease due to its complexation with tetracyclines (TC) was monitored. The detector used was a homogeneous crystalline CuS/Ag2S double membrane tubular electrode with increased sensitivity. The present system allows tetracyclines determinations within a 48.1-4.8 x 10(3) ppm for TCH, 49.1-4.9 x 10(3) ppm for OTCH and 51.5-5.1 x 10(3) ppm for CTCH and a precision better than 0.4% for the three TC species. This procedure accomplishes 150-200 samples h(-1) with a Cu(II) consumption of about 13 microg determination(-1).

  5. Immunosuppressant Effect of Long Term and Subtherapeutic Chlortetracycline in Broiler

    Institute of Scientific and Technical Information of China (English)

    TONG Jian-ming; ZHANG Ri-jun; SA Ren-na; PAN Shu-yuan; HUANG Yan

    2002-01-01

    Experiments were conducted with 270 Abore Acre male broilers to study the influence of dietary chlortetracycline (CTC) on the bursal, thymus and spleen weight index and T, B cell proliferation and antibody responses of broilers. Results show that 50mg/kg CTC had no significant inhibitory effect on thymus and spleen weight index, and T, B cell proliferation of broilers at 21 days old. 150mg/kg CTC had significant inhibitory effect on bursal, thymus and spleen, and T, B cell proliferation (P<0.05). Dietary CTC had a direct inhibitory effect on lymphocyte proliferation in vitro. 50mg/kg and 150mg/kg CTC had a significant inhibitory effect on the antibody responses to BSA (P<0.01). Results demonstrated that subtherapeutic and long term CTC had significant inhibitory effects on immune organs and antibody response. The immunosuppressant effect became higher when the level of CTC increased. 150mg/kg CTC increased the growth rate of broilers significantly.

  6. Kinetics of tetracycline, oxytetracycline, and chlortetracycline adsorption and desorption on two acid soils

    DEFF Research Database (Denmark)

    Fernandez Calviño, David; Bermúdez-Couso, Alipio; Arias-Estévez, Manuel;

    2015-01-01

    The purpose of this work was to quantify retention/release of tetracycline, oxytetracycline, and chlortetracycline on two soils, paying attention to sorption kinetics and to implications of the adsorption/desorption processes on transfer of these pollutants to the various environmental compartments...

  7. Quantification of bioavailable chlortetracycline in pig feces using a bacterial whole-cell biosensor

    DEFF Research Database (Denmark)

    Hansen, L. H.; Aarestrup, Frank Møller; Sørensen, S. J.

    2002-01-01

    Bacterial whole-cell biosensors were used to measure the concentration of chlortetracycline (CTC) in the feces of pigs. In this study, the Escherichia coli biosensor used has a detection limit of 0.03 mg/kg CTC in pig feces. The tetracycline concentration was correlated with the appearance...

  8. Quantification of bioavailable chlortetracycline in pig feces using a bacterial whole-cell biosensor

    DEFF Research Database (Denmark)

    Hansen, Lars Hestbjerg; Aarestrup, Frank; Sørensen, Søren Johannes

    2002-01-01

    Bacterial whole-cell biosensors were used to measure the concentration of chlortetracycline (CTC) in the feces of pigs. In this study, the Escherichia coli biosensor used has a detection limit of 0.03 mg/kg CTC in pig feces. The tetracycline concentration was correlated with the appearance...

  9. Development of an automatic measuring device for total sugar content in chlortetracycline fermenter based on STM32

    Science.gov (United States)

    Liu, Ruochen; Chen, Xiangguang; Yao, Minpu; Huang, Suyi; Ma, Deshou; Zhou, Biao

    2017-01-01

    Because fermented liquid in chlortetracycline fermenter has high viscosity and complex composition, conventional instruments can't directly measure its total sugar content of fermented liquid. At present, offline artificial sampling measurement is usually the way to measuring total sugar content in chlortetracycline Fermenter. it will take too much time and manpower to finish the measurement., and the results will bring the lag of control process. To realize automatic measurement of total sugar content in chlortetracycline fermenter, we developed an automatic measuring device for total sugar content based on STM32 microcomputer. It can not only realize the function of automatic sampling, filtering, measuring of fermented liquid and automatic washing of the device, but also can make the measuring results display in the field and finish data communication. The experiment results show that the automatic measuring device of total sugar content in chlortetracycline fermenter can meet the demand of practical application.

  10. Effect of Vancomycin, Tylosin, and Chlortetracycline on Vancomycin-Resistant Enterococcus faecium Colonization of Broiler Chickens During Grow-Out.

    Science.gov (United States)

    Hume, Michael E; Donskey, Curtis J

    2017-01-27

    Broiler chickens may serve as reservoirs for human colonization by vancomycin-resistant Enterococcus (VRE). We examined the effects of vancomycin and two commonly used antimicrobial feed additives on VRE colonization in broiler chickens during grow-out. Chicks received unsupplemented feed or feed containing vancomycin, chlortetracycline, or tylosin from day of hatch to grow-out at 6 weeks. At 3 days of age, chicks received by crop gavage 10(7) colony-forming units (CFUs) of a human or poultry VRE isolate. Cecal contents were monitored weekly for VRE, short-chain fatty acids (SCFAs), and bacterial denaturing gradient gel electrophoresis (DGGE) profile methods. Vancomycin promoted persistent and high-level colonization with human- and poultry-derived VRE to grow-out in comparison with controls, while treatment with chlortetracycline and tylosin did not. Colonization by the poultry isolate in control, chlortetracycline, and tylosin groups persisted throughout the grow-out period with low concentrations present at 6 weeks, whereas the human isolate decreased to an undetectable level by week 6. Vancomycin resulted in significant reductions in cecal acetic acid and butyric acid in comparison with controls, but chlortetracycline and tylosin did not. DGGE profiles contained two main clusters with all vancomycin profiles in a smaller cluster and all other profiles in a larger cluster. These results demonstrate that vancomycin, but not chlortetracycline or tylosin, disrupted the indigenous microbiota and SCFA patterns of broiler chickens and promoted colonization by VRE.

  11. The efficacy of three chlortetracycline regimens in the treatment of persistent Anaplasma marginale infection.

    Science.gov (United States)

    Reinbold, James B; Coetzee, Johann F; Hollis, Larry C; Nickell, Jason S; Riegel, Casey; Olson, K C; Ganta, Roman R

    2010-09-28

    Chemosterilization is reported in cattle fed chlortetracycline hydrochloride (CTC) at dosages ranging from 1.1mg/kg for 120 days to 11 mg/kg for 30-60 days. The relationship between plasma CTC drug concentration and carrier clearance has not been described. Chronic carrier status was established in 21 steers with a Virginia isolate of Anaplasma marginale and confirmed by cELISA and an A. marginale-specific RT-PCR. Four negative, splenectomized steers served as active disease transmission sentinels. Steers were randomized to receive 4.4 mg/kg/day (LD); 11 mg/kg/day (MD); or 22 mg/kg/day (HD) of oral chlortetracycline; or placebo (CONTROL) for 80 days. The LD, MD and HD treatment groups consisted of 5 infected steers and 1 splenectomized steer; CONTROL group had six infected steers and 1 splenectomized steer. The daily treatments and ration were divided equally and fed twice daily. Blood samples were collected semi-weekly for determining plasma drug concentration by ultrahigh performance liquid chromatography-mass spectrometry/mass spectrometry method and assessment of disease status by both cELISA and RT-PCR. Mean (CV%) chlortetracycline plasma drug concentrations in the LD, MD, and HD groups were 85.3 (28%), 214.5 (32%) and 518.9 (40%)ng/mL during days 4 through 53 of treatment. A negative RT-PCR assay result was confirmed in all CTC-treated groups within 49 days of treatment; however, cELISA required an additional 49 to 88 days before similar results. Subinoculation of splenectomized steers confirmed chemosterilization. These results are important for influencing future chemosterilization strategies and impacting free trade policy among countries and regions of contrasting endemicity.

  12. Reduction of chlortetracycline-resistant Escherichia coli in weaned piglets fed fermented liquid feed.

    Science.gov (United States)

    Kobashi, Yuri; Ohmori, Hideyuki; Tajima, Kiyoshi; Kawashima, Tomoyuki; Uchiyama, Hiroo

    2008-10-01

    We investigated the change in chlortetracycline resistance in 360 Escherichia coli strains separated from the feces of piglets fed fermented liquid feed (FLF) in comparison with those fed dry feed (control). The total amount of lactic acid bacteria in feces was 8.77 x 10(8) CFU/g DM at weaning, which increased to 1.23 x 10(12) CFU/g DM (FLF) at 28 days after weaning (Pfeeding of FLF can possibly reduce antibiotic-resistance bacteria.

  13. Degradation and toxicity assessment of sulfamethoxazole and chlortetracycline using electron beam, ozone and UV.

    Science.gov (United States)

    Kim, Tae-Hun; Kim, Sang Don; Kim, Hyun Young; Lim, Seung Joo; Lee, Myunjoo; Yu, Seungho

    2012-08-15

    Recently, the occurrence of antibiotics in sewage treatment plant effluent, as well as drinking water, has raised concern about their potential impacts on the environment and public health. Antibiotics are found in surface and ground waters, which indicate their ineffective removal by conventional wastewater treatment processes. Therefore, advanced oxidation processes (AOPs) have received considerable attention for the removal of antibiotics. This study was conducted to evaluate the degradation and mineralization of antibiotics (sulfamethoxazole and chlortetracycline) using an electron beam, ozone and UV, and the change of toxicity. Also, the electrical energy consumption based on the EE/O parameter (the electrical energy required per order of pollutants removal in 1 m(3) wastewater) was used to quantify the energy cost associated with the different AOPs (electron beam, ozone and UV) for the degradation of antibiotics. The results showed that an electron beam effective for the removals of both sulfamethoxazole and chlortetracycline in aqueous solutions. However, degradation of the target compounds by ozone and UV showed different trends. The oxidation efficiency of each organic compound was very dependent upon the AOP used. Algal toxicity was significantly reduced after each treatment. However, based on the electrical energy, the electron beam was more efficient than ozone and UV. Electron beam treatment could be an effective and safe method for the removal of antibiotic compounds.

  14. Effects of in-feed chlortetracycline prophylaxis of beef cattle on animal health and antimicrobial-resistant Escherichia coli

    Science.gov (United States)

    Concerns have been raised that in-feed chlortetracycline (CTC) may increase antimicrobial resistance (AMR), specifically tetracycline-resistant (TETr) Escherichia coli, and third-generation cephalosporin-resistant (3GCr) E. coli. We evaluated the impact of a 5-day in-feed CTC prophylaxis on animal h...

  15. Distribution of sulfamethazine, chlortetracycline and tylosin in manure and soil of Canadian feedlots after subtherapeutic use in cattle

    Energy Technology Data Exchange (ETDEWEB)

    Aust, Marc-Oliver [University of Trier, FB VI: Geography/Geosciences, Soil Science Section, Campus II, 54286 Trier (Germany); University of Rostock, Institute for Land Use/Soil Science Section (Germany)], E-mail: aust@uni-trier.de; Godlinski, Frauke [University of Rostock, Institute for Land Use/Soil Science Section (Germany); Agriculture and Agri-Food Canada, Research Centre Lethbridge, AB (Canada); Travis, Greg R.; Hao, Xiying; McAllister, Tim A. [Agriculture and Agri-Food Canada, Research Centre Lethbridge, AB (Canada); Leinweber, Peter [University of Rostock, Institute for Land Use/Soil Science Section (Germany); Thiele-Bruhn, Soeren [University of Trier, FB VI: Geography/Geosciences, Soil Science Section, Campus II, 54286 Trier (Germany); University of Rostock, Institute for Land Use/Soil Science Section (Germany)], E-mail: thiele@uni-trier.de

    2008-12-15

    Feedlots are potential point sources for the flow of antibiotics into the environment due to common use of antibiotics such as sulfamethazine, chlortetracycline and tylosin. Hence soils and manures originating from a grassland control, an experimental and a commercial feedlot were analyzed and mass balances were calculated for these antibiotics. Up to 9990 {mu}g kg{sup -1} sulfamethazine and 401 {mu}g kg{sup -1} chlortetracycline on a dry matter basis were determined in feedlot manure. Soil concentrations were two orders of magnitude smaller. This corresponds to 7-40% of the calculated residual amount. In the commercial feedlot chlortetracycline was found down to soil depths of -40 cm; sulfamethazine was still detectable 1 year after medication. Sulfamethazine and chlortetracycline were additionally determined in manure of a control treatment in the experimental feedlot where cattle never received antibiotics. This was attributed to runoff from upslope pens. Consequently, antibiotics partially persist within feedlots and may be dislocated into the surrounding environment by vertical transport and runoff. - Veterinary antibiotics partially persist within feedlots and may be dislocated to the environment by transport processes following subtherapeutic application to cattle.

  16. Flow-injection chemiluminescence determination of chlortetracycline using on-line electrogenerated [Cu(HIO(6))(2)](5-) as the oxidant.

    Science.gov (United States)

    Li, B; Zhang, Z; Liu, W

    2001-12-24

    A novel chemiluminescence (CL) flow system for the determination of chlortetracycline is described. It is based on the direct CL reaction of chlortetracycline and [Cu(HIO(6))(2)](5-) in KOH medium. The unstable [Cu(HIO(6))(2)](5-) was on-line electrogenerated by constant-current electrolysis. The CL intensity was linear with chlortetracycline concentration in the range of 0.1-100 mug ml(-1). The determination limit was 5.3x10(-8) g ml(-1). The whole process could be completed in 1 min. The proposed method is suitable for automatic and continuous analysis, and has been applied satisfactorily to analysis of chlortetracycline in biological fluid.

  17. Tylosin and chlortetracycline effects during swine manure digestion: influence of sodium azide.

    Science.gov (United States)

    Stone, James J; Clay, Sharon A; Spellman, Garth M

    2010-12-01

    The antibiotics tylosin and chlortetracycline (CTC), which are commonly used in pig production, were studied to determine their effects on swine manure digestion in the presence and absence of biocide sodium azide. CTC enhanced initial hydrolysis reactions through volatile suspended solids production, while inhibiting methane and carbon dioxide production. Tylosin did not affect methane and carbon dioxide production; however, the relative abundance of both hydrogen utilizing and acetate-only utilizing microbial populations was significantly compromised. Sodium azide in the absence of antibiotics enhanced metabolic output and initial biomass production, and this observation suggests that populations of Methanobacteriales and Methanosaetaceae spp. appeared to contain sufficient periplasmic bound reductase to effectively utilize acetate and hydrogen in the presence of sodium azide. However, the combination of sodium azide and either CTC or tylosin was a very effective metabolic inhibitor, inhibiting methane and carbon dioxide production and VSS consumption compared to their no-azide counterpart.

  18. Impact of subtherapeutic administration of tylosin and chlortetracycline on antimicrobial resistance in farrow-to-finish swine.

    Science.gov (United States)

    Holman, Devin B; Chénier, Martin R

    2013-07-01

    The use of antimicrobial agents in swine production at subtherapeutic concentrations for the purpose of growth promotion remains controversial due to the potential impact on public health. Beginning at weaning (3 weeks), pigs received either nonmedicated feed or feed supplemented with subtherapeutic levels of either tylosin (11-44 ppm) or chlortetracycline (5.5 ppm). After only 3 weeks, pigs given feed supplemented with tylosin had significantly higher levels of tylosin-resistant anaerobes (P Tylosin-fed pigs had a higher incidence of detection for erm(A), erm(F), and erm(G), as well as significantly (P 0.05). The resistance genes tet(O), tet(Q), and erm(B) were detected in all pigs at each sampling time, while tet(G), tet(L), and tet(M) were also frequently detected. Neither chlortetracycline nor tylosin increased the growth rate of pigs.

  19. Online prediction for contamination of chlortetracycline fermentation based on Dezert-Smarandache theory

    Institute of Scientific and Technical Information of China (English)

    Jianwen Yang; Xiangguang Chen; Huaiping Jin

    2015-01-01

    Fermentative production of chlortetracycline is a complex fed-batch bioprocess. It generally takes over 90 h for cultivation and is often contaminated by undesired microorganisms. Once the fermentation system is contami-nated to certain extent, the product quality and yield wil be seriously affected, leading to a substantial economic loss. Using information fusion based on the Dezer–Smarandache theory, self-recursive wavelet neural network and unscented kalman filter, a novel method for online prediction of contamination is developed. Al state vari-ables of culture process involving easy-to-measure and difficult-to-measure variables commonly obtained with soft-sensors present their contamination symptoms. By extracting and fusing latent information from the chang-ing trend of each variable, integral and accurate prediction results for contamination can be achieved. This makes preventive and corrective measures be taken promptly. The field experimental results show that the method can be used to detect the contamination in time, reducing production loss and enhancing economic efficiency.

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

    Science.gov (United States)

    Huang, Lu; Wen, Xin; Wang, Yan; Zou, Yongde; Ma, Baohua; Liao, Xindi; Liang, Juanboo; Wu, Yinbao

    2014-10-01

    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 0.22mg CTC/g dry manure). The results showed that CTC degradation rate in which manure was directly added at 0.55mg CTC/g (HSPIKE treatment) was lower than the control values and the rest of the treatment groups. Methane production from the HSPIKE treatment was reduced (pmethane 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.

  1. Chlortetracycline and tylosin runoff from soils treated with antimicrobial containing manure.

    Science.gov (United States)

    Hoese, A; Clay, S A; Clay, D E; Oswald, J; Trooien, T; Thaler, R; Carlson, C G

    2009-05-01

    This study assessed the runoff potential of tylosin and chlortetracycline (CTC) from soils treated with manure from swine fed rations containing the highest labeled rate of each chemical. Slurry manures from the swine contained either CTC at 108 microg/g or tylosin at 0.3 microg/g. These manures were surface applied to clay loam, silty clay loam, and silt loam soils at a rate of 0.22 Mg/ha. In one trial, tylosin was applied directly to the soil surface to examine runoff potential of water and chemical when manure was not present. Water was applied using a sprinkler infiltrometer 24-hr after manure application with runoff collected incrementally every 5 min for about 45 min. A biofilm crust formed on all manure-treated surfaces and infiltration was impeded with > 70% of the applied water collected as runoff. The total amount of CTC collected ranged from 0.9 to 3.5% of the amount applied whereas tylosin ranged from 8.4 to 12%. These data indicate that if surface-applied manure contains antimicrobials, runoff could lead to offsite contamination.

  2. Effect of antimicrobial compounds tylosin and chlortetracycline during batch anaerobic swine manure digestion.

    Science.gov (United States)

    Stone, James J; Clay, Sharon A; Zhu, Zhenwei; Wong, Kwok L; Porath, Laura R; Spellman, Garth M

    2009-10-01

    Tylosin and chlortetracycline (CTC) are antimicrobial chemicals that are fed to >45% of the US swine herds at therapeutic and sub-therapeutic dosages to enhance growth rates and treat swine health problems. These compounds are poorly absorbed during digestion so that the bioactive compound or metabolites are excreted. This study investigated the degradation and stabilization of swine manure that contained no additives and compared the observed processes with those of manure containing either tylosin or CTC. The batch anaerobic incubation lasted 216 days. The breakdown of insoluble organic matter through anaerobic hydrolysis reactions was faster for manure containing CTC compared with tylosin or no-antimicrobial treatments. Volatile fatty acid (VFA) accumulation, including acetate, butyrate, and propionate, was greater for CTC-containing manure compared to tylosin and no-antimicrobial treatments. The relative abundance of two aceticlastic methanogens, Methanosaetaceae and Methanosarcinaceae spp., were less for CTC manure than manure with no-antimicrobial treatment. In addition, generation of methane and carbon dioxide was inhibited by 27.8% and 28.4%, respectively, due to the presence of CTC. Tylosin effects on manure degradation were limited, however the relative abundance of Methanosarcinaceae spp. was greater than found in the CTC or no-antimicrobial manures. These data suggest that acetate and other C-1 VFA compounds would be effectively utilized during methanogenesis in the presence of tylosin.

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

    Institute of Scientific and Technical Information of China (English)

    Bei Wen; Yu Liu; Peng Wang; Tong Wu; Shuzhen Zhang; Xiaoquan Shan; Jingfen Lu

    2012-01-01

    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 EC10 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.

  4. Removal of chlortetracycline from spiked municipal wastewater using a photoelectrocatalytic process operated under sunlight irradiations

    Energy Technology Data Exchange (ETDEWEB)

    Daghrir, Rimeh, E-mail: rimeh.daghrir@ete.inrs.ca [Institut National de la Recherche Scientifique, Centre Eau, Terre et Environnement, 490 rue de la Couronne, Québec, Qc G1K 9A9 (Canada); Drogui, Patrick, E-mail: patrick.drogui@ete.inrs.ca [Institut National de la Recherche Scientifique, Centre Eau, Terre et Environnement, 490 rue de la Couronne, Québec, Qc G1K 9A9 (Canada); Delegan, Nazar, E-mail: delegan@emt.inrs.ca [Institut National de la Recherche Scientifique, INRS-Énergie, Matériaux et Télécommunications, 1650 Blvd. Lionel-Boulet, Varennes, Qc J3X 1S2 (Canada); El Khakani, My Ali, E-mail: elkhakani@emt.inrs.ca [Institut National de la Recherche Scientifique, INRS-Énergie, Matériaux et Télécommunications, 1650 Blvd. Lionel-Boulet, Varennes, Qc J3X 1S2 (Canada)

    2014-01-01

    The degradation of chlortetracycline in synthetic solution and in municipal effluent was investigated using a photoelectrocatalytic oxidation process under visible irradiation. The N-doped TiO{sub 2} used as photoanode with 3.4 at.% of nitrogen content was prepared by means of a radiofrequency magnetron sputtering (RF-MS) process. Under visible irradiation, higher photoelectrocatalytic removal efficiency of CTC was recorded using N-doped TiO{sub 2} compared to the conventional electrochemical oxidation, direct photolysis and photocatalysis processes. The photoelectrocatalytic process operated at 0.6 A of current intensity during 180 min of treatment time promotes the degradation of 99.1 ± 0.1% of CTC. Under these conditions, removal rates of 85.4 ± 3.6%, 87.4 ± 3.1% and 55.7 ± 2.9% of TOC, TN and NH{sub 4}{sup +} have been recorded. During the treatment, CTC was mainly transformed into CO{sub 2} and H{sub 2}O. The process was also found to be effective in removing indicator of pathogens such as fecal coliform (log-inactivation was higher than 1.2 units). - Highlights: •PECO process is a feasible technology for the treatment of MWW contaminated by CTC. •99.1% ± 0.1% of CTC was degraded by PECO using N-doped TiO{sub 2}. •85.4% ± 3.6% of TOC removal and 97.5% ± 1.2% of COD removal were achieved. •87.4% ± 3.1% of TN removal and 55.7% ± 2.9% of NH{sub 4}{sup +} removal were recorded. •More than 94% of fecal coliform was removed (abatement > 1.2-log units)

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

    Science.gov (United States)

    Lu, Lei; Wu, Yixiao; Ding, Huijun; Zhang, Weihao

    2015-07-01

    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.

  6. Inhibition of resistance plasmid transfer in Escherichia coli by ionophores, chlortetracycline, bacitracin, and ionophore/antimicrobial combinations.

    Science.gov (United States)

    Mathers, Jeremy J; Clark, Steven R; Hausmann, Denny; Tillman, Paul; Benning, Valerie R; Gordon, Shelly K

    2004-01-01

    Medicinal feed additives bacitracin, chlortetracycline (CTC), laidlomycin, lasalocid, and salinomycin inhibited the transfer of multiresistance-conferring plasmid pBR325 (Tet(r) Amp(r) Cp(r), 6.0 kb) into selected gram-negative strains with the use of an in vitro model. High concentrations of ampicillin-sensitive competence-pretreated Escherichia coli HB 101 cells were exposed to 10% (v/v) of 1:10 dimethyl sulfoxide/agent : water containing test mixtures for 0.5 hr prior to plasmid addition and transforming conditions. Transformation was inhibited for all antimicrobials and showed a positive association wich higher concentration. Additional testing of ionophore compounds separately and in combination with bacitracin, chlortetracycline, lincomycin, roxarsone, tylosin, and virginiamycin at representative feed concentrations demonstrated 80.6% to >99.9% inhibition (P < 0.001) of resistance transfer. Bacitracin alone inhibited transformation within the range of 50-500 ppm. No increase in resistance transfer was observed when poultry-derived and reference gram-negative isolates having low or no transformation efficiency were additionally tested. The results suggest that these compounds, at relevant concentrations used in animal feed, may interfere with cell envelope-associated DNA uptake channels or other transformation competence mechanisms. Through these mechanisms, ionophores and cell membrane-interactive feed agents such as CTC and bacitracin may act to inhibit resistance transfer mechanisms within poultry and livestock.

  7. Risk assessment of chlortetracycline, oxytetracycline, sulfamethazine, sulfathiazole, and erythromycin in aquatic environment: are the current environmental concentrations safe?

    Science.gov (United States)

    Ji, Kyunghee; Kim, Sunmi; Han, Sunyoung; Seo, Jihyun; Lee, Sangwoo; Park, Yoonsuk; Choi, Kyunghee; Kho, Young-Lim; Kim, Pan-Gyi; Park, Jeongim; Choi, Kyungho

    2012-10-01

    To understand potential risks of major pharmaceutical residues in waters, we evaluated ecotoxicities of five major veterinary pharmaceuticals, i.e., chlortetracycline, oxytetracycline, sulfamethazine, sulfathiazole, and erythromycin, which have been frequently detected in freshwater environment worldwide. We conducted acute and chronic toxicity tests using two freshwater invertebrates (Daphnia magna and Moina macrocopa) and a fish (Oryzias latipes). In general, D. magna exhibited greater sensitivity than M. macrocopa, and chronic reproduction was the most sensitive endpoints for both organisms. The population growth rate was adversely influenced by exposure to chlortetracycline, sulfamethazine, or sulfathiazole in water fleas, but reduction in population size was not expected. In O. latipes, the tested pharmaceuticals affected several reproduction related endpoints including time to hatch and growth. Based on the toxicity values from the present study and literature, algae appeared to be the most sensitive organism, followed by Daphnia and fish. Hazard quotients derived from measured environmental concentrations (MECs) and predicted no effect concentrations (PNECs) for erythromycin and oxytetracycline exceeded unity, suggesting that potential ecological effects at highly contaminated sites cannot be ruled out. Long-term consequences of veterinary pharmaceutical contamination in the environment deserve further investigation.

  8. Electrochemical degradation of chlortetracycline using N-doped Ti/TiO2 photoanode under sunlight irradiations.

    Science.gov (United States)

    Daghrir, Rimeh; Drogui, Patrick; Delegan, Nazar; El Khakani, My Ali

    2013-11-01

    The appearance and the persistence of pharmaceutical products in the aquatic environment urgently call for the development of an innovative and practical water treatment technology. This study deals with the development of nanostructured nitrogen-doped TiO2 photoanodes and their subsequent use for chlortetracycline (CTC) photoelectrocatalytic oxidation under visible light. The N-doped TiO2 photoanodes with different nitrogen contents were prepared by means of a radiofrequency magnetron sputtering (RF-MS) process, with the objective to tune shift their optical absorption from the UV towards the visible. The N-doped TiO2 consist of nanostructured anatase phase with average TiO2 nanocrystallite size of 29 nm. The nitrogen doping is clearly shown to produce the desired red shift of the absorption onset of the TiO2 coatings (from ~380 nm to ~550 nm). Likewise, the N-doped TiO2 are found to be highly photo-electroactive not only under the UV light but most interestingly under the visible light as well. Using the optimal N-doped photoanodes, 99.6% of CTC (100 μg/L) was successfully degraded after 180 min of treatment time with a current intensity of 0.6 A. Under these conditions, a relatively high mineralization of CTC (92.5% ± 0.26% of TOC removal and 90.3% ± 1.1% of TN removal) was achieved.

  9. A survey of chlortetracycline concentration in feed and its residue in chicken egg in commercial layer farms

    Indian Academy of Sciences (India)

    K Kodimalar; R A Rajini; S Ezhivalavan; G Sarathchandra

    2014-06-01

    The worldwide increase in the use of antibiotics as an integral part of poultry and livestock production industry has recently received increasing attention as a contributory factor in the international emergence of antibiotic-resistant bacteria in human beings. To gauge the presence of the aforementioned scenario in the Indian context, a preliminary survey was conducted to assess the use of chlortetracycline (CTC) in 12 commercial layer farms and to quantify and confirm its residue in the egg. Samples of feed and eggs were collected at day 0 (prior to CTC addition), 3rd, 5th and 7th day during treatment and on the 9th and 14th day (2nd and 7th day after withdrawal of CTC) from each of the 12 commercial poultry farms studied. Concentration of CTC in feed was significantly ( < 0.01) high on the 3rd, 5th and 7th day. On the 9th day and 14th day CTC concentration in feed was significantly ( <0.01) lower compared to the earlier 3 days studied. A highly significant difference ( <0.01) of the antibiotic residue in egg was observed in all the 5 days with high residual levels of CTC in egg. CTC in feed and its residue in egg were detected even on the 9th and 14th day respectively.

  10. Effects of In-Feed Copper, Chlortetracycline, and Tylosin on the Prevalence of Transferable Copper Resistance Gene, tcrB, Among Fecal Enterococci of Weaned Piglets.

    Science.gov (United States)

    Amachawadi, Raghavendra G; Scott, H Morgan; Vinasco, Javier; Tokach, Mike D; Dritz, Steve S; Nelssen, Jim L; Nagaraja, Tiruvoor G

    2015-08-01

    Heavy metals, such as copper, are increasingly supplemented in swine diets as an alternative to antibiotics to promote growth. Enterococci, a common gut commensal, acquire plasmid-borne, transferable copper resistance (tcrB) gene-mediated resistance to copper. The plasmid also carried resistance genes to tetracyclines and macrolides. The potential genetic link between copper and antibiotic resistance suggests that copper supplementation may exert a selection pressure for antimicrobial resistance. Therefore, a longitudinal study was conducted to investigate the effects of in-feed copper, chlortetracycline, and tylosin alone or in combination on the selection and co-selection of antimicrobial-resistant enterococci. The study included 240 weaned piglets assigned randomly to 6 dietary treatment groups: control, copper, chlortetracycline, tylosin, copper and chlortetracycline, and copper and tylosin. Feces were collected before (day 0), during (days 7, 14, 21), and after (days 28 and 35) initiating treatment, and enterococcal isolates were obtained from each fecal sample and tested for genotypic and phenotypic resistance to copper and antibiotics. A total of 2592 enterococcal isolates were tested for tcrB by polymerase chain reaction. The overall prevalence of tcrB-positive enterococci was 14.3% (372/2592). Among the tcrB-positive isolates, 331 were Enterococcus faecium and 41 were E. faecalis. All tcrB-positive isolates contained both erm(B) and tet(M) genes. The median minimum inhibitory concentration of copper for tcrB-negative and tcrB-positive enterococci was 6 and 18 mM, respectively. The majority of isolates (95/100) were resistant to multiple antibiotics. In conclusion, supplementing copper or antibiotics alone did not increase copper-resistant enterococci; however, supplementing antibiotics with copper increased the prevalence of the tcrB gene among fecal enterococci of piglets.

  11. Determination of the inhibitory concentration 50% (IC50) of four selected drugs (chlortetracycline, doxycycline, enrofloxacin and difloxacin) that reduce in vitro the multiplication of Chlamydophila psittaci.

    Science.gov (United States)

    Failing, K; Theis, P; Kaleta, E F

    2006-11-01

    A total of 18 chlamydial isolates from various psittacine birds, one isolate from a domestic pigeon and one isolate from a Pekin duck were isolated in continuous Buffalo Green Monkey (BGM) kidney cell cultures. All 20 isolates were identified by nested multiplex polymerase chain reaction as Chlamydophila psittaci. These isolates were multiplied to high titres and subsequently tested for in vitro sensitivity against two tetracyclines (chlortetracycline and doxycycline) and two quinolones (enrofloxacin and difloxacin) at concentrations of 0.0, 0.25, 0.50, 1.00, and 10.00 microg/ml. Replication of chlamydia in BGM cell cultures is assayed on the basis of formation of intracytoplasmic inclusions that are visualized by Giménez staining. All isolates, although to variable degrees, are sensitive to all four drugs. The number of chlamydial inclusions decreases gradually over a broad range of increasing concentrations of the drugs. The variation in the number of inclusions between isolates is remarkably high for chlortetracycline less for doxycycline and minimal for both fluoroquinolones, the enrofloxacin and difloxacin. The decline in numbers of inclusions is highly dose-dependend and the observed reduction stretches over a wide range of drug dilutions. Therefore, it is proposed to calculate drug sensitivity values in terms of inhibitory concentration 50%, (IC5). Its calculation includes all tested drug dilutions instead of the hitherto more common minimal inhibitory concentration, MIC, which is based on results of serial dilution tests for cell-free growing bacteria. Using a logistic regression model for the calculation of the inhibitory concentration 50% of all 20 chlamydial isolates, the IC50 is 0.807 microg/ml for tetracycline, 0.497 microg/ml for doxycycline, 0.180 microg/ml for enrofloxacin and 0.168 microg/ml for difloxacin. Complete prevention of inclusion formation was already seen for enrofloxacin at a concentration of 1.0 microg/ml in 12 out of 20 and for

  12. Effects of Chlortetracycline and 4-epi-chlortetracycline on the Growth of Rape and Its Accumulation in Seedling%金霉素和4-差向金霉素对油菜生长的影响及其在幼苗体内的积累

    Institute of Scientific and Technical Information of China (English)

    魏瑞成; 邵明诚; 陈明; 王冉

    2012-01-01

    以油菜为材料,研究不同浓度金霉素(chlortetracycline,CTC)和代谢物差向金霉素(4-epi-chlortetracycline,ECTC)胁迫对其发芽、生长和抗氧化酶活性的影响,以及CTC和ECTC在幼苗体内的积累.结果表明:随着处理浓度的升高,种子发芽率、发芽势没有显著差异,但株高和根长受到明显抑制;在1~30 mg·L-1处理浓度下,CTC和ECTC诱导的幼苗体内抗氧化酶SOD、POD、CAT活性均高于对照组;幼苗体内CTC、ECTC积累含量随着处理浓度的升高而增加,表明CTC和ECTC可在油菜幼苗体内积累,这可能产生潜在食品安全风险.%Effects of different concentrations of chlortetracycline (CTC ) and 4-epi-chlortetracycline(ECTC) on the growth and the activities of antioxidant enzymes in rape and the accumulation of CTC and ECTC in seedlings were studied by using water culture method. The results showed that seedling height and root length significantly reduced, compared to the control, and the root elongation could be recommended as a sensitive indicator to assess crops eco-toxicity of CTC and ECTC. The activities of antioxidant enzymes as superoxide dismutase(SOD), peroxidase(POD) and catalase(CAT) induced by CTC and ECTC at concentration ranges of l~30 mg·L‐1 were higher than the control. When rape seedling was stressed by 1 mg · L‐1 and 10 mg · L‐1 of CTC and ECTC, the activities of SOD and POD were enhanced, but declined by 30 mg·L‐1. SOD, POD removed oxygen free radical in coordination with CAT at concentration levels of 1 mg·L‐1 and 10 mg·L‐1 of CTC and ECTC. However, at a concentration of 30 mg · L‐1, CTC and ECTC would cause certain oxidative damage on rape seedling. The difference of antioxidant enzyme system suggested that ability of rape seedling anti-CTC stress was stronger than anti-ECTC. With the concentration of CTC and ECTC increased, the contents of them accumulated in seedlings were increased, which could result in a potential food safety

  13. [A dynamic study of drug resistance in populations of fecal "Escherichia coli" from axenic mice, fed chlortetracycline continuously and associated with flora from infant, calf and piglet (author's transl)].

    OpenAIRE

    Corpet, D E

    1980-01-01

    International audience; In order to prove the effect of low doses of antibiotics on the level of drug resistance of the gastrointestinal flora, gnotoxenic mice, harbouring various microflora from newborn calf, piglet or infant, were given continuously water containing 20 microgram/ml chlortetracycline. Control mice, harbouring the same flora, were maintained as well in isolators. Sensible and resistant fecal Escherichia coli were counted daily. Results showed that even before the introduction...

  14. Complexação da tetraciclina, da oxitetraciclina e da clortetraciclina com o catião cobre (II. Estudo potenciométrico Tetracycline, oxytetracycline and chlortetracycline complexation with copper (II. Potentiometric study

    Directory of Open Access Journals (Sweden)

    Cristina M. C. M. Couto

    2000-08-01

    Full Text Available Stability constants of complexes formed by copper (II with three different tetracyclines (tetracycline, oxytetracycline and chlortetracycline have been determined potentiometrically with an automatic system in aqueous medium at 25,0 ± 0,2 ºC and I = 0,1 mol L-1 NaNO3. The protonation constants of the three tetracyclines were also determined under the same conditions. The distribution of the complexes was then simulated at therapeutic levels of the drugs.

  15. Development and validation of a liquid chromatographic/ tandem mass spectrometric method for determination of chlortetracycline, oxytetracycline, tetracycline, and doxycycline in animal feeds.

    Science.gov (United States)

    Guo, Liang; Chen, Yiqiang; Zhang, Liying; Yang, Wenjun; He, Pingli

    2012-01-01

    A selective and accurate LC/MS/MS method for the simultaneous determination of chlortetracycline (CTC), oxytetracycline (OTC), tetracycline (TC), and doxycycline (DC) in animal feeds was developed. Samples were extracted with Na2EDTA-McIlvaine buffer and further purified with Oasis HLB SPE columns. The purified extract was separated on an Xbridge C18 column and detected by LC/MS/MS with positive electrospray ionization in the multiple reaction monitoring mode. This method provided average recoveries of 80.9 to 119.5%, with CVs of 1.7 to 9.8% in the range of 0.5 to 50 mg/kg CTC, OTC, TC, and DC in feeds, except the average recovery of CTC was 76.0%, with a CV of 14.6% in pig feed spiked with 0.5 mg/kg CTC. The linear ranges for the four TCs determined by LC/MS/MS ranged from 0.005 to 2.5 microg/mL with a linear correlation coefficient (R2) >0.99. The LOD and LOQ for CTC, OTC, TC, and DC in pig and poultry feeds ranged from 0.003 to 0.02 and 0.01 to 0.05 microg/g, respectively. The method was successfully applied for the analysis of 30 real feed samples, and no illegal use was detected.

  16. 21 CFR 558.128 - Chlortetracycline.

    Science.gov (United States)

    2010-04-01

    ... (reducing the incidence of abortion and shedding of leptospirae) caused by Leptospira pomona susceptible to... (i) 20 to 50 g/ton Growing sheep; increased rate of weight gain and improved feed efficiency. 046573,048164, 066104. (ii) 80 mg/head/day Breeding sheep; reducing the incidence of (vibrionic) abortion...

  17. Otimização e validação de método para determinação de resíduos de oxitetraciclina, tetraciclina e clortetraciclina em leite por cromatografia líquida de alta eficiência Optimization and validation of a method for determination of oxytetracycline, tetracycline and chlortetracycline residues in milk by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Iara C. A. Ruela

    2005-03-01

    Full Text Available Um método para determinação de resíduos de oxitetraciclina, tetraciclina e clortetraciclina em leite por cromatografia líquida de alta eficiência foi otimizado e validado. Ensaios intralaboratoriais empregando soluções padrão, brancos de amostras e amostras adicionadas de soluções padrão dos antimicrobianos foram realizados para avaliação da linearidade, seletividade, exatidão, precisão, limites de detecção e de quantificação do método. As amostras foram homogeneizadas, tratadas com solução tampão de succinato de sódio pH 4,0 e centrifugadas, descartando-se o precipitado. As tetraciclinas foram removidas do sobrenadante por quelação com íons cobre reversivelmente ligados à resina quelante e eluídas com tampão McIlvaine-EDTA pH 4,0. As médias de recuperação para as tetraciclinas foram de 76,2 a 104,9% com coeficiente de variação na faixa de 1,4 a 18,9%. Limites de detecção de 10, 15 e 40ng/mL e limites de quantificação de 10, 20 e 50ng/mL foram determinados para oxitetraciclina, tetraciclina e clortetraciclina, respectivamente.A method for determination of oxytetracycline, tetracycline and chlortetracycline residues in milk by high performance liquid chromatography was optimized and validated. Linearity, selectivity, trueness, precision, limits of detection and quantitation were evaluated by in-house validation procedures employing reference materials, sample blanks and spiked samples. Milk samples were homogenized, treated with sodium succinate pH 4.0 buffer and centrifuged, discarding the precipitate. Tetracyclines were removed from the supernatant by chelating with metal ions bound to a chelating resin and eluted with McIlvaine-EDTA pH 4.0 buffer. Mean recoveries for tetracyclines ranged from 76.2 to 104.9% with coefficients of variation from 1.4 to 18.9%. Detection limits of 10, 15 and 40ng/mL and quantitation limits of 10, 20 and 50ng/mL were determined for oxytetracycline, tetracycline and

  18. 21 CFR 520.445b - Chlortetracycline powder.

    Science.gov (United States)

    2010-04-01

    ... bacterial enteritis (scours) caused by E. coli and Salmonella spp. and bacterial pneumonia associated with.... Control and treatment of bacterial enteritis (scours) caused by E. coli and Salmonella spp. and bacterial... treatment of bacterial enteritis (scours) caused by E. coli and Salmonella spp., and bacterial...

  19. 21 CFR 520.445c - Chlortetracycline tablets and boluses.

    Science.gov (United States)

    2010-04-01

    ... enteritis (scours) caused by E. coli and Salmonella spp. and bacterial pneumonia associated with Pasteurella... Salmonella spp., and bacterial pneumonia associated with Pasteurella spp., Hemophilus spp., and Klebsiella... associated with Pasteurella spp., Klesbsiella spp., and Hemophilus spp. (ii) Limitations. Administer...

  20. 75 FR 9334 - New Animal Drugs for Use in Animal Feeds; Chlortetracycline

    Science.gov (United States)

    2010-03-02

    ... HUMAN SERVICES Food and Drug Administration 21 CFR Part 558 New Animal Drugs for Use in Animal Feeds... Administration (FDA) is amending the animal drug regulations to reflect approval of a supplemental new animal drug application (NADA) filed by ADM Alliance Nutrition, Inc. The supplemental NADA provides for use...

  1. 77 FR 5700 - New Animal Drugs; Change of Sponsor; Chlortetracycline Powder

    Science.gov (United States)

    2012-02-06

    ...: Steven D. Vaughn, Center for Veterinary Medicine (HFV-100), Food and Drug Administration, 7520 Standish... to the Center for Veterinary Medicine, 21 CFR parts 510 and 520 are amended as follows: PART 510--NEW..., LLC, 277 Faison McGowan Rd., Kenansville, NC 28349 * * * * * PART 520--ORAL DOSAGE FORM NEW...

  2. Land application of tylosin and chlortetracycline swine manure: Impacts to soil nutrients and soil microbial community structure.

    Science.gov (United States)

    Stone, James J; Dreis, Erin K; Lupo, Christopher D; Clay, Sharon A

    2011-01-01

    The land application of aged chortetracycle (CTC) and tylosin-containing swine manure was investigated to determine associated impacts to soil microbial respiration, nutrient (phosphorus, ammonium, nitrate) cycling, and soil microbial community structure under laboratory conditions. Two silty clay loam soils common to southeastern South Dakota were used. Aerobic soil respiration results using batch reactors containing a soil-manure mixture showed that interactions between soil, native soil microbial populations, and antimicrobials influenced CO(2) generation. The aged tylosin treatment resulted in the greatest degree of CO(2) inhibition, while the aged CTC treatment was similar to the no-antimicrobial treatment. For soil columns in which manure was applied at a one-time agronomic loading rate, there was no significant difference in soil-P behavior between either aged CTC or tylosin and the no-antimicrobial treatment. For soil-nitrogen (ammonium and nitrate), the aged CTC treatment resulted in rapid ammonium accumulation at the deeper 40cm soil column depth, while nitrate production was minimal. The aged CTC treatment microbial community structure was different than the no-antimicrobial treatment, where amines/amide and carbohydrate chemical guilds utilization profile were low. The aged tylosin treatment also resulted in ammonium accumulation at 40 cm column depth, however nitrate accumulation also occurred concurrently at 10 cm. The microbial community structure for the aged tylosin was also significantly different than the no-antimicrobial treatment, with a higher degree of amines/amides and carbohydrate chemical guild utilization compared to the no-antimicrobial treatment. Study results suggest that land application of CTC and tylosin-containing manure appears to fundamentally change microbial-mediated nitrogen behavior within soil A horizons.

  3. Effect of vancomycin, tylosin, and chlortetracycline on vancomycin-resistant Enterococcus faecium colonization of broiler chickens during grow-out

    Science.gov (United States)

    Broiler chickens may serve as reservoirs for human colonization by vancomycin-resistant Enterococcus (VRE). We examined the effects of vancomycin and two commonly-used antimicrobial feed additives on VRE colonization in broiler chickens during grow-out. Chicks received unsupplemented feed or feed ...

  4. Effect of anaerobic digestion temperature on odour, coliforms and chlortetracycline in swine manure or monensin in cattle manure

    Science.gov (United States)

    Antibiotics used in animal feeding operations have been detected in the environment. There is a growing concern about the impact of these pharmaceutical compounds in the manure and the effect they may have on aquatic and terrestrial organisms, and the potential development of antibiotic resistant m...

  5. Application and prospects of chlortetracycline and salinomycin as feed additive%金霉素和盐霉素饲料添加剂的应用现状与展望

    Institute of Scientific and Technical Information of China (English)

    王统石

    2010-01-01

    本文探讨了抗生素在饲料中作为添加剂使用的原理,从肠道内营养再分配的角度,探讨其作为饲料添加剂的经济效益和价值,并分析了金霉素和盐霉素在饲料中使用的现状及应用前景.

  6. 金霉素眼膏对胃肠减压患者鼻咽舒适度的影响%The Effect of Chlortetracycline Eye Ointment on Nasopharyngeal Comfort of Patients with Gastrointestinal Decompression

    Institute of Scientific and Technical Information of China (English)

    张锦莲; 张柳华; 杨璐晖; 曾芝云

    2014-01-01

    Objective:To investigate the effect of chlorotetracycline eye ointment on nasopharyngeal comfort of patients with gastroenteric decompression.Method:182 patients with indwelling gastrointestinal decompression tube in our department from July 2012 to October 2013 were selected,they were divided into the control group for 90 cases and the intervention group for 92 cases according to the random number table method.The control group was treated with routine requirements including oral care,gargling with collutory,blowing the nasal with normal saline cotton stick,while the intervention group was treated with chlorotetracycline eye ointment in the local administration of a gastric tube side nasal cavity.The degree of comfort of pars nasalis pharyngis and the acceptance of stomach tube of two groups were compared. Result:The nasopharyngeal discomfort incidence of intervention group was 67.40%(62/92),it was significantly lower than 96.67%(87/90)of control group,the acceptance of stomach tube in intervention group was significantly higher than the control group,the differences were statistically significant(P<0.01).Conclusion:Topically treatment of chlorotetracycline eye ointment to patients with gastroenteric decompression can reduce the discomfort of pars nasalis pharyngis,increase the patients’acceptance of the stomach tube,improve degree of comfort and compliance of treatment,is worthy of clinical promotion.%目的:探讨金霉素眼膏鼻腔局部用药对胃肠减压患者鼻咽部舒适度的影响。方法:选取2012年7月-2013年10月本科留置胃肠减压管的182例患者,按照随机数字表法将其分为对照组90例和干预组92例。对照组按常规要求进行口腔护理、漱口液漱口、生理盐水棉签清洁鼻腔;干预组在对照组的基础上使用金霉素眼膏在插胃管侧鼻腔局部用药。比较两组患者鼻咽部舒适度及患者对胃管的接受程度。结果:干预组的鼻咽部不适发生率67.40%(62/92)明显低于对照组的96.67%(87/90),对胃管接受程度明显高于对照组,差异均有统计学意义(P<0.01)。结论:胃肠减压患者采用金霉素眼膏鼻腔局部用药可减轻鼻咽部不适,增加患者对胃管的接受程度,提高患者舒适度与治疗依从性,值得临床推广。

  7. 半乳甘露寡糖和金霉素在生长育肥猪上效果对比试验%A comparative trial on galactmannan-oligosaccharides and chlortetracycline in diets for growing and fattening pigs

    Institute of Scientific and Technical Information of China (English)

    王彬; 张军; 黄瑞林; 李铁军; 印遇龙; 李长明

    2005-01-01

    将48头生长猪随机分为2组,在日粮中分别添加0.1 %半乳甘露寡糖(GMOS组)和50 mg/kg金霉素(ANT组),进行了96 d的对比试验,结果表明,1)生长性能GMOS组与ANT组相比生长期内平均日增重提高3.15 %,平均日采食量略高,料肉比下降2.82 %, 但差异均不显著;育肥期内平均日增重提高10.2 %, 差异显著,平均日采食量降低25.2 %,差异极显著,料肉比下降13.2 %, 差异极显著;全期内,平均日增重提高6.66 %,差异不显著,平均日采食量降低1.83 %,差异不显著,料肉比下降8.01 %,差异显著;2)抗病效果GMOS组比ANT组发生腹泻等消化道病的头日数要少一点,感染气喘等呼吸道病的头日数却多一点,但差异均不显著; 3)屠宰性能GMOS组与ANT组相比背膘厚下降了15.6 %, 皮厚下降了11.5 %, 瘦肉率提高了2.94 %, 但差异均不显著.

  8. Multi-class determination and confirmation of antibiotic residues in honey using LC-MS/MS

    Science.gov (United States)

    A multi-class method was developed for the determination and confirmation in honey of tetracyclines (chlortetracycline, doxycycline, oxytetracycline, and tetracycline), fluoroquinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, and sarafloxacin), macrolides (tylosin), lincosamides (lin...

  9. Toxicity of tetracyclines and tetracycline degradation products to environmentally relevant bacteria, including selected tetracycline-resistant bacteria

    DEFF Research Database (Denmark)

    Halling-Sørensen, B.; Sengeløv, G.; Tjørnelund, J.

    2002-01-01

    Tetracyclines used in veterinary therapy invariably will find their way as parent compound and degradation products to the agricultural field. Major degradation products formed due to the limited stability of parent tetracyclines (tetracycline, chlortetracycline, and oxytetracycline) in aqueous s...

  10. The application of the treatment of resina draconis combined with chlortetracycline eye ointment and nursing intervention for the advanced cancer patients with Ⅱ period of pressure ulcers%龙血竭联合金霉素眼膏治疗晚期癌症患者Ⅱ期压疮临床观察及护理体会

    Institute of Scientific and Technical Information of China (English)

    梁丽云; 刘美荣; 张世永

    2016-01-01

    目的:探讨龙血竭联合金霉素眼膏及护理干预治疗晚期癌症患者Ⅱ期压疮的临床效果.方法:选择40例晚期癌症Ⅱ期压疮患者分为对照组和观察组,各20例,均给予0.9%氯化钠注射液棉球清洗创面,对照组采用常规护理方法及0.5%碘伏纱外敷治疗,观察组采用龙血竭联合金霉素眼膏及护理干预方法治疗.换药1次/d,1个疗程为7d,3个疗程后比较两组的临床疗效.结果:观察组出现新生健康肉芽组织时间(5.8±0.3)d,对照组出现新生健康肉芽组织时间为(7.1±0.6)d,组间比较,差异具有统计学意义(P<0.05);观察组愈合时间(14.7±0.6)d,对照组愈合时间(18.2±0.9)d,组间比较,差异具有统计学意义(P<0.05);两组治疗效果比较,差异具有统计学意义(P<0.05).结论:应用龙血竭联合金霉素眼膏及护理干预治疗晚期癌症患者Ⅱ期压疮具有显著疗效,能明显促进患者压疮愈合,缩短愈合时间.

  11. 半胱胺与金霉素对肉鸭的促生长效果及相关代谢激素水平影响的比较%Comparison of Cysteamine and Chlortetracycline on Production and Metabolic Hormones Levels in Meat Duck

    Institute of Scientific and Technical Information of China (English)

    陈晓生; 朱锦兰; 刘为民; 张辉华; 田允波; 罗锦标

    2005-01-01

    240只1日龄肉鸭苗随机分成4个处理组,分别为高、低剂量半胱胺的2个试验组,每吨日粮加50克金霉素的金霉素组,使用基础日粮的对照组.结果表明:高、低2个半胱胺剂量组的增重效果均达到金霉素组的水平(P>0.05),但与对照组比,日粮中按80g/T使用的高剂量组差异显著(P<0.05),30g/T使用的低剂量组没有显著差异(P>0.05);血清中3种主要代谢激素活动增强,其中高剂量试验组类胰岛素生长因子IGF-I水平显著高于其他3组(P<0.05).

  12. The action of certain antibiotics and ether on swine enzootic pneumonia.

    Science.gov (United States)

    Huhn, R G

    1971-01-01

    The susceptibility of Mycoplasma hyopneumoniae to the action of three antibiotics and diethyl ether was determined. Infected swine were used in an in vivo sensitivity detection system. The parameter of susceptibility was lesion prophylaxis. In vivo, Mycoplasma hyopneumoniae appeared to be resistant to diethyl ether, tylosin tartrate, and erythromycin, but was susceptible to the action of chlortetracycline. Chlortetracycline was effective in preventing the development of lesions when given at levels which would be practical in commercial swine operations.

  13. Experimental infection of cattle with ovine Dichelobacter nodosus isolates

    DEFF Research Database (Denmark)

    Knappe-Poindecker, Maren; Jørgensen, Hannah Joan; Jensen, Tim Kåre

    2015-01-01

    be transferred to the interdigital skin of cattle under experimental conditions. Further, we wanted to observe the impact of such infection on bovine foot health, and test the effect of topical chlortetracycline (Cyclo spray(®): Eurovet) on the infection. Six heifers were included in the study. After an initial...... developed interdigital dermatitis. In five of the heifers D. nodosus organisms were demonstrated within the epidermis. Twenty-four days after treatment with chlortetracycline all heifers were negative by cultivation, but tested positive for D. nodosus by polymerase chain reaction (PCR). Two of the six......, the study supports the hypothesis that infections with virulent D. nodosus in cattle are associated with interdigital dermatitis. No conclusion regarding the treatment of D. nodosus infection with chlortetracycline was possible....

  14. 21 CFR 556.500 - Oxytetracycline.

    Science.gov (United States)

    2010-04-01

    ... kilogram of body weight per day. (b) Beef cattle, dairy cattle, calves, swine, sheep, chickens, turkeys... chlortetracycline, oxytetracycline, and tetracycline, in tissues and milk as follows: (1) 2 parts per million (ppm) in muscle. (2) 6 ppm in liver. (3) 12 ppm in fat and kidney. (4) 0.3 ppm in milk....

  15. 21 CFR 333.110 - First aid antibiotic active ingredients.

    Science.gov (United States)

    2010-04-01

    ... gram, 30 milligrams of chlortetracycline hydrochloride in a suitable ointment base. (d) Neomycin sulfate ointment containing, in each gram, 3.5 milligrams of neomycin in a suitable water soluble or oleaginous ointment base. (e) Neomycin sulfate cream containing, in each gram, 3.5 milligrams of neomycin...

  16. Fate of antimicrobials and antimicrobial resistance genes in simulated swine manure storage

    Science.gov (United States)

    The behavior of three antibiotics (bacitracin, chlortetracycline, and tylosin) and two classes of antibiotic resistance genes (ARGs), tet and erm, were monitored in swine manure slurry under anaerobic conditions. First-order decay rates were determined for each antibiotic with half-lives ranging fr...

  17. 21 CFR 558.55 - Amprolium.

    Science.gov (United States)

    2010-04-01

    ... (air-sac infection) and blue comb (nonspecific infectious enteritis) As bacitracin methylene... aid in the control of necrotic enteritis caused or complicated by Clostridium spp. or other organisms... infectious enteritis) As bacitracin methylene disalicylate, or zinc bacitracin Chlortetracycline 100 to...

  18. 21 CFR 558.305 - Laidlomycin.

    Science.gov (United States)

    2010-04-01

    ... Echerichia coli and bacterial pneumonia caused by Pasteurella multocida organisms susceptible to... for breeding. (e) Conditions of use. It is used in cattle being fed in confinement for slaughter as... associated with shipping fever complex caused by Pasteurella spp. susceptible to chlortetracycline....

  19. Dissipation of Three Veterinary Antimicrobials in Beef Cattle Feedlot Manure Stockpiled over Winter.

    Science.gov (United States)

    Sura, Srinivas; Degenhardt, Dani; Cessna, Allan J; Larney, Francis J; Olson, Andrew F; McAllister, Tim A

    2014-05-01

    Dissipation of veterinary antimicrobials is known to occur during aerated windrow composting of beef cattle manure. However, it is unclear if a similar dissipation occurs during stockpiling. Chlortetracycline, tylosin, and sulfamethazine are three of the most commonly used veterinary antimicrobials in beef cattle production in western Canada. Their dissipation in stockpiled manure was investigated over 140 d during winter in Alberta, Canada. Beef cattle housed in pens were administered 44 mg of chlortetracycline kg feed (dry weight), 44 mg of chlortetracycline + 44 mg sulfamethazine kg feed, 11 mg of tylosin kg feed, or feed without antimicrobials (control). Manure samples were extracted using pressurized liquid extraction, and the extracts were analyzed for chlortetracycline, sulfamethazine, and tylosin by LC-MS-MS. Dissipation of all three antimicrobials in the manure was explained by exponential decay kinetics. Times for 50% dissipation (DT) were 1.8 ± 0.1 d for chlortetracycline alone or 6.0 ± 0.8 d when mixed with sulfamethazine, 20.8 ± 3.8 d for sulfamethazine, and 4.7 ± 1.2 d for tylosin. After 77 d, manure did not inhibit microbial activity, as indicated by temperature and mass losses of carbon (C) and nitrogen (N). The C/N ratio in the manure decreased over the stockpiling period, indicating decomposition of manure to a more stable state. Dissipation of excreted residues with DT values 1.8 to 20.8 d showed that stockpiling can be as effective as windrow composting in mitigating the transfer of these three veterinary antimicrobials into the environment during land application of processed manure.

  20. In vitro antibiotic susceptibility of field isolates of Mycoplasma synoviae in Argentina.

    Science.gov (United States)

    Cerdá, R O; Giacoboni, G I; Xavier, J A; Sansalone, P L; Landoni, M F

    2002-01-01

    Minimum inhibitory concentrations (MICs) were determined in vitro for 7 antibiotics (aivlosin, enrofloxacine, tylosin, tiamulin, kitasamycin, chlortetracycline, and oxytetracycline) against eight recent local Argentinean isolates and two standard strains of Mycoplasma synoviae. Aivlosin (3-acetyl-4"-isovaleryl tylosin tartrate), tylosin, and tiamulin showed the lowest MICs with MIC90s of 0.006, 0.012, and 0.05 microg/ml, respectively. Except one strain that showed resistant values to chlortetracycline (> or = 12.5 microg/ml), all the analyzed strains were susceptible in different degrees to all the antibiotics tested. In this study, the improved activity of the tylosin-derived drug, aivlosin, was confirmed because it showed, in most strains, MIC values half those for tylosin.

  1. Evidence on Possible Mycoplasma Etiology of Aster Yellows Disease II. Suppression of Aster Yellows in Insect Vectors.

    Science.gov (United States)

    Whitcomb, R F; Davis, R E

    1970-08-01

    Chlortetracycline or chloramphenicol (but not kanamycin, penicillin, or erythromycin), when administered in hydroponic solution to diseased aster, reduced the availability of the aster yellows (AY) agent to nymphs of Macrosteles fascifrons (Stål). Insects exposed to healthy plants whose roots were immersed in chlortetracycline were able to acquire AY agent from diseased plants the day after removal from the antibiotic-treated plants, but the latent period of the ensuing disease in the insects was prolonged. Chlortetracycline or tylosin tartrate blocked AY infection in nymphs injected with a mixture of antibiotic and the AY agent, but polymyxin, neomycin, vancomycin, penicillin, carbomycin, or chloramphenicol did not. All tetracyclines tested, methacycline, oxytetracycline, and chlortetracycline, produced a dramatic reduction in the ability of infected vectors to transmit AY agent. Tylosin tartrate also reduced transmission when injected into AY-transmitting vectors, but carbomycin, spectinomycin, cycloserine, penicillin, erythromycin, or kanamycin had no such effect. During the first 10 days after injection of tylosin tartrate or oxytetracycline into transmitting vectors, ability of the insects to transmit AY decayed rapidly. Transmission by insects injected with buffer alone, after decreasing the first day after injection, gradually returned to its normal level in less than 1 week. By 2 to 3 weeks after injection with tylosin or oxytetracycline, ability to transmit AY was regained by vectors. The results suggest that tetracycline antibiotics and tylosin tartrate inhibit multiplication of AY agent in the insect. The spectrum of antibiotic activity in the insect is consistent with the hypothesis that AY and other plant yellows diseases are caused by mycoplasma-like organisms.

  2. In Vivo Transmission of an IncA/C Plasmid in Escherichia coli Depends on Tetracycline Concentration, and Acquisition of the Plasmid Results in a Variable Cost of Fitness.

    Science.gov (United States)

    Johnson, Timothy J; Singer, Randall S; Isaacson, Richard E; Danzeisen, Jessica L; Lang, Kevin; Kobluk, Kristi; Rivet, Bernadette; Borewicz, Klaudyna; Frye, Jonathan G; Englen, Mark; Anderson, Janet; Davies, Peter R

    2015-05-15

    IncA/C plasmids are broad-host-range plasmids enabling multidrug resistance that have emerged worldwide among bacterial pathogens of humans and animals. Although antibiotic usage is suspected to be a driving force in the emergence of such strains, few studies have examined the impact of different types of antibiotic administration on the selection of plasmid-containing multidrug resistant isolates. In this study, chlortetracycline treatment at different concentrations in pig feed was examined for its impact on selection and dissemination of an IncA/C plasmid introduced orally via a commensal Escherichia coli host. Continuous low-dose administration of chlortetracycline at 50 g per ton had no observable impact on the proportions of IncA/C plasmid-containing E. coli from pig feces over the course of 35 days. In contrast, high-dose administration of chlortetracycline at 350 g per ton significantly increased IncA/C plasmid-containing E. coli in pig feces (P IncA/C plasmid to other indigenous E. coli hosts. There was no evidence of conjugal transfer of the IncA/C plasmid to bacterial species other than E. coli. In vitro competition assays demonstrated that bacterial host background substantially impacted the cost of IncA/C plasmid carriage in E. coli and Salmonella. In vitro transfer and selection experiments demonstrated that tetracycline at 32 μg/ml was necessary to enhance IncA/C plasmid conjugative transfer, while subinhibitory concentrations of tetracycline in vitro strongly selected for IncA/C plasmid-containing E. coli. Together, these experiments improve our knowledge on the impact of differing concentrations of tetracycline on the selection of IncA/C-type plasmids.

  3. Quantitative Campylobacter spp., antibiotic resistance genes, and veterinary antibiotics in surface and ground water following manure application: Influence of tile drainage control.

    Science.gov (United States)

    Frey, Steven K; Topp, Edward; Khan, Izhar U H; Ball, Bonnie R; Edwards, Mark; Gottschall, Natalie; Sunohara, Mark; Lapen, David R

    2015-11-01

    This work investigated chlortetracycline, tylosin, and tetracycline (plus transformation products), and DNA-based quantitative Campylobacter spp. and Campylobacter tetracycline antibiotic resistant genes (tet(O)) in tile drainage, groundwater, and soil before and following a liquid swine manure (LSM) application on clay loam plots under controlled (CD) and free (FD) tile drainage. Chlortetracycline/tetracycline was strongly bound to manure solids while tylosin dominated in the liquid portion of manure. The chlortetracycline transformation product isochlortetracycline was the most persistent analyte in water. Rhodamine WT (RWT) tracer was mixed with manure and monitored in tile and groundwater. RWT and veterinary antibiotic (VA) concentrations were strongly correlated in water which supported the use of RWT as a surrogate tracer. While CD reduced tile discharge and eliminated application-induced VA movement (via tile) to surface water, total VA mass loading to surface water was not affected by CD. At both CD and FD test plots, the biggest 'flush' of VA mass and highest VA concentrations occurred in response to precipitation received 2d after application, which strongly influenced the flow abatement capacity of CD on account of highly elevated water levels in field initiating overflow drainage for CD systems (when water level <0.3m below surface). VA concentrations in tile and groundwater became very low within 10d following application. Both Campylobacter spp. and Campylobacter tet(O) genes were present in groundwater and soil prior to application, and increased thereafter. Unlike the VA compounds, Campylobacter spp. and Campylobacter tet(O) gene loadings in tile drainage were reduced by CD, in relation to FD.

  4. Use estimates of in-feed antimicrobials in swine production in the United States.

    Science.gov (United States)

    Apley, Michael D; Bush, Eric J; Morrison, Robert B; Singer, Randall S; Snelson, Harry

    2012-03-01

    When considering the development of antimicrobial resistance in food animals, comparing gross use estimates of different antimicrobials is of little value due to differences in potencies, duration of activity, relative effect on target and commensal bacteria, and mechanisms of resistance. However, it may be valuable to understand quantities of different antimicrobials used in different ages of swine and for what applications. Therefore, the objective of this project was to construct an estimate of antimicrobial use through the feed in swine production in the United States. Estimates were based on data from the National Animal Health Monitoring System (NAHMS) Swine 2006 Study and from a 2009 survey of swine-exclusive practitioners. Inputs consisted of number of pigs in a production phase, feed intake per day, dose of the antimicrobial in the feed, and duration of administration. Calculations were performed for a total of 102 combinations of antimicrobials (n=17), production phases (n=2), and reasons for use (n=3). Calculations were first conducted on farm-level data, and then extrapolated to the U.S. swine population. Among the nursery phase estimates, chlortetracycline had the largest estimate of use, followed by oxytetracycline and tilmicosin. In the grower/finisher phase, chlortetracycline also had the largest use estimate, followed by tylosin and oxytetracycline. As an annual industry estimate for all phases, chlortetracycline had the highest estimated use at 533,973 kg. The second and third highest estimates were tylosin and oxytetracycline with estimated annual uses of 165,803 kg and 154,956 kg, respectively. The estimates presented here were constructed to accurately reflect available data related to production practices, and to provide an example of a scientific approach to estimating use of compounds in production animals.

  5. Magnetic separation of antibiotics by electrochemical magnetic seeding

    Energy Technology Data Exchange (ETDEWEB)

    Ihara, I; Toyoda, K [Department of Agricultural Engineering and Socio Economics, Kobe University, Nada, Kobe 657-8501 (Japan); Beneragama, N; Umetsu, K [Department of Animal Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555 (Japan)], E-mail: ihara@port.kobe-u.ac.jp

    2009-03-01

    Magnetic separation of several classes of antibiotics was investigated using electrochemical magnetic seeding. Electrocoagulation with a sacrificial anode followed by addition of magnetite particles was applied for the magnetic seeding of antibiotics. With electrochemical magnetic seeding using an iron anode, tetracycline antibiotics (oxytetracycline, chlortetracycline, doxycycline and tetracycline) and cephalosporin antibiotic (cefdinir) were rapidly removed from synthetic wastewater by magnetic separation using a neodymium magnet. Iron and aluminium anodes were suitable for magnetic seeding of the antibiotics. The results indicated that the ability of antibiotics to form strong complex with iron and aluminium allowed the higher removal by magnetic separation. This method would be appropriate for rapid treatment of antibiotics in wastewater.

  6. Effects of disinfectants and antibiotics on the anaerobic digestion of piggery waste

    Energy Technology Data Exchange (ETDEWEB)

    Poels, J.; Van Assche, P.; Verstraete, W.

    1984-01-01

    The effects of disinfectants and antibiotics on the anaerobic digestion of piggery waste were investigated. The disinfectants Tego 51, Dettol, NaOCl and Creolin, and the antibiotics, chlortetracyclin, tylosin, erythromycin, chloramphenicol, bacitracin and virginiamycin were tested at different concentrations. At concentrations normally used in practice, no inhibitory effect on methanisation process was detected. However, higher concentrations of the antimicrobial agents, Dettol, Creolin, bacitracin and virginiamycin, markedly inhibited biogas production. In order to minimize possible digester failures, farmers are advised to respect the normal recommended dose and to use low-toxicity antimicrobial agents. (Refs. 8).

  7. A study on monitoring of pharmaceuticals and personal care products (PPCPs) in livestock wastewater and treatment by radiation techniques

    Energy Technology Data Exchange (ETDEWEB)

    Myung, Seung Woon [KAERI, Daejeon (Korea, Republic of)

    2010-05-15

    In this study, and effective monitoring and the investigation of treatment efficiency of pharmaceuticals from the influent and effluent of livestock wastewater treatment plant (WWTPs) and by-product with radiation processing by LC/ESI-MS/MS was performed. Thirteen pharmaceuticals including antibiotics, growth promoters and disinfectants were assayed from twelve WWTPs in South Korea. The established method could be used to determine low concentration levels of pharmaceuticals in environmental samples. From few influents of live-stock WWTPs, chlortetracycline and acetaminophen were detected with the highest concentration among the monitoring pharmaceuticals. And also lincomycin, sufathiazole, sulfamethazine, trimethoprim, acetyl salicylic acid, tylosin, glutaraldehyde and formaldehyde were detected from the influents of WWTPs

  8. Construction of an extended range whole-cell tetracycline biosensor by use of the tet(M) resistance gene

    DEFF Research Database (Denmark)

    Bahl, Martin Iain; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2005-01-01

    An extended range whole-cell tetracycline biosensor strain was constructed by insertion of the tet(M) gene, encoding tetracycline resistance by ribosomal protection, into plasmid pTGFP2, which contains a transcriptional fusion between a tetracycline regulated promoter and the green fluorescent...... protein gene. Tetracycline, oxytetracycline, chlortetracycline and minocycline all effectively induced the resulting Escherichia coli MC4100/pTGM biosensor and similar dose-response characteristics were recorded by flow cytometry for all four compounds. The novel tetracycline biosensor was responsive...

  9. A study of fluctuations in Escherichia coli sensitivity patterns from pigs fed a halquinol supplemented diet.

    Science.gov (United States)

    Cosgrove, R F; Forster, T C; Jones, G T; Pickles, R W

    1981-03-01

    Escherichia coli isolated from pigs fed on a medicated diet containing 120 p.p.m. halquinol did not develop any resistance to this addition over a 6-week period. Sensitivity patterns of the E. coli isolates to eight antimicrobial substances, although fluctuating slightly during the test period (but no more than a control group), did not significantly alter. However, the patterns did change significantly when for 17 days after the completion of the halquinol trial the pigs were fed a normal commercial ration medicated with a commonly used feed additive containing chlortetracycline hydrochloride, procaine penicillin and sulphadimidine.

  10. Fate of tetracycline, sulfonamide and fluoroquinolone resistance genes and the changes in bacterial diversity during composting of swine manure.

    Science.gov (United States)

    Selvam, Ammaiyappan; Xu, Delin; Zhao, Zhenyong; Wong, Jonathan W C

    2012-12-01

    This study monitored the abundance of antibiotic resistant genes (ARGs) and the bacterial diversity during composting of swine manure spiked with chlortetracycline, sulfadiazine and ciprofloxacin at two different levels and a control without antibiotics. Resistance genes of tetracycline (tetQ, tetW, tetC, tetG, tetZ and tetY), sulfonamide (sul1, sul2, dfrA1 and dfrA7) and fluoroquinolone (gyrA and parC) represented 0.02-1.91%, 0.67-10.28% and 0.00005-0.0002%, respectively, of the total 16S rDNA copies in the initial composting mass. After 28-42 days of composting, these ARGs, except parC, were undetectable in the composting mass indicating that composting is a potential method of manure management. Polymerase chain reaction-denaturing gradient gel electrophoresis analysis of bacterial 16S rDNA of the composting mass indicated that the addition of antibiotics up to 100, 20 and 20mg/kg of chlortetracycline, sulfadiazine and ciprofloxacin, respectively, elicited only a transient perturbation and the bacterial diversity was restored in due course of composting.

  11. Environmental monitoring study of selected veterinary antibiotics in animal manure and soils in Austria

    Energy Technology Data Exchange (ETDEWEB)

    Martinez-Carballo, Elena [Department of Hazardous Substances and Metabolites, Umweltbundesamt GmbH - Austrian Federal Environment Agency, Spittelauer Laende 5, A-1090 Vienna (Austria); Gonzalez-Barreiro, Carmen [Department of Hazardous Substances and Metabolites, Umweltbundesamt GmbH - Austrian Federal Environment Agency, Spittelauer Laende 5, A-1090 Vienna (Austria); Scharf, Sigrid [Department of Hazardous Substances and Metabolites, Umweltbundesamt GmbH - Austrian Federal Environment Agency, Spittelauer Laende 5, A-1090 Vienna (Austria); Gans, Oliver [Department of Hazardous Substances and Metabolites, Umweltbundesamt GmbH - Austrian Federal Environment Agency, Spittelauer Laende 5, A-1090 Vienna (Austria)]. E-mail: oliver.gans@umweltbundesamt.at

    2007-07-15

    LC-MS/MS was used for determination of selected tetracyclines, sulfonamides, trimethoprim, and fluoroquinolones in manure samples of pig, chicken and turkey, as well as arable soils fertilized with manure. Recoveries from spiked samples ranged from 61 to 105%. Method quantification limits were set to 100 {mu}g/kg for all substances. Analysis of 30 pig manure, 20 chicken and turkey dung, and 30 lyophilized soil samples taken in Austria revealed that in pig manure up to 46 mg/kg chlortetracycline, 29 mg/kg oxytetracycline and 23 mg/kg tetracycline could be detected. As representatives of the group of sulfonamides, sulfadimidine in pig manure and sulfadiazine in chicken and turkey dung were detected in significant amounts (maximum concentration, 20 and 91 mg/kg, respectively). Enrofloxacin was particularly observed in chicken and turkey samples. Positive detection of chlortetracycline, enrofloxacin, and ciprofloxacin, in soil samples should be outlined as most important results of this study. - Specific exposure data of selected veterinarian antibiotics in manure and samples of agriculturally used soils are reported for the first time in Austria.

  12. Degradation of typical antibiotics during human feces aerobic composting under different temperatures.

    Science.gov (United States)

    Shi, Honglei; Wang, Xiaochang C; Li, Qian; Jiang, Shanqing

    2016-08-01

    Four typical antibiotics were added to human feces for aerobic composting using batch reactors with sawdust as the bulk matrix. Under three composting temperatures (room temperature, 35 ± 2 °C and 55 ± 2 °C), decreases in the extractable concentrations of antibiotics in the compost were monitored for 20 days. As a result, the removals of extractable tetracycline and chlortetracycline were found to be more temperature-dependent than the removals of sulfadiazine and ciprofloxacin. However, more than 90 % of all of the extractable antibiotics were removed at 55 ± 2 °C. Three specific experiments were further conducted to identify the possible actions for antibiotic removal, including self-degradation in aqueous solution, composting with a moist sterile sawdust matrix without adding feces and composting with human feces and moist sterile sawdust. As a result, it was found that the removal of tetracycline and chlortetracycline was mainly due to chemical degradation in water, whereas the removal of sulfadiazine was mainly attributed to adsorption onto sawdust particles. The microbial activity of compost varied with temperature to a certain extent, but the differences were insignificant among different antibiotics. Although microbial action is important for organic matter decomposition, its contribution to antibiotic degradation was small for the investigated antibiotics, except for ciprofloxacin, which was degraded by up to 20 % due to microbial action.

  13. Characterization of Antibiotics and Antibiotic Resistance Genes on an Ecological Farm System

    Directory of Open Access Journals (Sweden)

    Songhe Zhang

    2015-01-01

    Full Text Available There is a growing concern worldwide about the prevalence of antibiotics and antibiotic resistance genes (ARGs on the farm. In this study, we investigated the distribution of seven antibiotics and ten ARGs in fresh and dried pig feces, in biogas slurry, and in grape-planting soil from an ecological farm. Antibiotics including sulfamethazine, norfloxacin, ofloxacin, tetracycline, oxytetracycline, and chlortetracycline were detected in these samples (except for sulfamethoxazole in dried feces. In general, antibiotics levels in samples were in the sequence: biogas slurry > fresh feces > soil or dried feces. Results of ecological risk assessments revealed that among the seven antibiotics chlortetracycline showed the highest ecological risk. Among the ten ARGs, sulI and tetO were the most prevalent on this ecological farm. There were positive correlations between certain ARGs and the corresponding antibiotics on this ecological farm. Therefore, continuous monitoring of antibiotics and their corresponding ARGs should be conducted in the agroecosystem near the concentrated animal farming operation systems.

  14. Microbial inhibition by pharmaceutical antibiotics in different soils--dose-response relations determined with the iron(III) reduction test.

    Science.gov (United States)

    Thiele-Bruhn, Sören

    2005-04-01

    Soil contamination from pharmaceuticals is an emerging problem, though quantitative data on their microbial effects are lacking. Thus, nine pharmaceutical antibiotics were tested for their effects on the microbial iron(III) reduction in six different topsoils. Complete dose-response curves were obtained and best-fit by sigmoidal Logit, Weibull, Box-Cox Logit, and Box-Cox Weibull equations (r2 0.73-1.00). The derived effective doses (ED [micromol/kg soil]) for the different antibiotics increased in the order (average ED50 in parentheses) chlortetracycline (53) fenbendazole at doses up to 5,800 and 3,300 micromol/kg, respectively. Due to a strong soil adsorption, especially of the tetracyclines, the corresponding effective concentrations in the soil solution (EC50), derived from sorption experiments, were considerably smaller and ranged from 0.004 micromol/L (chlortetracycline) to 120 micromol/L (sulfapyridine). The effects of the antibiotics were governed by soil sorptive properties, especially the concentration of soil organic matter. The microbial inhibition was influenced indirectly by the soil pH, which affects the ionization status of the amphoteric antibiotics.

  15. Development of adsorptive membranes by confinement of activated biochar into electrospun nanofibers

    Directory of Open Access Journals (Sweden)

    Mehrdad Taheran

    2016-11-01

    Full Text Available Adsorptive membranes have many applications in removal of contaminants, such as heavy metals and organic contaminants from water. Recently, increasing concentrations of pharmaceutically active compounds, especially antibiotics, such as chlortetracycline in water and wastewater sources has raised concerns about their potentially adverse impacts on environment and human health. In this study, a series of polyacrylonitrile (PAN/activated biochar nanofibrous membranes (NFMs with different loadings of biochar (0–2%, w/w were fabricated using electrospinning. The morphology and structure of fabricated membranes was investigated by scanning electron microscopy, Fourier transform infrared and thermogravimetric analysis. The results showed that at 1.5% of biochar loading, the surface area reached the maximum value of 12.4 m2/g and beyond this loading value, agglomeration of particles inhibited fine interaction with nanofibrous matrix. Also, the adsorption tests using chlortetracycline showed that, under environmentally relevant concentrations, the fabricated adsorptive NFMs had a potential for removal of these types of emerging contaminants from water and wastewaters.

  16. Liquid chromatography – tandem mass spectrometry method for the determination of ten tetracycline residues in muscle samples

    Directory of Open Access Journals (Sweden)

    Gajda Anna

    2015-09-01

    Full Text Available A liquid chromatography – tandem mass spectrometry (LC-MS/MS method for the determination of oxytetracycline (OTC, 4-epi oxytetracycline (4-epi OTC, tetracycline (TC, 4-epi tetracycline (4-epi TC, chlortetracycline (CTC, 4-epi chlortetracycline (4-epi CTC, doxycycline (DC, minocycline (MINO, methacycline (META and rolitetracycline (ROLI residues in muscles was developed. The procedure consisted of an oxalic acid extraction followed by protein removal with trichloroacetic acid. Further solid phase clean-up on polymeric (Strata X reversed phase columns was performed to obtain an extract suitable for LC-MS/MS analysis. The tetracyclines were separated on a C 18 analytical column with mobile phase consisting of 0.01% formic acid in acetonitrile and 0.01% formic acid in water in gradient mode. The method was validated according to the Commission Decision 2002/657/EC. The recoveries of all target compounds were 91.8% – 103.6%. The decision limits were from 109.0 to 119.8 μg/kg and detection capability varied within the range of 122.2 to 137.6 μg/kg, depending on the analyte.

  17. Tetracycline residues in meat and bone meals. Part 1: methodology and examination of field samples.

    Science.gov (United States)

    Körner, U; Kühne, M; Wenzel, S

    2001-04-01

    The incidence of tetracycline residues in commercially available samples of meat meals and meat and bone meals was investigated. Four different methods were used on each of the 87 samples from nine different rendering plants in Germany: examination using a screening fluorescence test; examination by high performance liquid chromatography analysis using three different extraction procedures (succinate buffer, hydrochloric acid, hydrochloric acid after sedimentation of bone particles). Tetracyclines were found in 100% of the samples by one or more of the extraction procedures. The highest concentrations found in meat meals were 2048 microgkg(-1), 1393 microgkg(-1) and 608 microgkg(-1) for oxytetracycline, tetracycline and chlortetracycline, respectively. In meat and bone meals the highest concentrations were 2295 microg oxytetracycline kg(-1) 848 microg tetracycline kg(-1) and 1274 microg chlortetracycline kg(-1). The extraction after sedimentation was the most effective of the applied extraction procedures and exposed the highest total tetracycline concentrations. The results of this investigation showed that considerable amounts of tetracyclines have to be expected in field samples. Further research has to be done on the heat stability of bound tetracycline residues.

  18. 全麻下头面部手术患者眼保护方法的对比研究%Eye-protection for operative patients with craniofacial surgery undergoing general anesthesia.

    Institute of Scientific and Technical Information of China (English)

    谭云燕; 宁传艺; 刘丽清; 韦江平

    2012-01-01

    Objective:To compare and assess the efficacy of eye ointment under general anesthesia eye -protection care, aim to find out the more effective measure for clinical nursing. Methods:200 general anesthesia patients were randomly divided into chlortetracycline group and tetracycline group. After the 1th ,3rd, 5th days of postoperative, we observed the clinical outcome including the availability of eye irritation, photophobia, tearing the cornea, conjunctivitis symptoms, and trichiasis, conjunctival hyperemia, corneal ulcers, vision loss and other concurrent disease and the complaints. Results:Observation outcome indicates that chlortetracycline group with 2 complications,tetracycline group had no ocular complications,no significant difference between groups( P >0. 05 ). Chlortetracycline group had photophobia, tearing and foreign body sensation with tingling in 15 cases,there was only 1 case of tetracycline group with tearing and photophobia,and 3 cases with foreign body sensation, was statistically significant in the results. Conclusions:There is no difference in the effective of eye - protection in chlortetracycline ointment and tetracycline ointment in general anesthesia patients. And the tetracycline ointment can reduce the uncomfortable symptoms more than the chlortetracycline ointment.%目的:对患者分别用金霉素软膏和四环素软膏进行眼保护护理比较,旨在寻求有效保护全麻患者眼睛的方法.方法:分别对我院需行全麻下头面部手术患者(200例)随机分成金霉素和四环素两组各100例,按照术中眼保护护理方法分别进行护理,在术后第1,3,5 d对两组患者的眼部表现及患者主诉,包括有无眼部刺痛、畏光、流泪等角膜、结膜炎症状,以及倒睫、结膜充血、角膜溃疡、视力减退等并发症及不适主诉进行比较.结果:金霉素组出现并发症2例,分别为角膜溃疡和视力减退,四环素组无眼部并发症,组间比较无统计学意义(P>0.05);金

  19. Photohemolytic potency of tetracyclines

    Energy Technology Data Exchange (ETDEWEB)

    Bjellerup, M.; Ljunggren, B.

    1985-04-01

    Hemolysis induced by long-wave ultraviolet radiation (UVA) and 8 different commercial tetracycline derivatives was studied in a model using human red blood cells. Demethylchlortetracycline and doxycycline were shown to have pronounced hemolytic properties causing 88% and 85% hemolysis, respectively, at a concentration of 50 micrograms/ml and 72 J/ cm2 of UVA. Tetracycline, oxytetracycline, and chlortetracycline caused maximally 18% hemolysis at 200 micrograms/ml and lymecycline only 7% at 100 micrograms/ml. Methacycline showed intermediate hemolytic effect of 36% at 200 micrograms/ml. Minocycline had no hemolytic effect whatsoever. These experimental data correlate very well with clinical reports and comparative phototoxicity trials in humans. Photohemolysis may thus be of value for predicting tetracycline phototoxicity.

  20. Chemiluminescence determination of tetracyclines using Fenton system in the presence europium(III) ions

    Energy Technology Data Exchange (ETDEWEB)

    Kaczmarek, Malgorzata [Department of Rare Earths, Faculty of Chemistry, Adam Mickiewicz University, Grunwaldzka 6, 60 - 780 Poznan (Poland); Lis, Stefan, E-mail: blis@amu.edu.pl [Department of Rare Earths, Faculty of Chemistry, Adam Mickiewicz University, Grunwaldzka 6, 60 - 780 Poznan (Poland)

    2009-04-20

    A new simple chemiluminescent method for the determination of chlortetracycline (Chlor-TC), oxytetracycline (Oxy-TC) and doxycycline (Doxy-TC) is described. This method is based on the europium(III) emission as a result of the energy transfer process from the excited product of the tetracyclines oxidation to the uncomplexed Eu(III). Under the optimum conditions, calibration graphs were obtained for 4 x 10{sup -7} to 2 x 10{sup -5} mol L{sup -1} of Chlor-TC; 2 x 10{sup -7} to 2 x 10{sup -5} mol L{sup -1} of Oxy-TC and 1 x 10{sup -7} to 3 x 10{sup -5} mol L{sup -1} of Doxy-TC. The method was successfully applied to the determination of these drugs in pharmaceutical and veterinary formulation and honey.

  1. In vitro sensitivities to antimicrobial drugs of ureaplasmas isolated from the bovine respiratory tract, genital tract and eye.

    Science.gov (United States)

    Kishima, M; Hashimoto, K

    1979-09-01

    The sensitivity to 18 antimicrobial drugs was examined for 66 strains of Ureaplasma sp isolated from respiratory tracts of calves suffering from enzootic pneumonia, urinary tracts of bulls and eyes of cows suffering from infectious bovine kerato-conjunctivitis. Furamizole, tiamulin fumarate, erythromycin lactobionate, malidomycin C, doxycycline hydrochloride, kitasamycin tartrate, tylosin tartrate, T-2636C, tetracycline hydrochloride, oxytetracycline hydrochloride, chlortetracycline hydrochloride, oleandomycin phosphate, furazolidone, spiramycin adipate, chloramphenicol and thiophenicol showed strong inhibiting activity on all the test strains. Among them, furamizole, tiamulin fumarate and erythromycin lactobionate were most active. Kanamycin sulphate showed weak activity on all the strains tested. The differences in origin of the test strains did not affect their sensitivity to any of the drugs.

  2. Effects of ubiquitin-protea some pathway on mouse sperm capacitation, acrosome reaction and in vitro fertilization

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Chlortetracycline (CTC) fluorescence patterns were used to study changes in the patterns B and AR of mouse sperm after incubation with reagents that would block the UPP. They were the monoclonal antibody against ubiquitinated proteins-UCP1; the polyclonal antibody against ubiquitin anti-Ub, and a special inhibitor against proteasome ALLN. Furthermore, we treated the capacitated sperm or the eggs with these reagents separately and tested whether the normal in vitro fertilization was blocked or not. Results illustrate that UCP1, anti-Ub, and ALLN have little effects on sperm capacitation and acrosome reaction,but they do inhibit fusion of mouse sperm with eggs, which suggests that UPP play an important role in mouse in vitro fertilization.``

  3. 应用发光细菌法检测饲用抗生素单一及联合毒性的研究%Primary Research on Single and Combined Toxicity of Antibiotics in Feeds by Luminescent Bacteria

    Institute of Scientific and Technical Information of China (English)

    任皓; 王金荣; 陈行杰; 刘俊功; 娄鹏

    2011-01-01

    以HgCl2为毒性参照,以等毒性配比法研究了饲用盐酸金霉素、吉他霉素、盐霉素和黄霉素对发光细菌的单一及联合毒性.结果表明:盐酸金霉素、吉他霉素、盐霉素、黄霉素对明亮发光杆菌的急性毒性EC50分别为9.21、146.13、4.24 mg/L和134.68mg/L,发光细菌的相对发光强度(RLU)随着抗生素浓度的提高而降低,并分别在2~16、25~200、1~8 mg/L和25~200mg/L范围内呈良好线性关系;多元饲用抗生素对发光细菌具有联合毒性的作用,吉他霉素与盐酸金霉素、盐霉素和黄霉素3种抗生素的二元组合对发光细菌的联合毒性表现为较强的协同作用,而盐酸金霉素、盐霉素和黄霉素两两分别组合对发光细菌的联合毒性表现为拮抗作用;三元及四元抗生素混合体系对发光细菌的联合毒性均表现为拮抗作用.%Using the equitoxic ratio analysis, the single and combined toxicity effects of Chlortetracycline, Kitasamycin, Salinomycin and Flavomycin were studied by luminescent bacteria based on the toxicity reference of HgC12. Results showed that the acute toxicity EC50 of Chlortetracycline, Kitasamycin, Flavomycin and Salinomycin to Photosbacterium phosphoreum were 9.21, 146.13, 4.24 and 134.68 mg/L, respectively. The relative luminous intensity of luminous bacteria (RLU) were decreased with increasing antibiotic concentration, and there were well linerly for these four antibiotics at the range of 2~16 mg/L, 25~200 mg/L, 1-8 mg/L and 25-200 mg/L, respectively. The combined toxicity effects of Kitasamycin with one of the Chlortetracycline, Salinomycin and Flavomycin were showed synergistic effect. Any two of Chlortetracycline, Salinomycin and Flavomycin were showed antagonistic combined toxicity effects, and the combined toxicity of three and four antibiotic mixtures on luminescent bacteria were showed antagonistic effects.

  4. Occurrence of antibiotics in soils and manures from greenhouse vegetable production bases of Beijing, China and an associated risk assessment.

    Science.gov (United States)

    Li, Cheng; Chen, Jiayi; Wang, Jihua; Ma, Zhihong; Han, Ping; Luan, Yunxia; Lu, Anxiang

    2015-07-15

    The occurrence of 15 antibiotics in soil and manure samples from 11 large-scale greenhouse vegetable production (GVP) bases in Beijing, China was investigated. Results showed that the greenhouse soils were ubiquitously contaminated with antibiotics, and that antibiotic concentrations were significantly higher in greenhouses than in open field soils. The mean concentrations of four antibiotic classes decreased in the following order: tetracyclines (102μg/kg)>quinolones (86μg/kg)>sulfonamides (1.1μg/kg)>macrolides (0.62μg/kg). This investigation also indicated that fertilization with manure and especially animal feces might be the primary source of antibiotics. A risk assessment based on the calculated risk quotients (RQs) demonstrated that oxytetracycline, chlortetracycline, norfloxacin, ciprofloxacin and enrofloxacin could pose a high risk to soil organisms. These results suggested that the ecological effects of antibiotic contamination in GVP bases and their potential adverse risks on human health need to be given special attention.

  5. Rapid detection of tetracyclines and their 4-epimer derivatives from poultry meat with bioluminescent biosensor bacteria.

    Science.gov (United States)

    Virolainen, Nina E; Pikkemaat, Mariël G; Elferink, J W Alexander; Karp, Matti T

    2008-12-10

    Tetracycline (TC) specific luminescent bacterial biosensors were used in a rapid TC residue assay sensitized to meet the EU maximum residue limit (MRL) for TC residues in poultry muscle tissue (100 microg kg(-1)) by membrane-permeabilizing and chelating agents polymyxin B and EDTA. Sensitivities of 5 ng g(-1) for doxycycline, 7.5 ng g(-1) for chlortetracycline, and 25 ng g(-1) for tetracycline and oxytetracycline were reached. Except for doxycycline, the MRLs of these tetracyclines include their 4-epimer metabolites. In the biosensor assay, all four 4-epimers showed induction capacity and antimicrobial activity, and antimicrobial activity was also observed in the inhibition assay, although with lower efficiency than that of the corresponding parent compound in both assays. The biosensor assay is an inexpensive and rapid high-throughput screening method for the detection of 4-epimer TC residues along with their parent compounds.

  6. Immunolocation of antisperm monoclonal antibody 6B10 and corresponding antigen

    Institute of Scientific and Technical Information of China (English)

    高绍荣; 胡国俊; 段崇文; 刘辉; 韩之明; 宋祥芬; 陈大元

    1999-01-01

    An antisperm monoclonal antibody 6B10 was produced by hybridoma technique of the isotype IgG. The monoclonal antibody was purified by means of ammonium sulfate precipitation and protein A-Sepharose Cl-4B affinity chromatography. SDS polyacrylamide gel electrophoresis was used to evaluate the purity of the antibody. Evaluation of the sperm acrosomal status was determined by chlortetracycline (CTC) staining. It was found that monoclonal antibody 6B10 can inhibit the sperm acrosome reaction induced by progesterone. The corresponding antigen recognized by monoclonal antibody 6B10 was located on the plasma membrane of the sperm acrosome by indirect immunofluorescent microscopy and immunoelectronmicroscopy. Sperm protein was extracted by 1% Triton X-100. The molecular weight of the antigen is 50 ku, detected by Western blot. The antigen is a key protein in the sperm acrosome reaction and may be the receptor of progesterone on the sperm acrosome. It may either be developed as a candidate contraceptive vaccine

  7. Hierarchical La0.7Ce0.3FeO3/halloysite nanocomposite for photocatalytic degradation of antibiotics

    Science.gov (United States)

    Li, Xiazhang; Zhu, Wei; Yan, Xiangyu; Lu, Xiaowang; Yao, Chao; Ni, Chaoying

    2016-08-01

    The hierarchical La0.7Ce0.3FeO3/halloysite nanotubes (HNTs) composites have been successfully prepared via sol-gel method. XRD and TEM characterizations indicated that the sheet-like La0.7Ce0.3FeO3 coupled with the co-precipitated CeO2 were evenly deposited onto the surface of halloysite. The photocatalytic degradation of chlortetracycline under visible light irradiation using La0.7Ce0.3FeO3/HNTs as catalyst was evaluated by high-performance liquid chromatography, which exhibited remarkable photocatalytic activity with the removal rate up to 99 % in 90 min, due to the formation of "solid solution/co-precipitation" heterostructure as well as the excellent adsorptive capability of halloysite for antibiotics.

  8. [Sensitivity of Pseudomonas chlororaphis to antibiotics and chemical tools of plant protection].

    Science.gov (United States)

    Shepelevich, V V; Kiprianova, E A; Iaroshenko, L V; Avdeeva, L V

    2012-01-01

    Examination of sensitivity of 10 Pseudomonas chlororaphis strains belonging to different subspecies to 54 antibiotics has shown that all studied representatives of Pchlororaphis subsp. chlororaphis, P. chlororaphis subsp. aureofaciens and Pchlororaphis subsp. aurantiaca were sensitive to aminoglycoside antibiotics and fluoroquinolones derivatives. Only part of studied strains has shown sensitivity to some beta-lactam antibiotics, imipeneme and meropeneme. In contrast to representatives of two other subspecies both strains of P. chlororaphis subsp. chlororaphis proved to be sensitive to chlortetracycline and cefepime that allows to consider this difference as the characteristic useful for differentiation. All studied P. chlororaphis strains were resistant to chemical fungicides (Scor and Svitch) and the insect growth regulators (Match, Lufox, Engio, Actellik). Resistance of bacteria to these herbicides gives evidence that their combined use is possible.

  9. A quantum dot-based immunoassay for screening of tetracyclines in bovine muscle.

    Science.gov (United States)

    García-Fernández, Jenifer; Trapiella-Alfonso, Laura; Costa-Fernández, José M; Pereiro, Rosario; Sanz-Medel, Alfredo

    2014-02-19

    A reliable and robust direct screening methodology based on a quantum dot (QD) fluorescent immunoassay has been developed to detect trace levels of different antibiotic species from the family of the tetracyclines (e.g., oxytetracycline, tetracycline, chlortetracycline, and doxycycline) in contaminated bovine muscle tissues. First, the synthesis and characterization of a new immunoprobe (oxytetracycline-bovine serum albumin-QD) has been carried out for its further application in the development of a competitive fluorescent QD-immunoassay. The developed fluoroimmunoassay provides sensitive and binary "yes/no" responses being appropriate for the screening of this family of antibiotics above or below a preset concentration threshold. The detection limit achieved with this strategy, 1 μg/L in aqueous media and 10 μg/kg in bovine muscle samples, is 10-fold lower than the maximum level concentration allowed by International Legislation in muscle tissue, enabling suitable and efficient screening of the antibiotics.

  10. A confirmatory method for the simultaneous extraction, separation, identification and quantification of Tetracycline, Sulphonamide, Trimethoprim and Dapsone residues in muscle by ultra-high-performance liquid chromatography-tandem mass spectrometry according to Commission Decision 2002/657/EC.

    Science.gov (United States)

    McDonald, Mark; Mannion, Celine; Rafter, Paul

    2009-11-13

    A rapid confirmatory multi-residue method for the analysis of tetracyclines, sulphonamides, trimethoprim and dapsone by UPLC-MS/MS is described. The method is able to quantify and confirm the following 19 compounds, oxytetracycline, tetracycline, chlortetracycline, doxycycline, sulfadiazine, sulfathiazole, sulfapyridine, trimethoprim, sulfamerazine, sulfamethizole, sulfamethazine, sulfamethoxypyridazine, sulfamonomethoxine, sulfachlorpyridazine, dapsone, sulfamethoxazole, sulfisoxazole, sulfaquinoxaline and sulfadimethoxine. Samples are extracted with 0.1M EDTA and acetonitrile, which is then evaporated under a stream of nitrogen and reconstituted in water. Following centrifugation and filtering, an aliquot is analysed by UPLC-MS/MS using positive electrospray ionisation and multiple reaction monitoring. The method is deemed rapid as all analytes are extracted by a single extraction technique, with no solid-phase extraction clean up required. Validation is according to Commission Decision 2002/657/EC and was carried out for bovine, porcine, ovine and poultry species. Specificity, recovery, repeatability, reproducibility, CCalpha and CCbeta data is presented.

  11. Keutuhan Membran Spermatozoa Disekuensing Sentrifugasi Gradien Densitas Percoll Berpengencer Andromed dan CEP-2 yang Ditambahkan Kuning Telur (MEMBRANE INTACT OF SPERMATOZOA FOLLOWING SEXING USING PERCOLL DENSITY GRADIENT CENTRIFUGATION IN ANDROMED AND C

    Directory of Open Access Journals (Sweden)

    Yudha Fika Diliyana

    2014-05-01

    Full Text Available The aim of this study was to observe the best extender in protecting the membrane of bovine spermatozoafollowing sexing by percoll density gradient centrifugation. Freshly collected semen were obtained fromBalai Besar Inseminasi Buatan Singosari-Malang. The semen were diluted in andromed and CaudalEpididymal Plasma-2 (CEP-2 added with 10% egg yolk extenders.The sperm membrane integrity wasobserved using Hypo-osmotic Swelling Test (HOST. Sperm capacitation and acrososome reaction wereassessed using Chlortetracycline Fluorescence Assay.The results showed that andromed and CEP-2 addedwith 10% egg yolk were able to retain the sperm membrane integrity, whereas sperm capacitation andacrosome reaction were kept low. Caudal Epididymal Plasma-2 (CEP-2 added with 10% egg yolk seemedto give better protection towards the sperm membrane intact in comparison to andromed extender.

  12. Effect of tetracycline residues in pig manure slurry on tetracycline-resistant bacteria and resistance gene tet(M) in soil microcosms

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Wulff, Gitte; Vaclavik, Elvira;

    2006-01-01

    manure slurry and a tetracycline-resistant Enterococcus faecalis, CG I 10, containing the tetracycline resistance gene tet(m) (on the conjugative transposon, Tn916). The prevalence of both tetracycline-resistant aerobic bacteria and tetracycline-resistant enterococci declined rapidly until day 45 where...... no significant differences in the levels of tetracycline-resistant bacteria in any of the four types of microcosms could be detected. tet(M) could be detected in microcosms supplemented with either pig manure slurry and/or E. faecalis CG 110 (tet(M)) for the whole period (152 days). tet(M) could be detected...... longer than tetracycline-resistant enterococci could be isolated (limit of detection 100 CFU/g soil) probably due to viable but not culturable (VBNC) bacteria with tet(M), horizontal gene transfer of tet(M) to indigenous soil bacteria or presence of "free" DNA. The concentration of chlortetracycline...

  13. Isolation of tetracyclines in milk using a solid-phase extracting column and water eluent.

    Science.gov (United States)

    Furusawa, Naoto

    2003-01-01

    An isolating method using a solid-phase extraction (SPE) ISOLUTE(R) C8 endcapped syringe-column for routine monitoring of residual tetracyclines (TCs) (oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), and doxycycline (DC)) in cow's milk is presented. In the simplest and most environmentally harmless method, milk samples could be applied directly to the SPE column, following which all TCs were eluted with water. No organic solvents were used at all. The purified sample was injected into a high-performance liquid chromatography (HPLC) with a photo-diode array detector (PDAD). For the HPLC determination/identification, a LiChrospher(R) 100 RP-8 endcapped column and a mobile phase of acetonitrile -7% (v v(-1)) acetic acid solution (in water) (35:65, v v(-1)) with a PDAD was used. The total time required for the analysis of one sample was 80 and <5%, respectively.

  14. Efficacy of salicylic acid in the treatment of digital dermatitis in dairy cattle

    DEFF Research Database (Denmark)

    Schultz, N.; Capion, N.

    2013-01-01

    Digital dermatitis (DD) is one of the most important causes of lameness in dairy cattle worldwide. The objective of this study was to evaluate the efficacy of salicylic acid in the treatment of the disease. A total of 201 DD lesions from 173 cows from four commercial dairy herds were evaluated...... at day 0 during routine hoof trimming and were allocated into two groups, namely, a control group given chlortetracycline spray, and a treatment group given 10 g of salicylic acid powder applied topically within a bandage. Pain, lesion size and clinical appearance (scored MO to M4) were evaluated on days...... the control group were 2.2 times more likely (P = 0.09) to have a pain score equal to 2 by day 14. The proportion of lesions getting smaller by days 14 and 34 was 2.5 times higher (P salicylic acid should be considered as an alternative...

  15. Expansion of Shiga Toxin-Producing Escherichia coli by Use of Bovine Antibiotic Growth Promoters.

    Science.gov (United States)

    Kim, Jong-Chul; Chui, Linda; Wang, Yang; Shen, Jianzhong; Jeon, Byeonghwa

    2016-05-01

    Antibiotics are routinely used in food-producing animals to promote growth and prevent infectious diseases. We investigated the effects of bovine antibiotic growth promoters (bAGPs) on the propagation and spread of Shiga toxin (Stx)-encoding phages in Escherichia coli. Co-culture of E. coli O157:H7 and other E. coli isolated from cattle in the presence of sublethal concentrations of bAGPs significantly increased the emergence of non-O157, Stx-producing E. coli by triggering the SOS response system in E. coli O157:H7. The most substantial mediation of Stx phage transmission was induced by oxytetracyline and chlortetracycline, which are commonly used in agriculture. bAGPs may therefore contribute to the expansion of pathogenic Stx-producing E. coli.

  16. Hierarchical La{sub 0.7}Ce{sub 0.3}FeO{sub 3}/halloysite nanocomposite for photocatalytic degradation of antibiotics

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xiazhang; Yao, Chao [Changzhou University, Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, School of Petrochemical Engineering, Changzhou (China); Chinese Academy of Science, R and D Center of Xuyi Attapulgite Applied Technology, Xuyi (China); Zhu, Wei; Yan, Xiangyu [Changzhou University, Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, School of Petrochemical Engineering, Changzhou (China); Lu, Xiaowang [Chinese Academy of Science, R and D Center of Xuyi Attapulgite Applied Technology, Xuyi (China); Ni, Chaoying [University of Delaware, Center for Advanced Microscopy and Microanalysis, Newark, DE (United States)

    2016-08-15

    The hierarchical La{sub 0.7}Ce{sub 0.3}FeO{sub 3}/halloysite nanotubes (HNTs) composites have been successfully prepared via sol-gel method. XRD and TEM characterizations indicated that the sheet-like La{sub 0.7}Ce{sub 0.3}FeO{sub 3} coupled with the co-precipitated CeO{sub 2} were evenly deposited onto the surface of halloysite. The photocatalytic degradation of chlortetracycline under visible light irradiation using La{sub 0.7}Ce{sub 0.3}FeO{sub 3}/HNTs as catalyst was evaluated by high-performance liquid chromatography, which exhibited remarkable photocatalytic activity with the removal rate up to 99 % in 90 min, due to the formation of ''solid solution/co-precipitation'' heterostructure as well as the excellent adsorptive capability of halloysite for antibiotics. (orig.)

  17. Determination of Tetracyclines in Honey Using Liquid Chromatography with Ultraviolet Absorbance Detection and Residue Confirmation by Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    LIU,Yan; XU,Jin-Zhong; DING,Tao; LI,Gong-Hai

    2007-01-01

    A determination method has been optimized and validated for the simultaneous analysis of tetracycline (TC),oxytetracycline (OTC), chlortetracycline (CTC) and doxycycline (DC) in honey. Tetracyclines (TCs) were removed from honey samples by chelation with metal ions bound to small Chelating Sepharose Fast Flow columns and eluted with Na2EDTA-Mcllvaine pH 4.0 buffers. Extracts were further cleaned up by Oasis HLB solid-phase extraction (SPE), while other solid-phase extraction cartridges were compared. Chromatographic separation was achieved using a polar end-capped C18 column with an isocratic mobile phase consisting of oxalic acid, acetonitrile,and methanol. LC with ultraviolet absorbance at 355 nm resulted in the quantitation of all four tetracycline residues from honey samples fortified at 15, 50, and 100 ng/g, with liner ranges for tetracyclines of 0.05 to 2 μg/mL. Mean recoveries for tetracyclines were greater than 50% with R.S.D. values less than 10% (n= 18). Detection limits of 5,5, 10, 10 ng/g for oxytetracycline, tetracycline, chlortetracycline and doxycycline, respectively and quantitation limits of 15 ng/g for all the four tetracyclines were determined. Direct confirmation of the four residues in honey (2-50 ng/g) was realized by liquid chromatography-tandem mass spectrometry (LC/MS/MS). The linear ranges of tetracyclines determined by LC/MS/MS were between 5 to 300 ng/mL, with the linear correlation coefficient r>0.995. The limits of detection of 1 to 2 ng/g were obtained for the analysis of the TCs in honey.

  18. Effect of subtherapeutic administration of antibiotics on the prevalence of antibiotic-resistant Escherichia coli bacteria in feedlot cattle.

    Science.gov (United States)

    Alexander, T W; Yanke, L J; Topp, E; Olson, M E; Read, R R; Morck, D W; McAllister, T A

    2008-07-01

    Antibiotic-resistant Escherichia coli in 300 feedlot steers receiving subtherapeutic levels of antibiotics was investigated through the collection of 3,300 fecal samples over a 314-day period. Antibiotics were selected based on the commonality of use in the industry and included chlortetracycline plus sulfamethazine (TET-SUL), chlortetracycline (TET), virginiamycin, monensin, tylosin, or no antibiotic supplementation (control). Steers were initially fed a barley silage-based diet, followed by transition to a barley grain-based diet. Despite not being administered antibiotics prior to arrival at the feedlot, the prevalences of steers shedding TET- and ampicillin (AMP)-resistant E. coli were >40 and <30%, respectively. Inclusion of TET-SUL in the diet increased the prevalence of steers shedding TET- and AMP-resistant E. coli and the percentage of TET- and AMP-resistant E. coli in the total generic E. coli population. Irrespective of treatment, the prevalence of steers shedding TET-resistant E. coli was higher in animals fed grain-based compared to silage-based diets. All steers shed TET-resistant E. coli at least once during the experiment. A total of 7,184 isolates were analyzed for MIC of antibiotics. Across antibiotic treatments, 1,009 (13.9%), 7 (0.1%), and 3,413 (47.1%) E. coli isolates were resistant to AMP, gentamicin, or TET, respectively. In addition, 131 (1.8%) and 143 (2.0%) isolates exhibited potential resistance to extended-spectrum beta-lactamases, as indicated by either ceftazidime or cefpodoxime resistance. No isolates were resistant to ciprofloxacin. The findings of the present study indicated that subtherapeutic administration of tetracycline in combination with sulfamethazine increased the prevalence of tetracycline- and AMP-resistant E. coli in cattle. However, resistance to antibiotics may be related to additional environmental factors such as diet.

  19. Implementation of the Bacillus cereus microbiological plate used for the screening of tetracyclines in raw milk samples with STAR protocol - the problem with false-negative results solved.

    Science.gov (United States)

    Raspor Lainšček, P; Biasizzo, M; Henigman, U; Dolenc, J; Kirbiš, A

    2014-01-01

    In antibiotic residue analyses the first step of screening is just as important as the following steps. Screening methods need to be quick and inexpensive, but above all sensitive enough to detect the antibiotic residue at or below the maximum residue limit (MRL). In the case of a positive result, the next step is conducted and further methods are used for confirmation. MRLs stated in European Union Regulation 37/2010 for tetracyclines in raw milk are: 100 µg kg(-1) for tetracycline, 100 µg kg(-1) for oxytetracycline, 100 µg kg(-1) for chlortetracycline and no limit for doxycycline because it is prohibited for use in animals from which milk is produced for human consumption. The current five-plate microbiological screening method for the detection of antibiotic residues in raw milk was found to be simple and inexpensive, but not specific, sensitive and reliable enough to detect tetracycline at MRL in routine raw milk screening procedures. Spiking samples with tetracycline at the MRL level and applying them on Bacillus cereus ATCC 11778 microbiological plates often gave false-negative results, indicating that tetracyclines may have to be inactivated or masked. Tetracyclines seem to bind to a certain component in milk. Consequently, when applying samples to the B. cereus microbiological plate the antibiotic cannot inhibit the growth of B. cereus which disables the formation of inhibition zones on the test plate. After adding the appropriate amount of citric acid into the milk samples, we solved the problem of false-negative results. During the validation 79 samples of milk were spiked with tetracyclines at different concentrations: 100 µg kg(-1) for tetracycline, 100 µg kg(-1) for oxytetracycline, 80 µg kg(-1) for chlortetracycline and 30 µg kg(-1) for doxycycline. Concentrations used in the validation matched the requirements for MRLs (they were either at or below the MRLs) stated in European Union Regulation 37/2010. The sensitivity of the validation was 100%.

  20. In vivo spread of macrolide-lincosamide-streptogramin B (MLSB) resistance--a model study in chickens.

    Science.gov (United States)

    Marosevic, D; Cervinkova, D; Vlkova, H; Videnska, P; Babak, V; Jaglic, Z

    2014-07-16

    The influence of specific and non-specific antibiotic pressure on in vivo spread of macrolide-lincosamide-streptogramin B (MLSB) resistance was evaluated in this study. Chickens repeatedly inoculated with Enterococcus faecalis harbouring the plasmid pAMβ1 carrying the erm(B) gene were perorally treated for one week with tylosin, lincomycin (both specific antibiotic pressure) and chlortetracycline (non-specific antibiotic pressure). Antibiotic non-treated but E. faecalis inoculated chickens served as a control. To quantify the erm(B) gene and characterise intestinal microflora, faecal DNA was analysed by qPCR and 454-pyrosequencing. Under the pressure of antibiotics, a significant increase in erm(B) was observed by qPCR. However, at the final stage of the experiment, an increase in erm(B) was also observed in two out of five non-treated chickens. In chickens treated with tylosin and chlortetracycline, the increase in erm(B) was accompanied by an increase in enterococci. However, E. faecalis was at the limit of detection in all animals. This suggests that the erm(B) gene spread among the gut microbiota other than E. faecalis. Pyrosequencing results indicated that, depending on the particular antibiotic pressure, different bacteria could be responsible for the spread of MLSB resistance. Different species of MLSB-resistant enterococci and streptococci were isolated from cloacal swabs during and after the treatment. PFGE analysis of MLSB-resistant enterococci revealed four clones, all differing from the challenge strain. All of the MLSB-resistant isolates harboured a plasmid of the same size as pAMβ1. This study has shown that MLSB resistance may spread within the gut microbiota under specific and non-specific pressure and even in the absence of any antimicrobial pressure. Finally, depending on the particular antibiotic pressure, different bacterial species seems to be involved in the spread of MLSB resistance.

  1. Distribution and ecological risk of antibiotics in a typical effluent-receiving river (Wangyang River) in north China.

    Science.gov (United States)

    Jiang, Yonghai; Li, Mingxiao; Guo, Changsheng; An, Da; Xu, Jian; Zhang, Yuan; Xi, Beidou

    2014-10-01

    In this study, the occurrence and distribution of sixteen antibiotics belonging to four groups in surface water, sediment and groundwater samples from the Wangyang River (WYR), a typical river receiving sewage discharges were investigated. Laboratory analyses revealed that antibiotics were widely distributed in the studied area. The aqueous samples were unavoidably contaminated with antibiotics, and the target antibiotics present in high levels were oxytetracycline, tetracycline, chlortetracycline, ofloxacin, sulfamethoxazole, and trimethoprim, with maximum concentrations of the individual contaminant at 3.6×10(5), 9.7×10(3), 6.9×10(4), 1.2×10(4), 4.8×10(3), and 1.1×10(3) ng L(-1), respectively. Oxytetracycline, tetracycline, ciprofloxacin and roxithromycin were the most frequently detected compounds in sediment samples, with maximum concentrations of the individual contaminant at 1.6×10(5), 1.7×10(4), 2.1×10(3) and 2.5×10(3) ng g(-1), respectively. The results also revealed that the high intensity of aquaculture activities could contribute to the increasing levels of antibiotics in the area. According to the ratios of measured environmental concentration (MEC) to predicted no-effect concentration (PNEC), chlortetracycline, tetracycline, ofloxacin, ciprofloxacin, erythromycin-H2O and sulfamethoxazole may present possible environmental risk to Pseudokirchneriella subcapitata, Synechococcus leopoliensis and M. aeruginosa. Attention should be given to the long-term ecological effects caused by the continuous discharge of antibiotics in the WYR area.

  2. Boar sperm changes after sorting and encapsulation in barium alginate membranes.

    Science.gov (United States)

    Spinaci, M; Bucci, D; Chlapanidas, T; Vallorani, C; Perteghella, S; Communod, R; Vigo, D; Tamanini, C; Galeati, G; Faustini, M; Torre, M L

    2013-09-15

    A routine use of boar-sexed semen is limited by the long sorting time necessary to obtain an adequate number of sexed spermatozoa for artificial insemination and by the high susceptibility of spermatozoa of this species to damages induced by sorting procedure and subsequent cryopreservation. The aim of this work was to study the impact of encapsulation in barium alginate membrane on sorted boar spermatozoa by evaluating membrane integrity, chlortetracycline staining patterns, protein tyrosine phosphorylation, and Hsp70 immunolocalization during storage over 72 hours in liquid or encapsulated form. The encapsulation procedure significantly (P < 0.05) decreased the overall membrane integrity of control unsorted semen (81.8 vs. 57.4, CTR vs. CPS), but did not negatively affect the overall viability and the chlortetracycline staining patterns of sorted encapsulated cells. Moreover, encapsulation significantly decreased (P < 0.05) the overall phosphotyrosin A pattern cell percentage in unsorted (98.4 vs. 92.6, CTR vs. CPS) but not in sorted semen (64.0 vs. 74.2; SORT CTR vs. SORT CPS). As for Hsp70, the overall percentage of cells displaying the different patterns was significantly influenced (P < 0.05) by treatment but not by storage time. The sorting procedure seems to induce the major changes, whereas encapsulation tends to exert a protective effect on sorted semen by increasing the percentage of spermatozoa displaying the T pattern (2.8 vs. 24.3; SORT CTR vs. SORT CPS). In conclusion, our data confirm that the damaging impact of the encapsulation in barium alginate capsules seems to be limited when compared with that of the sorting procedure and, moreover, the association of the two procedures does not result in an algebraic sum of the negative effects. These results suggest the possibility of a future utilization of the encapsulation technology in order to store sorted spermatozoa and permit their controlled release in the female genital tract.

  3. Antibiotic resistance of lactic acid bacteria isolated from Chinese yogurts.

    Science.gov (United States)

    Zhou, N; Zhang, J X; Fan, M T; Wang, J; Guo, G; Wei, X Y

    2012-09-01

    The aim of this study was to evaluate the susceptibility of 43 strains of lactic acid bacteria, isolated from Chinese yogurts made in different geographical areas, to 11 antibiotics (ampicillin, penicillin G, roxithromycin, chloramphenicol, tetracycline, chlortetracycline, lincomycin, kanamycin, streptomycin, neomycin, and gentamycin). The 43 isolates (18 Lactobacillus bulgaricus and 25 Streptococcus thermophilus) were identified at species level and were typed by random amplified polymorphic DNA analysis. Thirty-five genotypically different strains were detected and their antimicrobial resistance to 11 antibiotics was determined using the agar dilution method. Widespread resistance to ampicillin, chloramphenicol, chlortetracycline, tetracyclines, lincomycin, streptomycin, neomycin, and gentamycin was found among the 35 strains tested. All of the Strep. thermophilus strains tested were susceptible to penicillin G and roxithromycin, whereas 23.5 and 64.7% of Lb. bulgaricus strains, respectively, were resistant. All of the Strep. thermophilus and Lb. bulgaricus strains were found to be resistant to kanamycin. The presence of the corresponding resistance genes in the resistant isolates was investigated through PCR, with the following genes detected: tet(M) in 1 Lb. bulgaricus and 2 Strep. thermophilus isolates, ant(6) in 2 Lb. bulgaricus and 2 Strep. thermophilus isolates, and aph(3')-IIIa in 5 Lb. bulgaricus and 2 Strep. thermophilus isolates. The main threat associated with these bacteria is that they may transfer resistance genes to pathogenic bacteria, which has been a major cause of concern to human and animal health. To our knowledge, the aph(3')-IIIa and ant(6) genes were found in Lb. bulgaricus and Strep. thermophilus for the first time. Further investigations are required to analyze whether the genes identified in Lb. bulgaricus and Strep. thermophilus isolates might be horizontally transferred to other species.

  4. Characterization of the europium tetracycline complex as a biomarker for atherosclerosis

    Science.gov (United States)

    Courrol, Lilia C.; da Silva, Mônica N.; Sicchieri, Leticia B.

    2016-04-01

    Atherosclerosis is a narrowing of the arteries caused by an increase of atheromatous plaque: material formed by macrophage cells containing cholesterol and fatty acids, calcium and a variable amount of fibrous connective tissue. The elation between vulnerable plaques and cardiovascular events can be determined using plaque biomarkers. In this work, atherosclerotic plaques stained with different molar ratios of europium, in a potential plaque biomarker, europium tetracycline complex, were studied by fluorescence microscopy. The tetracycline antibiotic used was chlortetracycline. The growth of atherosclerotic plaque was followed during 60 days in New Zealand rabbits divided in two groups: an experimental group (EG), with nine animals and a control group (CG) with three animals. The animals in the EG received a diet with 1% of cholesterol and the animals of GC received a normal diet. The aortic arch of the animals with 60 days were cut in the vertical plane in 6 μm thick slices, which were mounted on glass slides and stained with hematoxylin an eosin and europium chlortetracycline complex (EuCTc). The fluorescence images were obtained exciting the EuCTc absorption band with a filter cube D (BP 355 - 425) and the emission was collected with a LP 470 suppression filter. Light intensity, detector gain and acquisition time were fixed for comparisons. The 20× magnified images were collected with 12 bit (or 4096 gray tones) resolution. The mean value of gray scale for each molar ratio of EuCTc was different, indicating that the complex interacts with the components of atherosclerotic plaque and the best molar ratio was 1.5 EuCTc. These results indicate the potential use of the EuCTc biomarker for atherosclerotic plaque characterization.

  5. Antimicrobial Consumption in Medicated Feeds in Vietnamese Pig and Poultry Production.

    Science.gov (United States)

    Van Cuong, Nguyen; Nhung, Nguyen Thi; Nghia, Nguyen Huu; Mai Hoa, Nguyen Thi; Trung, Nguyen Vinh; Thwaites, Guy; Carrique-Mas, Juan

    2016-09-01

    Antimicrobials are extensively used as growth promoters in animal feeds worldwide, but reliable estimates are lacking. We conducted an internet-based survey of commercial chicken and pig feed products officially approved for sale in Vietnam over the period March-June 2015. Information on the antimicrobial contents in feed products, alongside animal production data, was used to estimate in-feed antimicrobial consumption to produce one kilogram of live animal (chicken, pig), as well as to estimate country-wide antimicrobial consumption through animal feeds. A total of 1462 commercial feed formulations were examined. The survey-adjusted estimated antimicrobial contents were 25.7 and 62.3 mg/kg in chicken and pig feeds, respectively. Overall, it was estimated that 77.4 mg [95% CI 48.1-106.8] and 286.6 mg [95% CI 191.6-418.3] of in-feed antimicrobials were used to raise 1 kg of live chicken and pig, respectively. Bacitracin (15.5% feeds), chlortetracycline (11.4%), and enramycin (10.8%) were the most common antimicrobials present in chicken feed formulations, whereas bacitracin (24.8%), chlortetracycline (23.9%), and florfenicol (17.4%) were the most common in pig feed formulations. Overall, 57% of the total quantitative usage consisted of antimicrobials regarded by WHO of importance for human medicine, including amoxicillin, colistin, tetracyclines, neomycin, lincomycin, and bacitracin. These figures confirm a very high magnitude of in-feed consumption of antimicrobials, especially in pig production. Results from this study should encourage further monitoring of antimicrobials used in animal production, and foster discussion about existing policies on inclusion of antimicrobials in animal feed rations.

  6. Transport of three veterinary antimicrobials from feedlot pens via simulated rainfall runoff.

    Science.gov (United States)

    Sura, Srinivas; Degenhardt, Dani; Cessna, Allan J; Larney, Francis J; Olson, Andrew F; McAllister, Tim A

    2015-07-15

    Veterinary antimicrobials are introduced to wider environments by manure application to agricultural fields or through leaching or runoff from manure storage areas (feedlots, stockpiles, windrows, lagoons). Detected in manure, manure-treated soils, and surface and ground water near intensive cattle feeding operations, there is a concern that environmental contamination by these chemicals may promote the development of antimicrobial resistance in bacteria. Surface runoff and leaching appear to be major transport pathways by which veterinary antimicrobials eventually contaminate surface and ground water, respectively. A study was conducted to investigate the transport of three veterinary antimicrobials (chlortetracycline, sulfamethazine, tylosin), commonly used in beef cattle production, in simulated rainfall runoff from feedlot pens. Mean concentrations of veterinary antimicrobials were 1.4 to 3.5 times higher in surface material from bedding vs. non-bedding pen areas. Runoff rates and volumetric runoff coefficients were similar across all treatments but both were significantly higher from non-bedding (0.53Lmin(-1); 0.27) than bedding areas (0.40Lmin(-1); 0.19). In keeping with concentrations in pen surface material, mean concentrations of veterinary antimicrobials were 1.4 to 2.5 times higher in runoff generated from bedding vs. non-bedding pen areas. Water solubility and sorption coefficient of antimicrobials played a role in their transport in runoff. Estimated amounts of chlortetracycline, sulfamethazine, and tylosin that could potentially be transported to the feedlot catch basin during a one in 100-year precipitation event were 1.3 to 3.6ghead(-1), 1.9ghead(-1), and 0.2ghead(-1), respectively. This study demonstrates the magnitude of veterinary antimicrobial transport in feedlot pen runoff and supports the necessity of catch basins for runoff containment within feedlots.

  7. Efficacy of salicylic acid in the treatment of digital dermatitis in dairy cattle.

    Science.gov (United States)

    Schultz, N; Capion, N

    2013-11-01

    Digital dermatitis (DD) is one of the most important causes of lameness in dairy cattle worldwide. The objective of this study was to evaluate the efficacy of salicylic acid in the treatment of the disease. A total of 201 DD lesions from 173 cows from four commercial dairy herds were evaluated at day 0 during routine hoof trimming and were allocated into two groups, namely, a control group given chlortetracycline spray, and a treatment group given 10 g of salicylic acid powder applied topically within a bandage. Pain, lesion size and clinical appearance (scored M0 to M4) were evaluated on days 3, 14 and 34 post-treatment. A change to M0 was defined as healing, while changes of M2 or M4 to M1 or M3 were classified as clinical improvements. Healing rates did not differ significantly between treatment groups at days 3 and 14. By day 34 the healing rate was fivefold better (P=0.01) for the treatment vs. the control group, with healing rates of 13.6% and 3.1%, respectively. By day 3, the rate of improvement was 2.5-fold better (P=0.02) for the controls. By day 34 the overall positive effect (i.e. healing and improvement) was 1.75-fold better (P=0.05) for the treatment group. Lesions from the control group were 2.2 times more likely (P=0.09) to have a pain score equal to 2 by day 14. The proportion of lesions getting smaller by days 14 and 34 was 2.5 times higher (Psalicylic acid should be considered as an alternative to chlortetracycline for the treatment of DD as it appears more efficacious and would assist in reducing antibiotic use.

  8. Effects of ionophores and antibiotics on in vitro hydrogen sulfide production, dry matter disappearance, and total gas production in cultures with a steam-flaked corn-based substrate with or without added sulfur.

    Science.gov (United States)

    Quinn, M J; May, M L; Hales, K E; DiLorenzo, N; Leibovich, J; Smith, D R; Galyean, M L

    2009-05-01

    Effects of 3 ionophores and 2 antibiotics on in vitro H(2)S production, IVDMD, total gas production, and VFA profile with or without added S were examined. In Exp. 1, ruminal fluid from 2 ruminally cannulated steers fed a steam-flaked corn-based diet (75% concentrate) without ionophore and antibiotics for 28 d before collection was used to inoculate in vitro cultures. Treatments were control (no ionophore or antibiotic), 3 ionophores (lasalocid sodium and monensin sodium at 5 mg/L or laidlomycin propionate at 1.65 mg/L), and 2 antibiotics (chlortetracycline hydrochloride at 5 mg/L and tylosin tartarate at 1.25 mg/L). Cultures also had 0 or 1.75 mg of S/L (from sodium sulfate). No S x ionophore-antibiotic treatment interactions were noted (P > 0.53) for IVDMD, total gas production, and H(2)S production. Hydrogen sulfide (mumol/g of fermentable DM) was increased (P production tended (P = 0.09) to be increased with additional S; however, IVDMD was not affected by added S (P = 0.90). Production of H(2)S was not affected by ionophores or antibiotics (P > 0.18). On average, IVDMD (P = 0.05) was greater for ionophores than for antibiotics, whereas total gas production was less for ionophores than for control (P antibiotics (P 0.20) in acetate, propionate, or acetate:propionate between ionophores and control (S x treatment interaction, P = 0.03). In Exp. 2, the effects of ionophore-antibiotic combinations with added S were examined using the same procedures as in Exp. 1. Treatments were control, monensin plus tylosin (MT), and lasalocid plus chlortetracycline (LCTC), with concentrations of the ionophores and antibiotics as in Exp. 1. No differences were observed among treatments for H(2)S production (P > 0.55). Treatments MT and LCTC tended (P = 0.06) to increase IVDMD and decreased (P = 0.02) gas production vs. control. Proportion of acetate (P = 0.01) and acetate:propionate (P antibiotics we evaluated did not affect production of H(2)S gas in an in vitro rumen culture

  9. Distribution and characterization of ampicillin- and tetracycline-resistant Escherichia coli from feedlot cattle fed subtherapeutic antimicrobials

    Directory of Open Access Journals (Sweden)

    Yanke L Jay

    2011-04-01

    Full Text Available Abstract Background Feedlot cattle in North America are routinely fed subtherapeutic levels of antimicrobials to prevent disease and improve the efficiency of growth. This practice has been shown to promote antimicrobial resistance (AMR in subpopulations of intestinal microflora including Escherichia coli. To date, studies of AMR in feedlot production settings have rarely employed selective isolation, therefore yielding too few AMR isolates to enable characterization of the emergence and nature of AMR in E. coli as an indicator bacterium. E. coli isolates (n = 531 were recovered from 140 cattle that were housed (10 animals/pen in 14 pens and received no dietary antimicrobials (control - 5 pens, CON, or were intermittently administered subtherapeutic levels of chlortetracycline (5 pens-T, chlortetracycline + sulfamethazine (4 pens-TS, or virginiamycin (5 pens-V for two separate periods over a 9-month feeding period. Phenotype and genotype of the isolates were determined by susceptibility testing and pulsed field gel electrophoresis and distribution of characterized isolates among housed cattle reported. It was hypothesized that the feeding of subtherapeutic antibiotics would increase the isolation of distinct genotypes of AMR E. coli from cattle. Results Overall, patterns of antimicrobial resistance expressed by E. coli isolates did not change among diet groups (CON vs. antibiotic treatments, however; isolates obtained on selective plates (i.e., MA,MT, exhibited multi-resistance to sulfamethoxazole and chloramphenicol more frequently when obtained from TS-fed steers than from other treatments. Antibiograms and PFGE patterns suggested that AMR E. coli were readily transferred among steers within pens. Most MT isolates possessed the tet(B efflux gene (58.2, 53.5, 40.8, and 50.6% of isolates from CON, T, TS, and V steers, respectively whereas among the MA (ampicillin-resistant isolates, the tem1-like determinant was predominant (occurring in 50, 66

  10. Longitudinal characterization of antimicrobial resistance genes in feces shed from cattle fed different subtherapeutic antibiotics

    Directory of Open Access Journals (Sweden)

    Read Ronald R

    2011-01-01

    Full Text Available Abstract Background Environmental transmission of antimicrobial-resistant bacteria and resistance gene determinants originating from livestock is affected by their persistence in agricultural-related matrices. This study investigated the effects of administering subtherapeutic concentrations of antimicrobials to beef cattle on the abundance and persistence of resistance genes within the microbial community of fecal deposits. Cattle (three pens per treatment, 10 steers per pen were administered chlortetracycline, chlortetracycline plus sulfamethazine, tylosin, or no antimicrobials (control. Model fecal deposits (n = 3 were prepared by mixing fresh feces from each pen into a single composite sample. Real-time PCR was used to measure concentrations of tet, sul and erm resistance genes in DNA extracted from composites over 175 days of environmental exposure in the field. The microbial communities were analyzed by quantification and denaturing gradient gel electrophoresis (DGGE of PCR-amplified 16S-rRNA. Results The concentrations of 16S-rRNA in feces were similar across treatments and increased by day 56, declining thereafter. DGGE profiles of 16S-rRNA differed amongst treatments and with time, illustrating temporal shifts in microbial communities. All measured resistance gene determinants were quantifiable in feces after 175 days. Antimicrobial treatment differentially affected the abundance of certain resistance genes but generally not their persistence. In the first 56 days, concentrations of tet(B, tet(C, sul1, sul2, erm(A tended to increase, and decline thereafter, whereas tet(M and tet(W gradually declined over 175 days. At day 7, the concentration of erm(X was greatest in feces from cattle fed tylosin, compared to all other treatments. Conclusion The abundance of genes coding for antimicrobial resistance in bovine feces can be affected by inclusion of antibiotics in the feed. Resistance genes can persist in feces from cattle beyond 175 days

  11. Carnitine supplementation decreases capacitation-like changes of frozen-thawed buffalo spermatozoa.

    Science.gov (United States)

    Longobardi, Valentina; Salzano, Angela; Campanile, Giuseppe; Marrone, Raffaele; Palumbo, Francesco; Vitiello, Milena; Zullo, Gianluigi; Gasparrini, Bianca

    2017-01-15

    The aim of this study was to evaluate the effect of carnitine supplementation of semen extender on fertility parameters of frozen-thawed buffalo sperm. Buffalo semen was cryopreserved in BioXcell containing 0 (control group), 2.5 and 7.5-mM carnitine. After thawing, viability, motility, membrane integrity and capacitation status (assessed by localization of phosphotyrosine-containing proteins and chlortetracycline, chlortetracycline assay) were evaluated. Furthermore, total antioxidant capacity, reactive oxygen species (ROS) and lipid peroxidation levels, as well as adenosine triphosphate (ATP) content and phospholipids concentration were assessed. Finally, in vitro-fertilizing ability was evaluated after heterologous IVF. An increased post-thawing sperm motility and membrane integrity were recorded in both treated groups compared with the control (44.4 ± 3.5, 53.1 ± 3.9, and 52.5 ± 3.6%; P carnitine, respectively). Supplementation of carnitine to the freezing extender decreased (P carnitine). In agreement with this, carnitine also decreased (P carnitine). Interestingly, carnitine increased total antioxidant capacity and ATP content of buffalo frozen-thawed sperm (1.32 ± 0.02, 1.34 ± 0.01, 1.37 ± 0.01 mM/L and 4.1 ± 0.1, 5.3 ± 0.1 and 8.2 ± 0.4 nM × 10(8) sperm; P carnitine). Intracellular ROS decreased in carnitine-treated sperm compared with the control, as indicated by dihydroethidium (DHE) values (0.22 ± 0.01, 0.18 ± 0.01, and 0.14 ± 0.0 μM/100 μL dihydroethidium, respectively, with 0, 2.5-, and 7.5-mM carnitine; P carnitine improved buffalo sperm quality by increasing ATP generation and modulating ROS production, without affecting in vitro fertilizing ability.

  12. Effect of low-dose fenvalerate on semen quality capacitation in adult mice

    Institute of Scientific and Technical Information of China (English)

    SHI Xiao-dan; BI Huan-jing; FU He-ling; LI Liang-yun; LIU De-kang; LI Jian-min

    2011-01-01

    Background Fenvalerate (FEN) has been demonstrated to be a reproductive toxicant in humans and rodents. However,little is known about whether short-term exposure to low-dose FEN produces reproductive toxicity.Methods We administered FEN (0.009 375, 0.1875, 3.750, or 45.00 mg·kg-1d-1 by gavage for 30 days) to male ICR mice and compared reproductive toxicity parameters between groups receiving different concentrations of FEN.Reproductive toxicity was evaluated by computer-assisted semen quality analysis (CASA), chlortetracycline (CTC) assay,and histopathology.Results The sperm morphology and testis histology of FEN-exposed mice (all doses) were similar to that in controlling mice. Exposure to FEN at a concentration of 0.1875 mg·kg-1d-1 decreased sperm path straightness (STR) and linearity (LIN) (both P< 0.05), but had no significant impact on average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), lateral amplitude (ALH), beat cross frequency (BCF), or progressive motility (MOT). FEN reduced the rate of mouse sperm capacitation in a dose-dependent manner.Conclusion The present results demonstrate that exposure to low-dose FEN for 30 days reduces semen quality and sperm capacitation in adult mice.

  13. Effects of a mixture of tetracyclines to Lemna gibba and Myriophyllum sibiricum evaluated in aquatic microcosms

    Energy Technology Data Exchange (ETDEWEB)

    Brain, Richard A. [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada)]. E-mail: rbrain01@uoguelph.ca; Wilson, Christian J. [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada); Johnson, David J. [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada); Sanderson, Hans [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada); Bestari, Ketut [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada); Hanson, Mark L. [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada); Sibley, Paul K. [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada); Solomon, Keith R. [Centre for Toxicology, Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 (Canada)

    2005-12-15

    The impact of a mixture of oxytetracycline, chlortetracycline, tetracycline and doxycycline on Myriophyllum sibiricum and Lemna gibba was investigated using fifteen 12,000-L microcosms (k = 5, n = 3). Significant concentration-response relationships were only found for M. sibiricum, where dry mass was 69, 47, 30, and 7% of controls at respective treatment concentrations of 0.080, 0.218, 0.668, and 2.289 {mu}mol/L. Somatic endpoints were strongly and negatively correlated with percent light transmission, except plant length, which was positively correlated. Treated microcosms experienced a reduction in the percent of surface irradiance penetrating the water column as high as 99.8% at a depth of 70 cm, relative to controls. Position relative to the water column was likely responsible for the differential effects observed between floating (L. gibba) and submerged (M. sibiricum) species of macrophytes. A hazard quotient assessment of the lowest EC{sub 10} value indicated significant risk, exceeding the critical HQ value, but not the lowest EC{sub 25} value. - Mixtures of tetracycline antibiotics pose a risk to submerged but not floating aquatic plants.

  14. Antimicrobial resistance trends among Escherichia coli isolates obtained from dairy cattle in the northeastern United States, 2004-2011.

    Science.gov (United States)

    Cummings, Kevin J; Aprea, Victor A; Altier, Craig

    2014-01-01

    Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform risk analyses and guide public policy regarding antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Escherichia coli isolates from dairy cattle in the northeastern United States and to identify trends in resistance to selected antimicrobial agents over time. We collected data retrospectively for all bovine E. coli isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. We investigated temporal trends in the prevalence of resistant E. coli for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 3373 bovine E. coli isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 2.7% (enrofloxacin) to 91.3% (oxytetracycline). There was evidence of a significantly decreasing trend in prevalence of resistance to several agents: chlortetracycline, florfenicol, neomycin, oxytetracycline, spectinomycin, and trimethoprim/sulfamethoxazole. However, a significantly increasing trend in prevalence of resistance to enrofloxacin was also evident. These results do not support the idea that current antimicrobial use practices on dairy operations are driving a general increase in the emergence and dissemination of drug-resistant E. coli in the region served by the laboratory. However, resistance to some drugs remained consistently high during the study period, and increasing resistance to enrofloxacin is a key area of concern.

  15. Acrosome membrane integrity and cryocapacitation are related to cholesterol content of bull spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Srivastava N; Srivastava SK; Ghosh SK; Amit Kumar; Perumal P; Jerome A

    2013-01-01

    Objective: To evaluate the cryoinjury prediction of spermatozoa in relation to its cholesterol content at fresh and frozen-thaw stages. Methods: Ejaculates (n=12) were processed for cryopreservation, acrosome integrity (fluorescent and giemsa stains), cryoinjury (distribution of non capacitated, capacitated and acrosome reacted, pattern F, B and AR, respectively of Chlortetracycline, CTC assay), in vitro fertiltiy (IVF) and cholesterol content of spermatozoa at fresh, pre-freeze and frozen-thaw stages were evaluated. Values were fitted in prediction equation to predict acrosome integrity (AI) and cryoinjury. Results: Study indicated that cholesterol content of fresh spermatozoa can be used to predict cholesterol content of spermatozoa at pre-freeze and frozen-thaw stages of cryopreservation protocol with medium to high level of accuracy (P<0.05 and P<0.01, respectively). Cholesterol content of fresh spermatozoa can be used to predict AI, pattern B and AR and Penetration Index (PI) of IVF with medium level of accuracy (P<0.05) at frozen-thaw but not at pre-freeze stage. Similarly cholesterol content of frozen-thaw spermatozoa can be used to predict AI and pattern AR of frozen-thaw spermatozoa with medium level of accuracy (P<0.05). Conclusion: This study revealed strong evidence that cholesterol content of fresh as well as frozen-thaw bull spermatozoa can be a good predictor of level of cryoinjury following preservation at ultra low temperature.

  16. Temporal changes and the effect of subtherapeutic concentrations of antibiotics in the gut microbiota of swine.

    Science.gov (United States)

    Holman, Devin B; Chénier, Martin R

    2014-12-01

    The use of antibiotics in swine production for the purpose of growth promotion dates back to the 1950s. Despite this long history of use, the exact mechanism(s) responsible for the growth-promoting effects of antibiotics in swine remain largely unknown. It is believed, however, that growth promotion is due to antibiotics having a direct impact on the gut microbiota. In this study, the effect of two antibiotics on the swine gut microbiota over a 19-week monitoring period was investigated using Illumina-based sequencing. A shift in the relative abundance of several taxa and in 26 operational taxonomic units (OTUs) was observed in pigs fed subtherapeutic concentrations of tylosin (44-11 mg kg(-1) feed). Only minor alterations were noted with the administration of chlortetracycline at 5.5 mg kg(-1) feed. The most notable changes in the relative abundance of taxa and OTUs were noted between suckling piglets and postweaned pigs. Diversity was also reduced in the gut microbiota of suckling piglets as measured using the Shannon, Chao1, and phylogenetic diversity indices. These results show that the effect of antibiotics on the swine gut microbiota is variable based on dosage and duration and that the swine gut microbiota exhibits considerable resilience to long-term changes due to antibiotic perturbations.

  17. Antibiogram pattern of bacteria causing endometritis in cows

    Directory of Open Access Journals (Sweden)

    S. Udhayavel

    2013-04-01

    Full Text Available Aim : To find out the organisms causing endometritis in cattle and to determine their in vitro sensitivity to various antibiotics Materials and methods: Thirty uterine secretion samples, 9 from Holstein Friesian and 21 from Jersey cows were collected in and around Namakkal district of Tamil Nadu from clinical cases of endometritis. The bacteria isolated both aerobically and anaerobically from endometritis showed the characteristic colony, were gram stained and confirmed by standard biochemical tests. The invitro antibiotic sensitivity test with different antibiotic discs were carried out. Results and Discussion: Out of 30 samples, 25 contained different strains of bacteria and 5 were found bacteriologically sterile. Different species of bacteria isolated from these samples were, Escherichia coli (36.66%, Klebsiella spp (30%, Proteus spp (13.33%, Pseudomonas aeruginosa (6.66%, Clostridium spp (3.33%. The in vitro antibiotic sensitivity test indicated that the highest number of isolates (64% were sensitive to Ceftriaxone, followed by Gentamicin, Enrofloxacin and Chlortetracycline (32%. Chloramphenicol showed sensitivity in minimum number (8% of isolates. Conclusion : This study revealed that the bacteria E.coli and Klebisella spp are more commonly isolated in endometritis in cows and the drug Ceftriaxone is highly effective. [Vet World 2013; 6(2.000: 100-102

  18. Regucalcin is widely distributed in the male reproductive tract and exerts a suppressive effect on in vitro sperm capacitation in the water buffalo (Bubalus bubalis).

    Science.gov (United States)

    Pillai, Harikrishna; Shende, Ashish Mukunda; Parmar, Mehtab Sing; A, Anjaneya; L, Sreela; Kumaresan, Arumugam; G, Taru Sharma; Bhure, Sanjeev Kumar

    2017-03-01

    Regucalcin is a multi-functional, calcium-binding protein with roles in calcium homeostasis, apoptosis, cell proliferation, and free radical neutralization. Regucalcin is broadly expressed in the male reproductive organs of rat and bovine; here, we report its expression in the reproductive tract of male buffalo-especially in testis, epididymis, seminal vesicle, prostate, and bulbourethral gland of buffalo-as analyzed by real-time PCR, Western blot, and immunolocalization. Regucalcin degradation in seminal plasma, despite its high abundance in vesicular fluid, was demonstrated using recombinant regucalcin co-incubated with buffalo seminal plasma. This depletion of regucalcin appears to be related to its suppressive effect on in vitro sperm capacitation, observed using the chlortetracycline assay after treating buffalo spermatozoa with recombinant protein. Indeed, addition of recombinant regucalcin to capacitating media significantly reduced (P < 0.05) the percentage of capacitated spermatozoa to 6.1 ± 0.6 from 36.4 ± 1.8 in the untreated group. Taken together, the wide distribution of regucalcin in male buffaloes, versus its degradation in the seminal plasma and suppressive effects on in vitro capacitation of spermatozoa, indicate its possible anti-capacitation role in the reproductive tract. Mol. Reprod. Dev. 84: 212-221, 2017. © 2016 Wiley Periodicals, Inc.

  19. Effective Removal of Tetracycline from Aqueous Solution by Organic Acid-Coated Magnetic Nanoparticles.

    Science.gov (United States)

    Guo, Liang; Liang, Yuyan; Chen, Xuelan; Xu, Wei; Wu, Kesheng; Wei, Hua; Xiong, Yonghua

    2016-03-01

    Self-assembled iron oxide nanocomposites are good magnetic nano-adsorbents that can be prepared using simple methods. Four types of organic acid-functionalised (oleic acid, undecenoic acid, caprylic acid or hexanoic acid) magnetic nanoparticles (MNPs) were synthesised through a one-pot chemisorption method for the removal of tetracycline (TC) from aqueous solution. The undecenoic acid-coated MNPs (UA-MNPs) exhibited the highest adsorption efficiency and can be easily retrieved with a low-gradient magnetic separator (0.4 Tesla) at pH 5.0 aqueous solution. The TC adsorption process on the UA-MNPs followed the Langmuir isotherm and the maximum adsorption capacities increased from 86.96 mg g(-1) to 222.2 mg g(-1) with the increase in temperature from 288 K to 318 K. The kinetics of adsorption fits pseudo-second-order model perfectly with a rate constant, 5.946 g mg(-1) min(-1) at 298 K. The positive values of the enthalpy (AH) and the negative value of the free energy (AG) indicated an endothermic and spontaneous adsorption process of TC on the UA-MNPs. Moreover, the UA-MNPs possessed excellent ability to adsorb the other three major types of TC antibiotics, including chlortetracycline, oxytetracycline and doxycycline.

  20. Effects of ten antibiotics on seed germination and root elongation in three plant species.

    Science.gov (United States)

    Hillis, Derek G; Fletcher, James; Solomon, Keith R; Sibley, Paul K

    2011-02-01

    We applied a screening-level phytotoxicity assay to evaluate the effects of 10 antibiotics (at concentrations ranging from 1 to 10,000 μg/L) on germination and early plant growth using three plant species: lettuce (Lactuca sativa), alfalfa (Medicago sativa), and carrot (Daucus carota). The range of phytotoxicity of the antibiotics was large, with EC₂₅s ranging from 3.9 μg/L to >10,000 μg/L. Chlortetracycline, levofloxacin, and sulfamethoxazole were the most phytotoxic antibiotics. D. carota was the most sensitive plant species, often by an order of magnitude or more, followed by L. sativa and then M. sativa. Plant germination was insensitive to the antibiotics, with no significant decreases up to the highest treatment concentration of 10,000 μg/L. Compared with shoot and total length measurements, root elongation was consistently the most sensitive end point. Overall, there were few instances where measured soil concentrations, if available in the publicly accessible literature, would be expected to exceed the effect concentrations of the antibiotics evaluated in this study. The use of screening assays as part of a tiered approach for evaluating environmental impacts of antibiotics can provide insight into relative species sensitivity and serve as a basis by which to screen the potential for toxic effects of novel compounds to plants.

  1. Effect of Different Oligosaccharides on Immunity, and Cecal Microflora in Broilers

    Institute of Scientific and Technical Information of China (English)

    GAO Ying; SHAN An-shan

    2004-01-01

    One hundred and forty-four Avin broilers (1 d old) were randomly divided into six treatments, with three replicates of eight birds each to investigate the effect of different oligosaccharides on index of immune organ, cecal weight and microflora in broilers. The control group (Group Ⅰ) was fed with corn-soybean meal as basal diet, and the trial groups (group Ⅱ, Ⅲ, Ⅳ, Ⅴ and Ⅵ ) were fed with basal diet plus 0.1% manoligosaccharides (MOS), 0.3% soybean-oligosaccharides (SBOS), 0.3% fructooligosaccharides (FOS), 0.3% α -glucooligosaccharides(α -GOS) and 0.03% chlortetracycline (CTC), respectively. The results showed that oligosaccharides supplementation significantly (P<0.05) increased bifidobacteria concentration ofyoung broilers and did not affect on bifidobacteria or escherichia coli concentration of the birds ( P >0.05). Cecal weights of 21 d old broilers fed with FOS and α -GOS were significantly (P <0.05) increased, higher than those with MOS and SBOS, but addition of oligosaccharides had no effect on the birds. The effect of oligosaccharides on the cecal length was the same as on the cecal weights. Oligosaccharides supplement did not affect on indexes of immune organs.

  2. Escherichia coli tetracycline efflux determinants in relation to tetracycline residues in chicken

    Institute of Scientific and Technical Information of China (English)

    Al-Bahry SN; Al-Mashani BM; Al-Ansari AS; Elshafie AE; Mahmoud IY

    2013-01-01

    Objective: To screen for Escherichia coli (E. coli) resistant to tetracycline, followed by identification of tet efflux genes by polymerase chain reaction (PCR). In addition, detection of tetracycline residues in chicken livers and kidneys were conducted using high performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS-MS). Methods:Strains of E. coli were isolated from samples of chicken colon and screened for tetracycline resistance. Tetracycline genes conferring resistance (Tcr) were detected by polymerase chain reaction (PCR). Most of the isolates were resistant to tetracycline (97.9%). Results: PCR analysis indicated that Tcr E. coli R-plasmids contained tet(A), tet(B) and a combination of both efflux genes. None of the isolates contained other efflux tet genes tet (C, D, E and Y). High performance liquid chromatography-tandem quadrupole mass spectrometry (HPLC-MS-MS), a sensitive technique, was used to detect residues of chlortetracycline (CTC), oxytetracycline (OTC), doxycycline (DC) in chicken livers and kidneys. The samples containing tetracycline residues were at 0.13-0.65 pg/μL levels. Conclusions: Tetracycline and other antibiotics are commonly used in the poultry and meat production industry for prevention of microbial infections. Multiple antibiotic resistant bacteria in Oman have increased to alarming levels, threatening public health, domestic and may have adverse effect on environment.

  3. Fate of tetracyclines in swine manure of three selected swine farms in China

    Institute of Scientific and Technical Information of China (English)

    Min Qiao; Wangda Chen; Jianqiang Su; Bing Zhang; Cai Zhang

    2012-01-01

    Veterinary antibiotics can enter the environment due to the common practice of land application of manure from treated animals.The environmental fate of tetracyclines in swine manure after composting and field application remains largely unknown.This study analyzed the concentrations of tetracyclines in manure,manure-based compost and compost amended soil in selected swine farms from Beijing,Jiaxing and Putian,China to determine the dilution effects of antibiotics when released into the soil environment.The results demonstrate that residues of antibiotics were detected in all samples and chlortetracycline as well as its degradation products should be regarded critically concerning their potential ecotoxicity.Application of manure-based compost to soil could reduce the possible risk posed by antibiotic contamination,but the trigger value of 100 μg/kg was still exceeded in soil samples (776.1 μg/kg dw) from Putian City after application of compost.Field studies such as the present one can help to improve the routine administration of antibiotic-containing composted manure.

  4. Sperm volumetric dynamics during in vitro capacitation process in bovine spermatozoa.

    Science.gov (United States)

    García-Herreros, M; Leal, C L V

    2015-06-01

    Previous studies have demonstrated that sperm head morphometry can be used as a potential diagnostic tool for detecting biophysical changes associated with sperm viability in bovine spermatozoa. In this study, sperm head morphometry was used to investigate its value as a biophysical marker for detecting volumetric changes in bovine spermatozoa under in vitro capacitating and non-capacitating incubation conditions. To further test this hypotesis, aliquots of pooled, washed bovine sperm were incubated in either Tyrode's complete medium with heparin (TCMH; a capacitating medium containing Ca2+, NaHCO3 and heparin), Tyrode's complete medium heparin-free (TCM; a medium containing just Ca2+ and NaHCO3) or Tyrode's basal medium (TBM; a non-capacitating medium free of Ca2+, NaHCO3 and heparin, used as control). Aliquots of sperm were processed for morphometric analysis at different incubation-time intervals (0, 3 and 6 h at 38°C), and the chlortetracycline assay was used simultaneously to confirm the ability of the sperm to undergo capacitation (B pattern) and the acrosome reaction (AR pattern) status in each medium. After 3 h of incubation under TCMH conditions, a significant increase was observed in the percentage of B and AR patterns and a significant decrease was found in all sperm morphometric parameters (Psperm size and AR pattern (r=-0.875, Psperm head morphometry can be used as a potential biophysical marker for detecting volumetric changes during capacitation process in bovine spermatozoa.

  5. Relative efficacy of organic acids and antibiotics as growth promoters in broiler chicken

    Directory of Open Access Journals (Sweden)

    Vikrant Laxman Bagal

    2016-04-01

    Full Text Available Aim: The objective of this study was to evaluate the effect of organic acids as replacer to antibiotics in their various combinations on feed consumption, body weight gain, and feed conversion ratio (FCR in broiler chicks during different phases of growth. Materials and Methods: Antibiotics and organic acids were incorporated into boiler feed in different combinations to form 10 maize based test diets (T1 to T10. Each test diet was offered to four replicates of 10 birds each constituting a total of 400 birds kept for 45 days. Results: Significantly better effect in terms of body weight gain from supplementation of 1% citric acid and 1% citric acid along with antibiotic was observed throughout the entire study, whereas the effect of tartaric acid supplementation was similar to control group. Citric acid (1% along with antibiotic supplementation showed highest feed intake during the experimental period. Significantly better FCR was observed in groups supplemented with 1% citric acid and 1% citric acid along with antibiotic followed by antibiotic along with organic acids supplemented group. Conclusion: Growth performance of birds in terms of body weight, body weight gain, and FCR improved significantly in 1% citric acid which was significantly higher than antibiotic supplemented group. 1% citric acid can effectively replace antibiotic growth promoter (chlortetracycline without affecting growth performance of birds.

  6. A novel green chemistry method for nonaqueous extraction and high-performance liquid chromatography detection of first-, second-, and third-generation tetracyclines, 4-epitetracycline, and tylosin in animal feeds.

    Science.gov (United States)

    Granados-Chinchilla, Fabio; Sánchez, Jorge; García, Fernando; Rodríguez, César

    2012-07-25

    Although tetracyclines and macrolides are common additives for animal nutrition, methods for their simultaneous determination in animal feeds are nonexistent. By coupling an organic extraction and solid-phase extraction cleanup to a high-performance liquid chromatography separation and a nonaqueous postcolumn derivatization, we succeeded in detecting from 0.2 to 24.0 μg kg(-1) of tetracycline, oxytetracycline, chlortetracycline, doxycycline, tigecycline, and 4-epitetracycline in this complex and heterogeneous matrix. Minocycline and tylosin could also be detected with our procedure, but using UV spectrophotometry (1.5 ≤ LOD ≤ 1.9 mg kg(-1)). Linear responses with correlation coefficients between 0.996 and 0.999 were obtained for all analytes in the 0.5-10 mg kg(-1) concentration range. Average recoveries between 59 and 97% and between 98 and 102% were obtained for the tetracyclines and tylosin, respectively. Replicate standard deviations were typically below 5%. When this method was applied to 20 feeds marketed in Costa Rica, we detected labeling inconsistencies, banned mixtures of tetracyclines, and tetracycline concentrations that contravene international regulation.

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

    Energy Technology Data Exchange (ETDEWEB)

    Liu Feng [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Ying Guangguo, E-mail: guangguo.ying@gmail.co [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Tao Ran; Zhao Jianliang; Yang Jifeng [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, 511 Kehua Street, Tianhe District, Guangzhou 510640 (China); Zhao Lanfeng [College of Resource and Environmental Science, South China Agricultural University, Guangzhou 510642 (China)

    2009-05-15

    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. - Terrestrial ecotoxicological effects of antibiotics are related to their sorption and degradation behavior in soil.

  8. Multiple Antimicrobial Resistance of Escherichia coli Isolated from Chickens in Iran

    Directory of Open Access Journals (Sweden)

    Reza Talebiyan

    2014-01-01

    Full Text Available Antimicrobial agents are used extremely in order to reduce the great losses caused by Escherichia coli infections in poultry industry. In this study, 318 pathogenic Escherichia coli (APEC strains isolated from commercial broiler flocks with coli-septicemia were examined for antimicrobials of both veterinary and human significance by disc diffusion method. Multiple resistances to antimicrobial agents were observed in all the isolates. Resistance to the antibiotics was as follows: Tylosin (88.68%, Erythromycin (71.70%, Oxytetracycline (43.40%, Sulfadimethoxine-Trimethoprim (39.62%, Enrofloxacin (37.74%, Florfenicol (35.85%, Chlortetracycline (33.96%, Doxycycline (16.98%, Difloxacin (32.08%, Danofloxacin (28.30%, Chloramphenicol (20.75%, Ciprofloxacin (7.55%, and Gentamicin (5.66%. This study showed resistance against the antimicrobial agents that are commonly applied in poultry, although resistance against the antibiotics that are only applied in humans or less frequently used in poultry was significantly low. This study emphasizes on the occurrence of multiple drug resistant E. coli among diseased broiler chickens in Iran. The data revealed the relative risks of using antimicrobials in poultry industry. It also concluded that use of antibiotics must be limited in poultry farms in order to reduce the antibiotic resistances.

  9. Antimicrobial growth promoters modulate host responses in mice with a defined intestinal microbiota

    Science.gov (United States)

    Brown, Kirsty; Zaytsoff, Sarah J. M.; Uwiera, Richard R. E.; Inglis, G. Douglas

    2016-01-01

    Antibiotics can promote growth in livestock (antimicrobial growth promoters, AGPs), however lack of knowledge regarding mechanisms has hampered the development of effective non-antibiotic alternatives. Antibiotics affect eukaryotic cells at therapeutic concentrations, yet effects of AGPs on host physiology are relatively understudied, partially due to the complexity of host-microorganism interactions within the gastrointestinal tract. To determine the direct effects of AGPs on the host, we generated Altered Schaedler Flora (ASF) mice, and administered chlortetracycline (CTC) and tylosin phosphate (TYL) in feed. Mice were challenged with Citrobacter rodentium to determine how AGPs alter host responses to physiological stress. Although CTC and TYL had inconsistent effects on the ASF taxa, AGPs protected mice from weight loss following C. rodentium inoculation. Mice treated with either CTC or TYL had lower expression of βd1 and Il17a in the intestine and had a robust induction of Il17a and Il10. Furthermore, AGP administration resulted in a lower hepatic expression of acute phase proteins (Saa1, Hp, and Cp) in liver tissue, and ameliorated C. rodentium-induced reductions in the expression of genes involved in lipogenesis (Hmgcl and Fabp1). Collectively, this indicates that AGPs directly affect host physiology, and highlights important considerations in the development of non-antibiotic alternatives. PMID:27929072

  10. In vitro activity of five tetracyclines and some other antimicrobial agents against four porcine respiratory tract pathogens.

    Science.gov (United States)

    Pijpers, A; Van Klingeren, B; Schoevers, E J; Verheijden, J H; Van Miert, A S

    1989-09-01

    The minimal inhibitory concentrations (MIC) of five tetracyclines and ten other antimicrobial agents were determined for four porcine bacterial respiratory tract pathogens by the agar dilution method. For the following oxytetracycline-susceptible strains, the MIC50 ranges of the tetracyclines were: P. multocida (n = 17) 0.25-0.5 micrograms/ml; B. bronchiseptica (n = 20) 0.25-1.0 micrograms/ml; H. pleuropneumoniae (n = 20) 0.25-0.5 micrograms/ml; S. suis Type 2 (n = 20) 0.06-0.25 micrograms/ml. For 19 oxytetracycline-resistant P. multocida strains the MIC50 of the tetracyclines varied from 64 micrograms/ml for oxytetracycline to 0.5 micrograms/ml for minocycline. Strikingly, minocycline showed no cross-resistance with oxytetracycline, tetracycline, chlortetracycline and doxycycline in P. multocida and in H. pleuropneumoniae. Moreover, in susceptible strains minocycline showed the highest in vitro activity followed by doxycycline. Low MIC50 values were observed for chloramphenicol, ampicillin, flumequine, ofloxacin and ciprofloxacin against P. multocida and H. pleuropneumoniae. B. bronchiseptica was moderately susceptible or resistant to these compounds. As expected tiamulin, lincomycin, tylosin and spiramycin were not active against H. pleuropneumoniae. Except for flumequine, the MIC50 values of nine antimicrobial agents were low for S. suis Type 2. Six strains of this species showed resistance to the macrolides and lincomycin.

  11. Comparison of methods for in vitro testing of susceptibility of porcine Mycoplasma species to antimicrobial agents.

    Science.gov (United States)

    Ter Laak, E A; Pijpers, A; Noordergraaf, J H; Schoevers, E C; Verheijden, J H

    1991-02-01

    The MICs of 18 antimicrobial agents used against strains of three porcine Mycoplasma species were determined by a serial broth dilution method. Twenty field strains of M. hyorhinis, ten field strains of M. hyopneumoniae, six field strains of M. flocculare, and the type strains of these species were tested. Twelve field strains and the type strain of M. hyorhinis were also tested by an agar dilution method. Tests were read at various time points. When the broth dilution method was used, the final MIC had to be read 2 days after color changes had stopped. MICs of tetracycline, oxytetracycline, doxycycline, and minocycline were low for the three Mycoplasma species tested. MICs of chlortetracycline were 8 to 16 times higher than MICs of the other tetracyclines. Spiramycin, tylosin, kitasamycin, spectinomycin, tiamulin, lincomycin, and clindamycin were effective against all strains of M. hyorhinis and M. hyopneumoniae. The quinolones were highly effective against M. hyopneumoniae but less effective against M. hyorhinis. The susceptibility patterns for M. hyopneumoniae and M. flocculare were similar.

  12. Identification and expression of GABAc receptor in rat testis and spermatozoa

    Institute of Scientific and Technical Information of China (English)

    Shifeng Li; Yunbin Zhang; Haixiong Liu; Yuanchang Yan; Yiping Li

    2008-01-01

    Our previous studies showed that γ-aminobutyric acid (GABA)A and GABAB receptors are involved in rat sperm acrosome reaction induced by progesterone or GABA. Here,we report the presence of GABAc receptor in rat testis and spermatozoa.Full-length complementary DNA encoding the ρ1,ρ2 and ρ3 subunits of GABAc receptor were cloned from rat testis;their sequences are identical to those of rat GABAc receptor in retina.Reverse transcription-polymerase chain reaction analysis showed that during the development of rat testis,the transcript levels of the ρ1 and ρ2 subunits showed little change,while the expression of ρ3 was gradually up-regulated.Immunofluorescence analysis using an anti-ρ1 antibody revealed that GABAc receptor exists on the elongated spermatid and sperm.Using a chlortetracycline assay,we found that N(4)-chloroacetylcytosine arabinoside, a GABAc receptor agonist,triggered rat sperm acrosome reaction;whereas(1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid,a GABAc receptor antagonist, inhibited the ability of N(4)-chloroacetylcytosine arabinoside to induce acrosome reaction.These results suggested that GABAc receptors are also involved in rat sperm acrosome reaction.

  13. Biological effects of exogenous adenosine 5 prime -triphosphate on cultured mammalian cells: Evidence for a receptor mechanism and its regulation by desensitization

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez, F.A.

    1989-01-01

    Exogenous adenosine 5{prime}-triphosphate (ATP) mobilized intracellular calcium in human carcinoma A43l cells and in Swiss 3T3 and 3T6 mouse fibroblasts by increasing inositol trisphosphate similar to well down growth factors (platelet-derived growth factor (PDGF), epidermal growth factor (EGF), bradykinin (BK), serum). Calcium mobilization was examined by video imaging of fura-2 fluorescence is single cells, following the radioactive isotope {sup 45}Ca, and monitoring the decrease in fluorescence of cells loaded with chlortetracycline. Uridine 5{prime}-triphosphate, but not other nucleotides, mimicked ATP. Single-cell analysis revealed synchronous responses in 10 sec to ATP, BK or serum, while PDGF (3T3) and EGF (A431) produced slower signals with significant cell-to-cell variation. PDGF desensitized 3T3 cells to ATP and BK added 100 sec later but ATP or BK did not desensitized to PDGF. Homologous desensitization was seen with all agonists. Heterologous desensitization was also observed in A431 cells where ATP desensitized to serum, but serum did not to ATP. ATP-stimulated calcium entry was detected after 10 sec in A431 cells, but not in Swiss 3T6 cells. Entry started before significant efflux had occurred and did not fit the capacitance model of Putney. A 2-3 hr ATP pretreatment produced a homologous desensitization state that required 20 hr to disappear, probably due to down-regulation of the putative ATP receptors.

  14. Quantifying Attachment and Antibiotic Resistance of from Conventional and Organic Swine Manure.

    Science.gov (United States)

    Zwonitzer, Martha R; Soupir, Michelle L; Jarboe, Laura R; Smith, Douglas R

    2016-03-01

    Broad-spectrum antibiotics are often administered to swine, contributing to the occurrence of antibiotic-resistant bacteria in their manure. During land application, the bacteria in swine manure preferentially attach to particles in the soil, affecting their transport in overland flow. However, a quantitative understanding of these attachment mechanisms is lacking, and their relationship to antibiotic resistance is unknown. The objective of this study is to examine the relationships between antibiotic resistance and attachment to very fine silica sand in collected from swine manure. A total of 556 isolates were collected from six farms, two organic and four conventional (antibiotics fed prophylactically). Antibiotic resistance was quantified using 13 antibiotics at three minimum inhibitory concentrations: resistant, intermediate, and susceptible. Of the 556 isolates used in the antibiotic resistance assays, 491 were subjected to an attachment assay. Results show that isolates from conventional systems were significantly more resistant to amoxicillin, ampicillin, chlortetracycline, erythromycin, kanamycin, neomycin, streptomycin, tetracycline, and tylosin ( < 0.001). Results also indicate that isolated from conventional systems attached to very fine silica sand at significantly higher levels than those from organic systems ( < 0.001). Statistical analysis showed that a significant relationship did not exist between antibiotic resistance levels and attachment in from conventional systems but did for organic systems ( < 0.001). Better quantification of these relationships is critical to understanding the behavior of in the environment and preventing exposure of human populations to antibiotic-resistant bacteria.

  15. Expression, cellular localization, and involvement of the pentose phosphate pathway enzymes in the regulation of ram sperm capacitation.

    Science.gov (United States)

    Luna, C; Serrano, E; Domingo, J; Casao, A; Pérez-Pé, R; Cebrián-Pérez, J A; Muiño-Blanco, T

    2016-08-01

    Spermatozoa require substantially more ATP than other cells, not only for sustaining sperm motility but also for regulating protein phosphorylation during capacitation. In this study, we have reported for the first time the presence of the two key enzymes of the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in ovine spermatozoa by indirect immunofluorescence, Western blotting, in-gel activity, and reverse transcription polymerase chain reaction analysis. We found that the activity of both enzymes significantly increased after in vitro capacitation in the presence of high-cAMP levels, with a concomitant increase in protein tyrosine phosphorylation and in the proportion of sperm-capacitated pattern assessed by the chlortetracycline staining. These results suggest that PPP is related with the progress of capacitation and that a relationship between calcium compartmentalization, protein tyrosine phosphorylation and PPP seems to exist. This is the first report that shows a connection between the PPP, cAMP/PKA signaling pathways and sperm capacitation. These findings can be of high-biological importance to improve our knowledge of the biochemical mechanisms involved in the acquisition of mammalian sperm functional competence and, ultimately, fertility.

  16. Impact of medicated feed along with clay mineral supplementation on Escherichia coli resistance to antimicrobial agents in pigs after weaning in field conditions.

    Science.gov (United States)

    Jahanbakhsh, Seyedehameneh; Kabore, Kiswendsida Paul; Fravalo, Philippe; Letellier, Ann; Fairbrother, John Morris

    2015-10-01

    The aim of this study was to examine changes in antimicrobial resistance (AMR) phenotype and virulence and AMR gene profiles in Escherichia coli from pigs receiving in-feed antimicrobial medication following weaning and the effect of feed supplementation with a clay mineral, clinoptilolite, on this dynamic. Eighty E. coli strains isolated from fecal samples of pigs receiving a diet containing chlortetracycline and penicillin, with or without 2% clinoptilolite, were examined for antimicrobial resistance to 15 antimicrobial agents. Overall, an increased resistance to 10 antimicrobials was observed with time. Supplementation with clinoptilolite was associated with an early increase but later decrease in blaCMY-2, in isolates, as shown by DNA probe. Concurrently, a later increase in the frequency of blaCMY-2 and the virulence genes iucD and tsh was observed in the control pig isolates, being significantly greater than in the supplemented pigs at day 28. Our results suggest that, in the long term, supplementation with clinoptilolite could decrease the prevalence of E. coli carrying certain antimicrobial resistance and virulence genes.

  17. Sorption and Dissipation of Tetracyclines in Soils and Compost

    Institute of Scientific and Technical Information of China (English)

    LI Ling-Ling; HUANG Li-Dong; CHUNG Ren-Shih; FOK Ka-Hang; ZHANG Yong-Song

    2010-01-01

    Two agricultural soils were collected from Dahu and Pinchen counties and swine manure compost(SMC)from Ping-tung County in Taiwan,China to investigate the sorption and dissipation of three tetracyclines(TCs),i.e.,oxytetracycline(OTC),tetracycline(TC)and chlortetracycline(CTC),in compost,soils and soil/compost mixtures with different organic carbon(OC)contents.There were seven treatments in total.TCs were most strongly adsorbed to SMC in all treatments due to the high OC content.When SMC was present in the soils,the sorption of TCs was significantly enhanced,which might be attributed to the increased OC content and CEC.The adsorption of TCs showed non-linear adsorption isotherms and fitted well to the Preundlich model.After 49 d of incubation at 25 ℃ in soils and soil/compost mixtures in the dark,TCs elapsed in all substrates,with the time required for 50% degradation(DT50)between 20 and 41 d,and the time for90% degradation(DT90)between 68 and 137 d.Soil amended with compost enhanced the stability of TCs and reduced their mobility.The dissipation of TCs in a soil environment was slow,indicating that these compounds might be persistent in soil.

  18. A novel approach to modeling the reaction kinetics of tetracycline antibiotics with aqueous ozone.

    Science.gov (United States)

    Hopkins, Zachary R; Blaney, Lee

    2014-01-15

    Tetracycline antibiotics represent one of the most successful classes of pharmaceuticals and are extensively used around the world for human and veterinary health. Ozone-based processes have emerged as a selective water treatment process for many pharmaceuticals. The primary objective of this study was to determine the reaction kinetics for transformation of five tetracycline antibiotics (i.e., chlortetracycline, doxycycline, oxytetracycline, rolitetracycline, and tetracycline) by ozone across the pH2 to 9 range. The apparent second-order rate constant for tetracycline was on the order of 1-6 × 10(4) M(-1) s(-1) at low pH, and 0.6-2.0 × 10(6) M(-1) s(-1) at near neutral pH. The apparent second-order rate constants did not fit a conventional pKa-based model, presumably due to the complex acid/base speciation of tetracycline antibiotics. A model that considers the net charge on tetracycline molecules in solution provided a nice fit to experimental data for all five tetracyclines. The five tetracycline antibiotics demonstrated similar reaction kinetics with ozone, and a cumulative analysis of all kinetics data provides a baseline model for other tetracycline compounds. The ozone exposure required for complete transformation of tetracycline antibiotics (10(-5) M-s) is well below that achieved during ozone disinfection processes (10(-3) M-s), indicating that ozone is an effective treatment for tetracycline antibiotics.

  19. Simultaneous Determination of TetracyclinesResidues in Bovine Milk Samples by Solid Phase Extraction and HPLC-FL Method

    Directory of Open Access Journals (Sweden)

    Mehra Mesgari Abbasi

    2011-06-01

    Full Text Available Introduction:Tetracyclines (TCs are widely used in animal husbandry and their residues in milk may resultinharmful effects on human. The aim of this study was to investigate the presence of TCs residues in various bovine milk samples from local markets of Ardabil, Iran. Methods:One hundred and fourteen pasteurized, sterilized and raw milk samples were collected from markets of Ardabil. Tetracycline, Oxytetracycline and Chlortetracycline (TCs residues extraction carried out by Solid Phase Extraction method. Determination of TCs residues were performed by high performance liquid chromatography (HPLC method using Fluorescence detector.Results: The mean of total TCs residues in all samples (114 samples was 97.6 ±16.9ng/g and that of pasteurized, sterilized and raw milk samples were 87.1 ± 17.7, 112.0 ± 57.3 and 154.0 ± 66.3ng/g respectively. Twenty five point four percent of the all samples, and24.4%, 30% and 28.6% of the pasteurized, sterilized and raw milk samples, respectively had higher TCs residues than the recommended maximum levels (100ng/g. Conclusion:This study indicates the presence of tetracycline residues more than allowed amount. Regulatory authorities should ensure proper withdrawal period before milking the animals and definite supervisions are necessary on application of these drugs.

  20. Disinfection of swine wastewater using chlorine, ultraviolet light and ozone.

    Science.gov (United States)

    Macauley, John J; Qiang, Zhimin; Adams, Craig D; Surampalli, Rao; Mormile, Melanie R

    2006-06-01

    Veterinary antibiotics are widely used at concentrated animal feeding operations (CAFOs) to prevent disease and promote growth of livestock. However, the majority of antibiotics are excreted from animals in urine, feces, and manure. Consequently, the lagoons used to store these wastes can act as reservoirs of antibiotics and antibiotic-resistant bacteria. There is currently no regulation or control of these systems to prevent the spread of these bacteria and their genes for antibiotic resistance into other environments. This study was conducted to determine the disinfection potential of chlorine, ultraviolet light and ozone against swine lagoon bacteria. Results indicate that a chlorine dose of 30 mg/L could achieve a 2.2-3.4 log bacteria reduction in lagoon samples. However, increasing the dose of chlorine did not significantly enhance the disinfection activity due to the presence of chlorine-resistant bacteria. The chlorine resistant bacteria were identified to be closely related to Bacillus subtilis and Bacillus licheniformis. A significant percentage of lagoon bacteria were not susceptible to the four selected antibiotics: chlortetracycline, lincomycin, sulfamethazine and tetracycline (TET). However, the presence of both chlorine and TET could inactivate all bacteria in one lagoon sample. The disinfection potential of UV irradiation and ozone was also examined. Ultraviolet light was an effective bacterial disinfectant, but was unlikely to be economically viable due to its high energy requirements. At an ozone dose of 100 mg/L, the bacteria inactivation efficiency could reach 3.3-3.9 log.

  1. A conditionally lethal mutant of Salmonella Typhimurium induces a protective response in mice.

    Science.gov (United States)

    Hidalgo, Alejandro A; Villagra, Nicolás A; Jerez, Sebastián A; Fuentes, Juan A; Mora, Guido C

    2016-02-01

    Here we present the design of a conditionally lethal mutant of Salmonella enterica serovar Typhimurium (S. Typhimurium) which growth depends on tetracycline (Tet). Four mutants of S. Typhimurium, with Tet-conditional growth, were created by inserting the tetRA cassette. Three of the mutants presented a conditional-lethal phenotype in vitro. One mutant in the yabB gene remained conditional inside cells and did not persisted after 24 h in cell cultures. The capacity of S. Typhimurium yabB::tetRA to invade deep organs was investigated in intraperitoneally (IP) infected mice fed with or without chlortetracycline (CTet), a Tet analog with lower antibiotic activity. The yabB::tetRA mutant was undetectable in liver or spleen of animals under normal diet, while in mice under diet including CTet, yabB::tetRA invaded at a level comparable to the WT in mice under normal diet. Moreover, yabB::tetRA produced a strong humoral-immunoresponse after one IP immunization with 10(6) bacteria, measured as serum reactivity against S. Typhimurium whole cell extract. By contrast, oral immunization with 10(6) bacteria was weaker and variable on inducing antibodies. Consistently, IP infected mice were fully protected in a challenge with 10(4) oral S. Typhimurium, while protection was partial in orally immunized mice. Our data indicate that S. Typhimurium yabB::tetRA is a conditionally attenuated strain capable of inducing a protective response in mice in non-permissive conditions.

  2. Effects of Dietary Saccharicterpenin on Mucosal Structure of Small Intestine in Gushi Chickens%糖萜素对固始鸡小肠黏膜结构的影响

    Institute of Scientific and Technical Information of China (English)

    黄鹏; 赵珊珊; 张学武; 张婷婷; 肖传斌

    2011-01-01

    This experiment was conducted to study the effects of dietary saccharicterpenin level on mucosal structure of small intestine in Gushi chickens. A total of 300 Gushi chickens were randomly assigned to five dietary treatments; control (basal diet) group, 300 mg/kg saccharicterpenin group, 500 mg/kg saccharicterpenin group, 700 mg/kg saccharicterpenin group and chlortetracycline (200 mg/kg) group. Each group had three replicates and each replicate had twenty chickens. Six chickens at 21, 35 and 49 days of age were picked in each group and bled to death. Using the conventional paraffin sections and HE staining, the mucosal structure of the duodenum, jejunum and ileum were observed, and then the villus height, crypt depth and intestinal wall thickness were measured. The results showed as follows: 1) there were no significant differences among all groups in the length of duodenum, jejunum and ileum and the thickness of intestinal wall in Gushi chickens aged different days (P >0. 05). 2) At different ages, the duodenum villi height of Gushi chickens in the 500 mg/kg saccharicterpenin group and 700 mg/kg saccharicterpenin group was higher, and there was no significant difference between the two groups (P >0.05), but at 21 days of age, it was significantly higher than that in the other groups (P 0. 05). At different ages, the duodenum V/C value of Gushi chickens in the 500 mg/kg saccharicterpenin group and chlortetracycline (200 mg/kg) group was higher, and there was no significant difference between the two groups(P >0. 05), but at 21 to 35 days of age, it was significantly higher than that in the control group (P > 0.05). 3) At different ages, the jejunum villi height of Gushi chickens in the 500 mg/kg saccharicterpenin group was the highest, and at 21 to 35 days of age, it was significantly higher than that in the control group (P 0.05). At different ages, the jejunum V/C value of Gushi chickens in the 500 mg/kg saccharicterpenin group, 700 mg/kg saccharicterpenin group

  3. Sperm Oxidative Stress Is Detrimental to Embryo Development: A Dose-Dependent Study Model and a New and More Sensitive Oxidative Status Evaluation

    Directory of Open Access Journals (Sweden)

    Letícia S. de Castro

    2016-01-01

    Full Text Available Our study aimed to assess the impact of sperm oxidative stress on embryo development by means of a dose-dependent model. In experiment 1, straws from five bulls were subjected to incubation with increasing H2O2 doses (0, 12.5, 25, and 50 μM. Motility parameters were evaluated by Computed Assisted System Analysis (CASA. Experiment 2 was designed to study a high (50 μM and low dose (12.5 μM of H2O2 compared to a control (0 μM. Samples were incubated and further used for in vitro fertilization. Analyses of motility (CASA, oxidative status (CellROX green and 2’-7’ dichlorofluorescein diacetate, mitochondrial potential (JC-1, chromatin integrity (AO, and sperm capacitation status (chlortetracycline were performed. Embryos were evaluated based on fast cleavage (30 h.p.i., cleavage (D=3, development (D=5, and blastocyst rates (D=8. We observed a dose-dependent deleterious effect of H2O2 on motility and increase on the percentages of positive cells for CellROX green, capacitated sperm, and AO. A decrease on cleavage and blastocyst rates was observed as H2O2 increased. Also, we detected a blockage on embryo development. We concluded that sperm when exposed to oxidative environment presents impaired motility traits, prooxidative status, and premature capacitation; such alterations resulting in embryo development fail.

  4. Toxicity of fungicides to natural bacterial communities in wetland water and sediment measured using leucine incorporation and potential denitrification.

    Science.gov (United States)

    Milenkovski, Susann; Bååth, Erland; Lindgren, Per-Eric; Berglund, Olof

    2010-02-01

    We assessed potential toxicity of fungicides to natural bacterial communities from a constructed wetland, located in southern Sweden, and compared the sensitivity of two endpoints indicating bacterial activity, leucine incorporation, and potential denitrification, in detecting toxicity. The effects of eight fungicides (benomyl, carbendazim, carboxin, captan, cycloheximide, fenpropimorph, propiconazole, and thiram), two bactericides (bronopol and chlortetracycline) as controls, and one reference compound (3,5-dichlorophenol), were tested in a water-sediment microcosm set-up. Leucine incorporation was measured in both the water and sediment column, while potential denitrification was measured for the entire microcosm. The bactericides and the reference compound gave sigmoid concentration-response curves for both endpoints in all but one case. The fungicides thiram, captan, and benomyl, and to a lesser extent fenpropimorph and propiconazole had quantifiable toxic effects on leucine incorporation, with EC(50) values ranging from 3 to 70 mg l(-1), while carbendazim, carboxin, and cycloheximide had little effect at the investigated concentrations. Only thiram and captan inhibited potential denitrification; the other fungicides showed no quantifiable effect. A greater toxic effect on leucine incorporation was recorded for bacterial communities associated with the water column, compared to the sediment column, for all tested compounds. Leucine incorporation was the more sensitive method for toxicity assessment of bacterial communities, and also allowed for a rapid and simple way of comparing exposure in the sediment and water column, making it an attractive standard method for community based toxicological assays in aquatic environments.

  5. Tetracycline residues in meat and bone meals. Part 2: the effect of heat treatments on bound tetracycline residues.

    Science.gov (United States)

    Kühne, M; Körner, U; Wenzel, S

    2001-07-01

    The stability of bound tetracycline residues during heat treatments at 133 degrees C and 100 degrees C for up to 45 min was investigated. An intermediate product from a rendering plant was mixed with bone splinters that contained bound tetracycline (TC) and chlortetracycline (CTC) residues. The mixture was heated in an autoclave at 133 degrees C for 20, 30 and 45 min and at 100 degrees C for 20 and 30 min and subsequently dried at 103 degrees C for 4 h. Two different extraction procedures with hydrochloric acid were used, one with and one without the previous sedimentation of bone particles. Tetracycline concentrations were determined by HPLC analysis before and after the heat treatment. A complete destruction of tetracyclines during heat treatment at 133 degrees C could not be demonstrated, but there was a significant decrease of TC by about 50%. CTC was less resistant to the same temperature, which brought about a reduction of 90-100%. Treatment at 100 degrees C did not bring about any reduction, except for CTC after extraction without sedimentation. The possible toxicological relevance of the findings is discussed. Further research has to be done on possible degradation products of the tetracycline derivatives.

  6. Research on the Migration Regularity of 5 Antibiotics Residue from Swine and Chicken Feedstuff to Organic Fertilizer%五种抗菌药物从猪、鸡饲料至有机肥中的药物残留迁移规律研究

    Institute of Scientific and Technical Information of China (English)

    吴剑平; 张鑫; 李丹妮; 严凤; 周悦榕; 顾欣

    2015-01-01

    To determine the migration regularity of antibiotics residue from swine and chicken feedstuff to organic fertilizer, 40swines and 200 chickens were selected and divided into two groups to do the feeding tests and fermentation experiments. The blank group was fed with only complete feed, while the test group was fed with the complete feed with 50 mg/kg chlortetracycline hydrochloride, 50 mg/kg tylosin phosphate, 50 mg/kg sulfametoxydiazine, 150 mg/kg enrofloxacin and 10 mg/kg trimethoprim. The manure was collected in time every day, and the sample was extracted and purified,before it was analyzed by the LC-MS. The results indicated that the antibiotics residue could be tested in manure after the swine and chicken were fed with antibiotics feedstuff, and it was able to enter the manufacture of organic fertilizer. The manure models were established according to the top value of the antibiotics residue in test group manure. The fermentation experiments were done to determine the residue of antibiotics in manure. The results indicated that enrofloxacin, tylosin phosphate and trimethoprim would be degraded by the rise of temperature and oxidation with a half-life of 5 weeks and they would be lower than limit of detection after 10 weeks. But chlortetracycline hydrochloride and sulfametoxydiazine would be less degraded with a higher risk of residue migration.%为研究抗菌药物从饲料向畜禽粪肥转移的残留迁移规律,将40头猪和200羽鸡分为饲喂组和空白对照组进行饲喂对照实验和粪便堆肥发酵实验,其中空白对照组使用普通全价饲料,饲喂组在饲料中加入盐酸金霉素50 mg/kg、磷酸泰乐菌素50 mg/kg、磺胺对甲氧嘧啶各50 mg/kg、恩诺沙星150 mg/kg、二甲氧苄啶10 mg/kg,每日及时对各组实验动物粪便进行采集,样品经提取和净化后使用液相色谱-串联质谱进行分析定量。所得结果表明,猪、鸡饲喂含多种抗生素饲料后粪便中可以

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

    Institute of Scientific and Technical Information of China (English)

    ZHANG Zhi-qiang; WANG Xiu-bin; LI Chun-hua; HUANG Shao-wen; GAO Wei; TANG Ji-wei; JIN Ji-yun

    2015-01-01

    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 lfuvo-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 ifeld 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+1/4 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 beneift in the ifrst four seasons, but addition of straw (2/4 CN+1/4 MN+1/4 SN treatment) performed better in the subsequent two seasons, and the average yields of 2/4 CN+1/4 MN+1/4 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

  8. Abundance and Persistence of Tetracycline Resistance Genes in Fecal Depositsy from Beef Cattle Fed Different Subtherapeutic Antibiotics%饲喂不同抗生素对肉牛废弃粪便中四环素耐药基因数量及持久性的影响

    Institute of Scientific and Technical Information of China (English)

    金鑫; 张文广; 张燕军; 苏蕊; 王瑞军; 李金泉

    2012-01-01

    通过对肉牛饲喂低于治疗水平的不同种类抗生素,研究其对肉牛废弃粪便微生物群落中四环素耐药基因数量与持久性的影响.试验肉牛被分成不同抗生素处理组,即氯四环素组、氯四环素和磺胺甲嘧啶组及对照组.将每个围栏中所有动物的新鲜粪便混匀为一份混合样品作为模式样品(每个处理组3份),分别露天放置,在第7、14、28、42、56、70、84、98、112、126和175天时采样并提取DNA,利用Real-time PCR方法测定四环素耐药基因tet(B),tet( C),tet(L),tet(M),tet(W)及16S rRNA的浓度.结果显示,16S rRNA的浓度在不同处理间相似,在56 d内均有增加的趋势(P<0.05);总体上看,四环素组的耐药基因初始浓度较高(P<0.05);所有处理组的tet(B)和tet(C)浓度到56 d时均增长了1~2个对数级,到175 d时又降低到初始水平,而tet(M)与tet(W)的浓度与其他耐药基因相比较高.因此,四环素耐药基因可以在废弃粪便中持续存在超过175 d,而某些基因的最初数量可能会导致错估其后的变化,其浓度的暂时改变并不能归因于微生物群落数量的变化.%This study was investigated the effects of administering beef cattle antimicrobials at subtherapeutic concentrations on the abundance and persistence of tetracycline resistance genes within the whole microbial community of fecal waste . Cattl were administered chlortetracyclinei chlortetracycline plus sulfamethazine and no antimicrobials. Model fecal deposits (n-3) were prepared by mixing fresh feces from pens into a single composite sample. Real-time PCR was used to measure concentrations of tetracycline resistance genes tet(B) , tet(C) , tet(L) , tet(M) , tet(W) and 16S rRNA in DNA extracted from composite feces after 7, 14, 28, 42, 56, 70, 84, 98, 112, 126 and 175 days in the field. The concentrations of 16S rRNA in feces were similar across treatments and increased by day 56. Generally, initial concentrations of

  9. A reversed-phase high performance liquid chromatography coupled with resonance Rayleigh scattering detection for the determination of four tetracycline antibiotics

    Energy Technology Data Exchange (ETDEWEB)

    Wang Lifeng [School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Peng Jingdong [School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)], E-mail: hxpengjd@swu.edu.cn; Liu Limin [School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

    2008-12-07

    A new reversed-phase high performance liquid chromatography with resonance Rayleigh scattering detection (HPLC-RRS) was developed for simultaneous separation and determination of four tetracycline antibiotics (TCs). A good chromatographic separation among the compounds was achieved using a Synergi Fusion-RP column (150 mm x 4.6 mm; 4 {mu}m) and a mobile phase consisting of methanol-acetonitrile-oxalic acid (5 mM) at the flow rate of 0.8 mL min{sup -1}. Column temperature was 30 deg. C. The RRS signal was detected at {lambda}{sub ex} = {lambda}{sub em} = 370 nm. The recoveries of sample added standard ranged from 95.3% to 103.5%, and the relative standard deviation was below 2.79%. A detection limit of 2.12-5.12 {mu}g mL{sup -1} was reached and a linear range was found between peak height and concentration in the range of 10.36-518.0 {mu}g mL{sup -1} for oxytetracycline (OTC), 12.11-605.5 {mu}g mL{sup -1} for tetracycline (TC), 11.79-589.5 {mu}g mL{sup -1} for chlortetracycline (CTC) and 10.32-516.0 {mu}g mL{sup -1} for doxycycline (DC). The linear regression coefficients were all above 0.999. The method has been applied successfully to the determination of OTC, TC, CTC, DC in pharmaceutical formulations and in honey. The method was simple, rapid and showed a better linear relation and high repeatability.

  10. Diurnal variations in the occurrence and the fate of hormones and antibiotics in activated sludge wastewater treatment in Oslo, Norway

    Energy Technology Data Exchange (ETDEWEB)

    Plosz, Benedek Gy., E-mail: benedek.plosz@niva.no [Norwegian Institute for Water Research, NIVA, Gaustadalleen 21, NO-0349, Oslo (Norway); Leknes, Henriette [Norwegian Institute for Air Research NILU, 2027 Kjeller (Norway); Liltved, Helge; Thomas, Kevin V. [Norwegian Institute for Water Research, NIVA, Gaustadalleen 21, NO-0349, Oslo (Norway)

    2010-03-15

    We present an assessment of the dynamics in the influent concentration of hormones (estrone, estriol) and antibiotics (trimethoprim, sulfamethoxazole, tetracycline, ciprofloxacin) in the liquid phase including the efficiency of biological municipal wastewater treatment. The concentration of estradiol, 17-{alpha}-ethinylestradiol, doxycycline, oxytetracycline, demeclocycline, chlortetracycline, cefuroxime, cyclophosphamide, and ifosfamide were below the limit of detection in all of the sewage samples collected within this study. Two different types of diurnal variation pattern were identified in the influent mass loads of selected antibiotics and hormones that effectively correlate with daily drug administration patterns and with the expected maximum human hormone release, respectively. The occurrence of natural hormones and antimicrobials, administered every 12 hours, shows a daily trend of decreasing contaminant mass load, having the maximum values in the morning hours. The occurrence of antibiotics, typically administered every 8 hours, indicates a daily peak value in samples collected under the highest hydraulic loading. The efficiency of biological removal of both hormones and antibiotics is shown to be limited. Compared to the values obtained in the influent samples, increased concentrations are observed in the biologically treated effluent for trimethoprim, sulfamethoxazole and ciprofloxacin, mainly as a result of deconjugation processes. Ciprofloxacin is shown as the predominant antimicrobial compound in the effluent, and it is present at quantities approximately 10 fold greater than the total mass of the other of the compounds due to poor removal efficiency and alternating solid-liquid partitioning behaviour. Our results suggest that, to increase the micro-pollutant removal and the chemical dosing efficiency in enhanced tertiary treatment, significant benefits can be derived from the optimisation of reactor design and the development of control schemes that

  11. Diversity and distribution of commensal fecal Escherichia coli bacteria in beef cattle administered selected subtherapeutic antimicrobials in a feedlot setting.

    Science.gov (United States)

    Sharma, Ranjana; Munns, Krysty; Alexander, Trevor; Entz, Toby; Mirzaagha, Parasto; Yanke, L Jay; Mulvey, Michael; Topp, Edward; McAllister, Tim

    2008-10-01

    Escherichia coli strains isolated from fecal samples were screened to examine changes in phenotypic and genotypic characteristics including antimicrobial susceptibility, clonal type, and carriage of resistance determinants. The goal of this 197-day study was to investigate the influence of administration of chlortetracycline alone (T) or in combination with sulfamethazine (TS) on the development of resistance, dissemination of defined strain types, and prevalence of resistance determinants in feedlot cattle. Inherent tetracycline resistance was detected in cattle with no prior antimicrobial exposure. Antimicrobial administration was not found to be essential for the maintenance of inherently ampicillin-resistant and tetracycline-resistant (Tet(r)) E. coli in control animals; however, higher Tet(r) E. coli shedding was observed in animals subjected to the two treatments. At day 0, high tetracycline (26.7%), lower sulfamethoxazole-tetracycline (19.2%), and several other resistances were detected, which by the finishing phase (day 197) were restricted to ampicillin-tetracycline (47.5%), tetracycline (31.7%), and ampicillin-tetracycline-sulfamethoxazole (20.8%) from both treated and untreated cattle. Among the determinants, bla(TEM1), tet(A), and sul2 were prevalent at days 0 and 197. Further, E. coli from day 0 showed diverse antibiogram profiles and strain types, which by the finishing phase were limited to up to three, irrespective of the treatment. Some genetically identical strains expressed different phenotypes and harbored diverse determinants, indicating that mobile genetic elements contribute to resistance dissemination. This was supported by an increased linked inheritance of ampicillin and tetracycline resistance genes and prevalence of specific strains at day 197. Animals in the cohort shed increasingly similar genotypes by the finishing phase due to animal-to-animal strain transmission. Thus, characterizing inherent resistance and propagation of cohort

  12. 沼液中几种抗生素的检测与樱花细菌性穿孔病防效研究%Detection of Antibiotics in Biogas Slurry and Study on its Control Effect on Cherry Blossom Bacterial Shot Hole Disease

    Institute of Scientific and Technical Information of China (English)

    聂莹; 曾晓楠

    2014-01-01

    In order to find out the control mechanism of biogas slurry to plant diseases, the study determined the contents of antibiotics existed in biogas slurry with high performance liquid chromatography (HPLC), and then observed its prevention and control effect on cherry blossom bacterial shot-hole disease. Results showed that antibiotics existed in biogas slurry included oxytetracycline (OTC), tetracycline (TC) and chlortetracycline (CTC) with amount of 0.08347 mg/L, 0.04907 mg/L and 0.04229 mg/L respectively. The prevention and control effects indicated that application of biogas slurry for 4 times in Summer resulted in a similar efficiency as that of streptomycin, which is a common pesticide. But the efficiency was not good when biogas slurry was applied less than 3 times.%为探索沼液防治植物病害的机理,利用高效液相色谱检测沼液中几种抗生素的含量,并对樱花细菌性穿孔病防效进行研究。检测结果表明,抗生素含量分别为:土霉素(OTC)0.08347 mg/L、四环素(TC)0.04907 mg/L、金霉素(CTC)0.04229 mg/L。防效试验结果表明:沼液4次以上的连续叶面喷施与细菌性病害防治的常用农药链霉素效果相当,但3次以下的喷施防效不理想。

  13. Identification of soil contamination hotspots with veterinary antibiotics using heavy metal concentrations and leaching data--a field study in China.

    Science.gov (United States)

    Ostermann, Anne; Gao, Jing; Welp, Gerhard; Siemens, Jan; Roelcke, Marco; Heimann, Lisa; Nieder, Rolf; Xue, Qiaoyun; Lin, Xianyong; Sandhage-Hofmann, Alexandra; Amelung, Wulf

    2014-11-01

    In regions with high livestock densities, the usage of antibiotics and metals for veterinary purposes or as growth promoters poses a risk in manured soils. We investigated to which degree the concentrations and depth distributions of Cu, Zn, Cr and As could be used as a tracer to discover contaminations with sulfonamides, tetracyclines and fluoroquinolones. Besides, we estimated the potential vertical translocation of antibiotics and compared the results to measured data. In the peri-urban region of Beijing, China, soil was sampled from agricultural fields and a dry riverbed contaminated by organic waste disposal. The antibiotic concentrations reached 110 μg kg(-1) sulfamethazine, 111 μg kg(-1) chlortetracycline and 62 μg kg(-1) enrofloxacin in the topsoil of agricultural fields. Intriguingly, total concentrations of Cu, Zn, Cr and As were smaller than 65, 130, 36 and 10 mg kg(-1) in surface soil, respectively, therewith fulfilling Chinese quality standards. Correlations between sulfamethazine concentrations and Cu or Zn suggest that in regions with high manure applications, one might use the frequently existing monitoring data for metals to identify potential pollution hotspots for antibiotics in topsoils. In the subsoils, we found sulfamethazine down to ≥2 m depth on agricultural sites and down to ≥4 m depth in the riverbed. As no translocation of metals was observed, subsoil antibiotic contamination could not be predicted from metal data. Nevertheless, sulfonamide stocks in the subsoil could be estimated with an accuracy of 35-200 % from fertilisation data and potential leaching rates. While this may not be sufficient for precise prediction of antibiotic exposure, it may very well be useful for the pre-identification of risk hotspots for subsequent in-depth assessment studies.

  14. A membrane-associated adenylate cyclase modulates lactate dehydrogenase and creatine kinase activities required for bull sperm capacitation induced by hyaluronic acid.

    Science.gov (United States)

    Fernández, Silvina; Córdoba, Mariana

    2017-04-01

    Hyaluronic acid, as well as heparin, is a glycosaminoglycan present in the female genital tract of cattle. The aim of this study was to evaluate oxidative metabolism and intracellular signals mediated by a membrane-associated adenylate cyclase (mAC), in sperm capacitation with hyaluronic acid and heparin, in cryopreserved bull sperm. The mAC inhibitor, 2',5'-dideoxyadenosine, was used in the present study. Lactate dehydrogenase (LDH) and creatine kinase (CK) activities and lactate concentration were determined spectrophotometrically in the incubation medium. Capacitation and acrosome reaction were evaluated by chlortetracycline technique, while plasma membrane and acrosome integrity were determined by trypan blue stain/differential interference contrast microscopy. Heparin capacitated samples had a significant decrease in LDH and CK activities, while in hyaluronic acid capacitated samples LDH and CK activities both increased compared to control samples, in heparin and hyaluronic acid capacitation conditions, respectively. A significant increase in lactate concentration in the incubation medium occurred in hyaluronic acid-treated sperm samples compared to heparin treatment, indicating this energetic metabolite is produced during capacitation. The LDH and CK enzyme activities and lactate concentrations in the incubation medium were decreased with 2',5'-dideoxyadenosine treatment in hyaluronic acid samples. The mAC inhibitor significantly inhibited heparin-induced capacitation of sperm cells, but did not completely inhibit hyaluronic acid capacitation. Therefore, hyaluronic acid and heparin are physiological glycosaminoglycans capable of inducing in vitro capacitation in cryopreserved bull sperm, stimulating different enzymatic pathways and intracellular signals modulated by a mAC. Hyaluronic acid induces sperm capacitation involving LDH and CK activities, thereby reducing oxidative metabolism, and this process is mediated by mAC.

  15. The effect of an acidified, ionized copper sulphate solution on digital dermatitis in dairy cows.

    Science.gov (United States)

    Holzhauer, Menno; Bartels, Chris J; Bergsten, Christer; van Riet, Miriam M J; Frankena, Klaas; Lam, T J G M

    2012-09-01

    Digital dermatitis (DD) is the most important infectious claw disorder in dairy cattle and herd-based foot bathing with antibacterials, such as 4% formalin, is often used to prevent it. However, there is a lack of long-term studies of the effectiveness of such regimes and in this study the preventive and curative effect of 4% formalin was compared with that of an acidified, ionized copper sulphate solution over a 4-month period on a commercial 120-cow dairy farm. The cows were walked through a split-leg footbath where left claws were treated with an acidified copper solution twice daily for 5 days/week, while right claws were treated with 4% formalin twice daily for 1 day/every second week. Hind claws were scored for the presence and severity of DD in a trimming chute at the start of the study and every 4 weeks throughout the study period. At the start of the study 21/110 cows had ulcerative DD lesions with 10 on the left hind foot, 8 on the right and 3 on both. These lesions, as well as any new lesions which arose during the study, were treated with chlortetracycline spray. During the study 440 observations were made and seven new DD lesions were recorded on left feet (copper treated) and 20 on right feet (formalin treated). Based on survival analysis, the risk of developing a new ulcerative DD lesion on copper-treated (left hind) feet was almost three times less (RR=0.37, 95% CI 0.16-0.91) than formalin-treated (right hind) feet. Cure rates of DD lesions were not different between copper and formalin.

  16. Antimicrobial resistance trends among Salmonella isolates obtained from dairy cattle in the northeastern United States, 2004-2011.

    Science.gov (United States)

    Cummings, Kevin J; Perkins, Gillian A; Khatibzadeh, Sarah M; Warnick, Lorin D; Altier, Craig

    2013-04-01

    Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform public policy regarding appropriate antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Salmonella isolates from dairy cattle in the northeastern United States and to identify trends in resistance to various antimicrobial agents over time. Data were collected retrospectively for all bovine Salmonella isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Temporal trends in the prevalence of resistant Salmonella were investigated for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 2745 bovine Salmonella isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 0% (amikacin, ciprofloxacin, and nalidixic acid) to 72.0% (sulfadimethoxine). There was evidence of a significantly decreasing trend in prevalence of resistance to most agents: amoxicillin/clavulanic acid (AUG), ampicillin (AMP), cefoxitin (FOX), ceftiofur (TIO), ceftriaxone (AXO), chloramphenicol (CHL), chlortetracycline (CTET), florfenicol (FFN), kanamycin (KAN), neomycin (NEO), oxytetracycline (OXY), spectinomycin (SPE), streptomycin (STR), sulfadimethoxine (SDM), sulfisoxazole (FIS), and tetracycline (TET). Among the 265 isolates that were tested using the National Antimicrobial Resistance Monitoring System (NARMS) panel, the most common resistance patterns were pansusceptible (54.0%), AUG-AMP-FOX-TIO-AXO-CHL-KAN-STR-FIS-TET (18.1%), and AUG-AMP-FOX-TIO-AXO-CHL-STR-FIS-TET (12.1%). Increasing prevalence of S. enterica serovar Cerro over the course of the study period presumably had an impact on the observed resistance trends. Nevertheless, these results do not support the notion that the current level of antimicrobial

  17. Intracellular pH and calcium signaling as molecular targets of diclofenac-induced apoptosis against colon cancer.

    Science.gov (United States)

    Kaur, Jasmeet; Sanyal, Sankar Nath

    2011-07-01

    The role of intracellular pH and Ca2+ and their association with mitochondrial dysfunction and intracellular reactive oxygen species (ROS) are explored in the chemoprevention of colon cancer. 1,2-dimethylhydrazine dihydrochloride (DMH), a potent procarcinogen with selectivity for the colon, at a dose of 30 mg/kg body weight was used to induce initial stages of colon cancer when administered for 6 weeks in male Sprague-Dawley rats. Diclofenac, a preferential cyclooxygenase-2 inhibitor, was used at the anti-inflammatory dose (8 mg/kg body weight) for chemoprevention. The control group was administered vehicles for both DMH and diclofenac. A diclofenac-alone group with the same dose was also run simultaneously. Intracellular pH values as determined by biscarboxyethyl carboxyfluorescein fluorescence assay showed an alkaline pH in colonocytes from the DMH-treated group as compared with the control group. Moreover, the level of intracellular Ca2+ was also found to be decreased with DMH treatment, as shown by the fura-2 acetoxymethyl study and chlortetracycline assay. Apoptosis was studied by comet assay and Apaf-1 immunofluorescent expression and was found to be markedly decreased in this group, indicating that disturbances in pH and Ca2+ homeostasis promoted proliferation in colon and inhibited apoptosis. Changes in mitochondrial membrane potential and ROS levels were analyzed in isolated colonocytes by rhodamine 123 and 2,7-dichlorofluorescein diacetate labeling, respectively. DMH treatment promoted a higher mitochondrial membrane potential while reducing ROS levels. These parameters are known to be associated with pH and Ca2+ changes intracellularly and hence can be suggested to be linked with them in this study also. Diclofenac promoted apoptosis in colonocytes when coadministered with DMH and also ameliorated the changes observed in the above parameters, confirming these mechanisms as early events for the onset of apoptosis in cancer cells.

  18. Prevalence of veterinary antibiotics and antibiotic-resistant Escherichia coli in the surface water of a livestock production region in northern China.

    Directory of Open Access Journals (Sweden)

    Xuelian Zhang

    Full Text Available This study investigated the occurrence of 12 veterinary antibiotics (VAs and the susceptibility of Escherichia coli (E. coli in a rural water system that was affected by livestock production in northern China. Each of the surveyed sites was determined with at least eight antibiotics with maximum concentration of up to 450 ng L(-1. The use of VAs in livestock farming probably was a primary source of antibiotics in the rivers. Increasing total antibiotics were measured from up- to mid- and downstream in the two tributaries. Eighty-eight percent of the 218 E. coli isolates that were derived from the study area exhibited, in total, 48 resistance profiles against the eight examined drugs. Significant correlations were found among the resistance rates of sulfamethoxazole-trimethoprim, chloromycetin and ampicillin as well as between tetracycline and chlortetracycline, suggesting a possible cross-selection for resistance among these drugs. The E. coli resistance frequency also increased from up- to midstream in the three rivers. E. coli isolates from different water systems showed varying drug numbers of resistance. No clear relationship was observed in the antibiotic resistance frequency with corresponding antibiotic concentration, indicating that the antibiotic resistance for E. coli in the aquatic environment might be affected by factors besides antibiotics. High numbers of resistant E. coli were also isolated from the conserved reservoir. These results suggest that rural surface water may become a large pool of VAs and resistant bacteria. This study contributes to current information on VAs and resistant bacteria contamination in aquatic environments particularly in areas under intensive agriculture. Moreover, this study indicates an urgent need to monitor the use of VAs in animal production, and to control the release of animal-originated antibiotics into the environment.

  19. Prevalence of veterinary antibiotics and antibiotic-resistant Escherichia coli in the surface water of a livestock production region in northern China.

    Science.gov (United States)

    Zhang, Xuelian; Li, Yanxia; Liu, Bei; Wang, Jing; Feng, Chenghong; Gao, Min; Wang, Lina

    2014-01-01

    This study investigated the occurrence of 12 veterinary antibiotics (VAs) and the susceptibility of Escherichia coli (E. coli) in a rural water system that was affected by livestock production in northern China. Each of the surveyed sites was determined with at least eight antibiotics with maximum concentration of up to 450 ng L(-1). The use of VAs in livestock farming probably was a primary source of antibiotics in the rivers. Increasing total antibiotics were measured from up- to mid- and downstream in the two tributaries. Eighty-eight percent of the 218 E. coli isolates that were derived from the study area exhibited, in total, 48 resistance profiles against the eight examined drugs. Significant correlations were found among the resistance rates of sulfamethoxazole-trimethoprim, chloromycetin and ampicillin as well as between tetracycline and chlortetracycline, suggesting a possible cross-selection for resistance among these drugs. The E. coli resistance frequency also increased from up- to midstream in the three rivers. E. coli isolates from different water systems showed varying drug numbers of resistance. No clear relationship was observed in the antibiotic resistance frequency with corresponding antibiotic concentration, indicating that the antibiotic resistance for E. coli in the aquatic environment might be affected by factors besides antibiotics. High numbers of resistant E. coli were also isolated from the conserved reservoir. These results suggest that rural surface water may become a large pool of VAs and resistant bacteria. This study contributes to current information on VAs and resistant bacteria contamination in aquatic environments particularly in areas under intensive agriculture. Moreover, this study indicates an urgent need to monitor the use of VAs in animal production, and to control the release of animal-originated antibiotics into the environment.

  20. Microbiological bioassay using Bacillus pumilus to detect tetracycline in milk.

    Science.gov (United States)

    Tumini, Melisa; Nagel, Orlando Guillermo; Althaus, Rafael Lisandro

    2015-05-01

    The tetracyclines (TCs) are widely used in the treatment of several diseases of cattle and their residues may be present in milk. To control these residues it is necessary to have available inexpensive screening methods, user-friendly and capable of analysing a high number of samples. The purpose of this study was to design a bioassay of microbiological inhibition in microtiter plates with spores of Bacillus pumilus to detect TCs at concentrations corresponding to the Maximum Residue Limits (MRLs). Several complementary experiments were performed to design the bioassay. In the first study, we determined the concentration of spores that produce a change in the bioassay's relative absorbance in a short time period. Subsequently, we assessed the concentration of chloramphenicol required to decrease the detection limit (DL) of TCs at MRLs levels. Thereafter, specificity, DL and cross-specificity of the bioassay were estimated. The most appropriate microbiological inhibition assay had a B. pumilus concentration of 1.6 × 10(9) spores/ml, fortified with 2500 μg chloramphenicol/l (CAP) in Mueller Hinton culture medium using brilliant black and toluidine blue as redox indicator. This bioassay detected 117 μg chlortetracycline/l, 142 μg oxytetracycline/l and 105 μg tetracycline/l by means of a change in the indicator's colour in a period of 5 h. The method showed good specificity (97.9%) which decreased slightly (93.3%) in milk samples with high somatic cell counts (>250,000 cells/ml). Furthermore, other antimicrobials studied (except neomycin) must be present in milk at high concentrations (from >5 to >100 MRLs) to produce positive results in this assay, indicating a low cross specificity.

  1. Selective Pressure Promotes Tetracycline Resistance of Chlamydia Suis in Fattening Pigs

    Science.gov (United States)

    Wanninger, Sabrina; Donati, Manuela; Di Francesco, Antonietta; Hässig, Michael; Hoffmann, Karolin; Seth-Smith, Helena M. B.; Marti, Hanna; Borel, Nicole

    2016-01-01

    In pigs, Chlamydia suis has been associated with respiratory disease, diarrhea and conjunctivitis, but there is a high rate of inapparent C. suis infection found in the gastrointestinal tract of pigs. Tetracycline resistance in C. suis has been described in the USA, Italy, Switzerland, Belgium, Cyprus and Israel. Tetracyclines are commonly used in pig production due to their broad-spectrum activity and relatively low cost. The aim of this study was to isolate clinical C. suis samples in cell culture and to evaluate their antibiotic susceptibility in vitro under consideration of antibiotic treatment on herd level. Swab samples (n = 158) identified as C. suis originating from 24 farms were further processed for isolation, which was successful in 71% of attempts with a significantly higher success rate from fecal swabs compared to conjunctival swabs. The farms were divided into three treatment groups: A) farms without antibiotic treatment, B) farms with prophylactic oral antibiotic treatment of the whole herd consisting of trimethoprime, sulfadimidin and sulfathiazole (TSS), or C) farms giving herd treatment with chlortetracycline with or without tylosin and sulfadimidin (CTS). 59 isolates and their corresponding clinical samples were selected and tested for the presence or absence of the tetracycline resistance class C gene [tet(C)] by conventional PCR and isolates were further investigated for their antibiotic susceptibility in vitro. The phenotype of the investigated isolates was either classified as tetracycline sensitive (Minimum inhibitory concentration [MIC] < 2 μg/ml), intermediate (2 μg/ml ≤ MIC < 4 μg/ml) or resistant (MIC ≥ 4 μg/ml). Results of groups and individual pigs were correlated with antibiotic treatment and time of sampling (beginning/end of the fattening period). We found clear evidence for selective pressure as absence of antibiotics led to isolation of only tetracycline sensitive or intermediate strains whereas tetracycline treatment

  2. In vitro susceptibility of Mycoplasma hyosynoviae and M. hyorhinis to antimicrobial agents.

    Science.gov (United States)

    Kobayashi, H; Sonmez, N; Morozumi, T; Mitani, K; Ito, N; Shiono, H; Yamamoto, K

    1996-11-01

    Fifty-four Japanese strains of Mycoplasma hyosynoviae isolated from porkers during 1980 to 1995, and 107 Japanese strains of M. hyorhinis isolated from piglets with respiratory disease during 1991 to 1994 were investigated for the in vitro activities of 13 antimicrobial agents [josamycin, tylosin, spiramycin, kitasamycin, erythromycin, lincomycin (LCM), kanamycin (KM), chloramphenicol (CP), thiamphenicol (TP), tiamulin (TML), oxytetracycline (OTC), chlortetracycline (CTC), and enrofloxacin (ERFX)] by the agar dilution method. Of the drugs tested TML showed the highest activity with minimum inhibitory concentration (MIC) of 0.013 to 0.1 microgram/ m/ (MIC90; 0.05 microgram/ml) against strains of M. hyosynoviae, and 0.2 to 0.78 microgram/ml (MIC90; 0.39 microgram/ml) against strains of M. hyorhinis. ERFX, LCM, most of the 16-membered macrolide antibiotics and tetracyclines also showed low MICs against both mycoplasma species. The susceptibility of KM, CP and TP to the mycoplasmas was considered to be of a secondary grade. Two of 54 strains of M. hyosynoviae, and 11 of 107 strains of M. hyorhinis showed resistance to all 14- and 16-membered macrolide antibiotics tested. Tetracyclines (OTC and CTC) showed a relatively broad MIC distribution from 0.1 to 6.25 micrograms/ml against the M. hyosynoviae strains tested. All of the strains isolated during 1980 to 1984 were susceptible at the concentration of 0.78 microgram/ml or less (MIC90; 0.78 microgram/ml) to OTC and 1.56 micrograms/ml or less (MIC90; 1.56 micrograms/ml) to CTC, while the susceptibility of strains isolated recently, during 1994 to 1995, was more than 0.78 microgram/ml (MIC90; 3.13 micrograms/ml) to OTC, and more than 1.56 micrograms/ml (MIC90; 6.25 micrograms/ml) to CTC.

  3. Comparative study of three screening tests, two microbiological tube tests, and a multi-sulphonamide ELISA kit for the detection of antimicrobial and sulphonamide residues in eggs.

    Science.gov (United States)

    Gaudin, V; Hedou, C; Rault, A; Sanders, P; Verdon, E

    2009-04-01

    The screening of antimicrobial residues in eggs is an especially important subject. Three different commercial kits for the screening of sulphonamides and other antimicrobials in eggs were validated in accordance with Decision 2002/657/EC: one enzyme-linked immunoabsorbant assay (ELISA) kit multi-sulphonamides (from RAISIO Diagnostics) and two microbiological tests (a Premi test from DSM and an Explorer kit from Zeu-Inmunotec). The false-positive rates were lower than 2% for all kits. The detection capabilities (CCbeta) have to be as low as possible for banned substances and lower than the maximum residue limit (MRL) when MRLs have been set. The sensitivity of the Premi test was better than that of the Explorer test, probably because of the dilution of the eggs before the Explorer test was used. The CCbeta values towards most of the tested sulphonamides were satisfactory with the Premi test (sulphonamides in eggs with the Premi test confirmed these results. The detection capabilities of tetracycline and doxycycline were at the level of the MRL or twice the MRL maximum. The detection capabilities for chlortetracycline and oxytetracycline were higher (four to six times the MRL). The detection capabilities for amoxicillin, neomycin, tylosin and erythromycin were lower than their respective MRLs. Detection capabilities for sulphonamides were much lower for the ELISA kit than for microbiological tests. The ELISA kit could be recommended for the targeted screening of sulphonamides in eggs. On the other hand, the Explorer and Premi tests could be used as wide screening tests allowing the detection of most of the antimicrobial families.

  4. Cholesterol addition aids the cryopreservation of dromedary camel (Camelus dromedarius) spermatozoa.

    Science.gov (United States)

    Crichton, Elizabeth G; Pukazhenthi, Budhan S; Billah, M; Skidmore, Julian A

    2015-01-15

    The cryopreservation of dromedary camel (Camelus dromedarius) sperm has proved challenging with little success reported. The routine application of artificial insemination with frozen semen would assist the flow of valuable genetic material nationally and internationally. The current study sought to examine the effects of cholesterol (cholesterol-loaded cyclodextrin [CLC]) preloading on camel sperm cryosurvival. Ejaculates (n = 3 males; 3 ejaculates per male) were collected using an artificial vagina during the breeding season and extended in HEPES-buffered Tyrode's albumin lactate pyruvate (TALP) and allowed to liquefy in the presence of papain (0.1 mg/mL) before removal of the seminal plasma by centrifugation. Sperm pellets were resuspended (120 million/mL) in fresh TALP and incubated (15 minutes; 37 °C) with 0, 1.5, or 4.5 mg CLC/mL. Sperm suspensions were then centrifuged and reconstituted in INRA-96 containing 20% (v:v) egg yolk and 2.5% (v:v) methylformamide, loaded in 0.5-mL plastic straws, sealed, and cooled for 20 minutes at 4 °C. Straws were frozen over liquid nitrogen (4 cm above liquid; 15 minutes), plunged, and stored. Sperm motility, forward progressive status, and acrosomal integrity were recorded at 0 and 3 hours after thawing and compared with these same parameters before freezing. Aliquots also were stained with chlortetracycline hydrochloride to assess spontaneous sperm capacitation status before freezing and post-thaw. Pretreatment with CLC (1.5 and 4.5 mg/mL) enhanced cryosurvival. Post-thaw sperm motility was highest (P dromedary camel sperm benefit from exposure to CLC before cryopreservation; this may facilitate the routine collection and storage of sperm from this species.

  5. Selective Pressure Promotes Tetracycline Resistance of Chlamydia Suis in Fattening Pigs.

    Science.gov (United States)

    Wanninger, Sabrina; Donati, Manuela; Di Francesco, Antonietta; Hässig, Michael; Hoffmann, Karolin; Seth-Smith, Helena M B; Marti, Hanna; Borel, Nicole

    2016-01-01

    In pigs, Chlamydia suis has been associated with respiratory disease, diarrhea and conjunctivitis, but there is a high rate of inapparent C. suis infection found in the gastrointestinal tract of pigs. Tetracycline resistance in C. suis has been described in the USA, Italy, Switzerland, Belgium, Cyprus and Israel. Tetracyclines are commonly used in pig production due to their broad-spectrum activity and relatively low cost. The aim of this study was to isolate clinical C. suis samples in cell culture and to evaluate their antibiotic susceptibility in vitro under consideration of antibiotic treatment on herd level. Swab samples (n = 158) identified as C. suis originating from 24 farms were further processed for isolation, which was successful in 71% of attempts with a significantly higher success rate from fecal swabs compared to conjunctival swabs. The farms were divided into three treatment groups: A) farms without antibiotic treatment, B) farms with prophylactic oral antibiotic treatment of the whole herd consisting of trimethoprime, sulfadimidin and sulfathiazole (TSS), or C) farms giving herd treatment with chlortetracycline with or without tylosin and sulfadimidin (CTS). 59 isolates and their corresponding clinical samples were selected and tested for the presence or absence of the tetracycline resistance class C gene [tet(C)] by conventional PCR and isolates were further investigated for their antibiotic susceptibility in vitro. The phenotype of the investigated isolates was either classified as tetracycline sensitive (Minimum inhibitory concentration [MIC] MIC MIC ≥ 4 μg/ml). Results of groups and individual pigs were correlated with antibiotic treatment and time of sampling (beginning/end of the fattening period). We found clear evidence for selective pressure as absence of antibiotics led to isolation of only tetracycline sensitive or intermediate strains whereas tetracycline treatment resulted in a greater number of tetracycline resistant isolates.

  6. Persistence of antibiotic resistance: evaluation of a probiotic approach using antibiotic-sensitive Megasphaera elsdenii strains to prevent colonization of swine by antibiotic-resistant strains.

    Science.gov (United States)

    Stanton, Thad B; Humphrey, Samuel B

    2011-10-01

    Megasphaera elsdenii is a lactate-fermenting, obligately anaerobic bacterium commonly present in the gastrointestinal tracts of mammals, including humans. Swine M. elsdenii strains were previously shown to have high levels of tetracycline resistance (MIC=64 to >256 μg/ml) and to carry mosaic (recombinant) tetracycline resistance genes. Baby pigs inherit intestinal microbiota from the mother sow. In these investigations we addressed two questions. When do M. elsdenii strains from the sow colonize baby pigs? Can five antibiotic-sensitive M. elsdenii strains administered intragastrically to newborn pigs affect natural colonization of the piglets by antibiotic-resistant (AR) M. elsdenii strains from the mother? M. elsdenii natural colonization of newborn pigs was undetectable (pigs became colonized (4 × 10(5) to 2 × 10(8) CFU/g feces). In a separate study, 61% (76/125) of M. elsdenii isolates from a gravid sow never exposed to antibiotics were resistant to chlortetracycline, ampicillin, or tylosin. The inoculation of the sow's offspring with mixtures of M. elsdenii antibiotic-sensitive strains prevented colonization of the offspring by maternal AR strains until at least 11 days postweaning. At 25 and 53 days postweaning, however, AR strains predominated. Antibiotic susceptibility phenotypes and single nucleotide polymorphism (SNP)-based identities of M. elsdenii isolated from sow and offspring were unexpectedly diverse. These results suggest that dosing newborn piglets with M. elsdenii antibiotic-sensitive strains delays but does not prevent colonization by maternal resistant strains. M. elsdenii subspecies diversity offers an explanation for the persistence of resistant strains in the absence of antibiotic selection.

  7. Multiple Antibiotic Resistances of Vibrio Isolates from Coastal and Brackish Water Areas

    Directory of Open Access Journals (Sweden)

    S. Manjusha

    2005-01-01

    Full Text Available An experiment was designed to assess the occurrence of multiple antibiotic resistances in Vibrio spp. from different (brackish and marine environments. Water samples from nine marine landing sites and two coastal inland aquaculture farms were screened for the Vibrio spp. and assessed their resistance to twenty-two different antibiotics, which are commonly encountered in the aquatic ecosystem. Tissue samples (shrimp, mussel and sepia were tested from the sampling site with highest antibiotic resistance. Of the total 119 Vibrio isolates, 16. 8% were susceptible to all antibiotics. Of the resistant (83.19% Vibrio strains, 30.3% were resistant against three antibiotics, 55.5% were resistant against 4-10 antibiotics, 14.14% were resistant against more than 10 antibiotics and 54% have shown multiple antibiotics resistance (MAR. Antibiotic resistance index was higher in Coastal 3, 6, Aqua farm 2 in isolates from water samples and all the tissues tested. Interestingly, incidence of antibiotic resistance in isolates from water samples was comparatively lower in aquaculture farms than that observed in coastal areas. Highest incidence of antibiotic resistance was evident against Amoxycillin, Ampicillin, Carbencillin and Cefuroxime followed by Rifampicin and Streptomycin and lowest against Chloramphenicol, Tetracycline, Chlortetracycline, Furazolidone, Nalidixic acid, Gentamycin Sulphafurazole, Trimethoprim, Neomycin and Amikacin irrespective of the sampling sites. Results from various tissue samples collected from the sites of highest antibiotic resistance indicated that antibiotic resistance Vibrio spp. collected from fish and tissue samples were higher than that of water samples. Overall results indicated that persistent use of antibiotics against diseases in human beings and other life forms may pollute the aquatic system and their impact on developing antibiotic resistant Vibrio spp. may be a serious threat in addition to the use of

  8. Cholesterol-Loaded Cyclodextrin Increases the Cholesterol Content of Goat Sperm to Improve Cold and Osmotic Resistance and Maintain Sperm Function after Cryopreservation.

    Science.gov (United States)

    Salmon, Vianney M; Leclerc, Pierre; Bailey, Janice L

    2016-04-01

    The success of semen cryopreservation depends on sperm membrane integrity and function after thawing. Cholesterol-loaded cyclodextrin (CLC) is used for in vitro incorporation of cholesterol to protect cells against cold temperatures. We hypothesized that CLC treatment also enhances sperm cholesterol content to increase tolerance to osmotic shock and cryoresistance, thereby improving fertility. We confirmed the fact that treatment of goat semen with 3 mg/ml CLC increases sperm cholesterol content using both the Liebermann-Burchard approach and filipin III labeling of membrane cholesterol. Sperm were then treated with or without CLC and cryopreserved. After thawing, sperm cholesterol dramatically fell, even in the presence of CLC, which explains the mechanism of cryocapacitation. CLC treatment, however, maintained a normal prefreeze cholesterol level in sperm after cryopreservation. Furthermore, fresh sperm treated with CLC and subjected to either cold shock or incubated in hypo-, iso-, and hyperosmotic media, designed to mimic stresses associated with freezing/thawing, displayed increased temperature and osmotic tolerance. CLC treatment also improved sperm viability, motility, and acrosome integrity after thawing. Furthermore, CLC treatment did not affect the sperm's ability to undergo in vitro capacitation according to chlortetracycline fluorescence and protein tyrosine phosphorylation. A pilot field trial demonstrated that artificial insemination with sperm that underwent increased cholesterol levels following CLC treatment yielded higher fertility ( ITALIC! P< 0.1) and proliferation ( ITALIC! P< 0.05) rates in vivo than untreated semen from the same ejaculate samples. These observations suggest that CLC treatment could be used to improve cryoprotection during the freezing and thawing of goat sperm.

  9. Degradation of Tetracyclines in Pig Manure by Composting with Rice Straw.

    Science.gov (United States)

    Chai, Rushan; Huang, Lidong; Li, Lingling; Gielen, Gerty; Wang, Hailong; Zhang, Yongsong

    2016-02-24

    A holistic approach was followed for utilizing tetracyclines (TCs)-contaminated pig manure, by composting this with rice straw in a greenhouse for CO₂ fertilization and composted residue application. After composting, the composted residues can be applied to cropland as a supplemental source of synthetic fertilizers. The objective of this study was to determine the effect of pig manure-rice straw composting on the degradation of TCs in pig manure. The results showed that greenhouse composting significantly accelerated the degradation of TCs. Contents (150 mg·kg(-1)) of oxytetracycline (OTC), tetracycline (TC) and chlortetracycline (CTC) in the composting feedstock could be completely removed within 42 days for OTC and TC, and 14 days for CTC. However, in the control samples incubated at 25 °C in the dark, concentrations of OTC, TC and CTC only decreased 64.7%, 66.7% and 73.3%, respectively, after 49 days. The degradation rates of TCs in the composting feedstock were in the order of CTC > TC > OTC. During the composting process, CTC dissipated rapidly with the time required for 50% degradation (DT50) and 90% degradation (DT90) of 2.4 and 7.9 days, but OTC was more persistent with DT50 and DT90 values of 5.5 and 18.4 days. On the basis of the results obtained in this study, it could be concluded that pig manure-rice straw composting in a greenhouse can help to accelerate the degradation of TCs in pig manure and make composted residues safer for field application. This technology could be an acceptable practice for greenhouse farmers to utilize TCs-contaminated pig manure.

  10. Effect of Origanum chemotypes on broiler intestinal bacteria.

    Science.gov (United States)

    Betancourt, Liliana; Rodriguez, Fernando; Phandanouvong, Vienvilay; Ariza-Nieto, Claudia; Hume, Michael; Nisbet, David; Afanador-Téllez, German; Van Kley, Alexandra Martynova; Nalian, Armen

    2014-10-01

    Essential oils have been proposed as alternatives to antibiotic use in food animal production. This study evaluated 3 chemotypes of the Origanum genus, containing varying amounts of secondary metabolites carvacrol, thymol, and sabinene, in the broiler chicken diet. Aerial parts of Origanum vulgare L. (OL), O. vulgare L. ssp. hirtum (OH), and O. majorana (OM) were collected from a greenhouse located in the high altitude Sabana de Bogotá (Savanna of Bogotá) and O. vulgare L. ssp. hirtum (OG) produced and ground in Greece. Oregano essential oils (OEO) from these plants were obtained by steam distillation and analyzed by gas chromatography coupled to a mass spectrometer. Six treatments were evaluated: 200 mg/kg of OEO from OH, OL, and OM, 50 mg/kg of OEO from OG, 500 mg/kg of chlortetracycline, and without additives. Broiler chicks were maintained at 2,600 m above sea level, placed in brooder cages under a completely randomized design. Template DNA was isolated from duodenal, jejunal, ileal, and cecal contents in each group and bacterial 16S rDNA patterns were analyzed by denaturing gradient gel electrophoresis. Dendrograms of denaturing gradient gel electrophoresis band patterns revealed 2 main clusters, OEO-treated chicks and nontreated control chicks, in each intestinal segment. Band patterns from different gut compartments revealed major bacterial population shifts in the foregut (duodenum, jejunum, and ileum) compared with the hindgut (cecum and colon) at all ages evaluated (P < 0.05). The OEO groups showed less shift (62.7% similarity coefficient) between these 2 compartments versus the control groups (53.7% similarity coefficient). A reduction of 59% in mortality from ascites was seen in additive-supplemented groups compared with the control group. This study represents the first work to evaluate the effects of the 3 main chemotypes of Origanum genus in broilers.

  11. Simultaneous determination of 11 antibiotics and their main metabolites from four different groups by reversed-phase high-performance liquid chromatography-diode array-fluorescence (HPLC-DAD-FLD) in human urine samples.

    Science.gov (United States)

    Fernandez-Torres, R; Consentino, M Olías; Lopez, M A Bello; Mochon, M Callejon

    2010-05-15

    A new, accurate and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) as analytical method for the quantitative determination of 11 antibiotics (drugs) and the main metabolites of five of them present in human urine has been worked out, optimized and validated. The analytes belong to four different groups of antibiotics (sulfonamides, tetracyclines, penicillins and anphenicols). The analyzed compounds were sulfadiazine (SDI) and its N(4)-acetylsulfadiazine (NDI) metabolite, sulfamethazine (SMZ) and its N(4)-acetylsulfamethazine (NMZ), sulfamerazine (SMR) and its N(4)-acetylsulfamerazine (NMR), sulfamethoxazole (SMX), trimetroprim (TMP), amoxicillin (AMX) and its main metabolite amoxicilloic acid (AMA), ampicillin (AMP) and its main metabolite ampicilloic acid (APA), chloramphenicol (CLF), thiamphenicol (TIF), oxytetracycline (OXT) and chlortetracycline (CLT). For HPLC analysis, diode array (DAD) and fluorescence (FLD) detectors were used. The separation of the analyzed compounds was conducted by means of a Phenomenex Gemini C(18) (150mm x 4.6mm I.D., particle size 5microm) analytical column with LiChroCART LiChrospher C(18) (4mm x 4mm, particle size 5microm) guard column. Analyzed drugs were determined within 34min using formic acid 0.1% in water and acetonitrile in gradient elution mode as mobile phase. A linear response was observed for all compounds in the range of concentration studied. Two procedures were optimized for sample preparation: a direct treatment with methanol and acetonitrile and a solid phase extraction procedure using Bond Elut Plexa columns. The method was applied to the determination of the analytes in human urine from volunteers under treatment with different pharmaceutical formulations. This method can be successfully applied to routine determination of all these drugs in human urine samples.

  12. Bacteria, phages and pigs: the effects of in-feed antibiotics on the microbiome at different gut locations.

    Science.gov (United States)

    Looft, Torey; Allen, Heather K; Cantarel, Brandi L; Levine, Uri Y; Bayles, Darrell O; Alt, David P; Henrissat, Bernard; Stanton, Thaddeus B

    2014-08-01

    Disturbance of the beneficial gut microbial community is a potential collateral effect of antibiotics, which have many uses in animal agriculture (disease treatment or prevention and feed efficiency improvement). Understanding antibiotic effects on bacterial communities at different intestinal locations is essential to realize the full benefits and consequences of in-feed antibiotics. In this study, we defined the lumenal and mucosal bacterial communities from the small intestine (ileum) and large intestine (cecum and colon) plus feces, and characterized the effects of in-feed antibiotics (chlortetracycline, sulfamethazine and penicillin (ASP250)) on these communities. 16S rRNA gene sequence and metagenomic analyses of bacterial membership and functions revealed dramatic differences between small and large intestinal locations, including enrichment of Firmicutes and phage-encoding genes in the ileum. The large intestinal microbiota encoded numerous genes to degrade plant cell wall components, and these genes were lacking in the ileum. The mucosa-associated ileal microbiota harbored greater bacterial diversity than the lumen but similar membership to the mucosa of the large intestine, suggesting that most gut microbes can associate with the mucosa and might serve as an inoculum for the lumen. The collateral effects on the microbiota of antibiotic-fed animals caused divergence from that of control animals, with notable changes being increases in Escherichia coli populations in the ileum, Lachnobacterium spp. in all gut locations, and resistance genes to antibiotics not administered. Characterizing the differential metabolic capacities and response to perturbation at distinct intestinal locations will inform strategies to improve gut health and food safety.

  13. Protein phosphatases decrease their activity during capacitation: a new requirement for this event.

    Directory of Open Access Journals (Sweden)

    Janetti R Signorelli

    Full Text Available There are few reports on the role of protein phosphatases during capacitation. Here, we report on the role of PP2B, PP1, and PP2A during human sperm capacitation. Motile sperm were resuspended in non-capacitating medium (NCM, Tyrode's medium, albumin- and bicarbonate-free or in reconstituted medium (RCM, NCM plus 2.6% albumin/25 mM bicarbonate. The presence of the phosphatases was evaluated by western blotting and the subcellular localization by indirect immunofluorescence. The function of these phosphatases was analyzed by incubating the sperm with specific inhibitors: okadaic acid, I2, endothall, and deltamethrin. Different aliquots were incubated in the following media: 1 NCM; 2 NCM plus inhibitors; 3 RCM; and 4 RCM plus inhibitors. The percent capacitated sperm and phosphatase activities were evaluated using the chlortetracycline assay and a phosphatase assay kit, respectively. The results confirm the presence of PP2B and PP1 in human sperm. We also report the presence of PP2A, specifically, the catalytic subunit and the regulatory subunits PR65 and B. PP2B and PP2A were present in the tail, neck, and postacrosomal region, and PP1 was present in the postacrosomal region, neck, middle, and principal piece of human sperm. Treatment with phosphatase inhibitors rapidly (≤1 min increased the percent of sperm depicting the pattern B, reaching a maximum of ∼40% that was maintained throughout incubation; after 3 h, the percent of capacitated sperm was similar to that of the control. The enzymatic activity of the phosphatases decreased during capacitation without changes in their expression. The pattern of phosphorylation on threonine residues showed a sharp increase upon treatment with the inhibitors. In conclusion, human sperm express PP1, PP2B, and PP2A, and the activity of these phosphatases decreases during capacitation. This decline in phosphatase activities and the subsequent increase in threonine phosphorylation may be an important

  14. Involvement of intracellular calcium and src tyrosine-kinase in capacitation of cryopreserved bovine spermatozoa Participación del calcio intracelular y src tirosina quinasas en la capacitación del espermatozoide bovino criopreservado

    Directory of Open Access Journals (Sweden)

    M Satorre

    2010-06-01

    Full Text Available Increase of intracellular calcium concentration and tyrosine kinase involvement are pivotal in sperm capacitation. The aim was to determine the involvement of intracellular calcium and tyrosine kinases activity in frozen-thawed spermatozoa capacitated with heparin or quercetin. Genistein or PP2 (4-amino-5-(4-chlorophenyl-7-(t-butylpyrazolo3,4, pyrimidine were used to inhibit tyrosine kinases, and methoxyverapamil to inhibit voltage dependent calcium channels. Capacitation was determined by chlortetracycline and calcium by fl uorescence spectrophotometry. Protein tyrosine-phosphorylation was determined by western blot. Pooled frozen semen samples from four bulls were used. In presence of heparin or quercetin capacitated spermatozoa percentage and intracellular calcium were greater (PEl incremento de calcio intracelular [Ca]i y la participación de tirosina quinasa son pasos cruciales en la inducción de la capacitación. El objetivo fue determinar en espermatozoides criopreservados, la variación [Ca]i y la actividad de tirosina quinasa en la capacitación con heparina o quercetina. Genisteina o PP2(4-amino-5-(4-clorofenil-7-(t-butilpirazolo3,4,d pirimidina son inhibidores de tirosinas quinasas y de la isoforma SRC, respectivamente. Metoxiverapamil es un inhibidor de canales de calcio voltaje dependiente (CCVD. La capacitación, [Ca]i y fosforilación en tirosina se evaluaron con clorotetraciclina, espectrofl uorometría y western blot, respectivamente. El porcentaje de espermatozoides capacitados y [Ca]i fueron mayores (P<0.05 en muestras con heparina o quercetina respecto a sus controles. Genisteina o PP2 disminuyeron (P<0.05 la capacitación y el incremento de [Ca]i en las muestras con heparina pero no en las tratadas con quercetina. Genisteina o PP2 inhibió diferencialmente la fosforilación de proteínas espermáticas con ambos inductores. Methoxiverapamil bloqueó el incremento de [Ca]i sólo en la muestras con heparina. Siendo los

  15. 电解氧化法去除养殖废水中抗生素和激素研究%Removal of Antibiotics and Hormones by Electrolytic Oxidation in Livestock Wastewater

    Institute of Scientific and Technical Information of China (English)

    王志刚; 陈宏; 陈玉成; 李向前; 曾维友; 刁香维

    2013-01-01

    通过研究三维电极的电解氧化法对模拟养殖废水中抗生素、激素的去除影响和处理效果.结果表明:对抗生素、激素去除率的影响大小顺序表现为电解时间,初始pH和曝气时间.最优试验参数条件为电解电压5V,电解时间2 min,初始pH值为9,曝气时间3h.最优条件下,喹乙醇、土霉素、四环素、金霉素的去除率分别为99.1%,90.8%,97.7%和90.7%.少量的柠檬酸(0.02 mol/L)、乙酸(0.175 mol/L)能够提高去除效果;加入0.02 mol/L十二烷基磺酸钠(SDS)能够显著提高喹乙醇去除效果,但是会对土霉素、四环素、金霉素的去除效果产生不同程度的抑制.%A three-dimensional electrode reactor was employed to treat simulated livestock wastewater,and the influencing factors and the removal rate were studied.The results showed that the influence on the removal rate of antibiotics and hormones of the wastewater was in the order of electrolysis time > initial pH > aeration time.The optimal operating conditions were as follows:5 V,electrolysis time 2 min,initial pH 9,aeration time 3 h.Under these conditions,removal rates of 62.1%,90.4%,96.5%00 and 91% were achieved for oxytetracycline,teacycline,chlortetracycline and laquindox,respectively.Addition of a small amount of citric acid (0.02 mol/L) or acetic acid (0.175 mol/L) significantly improved the removal efficiency; SDS addition at 0.02 mol/L significantly improved olaquindox removal rate,but reduced,to some degree,the removal rate of the other three antibiotics studied.

  16. Effect of estrogens on boar sperm capacitation in vitro

    Directory of Open Access Journals (Sweden)

    Ded Lukas

    2010-07-01

    Full Text Available Abstract Background Mammalian sperm must undergo a series of controlled molecular processes in the female reproductive tract called capacitation before they are capable of penetrating and fertilizing the egg. Capacitation, as a complex biological process, is influenced by many molecular factors, among which steroidal hormone estrogens play their role. Estrogens, present in a high concentration in the female reproductive tract are generally considered as primarily female hormones. However, there is increasing evidence of their important impact on male reproductive parameters. The purpose of this study is to investigate the effect of three natural estrogens such as estrone (E1, 17beta-estradiol (E2 and estriol (E3 as well as the synthetical one, 17alpha-ethynylestradiol (EE2 on boar sperm capacitation in vitro. Methods Boar sperm were capacitated in vitro in presence of estrogens. Capacitation progress in control and experimental samples was analyzed by flow cytometry with the anti-acrosin monoclonal antibody (ACR.2 at selected times of incubation. Sperm samples were analyzed at 120 min of capacitation by CTC (chlortetracycline assay, immunocytochemistry and flow cytometry with anti-acrosin ACR.2 antibody. Furthermore, sperm samples and capacitating media were analyzed by immunocytochemistry, ELISA with the ACR.2 antibody, and the acrosin activity assay after induced acrosomal reaction (AR. Results Estrogens stimulate sperm capacitation of boar sperm collected from different individuals. The stimulatory effect depends on capacitation time and is highly influenced by differences in the response to estrogens such as E2 by individual animals. Individual estrogens have relatively same effect on capacitation progress. In the boar samples with high estrogen responsiveness, estrogens stimulate the capacitation progress in a concentration-dependent manner. Furthermore, estrogens significantly increase the number of acrosome-reacted sperm after zona

  17. Unravelling a vicious circle: animal feed marketed in Costa Rica contains irregular concentrations of tetracyclines and abundant oxytetracycline-resistant Gram-positive bacteria.

    Science.gov (United States)

    Granados-Chinchilla, Fabio; Alfaro, Margarita; Chavarría, Guadalupe; Rodríguez, César

    2014-01-01

    Diverse tetracyclines are used to prevent and control bacterial infections in livestock and farmed fish. These drugs are administered through the diet, but farmers seldom check whether feed contains antibiotic-resistant bacteria that may colonise their crops or transfer their resistance traits to species of veterinary relevance. To examine whether antibiotic dosage defines the abundance of antibiotic-resistant bacteria in animal feed, we determined the concentration of parental compounds and epimers of oxytetracycline (OTC), doxycycline, tetracycline and chlortetracycline, as well as the abundance and resistance level of OTC-resistant bacteria in samples of fish (n = 21), poultry (n = 21), swine (n = 21), and shrimp feed (n = 21) marketed in Costa Rica. Fish feed contained the highest amounts of tetracyclines (119-8365 mg kg(-1)) and the largest proportion of bacteria resistant to 10 μg ml(-1) (1.8-92.4%) or 100 μg ml(-1) of OTC (12.5-63.8%). Poultry (78-438 mg kg(-1)) and swine (41-1076 mg kg(-1)) feed had intermediate concentrations of tetracyclines and OTC-resistant bacteria (0.2-66% and 0.3-49%, respectively), whereas shrimp feed showed the lowest amounts of tetracyclines (21.5-50.3 mg kg(-1)), no OTC and no culturable OTC-resistant bacteria. In line with these results, the MIC50 of OTC for 150 isolates from fish and poultry feed was > 256 µg ml(-1), while that of 150 bacteria isolated from swine feed was 192 µg ml(-1). Phenotypic tests, fatty acid profiles and proteotypic analyses by matrix-assisted laser desorption/ionisation-time of flight mass-spectroscopy revealed that most OTC-resistant isolates were Gram-positive bacteria of low G+C% content from the genera Staphylococcus and Bacillus. Clear correlations between OTC dosage and feed colonisation with OTC-resistant bacteria were seen in medicated feed for fish (r = 0.179-0.651). Nonetheless, some unmedicated feed for fish, swine and poultry contained large populations of OTC-resistant bacteria

  18. Bandgap tailoring of in-situ nitrogen-doped TiO₂ sputtered films intended for electrophotocatalytic applications under solar light

    Energy Technology Data Exchange (ETDEWEB)

    Delegan, N.; El Khakani, M. A., E-mail: elkhakani@emt.inrs.ca [Institut National de la Recherche Scientifique, Centre Énergie, Matériaux et Télécommunications, 1650, Boulevard Lionel-Boulet, Varennes, Québec J3X-1S2 (Canada); Daghrir, R.; Drogui, P. [Institut National de la Recherche Scientifique, Centre Eau, Terre et Environnement, 490 Rue de la Couronne, Québec G1K-9A9 (Canada)

    2014-10-21

    We report on a reactive RF-sputtering process permitting the in-situ nitrogen doping of TiO₂ films in order to shift their photoactivity from UV to visible range. By carefully controlling the relative nitrogen-to-argon mass flow rate ratio (within the 0%–25% range) in the sputter deposition chamber, TiO₂:N films were grown with nitrogen contents ranging from 0 to 6.2 at. %, as determined by high-resolution X-ray spectroscopy measurements. A systematic investigation of the crystalline structure of the TiO₂:N films, as a function of their N content, revealed that low N contents (0.2–0.3 at. %) induce crystallization in the rutile phase while higher N contents (≥1.4 at. %) were accompanied with the recovery of the anatase structure with an average crystallite size of ~35 nm. By using both UV-Vis absorption and spectroscopic ellipsometry measurements, we were able to quantitatively determine the bandgap (E{sub g}) variation of the TiO₂:N films as a function of their N content. Thus, we have demonstrated that the E{sub g} of the TiO₂:N films effectively narrows from 3.2 eV down to a value as low as ~2.3 eV for the optimal N doping concentration of 3.4 at. % (higher N incorporation does not translate into further red shifting of the TiO₂:N films' E{sub g}). The photoactivity of the TiO₂:N films under visible light was confirmed through electro-photocatalytic decomposition of chlortetracycline (CTC, an emerging water pollutant) under standard 1.5AM solar radiation. Thus, CTC degradation efficiencies of up to 98% were achieved with 2 hours process cycles under simulated solar light. Moreover, the electro-photocatalytic performance of the TiO₂:N films is shown to be directly correlated to their optoelectronic properties (namely their bandgap narrowing).

  19. Direct screening of tetracyclines in water and bovine milk using room temperature phosphorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Traviesa-Alvarez, J.M. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain); Costa-Fernandez, J.M. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain); Pereiro, R. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain); Sanz-Medel, A. [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, c/Julian Claveria 8, 33006 Oviedo (Spain)]. E-mail: asm@uniovi.es

    2007-04-18

    A fast and simple flow-through optosensor was designed and characterized for the direct screening of four tetracycline (TCC) antibiotics (tetracycline, oxytetracycline, chlortetracycline and doxycycline) in water and bovine milk samples. The proposed optosensor provides rapid binary yes/no overall responses, being appropriate for the screening of this family of antibiotics above or below a pre-set concentration threshold. The experimental set-up is based on a flow-injection manifold coupled on-line to a phosphorescence detector. Aliquots of the samples are pretreated with Eu(III) to form room temperature phosphorescent metal chelates and injected in the flow manifold. Those chelates are then on-line retained on a conventional flow-cell (packed with polymeric Amberlite XAD-4 particles) which is placed inside the cell holder of the phosphorimeter. After the emission is registered, the antibiotic-metal complexes are eluted from the packed resin with 1 M HCl (for milk samples a second regeneration step, using methanol, should be performed). A sample throughput of about 20 samples per hour was obtained. Optimum experimental conditions include a pH 9, a Eu(III) concentration of 2 x 10{sup -4} M and 8 mM sodium sulphite as chemical deoxygenant. The phosphorescence emitted by the europium-TCC complexes was measured at 394 and 617 nm for excitation and emission wavelengths, respectively. The unreliability region, given by the probability of false positives and false negatives, respectively (set at 5% in both cases) was in the range between 0.2 and 11.6 nM for detection of tetracyclines in water samples (at a cut-off level of 4 nM) and in the range between 165 and 238 nM for detection of tetracyclines in milk (cut-off level fixed at the normative EU level of 200 nM). Finally, the applicability of the proposed screening optosensor was tested for the reliable control of tetracyclines in contaminated and uncontaminated water and milk samples.

  20. Tetracycline residues and tetracycline resistance genes in groundwater impacted by swine production facilities

    Science.gov (United States)

    Mackie, R.I.; Koike, S.; Krapac, I.; Chee-Sanford, J.; Maxwell, Susan; Aminov, R.I.

    2006-01-01

    Antibiotics are used at therapeutic levels to treat disease; at slightly lower levels as prophylactics; and at low, subtherapeutic levels for growth promotion and improvement of feed efficiency. Over 88% of swine producers in the United States gave antimicrobials to grower/finisher pigs in feed as a growth promoter in 2000. It is estimated that ca. 75% of antibiotics are not absorbed by animals and are excreted in urine and feces. The extensive use of antibiotics in swine production has resulted in antibiotic resistance in many intestinal bacteria, which are also excreted in swine feces, resulting in dissemination of resistance genes into the environment.To assess the impact of manure management on groundwater quality, groundwater samples have been collected near two swine confinement facilities that use lagoons for manure storage and treatment. Several key contaminant indicators-including inorganic ions, antibiotics, and antibiotic resistance genes-were analyzed in groundwater collected from the monitoring wells. Chloride, ammonium, potassium, and sodium were predominant inorganic constituents in the manure samples and served as indicators of groundwater contamination. Based on these analyses, shallow groundwater has been impacted by lagoon seepage at both sites. Liquid chromatography-mass spectroscopy (LC-MS) was used to measure the dissolved concentrations of tetracycline, chlortetracycline, and oxytetracycline in groundwater and manure. Although tetracyclines were regularly used at both facilities, they were infrequently detected in manure samples and then at relatively trace concentrations. Concentrations of all tetracyclines and their breakdown products in the groundwater sampled were generally less than 0.5 ??g/L.Bacterial tetracycline resistance genes served as distinct genotypic markers to indicate the dissemination and mobility of antibiotic resistance genes that originated from the lagoons. Applying PCR to genomic DNA extracted from the lagoon and

  1. Determination of nine antibiotics residues in organic fertilizer by high performance liquid chromatography-tandem mass spectrometry%高效液相色谱-串联质谱同时测定有机肥料中9种抗生素药物残留

    Institute of Scientific and Technical Information of China (English)

    史丽; 李昌伟; 赵鲁; 刘继远; 季卫; 邱静

    2014-01-01

    A sensitive high performance liquid chromatography-tandem mass spectrometric ( HPLC-MS/MS) method has been developed for simultaneous determination of nine antibiotics residues ( oxytetracycline, tetracycline, chlortetracycline, doxy-cycline, penicillin G, procaine penicillin, sulfadiazine, sulfadimidine and sulfathiazole) in organic fertilizer. It is of great significance to the rapid detection of antibiotic residues in organic fertilizers. In this paper, the samples were extracted by ul-trasonic oscillation with Na2 EDTA-McIlvaine buffer solution, then extracts was cleaned up by the Strata SDB-L solid phase ex-traction column. The contents of antibiotics were determined by high performance liquid chromatography-tandem mass spectrom-etry and calculated by external standard method. Multiple reactions monitoring mode was utilized. Most analytes were scanned by positive-ion electrospray ionization except penicillin G. Average recoveries of the nine analytes were in the range of 63. 1%to 93. 4% at spiking levels of 0. 1, 0. 5 and 1. 0 mg/kg. Relative standard deviations were between 1. 7% and 9. 5%. The method detection limits of tetracycline antibiotics were 0. 05 mg/kg, and the method detection limits of sulfonamides and peni-cillins were 0. 02 mg/kg. The method was convenient, fast, with good reproducibility, and could be used to simultaneously and fast confirm multi-residue of antibiotics in organic fertilizer.%采用高效液相色谱-串联质谱法( HPLC-MS/MS),同时测定有机肥料中3类9种抗生素(土霉素、四环素、金霉素、强力霉素、青霉素G和普鲁卡因青霉素、磺胺嘧啶、磺胺二甲基嘧啶、磺胺噻唑)残留,对于有机肥料中抗生素残留的快速检测具有重要的意义。样品用Na2 EDTA-McIlvaine缓冲液超声振荡提取后,采用固相萃取小柱净化,高效液相色谱-串联质谱法检测,标准曲线外标法定量。采用多反应监测模式检测,除负离子扫描青霉素G外其他均

  2. Effects of Cordyceps power on growth performance and immune function of broilers%冬虫夏草菌粉对肉鸡生长性能和免疫功能的影响

    Institute of Scientific and Technical Information of China (English)

    邵紫培; 尹艳军; 袁洪水

    2016-01-01

    试验旨在研究冬虫夏草菌粉对肉鸡生长性能和免疫功能的影响。将200羽1日龄AA肉鸡随机分为5组,每组4个重复。空白对照组仅饲喂基础日粮;抗生素对照组在基础日粮中添加350 mg/kg金霉素;冬虫夏草菌粉试验组设低、中、高3个剂量,分别在基础日粮中添加0.05%、0.1%、0.2%冬虫夏草菌粉。试验周期42 d,分为前期(1~21 d)、后期(22~42 d)两个阶段。结果表明,与空白对照组相比,中、高剂量组在增加肉鸡均重、平均日增重,降低肉鸡料重比,提高肉鸡胸腺指数和脾脏指数,增加机体外周血T淋巴细胞的转化率、增强肉鸡白细胞吞噬能力方面均呈显著水平(P<0.05);与抗生素对照组相比,可降低腹泻发生率。由此可知,冬虫夏草菌粉可作为饲料添加剂用来提高肉鸡的生长性能和免疫功能,替代抗生素预防腹泻发生。%The aim of this experiment is to study the effects of Cordyceps power on growth performance and immune function of broiler chicken. 200 1-day-old Arbor Acres broilers were randomly divided into 5 groups with 5 repeats. The blank control group was fed basal diet and the antibiotics control group was supplemented with 350 mg/kg chlortetracycline, the experimental groups were supplemented with 0.05%, 0.1%, 0.2% Cordyceps power respectively. The cycle of test were 42 days, including pro⁃phase (1~21 d) and anaphase (22~42 d). The results showed that: compared with control group, middle and high dose groups increased average weigh, average daily gain and reduced feed gain ratio signifi⁃cantly (P<0.05); improved thymus index and spleen index dramatically (P<0.05); increased the quanti⁃ty of T lymphocyte (P<0.05) and enhanced phagocytic function of leukocyte remarkably (P<0.05). Com⁃pared with antibiotics control group, middle and high dose groups can reduce incidence of diarrhea. This research indicated that growth performance and

  3. Effects of Allicin on Performance and Immunization of Broilers%大蒜素对肉鸡生产性能和免疫机能的影响

    Institute of Scientific and Technical Information of China (English)

    王飞; 章生根; 李勇

    2012-01-01

    选择1日龄爱拔益加(AA)商品肉鸡135只,随机分成3组(A、B、C),每组设3个重复,每个重复15只.A组为空白对照组,饲喂基础日粮;B组为在基础日粮中添加50 mg/kg洛克沙生和750 mg/kg 10%金霉素;C组在基础日粮中添加200 mg/kg大蒜素,研究大蒜素对肉鸡生产性能和免疫机能的影响.结果表明,与A组相比,大蒜素组在21、42日龄都能显著提高肉鸡的体重、日增重和成活率,显著降低21日龄料重比(P<0.05),但42日龄料重比降低不明显,采食量在42日龄明显增加(P<0.01);与B组相比.21日龄大蒜素组也能显著提高肉鸡体重(P<0.01)、日增重(P<0.05),降低料重比(P>0.05),在42日龄大蒜素组也使肉鸡体重、日增重增加,料重比下降(P>0.05);与A组相比,日粮中添加大蒜素后肉鸡胸腺指数和新城疫抗体效价显著提高(P<0.05),但法氏囊指数和脾脏指数有所下降(P>0.05);与B组相比,胸腺指数和新城疫抗体效价也有所增加(P>0.05).这说明大蒜素具有促进肉鸡生长和提高免疫功能的效果.%135 one-day-old AA broilers were randomly divided into three groups (A, B, C) , with three replicates each group and 15 birds each replicates. Group A was the negative control, feeding the basal diet. Group B was the positive control, adding 50 mg/kg roxarsone and 750 mg/kg 10% chlortetracycline to diet. Group C was the experiment, adding 200mg/kg allicin to diet. Effects of allicin on performance and immunization of broilers were studied. The results showed that: Compared with control A, body weight, daily gain and survival rate of broilers at 21 days old and 42 days old were significantly increased after adding allicin to the basal diet( P 0.05). Compared with control group, the thymas index and serun antibody titres to ND in adding allicin to the basal diet were significantly enhanced (P 0. 05). The thymas index and serun antibody titres to ND in allicin also were enhanced

  4. 大蒜素对肉鸡生产性能和消化酶活性的影响%Effect of Al!icin on Performance and Digestive Enzyme Activity in Broilers

    Institute of Scientific and Technical Information of China (English)

    胡忠泽; 张玉; 刘雷; 王传光

    2011-01-01

    To study the effects of allicin on performance and digestive enzyme activity of broiler chicken, 135 one day -old AA broilers were randomly divided into three groups, with three replicates in each group, and 15 birds in each replicate. Group A was the control group, which was fed with basic diet. Group B was antibiotic group, which was fed with basic diet added by 50mg/kg roxarsone and 750mg/kg 10% chlortetracycline. Group C was experimental group, which was fed with basic diet added by 160 mg/kg 25% allicin. The results showed the difference wasn't significant in the average body weight and the average daily gain among the three groups ( P 〉0.05 ) ; compared with the group A, F/G was significantly decreased in the groupC at 42 day -old( P 〈0.05). In contrast to group A, the amylase activity in the duodenum was obviously improved by 122.87% and 40.50% (P 〈0.01 ) in group C at 21 and 42 day -old respectively, but lipase and total proteinase activity weren't obvi- ously improved in group C at 21 and 42 day - old ( P 〉 0.05 ). It is suggested that allicin should improve FCR and amylase lactivity of the duodenum in broilers.%135只1日龄AA鸡随机分成A、B、C3组,每组设3个重复,每个重复15只。A为空白对照组,饲喂基础日粮,B为抗生素组,在基础日粮中添加50mg/kg洛克沙生和750mg/kg10%金霉素,C为试验组,在基础日粮中添加160mg/kg25%大蒜素,以研究大蒜素对肉鸡生产性能和十二指肠消化酶活性的影响。试验结果表明,A、B、C3组肉鸡平均体重和平均日增重差异不显著(P〉0.05),与A组相比,42日龄时C组料重比差异显著(P〈0.05)。在21日龄和42日龄时,C组淀粉酶活性较A组提高了122.87%(P〈0.01)和40.50%(P〈0.01),脂肪酶活性较A组提高了89.98%(P〉0.05)和33.33%(P〉0.05),总蛋白酶活性较A组提高了10.29%%(P〉0.05)和1.7

  5. Evaluation of the Charm maximum residue limit β-lactam and tetracycline test for the detection of antibiotics in ewe and goat milk.

    Science.gov (United States)

    Beltrán, M C; Romero, T; Althaus, R L; Molina, M P

    2013-05-01

    CCβ for tetracyclines was also lower than EU-MRL for chlortetracycline (ewe milk: ≤ 50 µg/kg; goat milk: 75 µg/kg), oxytetracycline (≤ 50 µg/kg), and tetracycline (≤ 50 µg/kg). Regarding the 4-epimers of these tetracyclines only 4-epioxytetracycline was detected by the Charm MRL BLTET test below EU-MRL (ewe milk: 75 µg/kg; goat milk: ≤ 50 µg/kg). Acidiol had no effect on the performance of the test. The Charm MRL BLTET test could be used routinely with adapted test procedure for the fast screening of ewe and goat milk.

  6. Antibiotic Susceptibility of Lactobacillus bulgaricus Isolated from Yoghurt%酸奶中保加利亚乳杆菌药物敏感性分析

    Institute of Scientific and Technical Information of China (English)

    周宁; 张建新; 樊明涛; 王静; 李子龙; 魏新元

    2012-01-01

    Lactobacillus bulgaricus strains were isolated from different yoghurt brands produced in different regions in China and analyzed for genetic diversity and antibiotic susceptibility.Further,antibiotic-resistant isolates were subjected to genetic analysis.A total of 18 Lactobacillus bulgaricus strains were obtained using modified MRS medium.They were subjected to RAPD typing and analyzed for susceptibility to 11 different antibiotics.Besides,their potential antibiotic-resistance genes were detected by PCR.The results showed that 18 Lactobacillus bulgaricus isolates had considerable genetic diversity and diverse antibiotic-resistance phenotypes.All of them were susceptible to roxithromycin and resistant to kanamycin.Meanwhile,they also could resist ampicillin,penicillin G,chlortetracycline,chloramphenicol,tetracycline,lincomycin,streptomycin,neomycin and gentamycin to different extents.tet(M) gene was detected in isolate B-8,and ant(6) gene in both B-8 and B-41,and aph(3')-Ⅲa gene in B-43,B-47,B-49 and B-51.These findings suggested that 18 Lactobacillus bulgaricus isolates had serious multiple antibiotic resistance.%从酸奶中分离保加利亚乳杆菌,对其遗传多样性和药物敏感性进行分析,并进一步对耐药菌株的抗性基因进行检测。利用改良MRS培养基,从国内不同品牌酸奶中分离获得18株保加利亚乳杆菌。18株分离菌先经RAPD分型后采用琼脂稀释法测定其对11种抗生素的药敏性,并通过PCR对耐药菌株中可能存在的抗性基因进行检测。结果显示,18株受试菌具有明显的遗传多样性和耐药表型多样性。18株菌全部对罗红霉素敏感,而全部对卡那霉素耐药;对氨苄青霉素、青霉素G、金霉素、氯霉素、四环素、林克霉素、链霉素、新霉素及庆大霉素等9种抗生素均表现出不同程度的耐药性。通过检测耐药菌株的抗性基因,从1株菌(B-8)中检出四环素抗性基因tet(M),从2株菌(B-8和B-41)

  7. Compost and residues from biogas plant as potting substrates for salt-tolerant and salt-sensitive plants

    Energy Technology Data Exchange (ETDEWEB)

    Cam Van, Do Thi

    2013-08-01

    proportions of compost and additives (Styromull or Perlite). The results show that the large percentage (> 50% by volume) of compost in the substrate had negative effects on plant growth and nutrient uptake (N, P, K, Mg and Na) because of its high salt content in compost-based substrates. However, both yield formation and nutrient uptake of the studied plants when grown in peat-based substrates significantly increased comparing to those of compost-based substrates and almost gained the level of the control. Especially, the growth of Salvia was significantly improved. Consequently, compost-based media (with more than 50% of compost) may not be recommended for salt sensitive ornamental plants, while less than 25% volume of compost incorporated with Peat creates favorable peat-based substrates which reasonably enhanced growth of Pelargonium and Salvia. Investigating antibiotic uptake by cabbage (Brassica oleracea var. capitata f. abba) from the manure-amended soils containing high and low carbon content spiked with the two antibiotics Chlortetracycline and Sulfamethazine was targeted as the third objective. The input concentrations of the studied antibiotics were 100, 200 and 400 {mu}g/kg regarded to their present concentration range in Chinese top soils. The antibiotics in plant materials were analyzed by HPLC-MS after extraction. The results reveal that the presence of available high carbon content in the soil increased crop yield of cabbage. However antibiotics were not detected in the cabbage materials according to the antibiotic employment with the initial studied concentrations. It may be concluded that with the small amounts of antibiotics applied to the soils, there is no risk of uptake of antibiotics by plants.

  8. 透明质酸面膜护理雀斑激光术后创面的临床评价

    Institute of Scientific and Technical Information of China (English)

    严蕾; 刘刚; 王芳; 谢军; 杨超; 袁晓慧; 张楠

    2014-01-01

    目的:观察透明质酸面膜对雀斑激光术后皮肤创面修复能力的临床疗效和安全性。方法:选择40例接受雀斑激光治疗的患者随机分为实验组和对照组,激光术后及观察期间按医生指导使用透明质酸面膜或金霉素软膏做为术后护理产品,分别于治疗后即刻,冷敷20分钟后、治疗后第3天以及第5天,记录两组患者的症状和体征,同时记录创面愈合情况。结果:雀斑激光术后20例患者经过透明质酸面膜冷敷后,术后刺痛、烧灼、紧绷感等主观症状较对照组得到效缓解,红斑、水肿等不良反应减轻,皮损创面修复较对照组快,痂壳脱落后创面光滑,整个治疗过程无过敏等不良反应。结论:透明质酸面膜可用于改善激光术后刺痛、烧灼等主观症状,减轻红斑、水肿等不良反应,可促进皮肤创面快速修复,缩短创面的愈合时间,疗效确切,是一种安全有效的激光术后修复治疗手段。%Objective To evaluate the clinical curative ef ect and safety of hyaluronic acid dressings applied after laser therapy for freckles.Methods Altogether 40 patients with freckles were enrol ed in this study.After the treatment with Q -switched Alexandrite Laser,the patients were randomly divided into two groups.The patients in the experimental group were smeared with hyaluronic acid dressings on the face for 20 minutes immediately after laser therapy,and then once per day for 7 days.Patients in the control group were treated by Chlortetracycline oint-ment.Symptoms,physical signs and wound healing of two groups were recorded immediately,20 minutes,three days and five days after treatment respectively.Results Compared with the con-trol group,the experimental group showed a bet er improvement of acute inflammatory responses,such as pricking,tight feeling and burning sensation.Adverse reactions,like erythema,edema were also reduced in the experimental group

  9. 芽孢杆菌类益生素对肉鸡血细胞及免疫器官组织结构的影响%Effect of the Bacillus Probiotics on Blood Cells and Structure of Immune Organs in Broilers

    Institute of Scientific and Technical Information of China (English)

    靳二辉; 陈耀星; 王群; 乔恩美; 吴国忠; 李升和

    2013-01-01

    In order to study the effect of the Bacillus probiotics on blood cells and tissue structure of broilers,480 newly hatched AA chicken were randomly divided into control group,antibiotic group,experiment Ⅰ and Ⅱ groups.The control group was fed with basal diet,and the antibiotic group was fed with 50 mg · kg-1 chlortetracycline in the basal diet,and the experimental Ⅰ and Ⅱ groups were respectively fed with 200 and 400 mg · kg-1 the Bacillus probiotics in the basal diet for 42 days.The results showed:compared with the control group,the quantity of red cells and the ratio of basophilic granulocytes and monocytes were significantly increased in 21-and 42-day-old broilers (P<0.05),and the weight and organ index of thymus and bursal of the 42-day-old broilers were respectively increased by 46.60% and 38.23%,66.67% and 47.62% (P<0.05) ; The weight and organ index of bursal were significantly increased by 35.40% and 36.92%,88.03% and 153.97% in the 21-and 42-day-old broilers (P<0.05),and the weight and organ index of thymus of the 42-day-old broilers were significantly increased by 46.27% and 28.13% in experiment Ⅰ group (P<0.05); the quantity of white cells was obviously decreased,and the weight and organ index of spleen were also obviously decreased in the antibiotic group at the 42 d (P<0.05).Compared the tissue structure of immune organs in experiment Ⅰ and Ⅱ group with the control group,the quantity of spleen node were increased,and periarterial lymphatic sheath were thickened; the thymic cortex were also thickened,and the quantity of thymic corpuscle were decreased; the bursal node were greatened with tightly rank lymphocytes,and which change were more obvious in the experiment Ⅰ than the experiment Ⅱ group.But the tissue structure of immune organs showed different degree atrophy in the antibiotic group.The results suggested that addition of 200 mg · kg-1 the Bacillus probiotics are better to improve development of

  10. The Effect of Replacing Antibiotics Feed Additive with Gluconic Acid, Glut amine and their Combination on Production Performance of the Piglets%葡萄糖酸、谷氨酰胺及其组合物替代饲用抗生素对仔猪生产性能的影响

    Institute of Scientific and Technical Information of China (English)

    于新明; 李虎; 赵小刚; 于青云; 杨开伦

    2012-01-01

    selected and randomly divided into four groups with 6 replicates per group,and the experiment lasted for 42 days. The control groupwas fed basal diet added antibiotic (60 mg/kg colistin, 75 mg/kg chlortetracycline) ; The GA group was fed the basal diet added 1. 5 g GA; The GN group was fed the basal diet added 2. 0 g GN; The GA + GN group was fed the basal diet with 1. 5 g GA +2.0 gGN. Piglets were weaned at 28 d of age. The litter was used as ■ replicate. Piglets were individually weighted at 1 d, 21 d, 28 d and 42 d of age, respectively. The weight of feed offered and feed remaining were recorded daily, while the amount of piglets, diarrhea and mortality were recorded in each litter. The average daily feed intake, average daily gain, feed intake to body weight gain, diarrhea ratio and death ratio were calculated at every stage and the whole trial period. [Result]From 21 d to 41 d, compared with the control group, the piglets of GA group, GN group, GA + GN group were decreased 8.00% (P >0.05), 16.00% (P 0. 05) , respectively; Diarrhea head times in each group were 37 , 31, 27 , 18 at the whole trial period, showing a gradually decreasing trend; About the average daily feed intake, the average body weight, the feed conversion ratio and the death ratio, the piglets in trial group and those in control group had no significant difference (P >0.05) . Among the piglets in trial groups, there was no significant difference (P>0. 05). [ Conclusion] Collectively, the piglets in the antibiotics -treated group had a better average daily gain than those in each replacing group from 21 d to 42 d, but compared with each replacing group, the average daily feed intake, live weight, feed intake to body weight gain ratio and other index of the antibiotics - treated group piglets were not significantly different. Compared with piglets of antibiotics - treated group, diarrhea frequency in each replacing group had a decreasing trend. This study provides a reference basis for replacing

  11. Pollution Characteristics of Antibiotics and Antibiotic-Resistant Bacteria and Genes in Wangyanggou River, Shijiazhuang, China%石家庄汪洋沟地区抗生素、抗性细菌和抗性基因污染特征

    Institute of Scientific and Technical Information of China (English)

    徐艳; 张远; 郭昌胜; 王红梅; 王丁明; 王凯; 李晓晨; 徐建

    2014-01-01

    chlortetracycline)and six sulfonamides(sulfadiazine, sulfamer-azine, sulfadimidine, sulfachinoxaline, sulfamethoxazole and trimethoprim)ranged from 5.81×101 to 3.87×105 ng·L-1 and 1.02×101 to 5.37× 103 ng·L-1 in water, and from 4.28×101 to 1.63×105 ng·g-1 and 1.18×101 to 1.68×104 ng·g-1 in sediments, respectively. The number of tetra-cycline and sulfonamide resistant bacteria in water was 4.00×101 to 2.13×104 CFU·mL-1 and 6.67×101 to 7.34×105 CFU·mL-1, respectively, 3 to 4 orders of magnitude lower than that in sediments. The abundance of five tetracycline resistant genes(tetA, tetB, tetE, tetW and tetZ), two sulfonamide(sul1and sul2)resistant genes and two integron genes(int1, int2)detected was relatively high in all samples. The genes, te-tA and sul1, were dominant resistant genes with relative abundance greater than 1.58 ×10-2 in Wangyanggou River. Principal component analysis indicated that the abundance of antibiotic-resistant genes might be affected by different pollution sources and water quality charac-teristics. The phylogenetic analysis of tet(B)showed that water quality change resulted in diverse resistant strains. The current results indi-cate that pollution of antibiotics, antibiotic-resistant genes in Wangyanggou River is more severe than in other rivers.

  12. Reduction of toxic effects of aflatoxin B1 by using baker yeast (Saccharomyces cerevisiae in growing broiler chicks diets Redução dos efeitos tóxicos da aflatoxina B1, utilizando-se levedura de panificação (Saccharomyces cerevisiae, na dieta de pintos de corte em crescimento

    Directory of Open Access Journals (Sweden)

    Kemal Çelýk

    2003-06-01

    Full Text Available This study was carried out to investigate the effects of adding baker yeast (BY, chlortetracycline (CTC and both BY + CTC to a control diet containing 200 ng/g of aflatoxin B1 (C + AFB1 on performance, serum parameters and pathologyc alterations of broilers. A total 100 chicks (Ross PM 3 were divided into five groups in individual cages and each containing 20 animals. BY, a rich source of protein and vitamin B complex, was mixed into the diets at 2.0 %, CTC was mixed into the diet at 2.5 ng/g. Feed consumption, body weight and feed efficiency were recorded weekly. Serum parameters and pathologyc alterations were determined at the end of the study. Dead animals were recorded daily. Liver changes were clearly apparent in the C+AFB1and C+ AFB1+CTC most of the livers were enlarged, yellow and had pethecial hemorrhages. Canalicula cholestosis was absent in group C+AFB1 and C+ AFB1+CTC, but not others. When compared to the control (C group, alkaline phosphatase (ALP, appear to be significantly increased in the C+AFB1 and C+CTC+ AFB1 groups. Serum glutamic oxalacetic transaminase (GOTwas increased in C+AFB1 birds. Serum alphaphetoprotein was not affected by the treatments. Feed consumption and body weight were significantly reduced in group AFB1. Birds receiving BY + AFB1, CTC + AFB1 and BY + CTC + AFB1 had a significantly higher body weight than group C+AFB1. Feed efficiency was better in group CTC + AFB1 than the others. The findings of this research suggest tha BY (2% can partly counteract some of the toxic effects of AFB1.Este estudo foi desenvolvido para avaliar os efeitos da adição de Levedura de panificação (BY e cortetraciclina (CTC e ambos BY+CTC a uma dieta controle © contendo 200 ng/g de aflatoxina B1 (C+AFB1 sobre desempenho, parâmetros séricos e alterações patológicas de frangos de corte. Um total de 100 pintinhos (Ross PM3 foi dividido em cinco grupos, em gaiolas individuais, contendo 20 animais para cada grupo. A levedura de

  13. 发酵床养猪废弃垫料的资源化利用评价%Evaluation on resource utilization of litters in pig-on-litter farming system

    Institute of Scientific and Technical Information of China (English)

    胡海燕; 于勇; 张玉静; 徐晶; 孙建光

    2013-01-01

    .其中有机质含量为42.62%~54.12%,全氮1.54%~2.12%,全磷(P2O5)2.24%~5.55%,全钾(K2O)0.57%~2.15%;Cu、Zn、Cr、As、Ni、Pb、Cd、Hg等8种重金属元素含量均符合国家有机肥料农业行业标准(NY 525-2012)和城镇垃圾农用控制标准(GB 8172-1987)的限量标准;土霉素(Oxytetracycline,OTC)、四环素(Tetracycline,TC)、金霉素(Chlortetracycline,CTC)等四环素类抗生素含量低,环境风险小;蛔虫卵和大肠杆菌含量均超过NY 525-2012和GB 8172-1987的规定限值,蛔虫卵死亡率范围为90%~94%,总大肠杆菌的数量范围为1.29×105~2.24×106cfus/g;垫料盐分含量达到22.11~45.71g/kg.评价结论:发酵床废弃垫料具有有机肥料的基本性质,但是盐分含量偏高;肠道寄生虫卵严重超标,具有生物安全隐患,施用前必须对其进行无害化处理.

  14. 2009-2011年广州市番禺区水产品污染状况分析%Monitoring of the pollution of aquatic products in Panyu, Guangzhou in 2009-2011

    Institute of Scientific and Technical Information of China (English)

    林东明; 吴利楠; 麦洁梅; 张琪; 杨福荣

    2012-01-01

    Objective To understand the exteni of pollution of aquatic products in Panyu Guangzhou and to provide the basic food safety supervision. Methods Aquatic products (e.g. freshwater fish, marine fish, shrimps, crabs, shellfish, chilled ready to eat aquatic products and other aquatic products) from different markets, meat and vegetable markets and catering units of Panyu Guangzhou, were randomly collected for the detection of heavy metals, antibiotics, pesticides, pathogens, liver fluke and other projects. Results All aquatic products had a high cadmium level, and 5.2% of the samples exceeded the limit. The average cadmium content of shrimps/crabs was 0.516 mg/kg, the highest content was 14.300 mg/kg; and the percentage of shellfish, marine fish and freshwater fish exceeding the limit was 4.5% , 3.7% and 2.5%, respectively. 99.5% of the samples had the lead content below the limit. The levels of mercury and arsenic were not exceeding the limit. Part of the aquatic products were found to have a high level of tetracycline, oxytetracycline and chlortetracycline, and the percentage of samples exceeding the limit for these antibiotics was ranged from 8.3% to 14.3%. The levels of BHC and DDT in all samples were below the limit. 98.1% of the samples had the content of formaldehyde below the limit. Among the 222 samples, 18.5% had Vibrio parahaemolyticus (VP). The percentage of shrimps/crabs and shellfish with Vibrio parahaemolyticus was 41.7% and 33.3%, respectively. There was a significant different in the degree of contamination with Vibrio parahaemolyticus in freshwater fish, marine fish, shrimps/crabs and shellfish (P0.05). Salmonella and Vibrio cholerae was not detected. Pathogens and liver fluke were not found in the ready to eat raw aquatic products. Conclusion There was a serious cadmium contamination of aquatic products. The level of Vibrio parahaemolyticus was high in the aquatic products in Panyu district. The abuse of antibiotics in aquaculture and by the food

  15. Effects of Chinese Herb Additive on the Pork Quality-related Gene Expression%中药添加剂对猪肉相关基因表达量的影响

    Institute of Scientific and Technical Information of China (English)

    胡广英; 曹日亮; 赵娟; 于纪宾; 孟冬霞; 任杰; 武果桃; 车向荣; 夏燕婷

    2014-01-01

    为研究复方中药对猪肉品质的影响,选择体重约15 kg的杜长大三元杂种猪72头,按体重和性别随机分为4个处理,每个处理3个重复,每个重复6头猪。对照组饲喂基础饲粮;抗生素组在生长猪每吨饲粮中添加金霉素50 g、肥育猪每吨饲粮中添加牛至油15 g;中药Ⅰ组在基础饲粮中添加1%中药配方Ⅰ;中药Ⅱ组在基础饲粮中添加1%中药配方Ⅱ。饲养试验预试期15 d,正试期120 d。结果表明,H-FABP基因表达量,3个试验组均极显著高于对照组(P<0.01)。CAST基因表达量,抗生素组是对照组的85%(P<0.01),中药Ⅰ组是对照组的1.10倍(P<0.01),中药Ⅱ组是对照组的1.24倍(P<0.01)。LPL基因表达量,抗生素组是对照组的1.65倍(P<0.01);中药Ⅰ组是对照组的1.92倍(P<0.01);中药Ⅱ组是对照组的1.20倍(P<0.01),并且中药Ⅱ组极显著低于抗生素组和中药Ⅰ组(P<0.01)。 MYOG基因表达量,抗生素组是对照组的55%(P<0.01),中药Ⅱ组是对照组的2.26倍(P<0.01),中药Ⅰ组是对照组的1.50倍(P<0.01),抗生素组极显著低于中药Ⅰ组和中药Ⅱ组(P<0.01)。%To study the effects of compound Chinese medicine on pork quality changes.Select Duroc ×Landrace × Yorkshire 72 which weighed about 15 kg,according to the weight and sex were randomly divided into four treatments with three replicates of six pigs.The control group was fed the basal diet;antibiotic group added in diets for growing pigs per ton chlortetracycline 50 g,per ton of feed finishing pigs add oregano oil 15 g;medicine groupⅠ1%on the basis of traditional Chinese medicine formula diets Ⅰ;medicine group Ⅱ1% on the basis of traditional Chinese medicine formula diets.Pre-trial feeding trial period 15 d,formal test 120 d.The results showed that,H-FABP gene,three experimental groups were significantly higher (P<0.01).CAST gene,antibiotic group was 85

  16. Antimicrobial Peptides: Effects on Small Intestinal Mucosal Morphology and Immune Active Cell Number in Hy-Line Brown Young Roosters%抗菌肽对海兰褐仔公鸡小肠黏膜形态结构及免疫活性细胞数量的影响

    Institute of Scientific and Technical Information of China (English)

    刘莉如; 杨开伦; 滑静; 王晓霞; 刘亭婷

    2013-01-01

    本试验旨在探讨饲粮中添加不同水平天蚕素抗菌肽对海兰褐仔公鸡小肠黏膜形态结构及免疫活性细胞数量的影响.选择336只1日龄健康海兰褐仔公鸡,随机分为7个处理,每个处理4个重复,每个重复12只鸡.7个处理分别为对照组(饲喂基础饲粮),抗生素组(150mg/kg金霉素),抗菌肽Ⅰ(饲喂基础饲粮+150 mg/kg天蚕素抗菌肽)、Ⅱ(饲喂基础饲粮+200mg/kg天蚕素抗菌肽)、Ⅲ(饲喂基础饲粮+250 mg/kg天蚕素抗菌肽)、Ⅳ(饲喂基础饲粮+300 mg/kg天蚕素抗菌肽)、V组(饲喂基础饲粮+350 mg/kg天蚕素抗菌肽),饲养试验期为42 d.结果表明:各处理肠黏膜结构与对照组相比较完整,除十二指肠隐窝深度、空肠肌层厚度各处理之间无显著性差异(P>0.05)外,不同添加水平的抗菌肽组绒毛高度、黏膜厚度、肌层厚度均在一定程度上高于对照组,隐窝深度均在一定程度上低于对照组,抗菌肽V组效果最为显著(P<0.05).抗菌肽V组空肠上皮内淋巴细胞数量显著高于对照组(P<0.05),抗菌肽Ⅲ、Ⅳ、Ⅴ组与对照组相比可显著提高十二指肠、空肠及回肠杯状细胞数量(P<0.05).由此可见,饲粮添加天蚕素抗菌肽可显著改善海兰褐仔公鸡小肠黏膜上皮细胞形态,同时也增加了小肠免疫活性细胞的数量.其中抗菌肽V组添加效果最好,天蚕素抗菌肽添加量为350mg/kg.%This experiment is conducted to study the effects of dietary antimicrobial peptides ( ABPs) - cecropin on small intestinal mucosal morphology and the number of immune active cells in Hy-Line Brown young roosters. Three hundred and thirty-six young roosters were selected, and randomly divided into seven treatments with four replicates in each treatment and twelve chicks in each replicate. The seven treatments were control group (fed a basal diet) , antibiotics group (fed basal diet + 150 mg/kg chlortetracycline) , and 150, 200, 250, 300, 350 mg/kg ABPs